<figure><img src="image_78.png" /></figure>

## S.28.2 Plant epigenetics: a contribution to understanding phenotypic variation in changing environments

Conchita Alonso¹, Mónica Medrano¹, Carlos M. Herrera¹

1 Estación Biológica de Doñana, EBD-CSIC, Sevilla, Spain

Epigenetics investigates phenotypic variation that is not accompanied by changes in DNA sequence, but instead involves the set of chromatin modifications that regulate the ability of a certain genome to produce diverse phenotypes. In plants, DNA cytosine methylation is a key epigenetic mechanism involved in genome stability and developmental change. In the last few decades we have learned that Angiosperms have evolved specific DNA methylation enzymes and pathways (e.g., chromomethylases, RNA-directed methylation) which target cytosines in any sequence context and genomic location, and that genome-wide cytosine methylation level is largely variable across species and has a strong phylogenetic signal and phenotypic correlates within and across plant individuals. Furthermore, changes in DNA methylation induced by environmental stress and transgenerational transmission of some methylation marks have been documented for many plant species. It remains unclear, however, the extent to which inter- and intra-specific variation in the ability of a given genotype to produce phenotypic variants depends on intrinsic genomic features and/or the ecological characteristics of their environments. In our group we have assessed epigenetic variation in wild plant populations at different spatial scales and adopted experimental approaches which combined a controlled alteration of epigenetic features (e.g., by applying demethylating agents) with exposure to selected biotic and abiotic stress factors. I summarize what we have learned from applying these two approaches to a set of non-model plant species. Understanding the relationships between phenotypic traits and changes in DNA methylation in contrasting environments is crucial to uncover the potential impact of epigenetics on fast plant adaptation to changing environments under current scenarios of climate change. Integration of molecular understanding gained from model species and ecological significance assessed in non-model plant species should contribute to better value the contribution of epigenetics in plant adaptation and evolution.

## S.28.3 Genetic and phenotypic differentiation in functional traits of Iris pseudacorus L. in native and introduced ranges

Blanca Gallego-Tévar¹, Brenda J. Grewell², John F. Gaskin³, Jesús M. Castillo¹

1 Departamento de Biología Vegetal y Ecología, Universidad de Sevilla, Sevilla, Spain. 2 USDA-ARS Invasive Species and Pollinator Health Research Unit, Department of Plant Sciences MS-4, University of California, Davis, CA, USA. 3 USDA ARS Pest Management Research Unit, Northern Plains Area, Sidney, MT, USA

Intraspecific variation in functional traits between native and alien invasive plant species may underlie resilience and invasiveness of aliens facing new physiological limits in response to climate change. Using a common garden experiment, we explored if observed phenotypic variation of *Iris pseudacorus* L. between tidal wetland populations in the native vs. invaded range may be explained by genetic differentiation and/or phenotypic plasticity. Iris seeds were collected from populations under Mediterranean climate along estuarine salinity and inundation gradients within four native and invasive populations in both Guadalquivir Estuary (Andalusia, Spain) and San Francisco Bay-Delta Estuary (California, USA). Germinants were potted (6 plants x 4 populations x 2 ranges; n = 48 experimental plants) and grown for 12 months under uniform environmental conditions. We then evaluated 23 functional traits including growth, biomass allocation, morphological and biochemical responses. Genetic analysis was performed on leaf tissue collected from plants in each seed donor population. Geographic range explained relative intraspecific trait variation segregating native from alien phenotypes. Native plants had lower SLA (-23%) and rhizome TNC concentrations (-63%) than introduced plants, providing evidence of genetic differentiation. Higher genetic diversity in the field and 27% higher phenotypic variation in the CGE for native compared to alien plants indicate longer-term adaptive processes in the native range, while genetic distance of alien field populations increased along with their phenotypic distance in the CGE suggesting rapid genetic differentiation. Phenotypic plasticity also explained some observed inter-range phenotypic differences in response to environmental conditions, since these field differences were not expressed by plants in the
<figure><img src="image_79.png" /></figure>

CGE. Management of alien *I. pseudacorus* in the invaded estuary should be established urgently since they represent novel genotypes with key functional traits that can support invasiveness through high competitive ability and physiological stress tolerance responses to sea level rise.

## S.28.4 Fitness and barriers to gene flow related to flower colour variation in a snapdragon hybrid zone

David L. Field¹, Sean Stankowski², Arka Pal³, Parvathy Surendranadh³, Desmond Bradley⁴, Enrico Coen⁴, Nick Barton³

¹ Macquarie University, Sydney, Australia. ² University of Sussex, UK. ³ Institute of Science and Technology, Klosterneuburg, Austria. ⁴ John Innes Centre, Norwich, UK

A fundamental question in evolutionary biology is how phenotypes and their underlying genes interact to generate reproductive barriers between populations. The fitness landscape has been an influential framework to model these connections, yet direct measures of how multiple genes interact to create fitness valleys between populations remains challenging to obtain in nature. I present some of the work we've been doing to address this gap by exploiting a natural hybrid zone between subspecies of *Antirrhinum majus* (snapdragons) with a well-defined genotype-phenotype map. This work includes investigations into the molecular genetic basis of flower colour, ecological experiments on the agents of selection, and the development of novel population genomic approaches to detect barriers in the genome. A key focus will be on our multi-generational study, where-by genotyping and phenotyping over 30,000 plants over 14 years, we've obtained a pedigree-based estimate of the fitness landscape separating these recently diverged subspecies. At deeper evolutionary time scales, using whole-genomes along geographic clines we find the absence of a genome-wide barrier to gene flow with only a small proportion of the genome resisting introgression. By linking recent ecological time-scales of fitness variation in nature to patterns of deeper genomic divergence, our goal is to provide a general model for how reproductive barriers arise during the early stages of divergence.

## S.28.5 Above- and below-ground trait covariation in perennial herbaceous species

Allison Miller¹,², Molly Hanlon², Matthew Rubin²

¹ Department of Biology, Saint Louis University, St. Louis, MO, USA. ² Danforth Plant Science Center, St. Louis, MO, USA

A fundamental question in plant evolutionary biology is how species that live for multiple years balance resource allocation over time to sexual reproduction (seed production) relative to vegetative growth. One component of this is understanding how reproductive output covaries with vegetative traits (e.g., stem thickness, leaf number, leaf thickness, physiology, root biomass, etc.), how these relationships shift from one year to the next, how they vary under different environmental conditions, and how they change in response to strong selection for increased reproduction output. Although the direction and magnitude of trait correlations can constrain or enhance response to artificial or natural selection, there are few comprehensive studies that describe relationships between traits expressed at early life stages (e.g., seed size and weight, germination timing, germination rate) with traits expressed later in life (e.g., reproductive output, root/shoot biomass ratio in Y1, Y2, Y3, etc.) in perennial herbaceous species. Here, we phenotyped seeds and seedlings of 1,650 plants representing eleven perennial, herbaceous species in the Asteraceae, Fabaceae, and Poaceae families, planted those individuals in a spaced plant nursery in Gray Summit, MO USA, and monitored height, flowering time, and yield over three years. We estimated root traits via soil cores in Y2 and Y3, and then excavated root crowns for further phenotyping. Analyses to date indicate strong correlations within and among life-stages, among reproductive and vegetative traits, and among above- and below-ground traits. These core questions can provide key insights into how perennial, herbaceous species evolve under nature and under artificial selection and may expedite pre-breeding and domestication of perennial grain crops through early stage selection of traits correlated expressed in seeds and seedling that correlate with desirable traits expressed at later life stages.
# S.29 AEROBIOLOGY. ADVANCES IN THE ATMOSPHERIC POLLEN RESEARCH AND CHALLENGES IN THE CONTEXT OF GLOBAL CHANGE. SESSION 1

## S.29.1 Metabarcoding and metagenomics techniques in monitoring airborne pollen

Rosa Pérez-Badia¹, Clara E. Pogner², Celia Antunes³, Godfrey P. Apangu⁴, Nicolas Bruffaets⁵, Sevcan Celenk⁶, Antonella Cristofori⁷, Nestor González-Roldán⁸, Agnieszka Grinn-Gofron⁹, Beatriz Lara¹, Mirela Lika¹⁰, Donat Magyar¹¹, Moises Martínez-Bracero¹², Lucia Muggia¹³, Babette Muyshondt⁴, David O’Connor¹², Alberto Pallavicini¹³, Alexandra M. Penha³, Helena Ribeiro¹⁴, Ana R. Costa.³, Zsófia Tischner¹¹, Merita Xhetani¹⁰, Carsten A. Skjøth¹⁵

¹ Institute of Environmental Sciences, University of Castilla-La Mancha, Toledo, Spain. ² Unit Bioresources, Center of Health and Bioresources, AIT Austrian Institute of Technology GmbH, Tulln, Austria. ³ Department of Medical and Health Sciences, School of Health and Human Development, University of Évora and Earth Sciences Institute (ICT), Évora, Portugal. ⁴ Protecting Crops and the Environment, Rothamsted Research, Harpenden, U.K. ⁵ Mycology and Aerobiology, Sciensano, Brussels, Belgium. ⁶ Bursa Uludag University, Arts and Science Faculty, Biology Department, Görükle-Bursa, Türkiye. ⁷ Research and Innovation Centre, Fondazione Edmund Mach, San Michele all'Adige, (TN), Italy. ⁸ Pollen Laboratory, Department of Biological and Environmental Sciences, University of Gothenburg, Gothenburg, Sweden. ⁹ Institute of Biology, University of Szczecin, Szczecin, Poland. ¹⁰ Department of Biology, Faculty of Natural Sciences, University of Tirana, Tirana, Albania. ¹¹ National Center for Public Health and Pharmacy, Budapest, Hungary. ¹² School of Chemical Sciences, Dublin City University, Dublin, Ireland. ¹³ Department of Life Sciences, University of Trieste, Trieste, Italy. ¹⁴ Department of Geosciences, Environment and Spatial Plannings, Faculty of Sciences, University of Porto and Earth Sciences Institute (ICT), Porto, Portugal.

15 Department of Environmental Science, iClimate, Aarhus University, Roskilde, Denmark

The importance of monitoring the concentration of pollen in the atmosphere has been widely demonstrated from the environmental, ecological and agronomic point of view, as well as from the perspective of human health, since allergenic pollen represents the most important cause of pollinosis globally. Traditionally, pollen concentrations in the air are estimated from analyses based on identification and quantification using optical microscopy. However, the classification of pollen types at the species level using this procedure is limited. This is due to the morphological characteristics of the pollen grains used for their recognition, often shared within genera, families, and even taxonomic orders. An alternative approach is the identification of pollen through molecular sequencing and DNA analysis. Different high-throughput sequencing (HTS) technologies have been used to massively analyse environmental DNA and therefore the airborne pollen. This process requires, before sequencing, the preparation and amplification of the specific library for each sequencing platform. "Ion Torrent" (ThermoFisher), "GenapSys" technology and above all "Ilumina", represent several platforms that sequence with the metabarcoding procedure short DNA fragments (reads) up to a few hundred base pairs in length. Other platforms called "third generation sequencing" such as "PromethION", "GridION" and "MinION" from "Oxford Nanopore Technology" and "PacBio CCS" from "Pacific Biosciences", generate much longer reads (LR) than those obtained in the previous ones, however they are still little used in aerobiology. This study presents a review and the state of the art of airborne pollen studies based on metabarcoding and metagenomics, as well as the limitations of these molecular analyses.

**Acknowledgments:** This study is conducted in the frame of Cost Action CA18226 (New approaches in detection of pathogens and aeroallergens) and has been carried out by the members of Working Group 2.
<figure><img src="image_80.png" /></figure>

## S.29.2 Airborne pollen trends in the Iberian Peninsula during the last three decades: flowering timing and intensity

Carmen Galán¹,², Jose Oteros¹,², Herminia García-Mozo¹,², Fátima Aguilera³, María Jesús Aira⁴, Jordina Belmonte⁵,⁶, Paloma Cariñanos⁷, Delia Fernández-González⁸,⁹, María Fernández-González¹⁰, Santiago Fernández-Rodríguez¹¹, Montserrat Gutiérrez-Bustillo¹², Beatriz Lara¹³, Concepción De Linares⁷, Stella Moreno-Grau¹⁴, José M. Moreno-Grau¹⁴, Rosa Pérez-Badía¹³, Antonio Picornell¹⁵, Alberto Rodríguez Fernández⁸, Javier Rodríguez-Rajo¹⁰, Jesús Rojo¹², Luís Ruiz-Valenzuela³, María del Mar Trigo¹⁵, Purificación Alcázar¹,²

1 Department on Botany, Ecology and Plant Physiology, Agrifood Campus of International Excellence (ceiaA3), University of Córdoba, Spain. 2 Inter-University Institute for Earth System Research, University of Córdoba, Spain. 3 Department of Animal Biology, Plant Biology and Ecology, University of Jaén, Spain. 4 Department of Botany, University of Santiago de Compostela, Spain. 5 Department of Animal Biology, Plant Biology and Ecology, Universitat Autònoma de Barcelona, Spain. 6 Institute of Environmental Sciences and Technology (ICTA), Universitat Autònoma de Barcelona, Spain. 7 Department of Botany, University of Granada, Spain. 8 Department of Biodiversity and Environmental Management (Botany), University of León, Spain. 9 Institute of Atmospheric Sciences and Climate, National Research Council, Bologna, Italy. 10 Department of Plant Biology and Soil Sciences, University of Vigo, Spain. 11 Department of Construction, University of Extremadura, Spain. 12 Department of Pharmacology, Pharmacognosy and Botany, Complutense University of Madrid, Spain. 13 Department of Environmental Sciences, University of Castilla-La Mancha, Toledo, Spain. 14 Department of Chemical and Environmental Engineering, Technic University of Cartagena, Spain. 15 Department of Botany and Plant Physiology, University of Málaga, Spain.

Airborne pollen significantly impacts air quality, human health, and ecosystems. Effective pollen monitoring is essential for understanding these effects and advancing in air protection strategies. Pollen allergies, affecting a substantial global population, lead to respiratory issues and decreased life quality. Pollen

monitoring also informs climate change research, agriculture, and forestry; shifts in pollen seasons indicate the climatic change impacts on vegetation and aiding in fruit production forecasting. Airborne pollen is a good indicator of floral phenology and intensity on anemophilous plants. Historical databases allow to study pollen season timing and intensity. The Mediterranean Climate is wet and mild during winter, and warm and dry in summer. During recent decades, the climate is changing on increasing temperatures and to the lack of water availability, due to a different precipitation pattern with more frequent floods and droughts. These trends are affecting to plant responding to extreme events. Woody plants response more to temperatures, especially those flowering during winter or early spring. Herbaceous plants respond more to water availability. Many recent studies have shown an advance in flowering start, specially on woody plants. However, winter is getting warmer, and the plants are not sufficiently exposed to chilling temperature for break dormancy and budburst; with lack of chilling requirements the forcing process at early spring will not be as effective, that together the lack of water, may cause a delay on flowering. Weeds and grasses respond more to water availability, although in different way, with an advance or delay depending on the species. For this reason, recently there are evidence of longer pollen seasons because more diversity on flowering timing. Regarding flowering intensity, recent studies also support the higher pollen season intensity, probably due to increasing CO2 in the air; however, in the Mediterranean climate it does not always occur due to lack of water.

## S.29.3 Effect of long-distance pollen transport on the local and regional airborne pollen evolution

José María Moreno¹, Luis Negra¹,
Francisco Aznar¹, Isabel Costa¹, Stella
Moreno-Grau¹

1 Department of Chemical and Environmental Engineering, Technical University of Cartagena, Cartagena, Spain

Long-distance pollen transport, primarily wind-facilitated, profoundly influences local and regional airborne pollen dynamics, impacting genetic diversity, species composition, allergen exposure, seasonal timing, ecological interactions, climate change implications, and biogeographical patterns. The transport introduces genetic material from distant populations, fostering in-
<figure><img src="image_81.png" /></figure>

creased genetic diversity within local plant populations for enhanced adaptability to environmental changes. Pollen transport alters the species composition of local and regional plant communities, potentially introducing new species or amplifying the abundance of certain ones, influencing regional biodiversity. The process significantly affects allergen exposure by introducing allergenic species, extending pollen seasons, and altering overall allergen composition, posing implications for public health. Understanding the impact of long-distance pollen transport on seasonal timing is crucial for predicting changes in allergen exposure patterns, with direct implications for public health. Ecological interactions arise, such as competition between introduced and native plant species, leading to cascading effects on local ecosystems and changes in insect and bird populations reliant on specific plant species for food. The complex interplay between long-distance pollen transport and climate change is pivotal for predicting ecological responses, managing biodiversity, and devising strategies to mitigate climate-driven environmental impacts. This interconnectedness contributes to biogeographical patterns, connecting plant populations across geographical regions, vital for maintaining genetic exchange and preventing isolation among plant populations. Monitoring pollen movement aids researchers in assessing the impact of global and regional environmental changes on plant populations. In summary, long-distance pollen transport is a valuable tool for ecological research and environmental monitoring, providing insights into plant dispersal, genetic connectivity, climate influences, and ecological dynamics, contributing to a broader understanding of interconnected processes shaping ecosystems.

**S.29.4 Response patterns of calendar and pollen intensity to climate at the southwestern distribution limit of Betula: a 30-year study**

Guillermo Guada¹, María Fernández-González¹, Rubén Amigo¹, Duarte A. Dias Lorenzo¹, Kenia C. Sánchez Espinosa¹, F. Javier Rodríguez-Rajo¹

1 Department of Vegetal Biology and Soil Sciences, Faculty of Sciences, University of Vigo, Ourense, Spain

We studied the relationship between the occurrence of *Betula* spp. airborne pollen and climate, during the main pollen season, as well as before

and in the preceding year. Our aim was to understand how environmental conditions can influence the main pollen season and the intensity of airborne pollen presence at the meridional boundary of the *Betula* forest distribution in Europe. For this purpose, pollen monitoring was conducted using Lanzoni VPPS 2000 volumetric trap and nearby meteorological data. The study area was located in the northwest of the Iberian Peninsula, specifically in three cities with diverse mesoclimate: Vigo (Atlantic), Ourense (Mediteranean) and Lugo (Cantabrian), which had a distinct oceanic influence. Aerobiological data was conducted based on the Spanish aerobiology network protocol. Weather variables precipitation, maximum, minimum and mean temperature were selected for three-time windows: 1st the main pollen season period, 2nd from 5 to 90 days before the onset of pollen presence, with a cumulative five-day interval, and 3rd the average monthly climate of the previous year. Precipitation delays the onset of the main pollination season and also delays the end of the season, increasing pollen counts in the post-peak interval at all three areas. Temperature during the pollen season affects the duration and pollen integral differently among area. The pollen integral shows an increasing trend over the last decades at all three sites, and the increase in the minimum temperature of the previous summer explains this increase at all three areas from a climatic point of view. In conclusion, the effect of weather before and during the main pollen season has varied effects on the date-related data across the mesoclimate, while the increase in temperature during the summer of the last few decades results in a spring with higher airborne pollen loads for all three locations.

**S.29.5 The aerobiology-floral phenology binomial. Phenoclimatic models for pollen forecasts.**

Giuseppe Frenguelli¹

1 Department of Agriculture Science, University of Perugia, Perugia, Italy

The release of pollen from a plant-source is characterized by a time distribution which is the consequence of the various flowering stages. The beginning of the release of pollen and the quantity
of pollen that each plant produces are in relation to a series of biotic factors such as the genotype, the age, the size of the plants, and environmental factors such as the climate, other than edafic and phytosanitary factors. In the aerobiological studies phenological models are used to obtain information useful for forecasting the beginning of pollination of many species, especially of agricoltural and allergo- logical interst, and to know what will be the severity of the pollination or the quantity of pollen which will be released into the atmosphere day by day. Aero- biological historical data bases on the pollen con- tent in the air, evidence how the climate change is affecting distinct species in different geographical areas. The aerobiology-floral phneology binomi- al can contribute to the improvement of bio-geo- graphical and ecological information of vegetation, the forest health and vitality being important for the protection of the plant biodiversity. When a predic- tive model on beginning of pollination is being de- veloped, it is necessary to take into consideration the different responses to environmental factors of the species considered. Temperature-based mod- els are commonly used to predict floral bud-burst as universal models disregarding that they are site- and species-specific. Models working well in some regions might not respond so well in other areas due to genetic characteristics that enable plants to show different tolerance to particular environmen- tal conditions. Modeling to predict the performance of the pollen season with the intention of giving a meaningful result as early as possible has led to the development of many analytical mathematical models, increasingly complex, like time series mod- els, multivariate regression models, Artificial Neural Networks and more.

S.29.6 Real-time airborne
pollen monitoring

B. Clot¹, B. Crouzy¹, S. Erb¹¹, G. Lieberherr¹,
F. Tummon¹, R. Pérez-Badia², EUMETNET
AutoPollen community

1 Federal Office of Meteorology and Climatology
MeteoSwiss, Payerne, Switzerland. 2 University of
Castilla-La Mancha, Toledo, Spain.

Between 20-30% of the European population suffers
from pollen allergy, resulting in costs ranging from
€50-150 billion per year. For decades, airborne pol-
len has been monitored using manual methods with
counting being done by hand under the microscope
and results reported as daily average values from

3-9 days after the measurement. New technologies
developed over the past years are revolutionising
this field. Combining artificial intelligence with ad-
vanced measurement methods, several devices
are now available on the market that provide re-
al-time observations of pollen as well as a consid-
erable potential to detect and identify a range of
other airborne particles. The EUMETNET AutoPollen
Programme has been established in 2018 to take full
advantage of the large potential for progress that
automatic observations provide. It brings together a
consortium from across Europe with the multidisci-
plinary expertise needed to address the challenges
along the entire information chain – from the initial
observation through to the final products and ser-
vices co-designed with end users. As an example,
the SwissPollen network, operated by MeteoSwiss,
comprises 15 monitoring stations and since the be-
ginning of 2023 provides hourly information in re-
al-time through its website and smartphone app.
Furthermore, the real-time observations are inte-
grated into the COSMO-ART model to provide fore-
casts to end-users.
# S.30 MERGING EVOLUTIONARY APPROACHES WITH ETHNOBOTANICAL KNOWLEDGE.
## SESSION 2

### S.30.1 Estimation of Activity Concentrations of naturally occurring radionuclides in selected infant-herbs grown in Igbokoda, Ondo state

Funmilola M. Ojo¹, Abiola O. Ilori², Kayode O. Karigidi³

1 Department of Biological Sciences, Olusegun Again University of Science and Technology, Okitipupa Ondo State, Nigeria. 2 Department of Physical Sciences, Olusegun Again University of Science and Technology, Okitipupa Ondo State, Nigeria. 3 Department of Chemical Sciences, Olusegun Again University of Science and Technology, Okitipupa Ondo State, Nigeria

The traditional use of medicinal herbs in treating infant ailments is entrenched in many cultures, including the coastal region of Ondo State, Nigeria. However, there is limited scientific information regarding their morphological characteristics and possible radiological health risks. This study aimed to investigate the morphological features and radiological health effects of six commonly consumed infant herbs. A comprehensive field survey was conducted to identify and collect representative samples of the selected infant herbs: Ananas comosus, Azadirachta indica, Carica papaya, Cymbopogon citratus, Jatropha curcas, Mangifera indica, Ocimum gratissimum, Psidium guajava, Vernonia amygdalina, and Zingiber officinale. Plant identification and classification were conducted morphologically to assess plants' growth habits, stem structure, leaf morphology, and flower characteristics. Radiation levels were measured to evaluate potential risks associated with the consumption of these herbs, using a portable radiation meter. The concentration of radionuclides (specifically, naturally occurring radioactive materials such as uranium, thorium, and potassium) was also determined using gamma spectroscopy techniques. The morphological studies revealed distinct features of each herb, aiding in their accurate identification and

classification. Additionally, the radiological health impact assessment indicated relatively low radiation levels in the selected infant herbs, with concentrations of radionuclides falling within acceptable limits set by international regulatory bodies. Thus, the consumption of these herbs was determined to be unlikely to pose significant radiological health risks to infants and young children. This study contributes to the documentation of the morphological characteristics of commonly consumed infant herbs in the coastal region of Ondo State, Nigeria. Furthermore, the radiological health impact assessment provides valuable insights into the safe use of herbs for infant care. The findings can inform healthcare providers, traditional medicine practitioners, and the general public about the potential benefits and risks associated with the consumption of these medicinal herbs, promoting safe and informed choices in infant healthcare practices.

### S.30.2 Market integration and changes in traditional practices in West Africa: the case of Shea

Nerea Turreira-García¹

1 Department of Food and Resource Economics, University of Copenhagen, Denmark

Commercial and development actors influence traditional ecological knowledge (TEK) practices among rural populations in the global south through complex mechanisms. This paper explores this rural transformation dynamic through the case of shea (Vitellaria paradoxa). For centuries, women have collected and processed shea nuts into dried kernels, and subsequently butter. Over the last decades, the international demand for kernels has increased as opposed to butter. The results of 1100 household surveys conducted in Ghana and Burkina Faso about women's current and past processing methods and end-use of shea products, reveal that 1) processing
<figure><img src="image_82.png" /></figure>

methods are being homogenized, and 2) women abandon the butter making process in favor of selling shea kernels. This trend is more visible in Ghana, where especially younger women follow processing methods promoted by big companies and development institutions, in contrast to Burkina Faso, where there remains higher processing diversity, and butter production. Loss of diversity in processing methods may reduce the ability to adapt to external changes, such as market demands and climate change. The decrease in butter production is leading to matrilinear knowledge loss and has implications for the household economy, as women substitute shea butter for industrial, more expensive cooking oils and skincare creams. The study offers a critical analysis of how the dynamic and multidirectional nature of rural transformations and TEK influence resource users and the entire value chain.

## S.30.3 Does scent guide Indigenous People's perception of medicinal plants?

Kimberly P. Castro¹, Rodrigo Cámara-Leret¹

1 Department of Systematic and Evolutionary Botany, University of Zurich, Zurich, Switzerland

Understanding the interactions between people and plants is of great relevance for both biodiversity and biocultural conservation efforts. The use of plants has been considerably investigated based on morphological traits, but explaining how people recognize a plant's usefulness based on chemistry has been challenging. Here, we explore to what degree the selection and utilization of medicinal plants is influenced by people's smell perception of plants. We assess this through a literature review of works published over 50 years on smell perception and Indigenous People's knowledge of tropical plants. We find that the sense of smell may very well be a nexus between human-plant interactions, but the concept has been relatively complex to establish due to the confines posed by disciplinary perspectives and methodological limitations in most studies. To advance the field, we propose integrating phytochemistry with standard ethnobotanical data collection within study communities to better understand the role of plant scents and smell perception in medicinal plant selection.

## S.30.4 Crop domestication is favored by higher competitiveness and resource-capture.

Rafael Rubio de Casas¹, Irene Martín-Brull¹,², Francisco J. Ocaña-Calahorro¹

1 Departamento de Ecología, Universidad de Granada, Granada, Spain. 2 EEAD-CSIC, Zaragoza, Spain.

One of the open questions regarding the relationship between humans and plants is why some taxa are domesticated and become crops while other ecologically and morphologically similar relatives are not. In all likelihood, this is mediated by functional differences between crops and their wild relatives. The identification of this key traits requires accounting for the divergence in ecological and demographic settings between natural populations and crop fields, in which interspecific density is much higher and diversity lower. Traits facilitating growth and survival under these conditions might have thus been relevant for domestication. Using a phylogenetically controlled experiment, we investigated whether domestication might be facilitated by functional traits that increase survival and growth under high density, hyper-competitive conditions. We picked three important annual legume crops, lentil (Lens sp.), grasspea (Lathyrus sp.) and vetch (Vicia sp.) and measured traits that determine competition (lodging; growth rate), resource acquisition (plant height; total aerial and root biomass) and yield (fruit no.). Bayesian models were fitted to these data to quantify the effects of evolutionary divergence between close wild and domestic relatives, as well as between natural and crop (landraces) populations of domesticated taxa. Our results showed important taxonomic differences, likely reflective of different selection pathways. Nevertheless, wild relatives consistently differed from their domesticated congeners in competition and resource acquisition. Never-domesticated taxa consistently produced smaller, slower-growing plants that were more prostrated and allocated less biomass to roots. Conversely, the effect of selection under domestication on vegetative traits was largely negligible, although it appeared to have led to an increase in yield. We conclude that, although domestication processes are always complex and case-dependent, they might indeed favor taxa that are particularly com-
<figure><img src="image_83.png" /></figure>

petitive and more effective in resource capture. Subsequent selection under domestication may have targeted reproductive traits (i.e., fruit and seed production).

## S.30.5 Molecular authentication, quantification, and in vitro production of metabolites from different Swertia species

Parthraj Kshirsagar¹ Dnyanesh Chinte² and Nikhil Gaikwad³

1 Department of Botany, Shri. Shivaji College Barshi, Dist-Solapur, India. 2 Department of Zoology, S. M. Dnyandeo Mohekar College, Kalamb, Dist- Osmanabad, India. 3 Department of Botany, Shivaji University Kolhapur, Dist-Kolhapur, India

Swertia is ethno-medicinally an important genus belonging to the family Gentianaceae. Swertia chirayita is used as an imperative medicinal plant in the Indian system of medicine. However, this species has been frequently adulterated due to its high demand and scarcity. Authentication of this species was needed to protect consumers and conservation measures and to find an alternative source. ITS region proves a robust molecular marker for differ-

ing the *S. chirayita* from its related adulterant species. All barcoding regions indicate that *S. chirayita* and *S. minor* both are more closely related than other *Swertia* species. Swertiamarin and mangiferin were estimated from different *Swertia* species and it was revealed that maximum swertiamarin and mangiferin content was found in *S. chirayita*, *S. minor*, and *S. angustifolia* species. Various extraction methods like static extraction (SE), continuous shaking extraction (CSE), and ultrasonic extraction (USE) were evaluated for increasing recovery % of swertiamarin and mangiferin from different *Swertia* species which revealed that SE was more proficient with 24 h extraction gave the maximum recovery in *S. chirayita* of swertiamarin (256.98 mg/g) and mangiferin (155.76 ± 7.78 mg/g). *In vitro* production of swertiamarin from cell suspension cultures of *S. minor* was studied chitosan-treated (25 ppm) cell biomass accumulated higher contents of swertiamarin than salicylic acid and methyl jasmonate. Findings showed that DNA barcoding is an efficient tool for the identification and authentication of *S. chirayita*. *S. minor* is remarkably the best match for *S. chirayita* as per molecular and phytochemical fingerprint is concerned, suggesting an alternative for chirayita. The described protocol can be effectively used for the large-scale propagation, and exploitation of active compounds and will serve as a potential alternative to *S. chirayita* for the fulfillment of over-growing industrial requirements.

## S.31 THE ANGIOSPERM REPRODUCTIVE SPECTRUM FROM FLOWERS TO SEEDS. SESSION 2

### S.31.1 Variation in fruit and seed dimensions is better explained by dispersal system than by leaf size in a tropical rainforest

Pablo R. Stevenson¹, Diana C. Acosta-Rojas¹, Sasha Cárdenas¹, L. Francisco Henao-Díaz¹,²

1 Centro de Investigaciones Ecológicas La Macarena, Departamento de Ciencias Biológicas, Universidad de Los Andes, Bogotá, Colombia. 2 Department of Ecology and Evolution, University of Chicago, Chicago, IL, USA

Variation in fruit and seed traits could originate from selection pressures exerted by frugivores or other ecological factors (adaptive hypotheses) and developmental constraints (by-product hypotheses) or chance. We evaluated fruit and leaf traits for nearly 850 plant species from a rainforest in Tinigua Park, Colombia. Through a series of linear regressions controlling for the phylogenetic signal of the traits (minimum N = 542), we
<figure><img src="image_84.png" /></figure>

tested (1) whether the allometry between seed width and length depends on seed dispersal system (Mazer and Wheelwright's adaptive hypothesis of allometry for species dispersed in the guts of animals = endozoochory) and (2) whether fruit length is associated with leaf length (i.e., Herrera's by-product hypothesis derived from the assumption that both organs develop from homologous structures). First, we discovered a robust negative allometric relationship between seed width and length in endozoochorous species, aligning with our expectations. Surprisingly, this association also held true for anemochorous species. In addition, we found a positive relationship between fruit and leaf length, but this relationship was not evident for zoochorous species. Our findings lead us to the conclusion that the allometric relationship between seed length and width exhibits variability among dispersal systems. This supports the notion that fruit and seed morphology has undergone modifications due to interactions with frugivores and the unique adaptability to rotate, particularly evident in certain wind-dispersed species.

## S.31.2 Morpho-anatomy of the flower-to-fruit transition in selected Andean Loranthaceae

Valentina Botero-Castaño¹, Favio González², Natalia Pabón-Mora¹*

1 Universidad de Antioquia, Medellín, Colombia. 2 Universidad Nacional de Colombia, Bogotá, Colombia *Author for correspondence: lucia.pabon@udea.edu.co

Loranthaceae (Santalales) is a family of hemiparasitic plants that exhibits unique structural modifications, such as: (1) structural reduction of the carpels, compensated by the formation of a massive hypanthium and (2) ategmic ovules, these reduced to the embryo sac; (3) migration of the embryo sac nuclei from the ovary to the style and ectopic fertilization in the style, followed by the repositioning of the proembryo(s) to the ovarian region; and (4) fruits developing from inferior ovaries fused with the hypanthium, with abundant viscine. Other atypical features recorded include the presence of dimorphic embryos (i.e., embryos have foliose or prismatic cotyledons) in the genus *Psittacanthus* and the absence of endosperm described in *Psittacanthus* and *Aetanthus*. Conventional serial sectioning of flowers and fruits at different developmental stages of *Aetanthus colombianus* A.C. Sm., *Gaiadendron punctatum* (Ruiz & Pav.) G. Don., and *Tristerix secundus* (Benth.) Kuijt,

allowed us to conclude that the flowers of these species go through: (1) migration of the nuclei of the megagametophyte, which reach the distal third portion of the style; (2) formation of a secretory parenchyma associated with the vascular bundles of petals and stamens; and (3) growth of a sclerenchymatic structure that delimits the megagametophyte in the flower and the proembryo in the fruit. Conversely, the fruit formation follows: (1) development of the pendular embryo, facing the sclerenchymatic tissue, with a conspicuous hypocotyl, and formation and expansion of the viscin in the epicarp; (2) development of two foliose cotyledons surrounded by massive endosperm (ruminate in Gaiadendron), and lack of seed coat; and 3) the fruit proper is delimited by the vascular bundles of the gynoecium, and surrounded by a thick epicarp with extra-carpelly viscin. This work lays the foundation for comparative evo-devo studies on the genetic bases for ovule reduction in hemiparasitic neotropical Santalales.

## S.31.3 Anatomy and micromorphology of the two genera with unusual fruit type in Apiaceae (Echinophora and Thecocarpus)

Golshan Zare¹, Aslı Doğru-Koca²

1 Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University, Ankara, Türkiye. 2 Laboratory of Plant Conservation & Phylogeny, Department of Biology, Faculty of Science, Hacettepe University, Ankara, Turkey

The genus *Echinophora*, with 10 species is represented in Türkiye by six species, three of which are endemic. *Thecocarpus carvifolius*, is the only representative of the genus. *Echinophora* and *Thecocarpus* are closely related genera with taxonomically controversial history, which are placed in *Echinophoreae*. This tribe is characterized by the remarkable structure of the fruiting umbellules. In this tribe the umbellules have sessile solitary female fruit in the centre, surrounded by the indurated pedicels of the male flower. In this group, different parts of the umbellules take part in fruit formation and in these infructescence structures. The anatomical characters of the fruit are widely used in the taxonomy of Apiaceae. In this study detailed anatomical and micromorphological futures of 7 taxa were done in detail including; E. tenuifolia, E. tournefortii, E. chrysantha, E. lamondiana, E. tricophylla, E. orientais, *T carvifolius*. The relationships of
<figure><img src="image_85.png" /></figure>

*Echinophora* and *Theocarpus* species grown in Türkiye were discussed based on the micromorphology and anatomy of fruits of 7 species. Plant material was collected from different regions of Türkiye and free hand sections were stained with astra blue and safranin. For scanning electron microscopy, fruits were coated with gold-palladium. We describe the range of variation in the infructescence, to discuss the available taxonomic characters and elucidate micro-morphological and anatomical fruit structure. *Echinophora* mericarps is generally pyriform, glabrous or hairy, located inside thickened structure called the cage or shell. Mericarps contain two carpels or monocarpel by abortion, 6vittae and carpophore. Stomata are absent in some species. *Theocarpus*, fruits called false nuts are hardened, glabrous with two carpels, 4vittae, and carpophore and stomata are present. Fruit structure such as presence and position of stomata, carpophore, vittae size, number and position provided valuable diagnostic characters in the taxonomy of these taxa.

### S.31.4 Pleurothallidinae inflorescences: Importance of correct morphology interpretation

Gustavo Rojas-Alvarado¹, Adam Karremans¹

1 Jardín Botánico Lankester, Universidad de Costa Rica, Cartago, Costa Rica

Despite its great relevance, the study of the inflorescence from a typological point of view generally goes unnoticed in taxonomy, which is fundamental for the comparison of structural elements of the same origin. Pleurothallidinae is the most diverse subtribe in Orchidaceae is not the exception, its typology inflorescence remains largely unexplored, causing incorrect interpretations of its structures and misapplication of terms. The morphology of Pleurothallidinae inflorescences is clearly illustrated by photographs and diagrams, and it is analyzed and discussed from a typological point of view, based on the detailed study of structural elements of living material, including more than 82% of genera of the subtribe. The study shows that the subtribe presents a generalized type of inflorescences formed by an abbreviated peduncle and branch system that cannot be seen with the naked eye, which is an uncommon condition in other orchid's groups. Each branch may produce coflorescences of different lengths with one or multiple flowers, also

presenting different patterns of succession that determine the general appearance of the plant. There are two main types of inflorescences in Pleurothallidinae, these with single-flowered coflorescences that are dominant in early divergent clades, and the multi-flowered coflorescences, which dominate higher clades. A general and practical classification is established for the different types of coflorescences according to the length and number of flowers produced

### S.31.5 Evolution of seed dispersal modes in the Orchidaceae: has the Vanilla mystery been solved?

Adam P. Karremans¹, Charlotte Watteyn², Daniela Scaccabarozzi³,⁴, Oscar A. Pérez-Escobar⁵ & Diego Bogarín¹,⁶

1 Lankester Botanical Garden, University of Costa Rica. P.O. Box 302-7050 Cartago, Costa Rica. 2 Department of Earth and Environmental Sciences, KU Leuven, Celestijnenlaan 200E, P.O. Box 2411, Leuven, Belgium. 3 Department of Ecology and Genetics, Uppsala University, Uppsala, Sweden. 4 School of Molecular and Life Sciences, Curtin University, Bentley, Australia. 5 Royal Botanic Gardens, Kew, Richmond, UK. 6 Evolutionary Ecology Group, Naturalis Biodiversity Center, CR Leiden, The Netherlands.

Orchidaceae seeds are said to have the smallest seeds among flowering plants. They lack an endosperm, and most have only a very small embryo within a thin, spindle-like, transparent seed coat. The seeds are predominantly wind-dispersed, often developed within dry, dehiscent fruits that typically release millions of dust-like seeds into the air. Wind dispersal appears to be a derived state in Orchidaceae and given its predominance, a trait likely associated with enhanced speciation rates. Animal-mediated seed dispersal is a lesser-known phenomenon in the family and predominantly occurs in groups belonging to early-diverging lineages bearing indehiscent, fleshy fruits with hard, rounded, dark seeds. Zoochory primarily occurs in groups derived from early-diverging lineages; occasional reversions suggest a link between dispersal mode and fruit and seed traits. Among the zoochrous groups of orchids, genus *Vanilla* is the most predominant. The more than 120 species of *Vanilla* are native to the Tropical regions of Asia, Africa and America where they grow as climbing vines. Vanillin, the organic com-
<figure><img src="image_86.png" /></figure>

pound responsible for the much appreciated vanilla aroma, is synthesized naturally only in the fruits of certain Neotropical species belonging to this genus. This, and related, aromatic compounds play a pivotal role in the multimodal seed dispersal. Ectozoochory occurs in dry, dehiscent fruits, whose seeds are dispersed by (i) male euglossine bees collecting the fruit's vanillin aromatic compounds and (ii) female stingless bees collecting the fruit's mesocarp. Endozoochory occurs in (iii) highly nutritious, indehiscent fruits consumed by terrestrial mammals or (iv) fleshy, dehiscent fruits whose mesocarp is consumed by arboreal mammals. Interestingly, fruits features, such as dehiscence and fleshiness lack phylogenetic signal in *Vanilla* despite their role in determining dispersal modes, suggesting potential environmental adaptability.

## S.31.6 Disentangling the Orchid-Euglossini interaction networks in the tropical forest in Costa Rica

Rengifo Alfonso Nicolás¹*, Fernández Otárola Mauricio²,³, Hanson, Paul². Karremans, Adam³

1 Graduate Biology Program, University of Costa Rica. 2 School of Biology, University of Costa Rica. 3 Center for Research in Biodiversity and Tropical Ecology, University of Costa Rica. 4 Lankester Bo-

tanical Garden Research Center, University of Costa Rica *Corresponding author: nicolas.rengifo@ucr.ac.cr

Orchids are one of the most diverse groups of plants, with sophisticated reproductive mechanisms that have allowed the diversification of niches and pollinators, such as Euglossini bees. The study of the reproductive phenology of orchids by means of bee body pollinaria provides a clear understanding of the interaction between plants and pollinators. Also, many bee species have a different phenology in certain ecosystems, which puts pressure on orchids that depend on these pollinators for reproduction. The objective of this work was to characterize the reproductive phenology of orchids and the Euglossini pollinator bee community. The work was carried out at La Selva Biological Station, Costa Rica. During one-year, monthly samplings of three days were carried out, using different chemical attractants to attract Euglossini males, and only those observed with pollinaria or accessory structures on their bodies were captured. The orchid pollinators were identified at the Lankester Botanical Garden and the bees at the School of Biology, University of Costa Rica. Gongora and Euglossa were the most abundant orchid and bee genera. During the dry season, the abundance of bees was higher than during the rainy season, and the species in flowering vary significantly from season to season, as do the species of bees. This work presents the first bipartite network between orchids and Euglossini bees, considering the two climatic seasons of the year.

## S.32 ADVANCES IN SEED CONSERVATION OF WILD SPECIES. SESSION 2

### S.32.1 Are seeds of today's endangered plants ready for tomorrow's climate change challenges?

Héctor E. Pérez¹

1 Environmental Horticulture Department, University of Florida, Gainesville, USA

Seeds operate in a world of physiological limits established by abiotic and biotic factors. These limits represent thresholds that govern not only in planta development of high-quality seeds but

also seed to seedling transitions via germination. Important plant life history stages such as seed development on mother plants and seedling establishment are crucial to sustaining life on Earth. Yet human induced climate change continues to influence thresholds regulating plant regeneration from seeds. Additionally, many of the predicted impacts of climate change may push seeds beyond limits established during their recent evolutionary history. This predicament may be magnified for seeds of endangered plants and raises a series of important questions. Are the seeds of today's endangered plants ready for the climate challenges of tomorrow? How will seeds have to adapt to climate change? Are we expecting too
much from seeds given the potential for increas-
ing levels of stress that may keep seeds at op-
erational limits? This presentation explores those
questions by first discussing projected impacts of
climate change. It then overlays a current syn-
thesis of seed responses to predicted climate-in-
duced stressors. We conclude with perspectives
that may contribute to development of adaptive
solutions for endangered species while calling for
more attention to seed biology especially in a cli-
mate change context.

S.32.2 Elevated spring temperatures promote premature germination in range-wide cork oak populations

Marta Benito Garzón¹, Fany Baillou¹, Filipe
Costa e Silva², Carla Faria², Maurizio
Marchi³, Giovanni Giuseppe Vendramin³,
Natalia Vizcaíno Palomar⁴

1 BIOGECO INRAE, UMR 1202, University of Bordeaux, Pessac, France. 2 Centro de Estudos Florestais, Instituto Superior de Agronomia, Universidade de Lisboa, Tapada da Ajuda, Lisbon, Portugal. 3 Florence Research Area, CNR-Institute of Biosciences and BioResources, Sesto Fiorentino, FI, Italy. 4 ICIFOR, INIA-CSIC, Madrid, Spain

Climate change is facilitating the northward mi-
gration of Mediterranean oak species, leading to
an expansion of their distribution at their leading
edges. Nevertheless, Mediterranean oaks possess
desiccation-sensitive acorns that lack the ability
to form seed banks, relying instead on post-seed
fall climate conditions and the interplay of genet-
ic determinants for germination. In this study, we
investigate Quercus suber populations' potential
adaptation to rising spring temperatures in germi-
nation timing and rates. A total of 701 acorns from 9
populations, with 10 mother trees each, were sown
at temperatures of 15, 20, and 25 °C, with daily mon-
itoring of germination over a 90-day period. Ger-
mination timing was modeled using Cox's propor-
tional-hazards models. Populations' adaptation to
the transfer distances associated with spring tem-
perature changes and germination climatic niches
under current and the RCP 8.5 scenario for the year
2080 were assessed using fixed-effects models.
Variations in germination timing were attributed
to both population origin and temperature treat-

ment, while germination rates exhibited sub-opti-
mal tendencies under temperatures exceeding the
species' native range. The timing of germination
decreased in correlation with rising spring tem-
peratures, with a projected germination occur-
rence in 2080 anticipated to be 12 days earlier than
the present timeframe in central Iberia. Elevated
spring temperatures significantly expedite the ger-
mination process of recalcitrant Mediterranean
species, potentially influencing the developmental
environment of seedlings and, consequently, the
regeneration and species composition of popula-
tions. Consequently, germination timing deserves
greater consideration in scientific and stakehold-
er discourse and should be integrated into forest
vulnerability assessments and assisted migration
programs designed to foster long-term forest ad-
aptation to climate change.

S.32.3 Functional traits at the service of seed conservation in Brazilian open ecosystems

Carlos A. Ordóñez-Parra¹,², Lara Amaral-Garcia², Rafael Figueiredo², Fernando A.O. Silveira²

1. Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil. 2 Centro de Síntese Ecológica e Conservação - CSEC, Departamento de Genética, Ecologia e Evolução, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.

Seed-based conservation and restoration are in-
creasingly recognised as cost-effective strategies
to protect biodiversity and restore degraded eco-
systems. Both strategies depend on seeds' ability
to remain viable in medium- to long-term storage,
which, in turn, is directly related to seeds' desicca-
tion tolerance —a trait whose assessment is highly
time-consuming and challenging to scale to vari-
ous species. To overcome this shortfall, trait-based
models to predict desiccation tolerance using easy-
to-measure traits, such as seed mass and moisture
content, have been proposed. Here, we used these
models to explore the potential of seed conserva-
tion in two hyperdiverse yet threatened open eco-
systems in Brazil: rock outcrop vegetation and the
Cerrado, with the ultimate goal of delivering knowl-
edge about native seed conservation and building
capacity for their use in conservation and resto-
ration projects. For rock outcrop vegetation, we built a database of 16 seed traits for 383 species. Unfortunately, data on desiccation tolerance was available for only 34 species, all of them producing orthodox seeds. Still, the available information for seed mass and moisture content for 87 species indicates a high potential for seed conservation, as suggested by the few empirical studies available for these and similar ecosystems worldwide. An ongoing project in the Cerrado is assessing seed desiccation tolerance in 84 species —56 of them with unknown storage behaviour— currently commercialised by local communities in Central Brazil. This species pool includes groups whose seed desiccation tolerance has been little studied, such as perennial grasses, which are a fundamental element of Cerrado plant communities. Ongoing data collection will be compared with the predictions of global trait-based models to assess whether these models produce accurate predictions of seed desiccation tolerance and, thus, used as a tool to facilitate the seed conservation work currently carried out by local communities.

### S.32.4 Enhancing Seed Conservation Strategies for Rare and Endangered Plant Species in Bavaria, Germany: Insights from Trait-Based Seed Longevity

Deshika Muthuthanthirige¹, Peter Poschlod¹

1 Institute of Plant Sciences, Faculty of Biology and Preclinical Medicine, University of Regensburg, Regensburg Germany

The rapid changes in the environment due to human activities and climate shifts pose a significant threat to the habitats of rare and endangered plant species, pushing them closer to extinction. It remains crucial to preserve these species to uphold the functioning of ecosystems. Our objective is to comprehend the impact of traits like seed morphology and physiology in endangered species on their capacity to endure extended storage periods. By investigating the relationship between these traits and seed longevity, especially in habitats like less fertile soils or transitional zones, our aim is to enhance the prioritization of in situ protection or alternative conservation cultures for certain species, as well as those suitable for long-term ex situ

storage in gene banks. Additionally, our analysis of habitat features and seed longevity data using Eltenberg indicator values (EIVs) aims to unveil why particular habitats favor short-lived or long-lived plant species. Our focus within Bavaria, Germany includes understanding how species, including those stored in projects like Bayern Arche establish their abundance in specific habitats based on seed longevity and environmental factors. This research endeavors to establish a direct connection between seed viability and essential traits, providing valuable insights to optimize seed conservation strategies and improve species selection for storage in seed banks.

### S.32.5 Green and fatty architecture: how chlorophyll, fatty acids and antioxidants modulate ageing rates in dry seeds

Daniel Ballesteros¹,², Rida Ajmal², Antonia Martin², Marina López-Pozo³, Davide Gerna², Christina Walters⁴, Hugh W. Pritchard⁵

1 University of Valencia, Burjassot, Spain. 2 Royal Botanic Gardens, Kew, Wakehurst Place, UK. 3 University of the Basque Country, Vizcaya, Spain. 4 USA-Agriculture Research Service, Fort Collins, USA. 5 Chinese Academy of Sciences, Kunming, China

Orthodox seeds, when dry, solidify their cell's aqueous cytoplasm forming a glass (i.e., an amorphous non-crystalline solid) where most biochemical reactions are inhibited. However, ageing is not completely stopped at this quiescent state and slowly progress mainly via oxidative reactions. Seed ageing kinetics is species dependent, but the causes for these differences are not well known. However, it has been well characterized that the seeds of some particular species age faster than the average. For example, "green seeds" - i.e., those containing chlorophyll at maturity, generally in fully developed chloroplasts - age fast at any storage temperature, like those from Salicaceae species. In addition, diverse "fatty (oily) seeds" age abnormally fast when stored to high subzero temperatures, as those standard in seeds bank (e.g., -20°C). The causes for the faster ageing rates in these two particular seed types have been related to specific reactions
<figure><img src="image_87.png" /></figure>

modulated by their "dry architecture" (Ballesteros et al., 2020). In other words, the particular physical-chemical properties of the cell cytoplasm of "green" or "fatty" seeds when dry and stored to low temperatures strongly influences their ageing kinetics. This influence is focused in two aspects: (1) the source of Reactive Oxygen Species (ROS), the target of oxidative reactions, and the antioxidant potential, and (2) the differential physical changes occurring in the glassy aqueous phase of the cytoplasm and the lipid oil droplets distributed therein. In this talk we review the state of art of ageing kinetics in dry seeds with green and fatty architectures. Moreover, we show recent advances on our understanding of this topic using both multicellular seeds but also fern spores as a unicellular model. References: Ballesteros D, Pritchard HW, Walters C. (2020). Dry architecture: towards the understanding of the variation of longevity in desiccation-tolerant germplasm. Seed Science Research 30(2): 142-155.

### S.32.6 Call for data sharing and an open invitation to join SeedArc, a global archive of primary seed germination data

Eduardo Fernández Pascual¹, Angelino Carta², Sergey Rosbakh³, Lydia Guja⁴,⁵, Shyam S. Phartyal⁶, Fernando A.O. Silveira⁷, Si-Chong Chen⁸,⁹, Julie E Larson¹⁰, Borja Jiménez-Alfaro¹

1 IMIB Biodiversity Research Institute (University of Oviedo - CSIC - Principality of Asturias), University of Oviedo, Spain. 2 Department of Biology, Botany Unit, University of Pisa, Pisa, Italy. 3 Department of Plant and Environmental Sciences, University of Copenhagen, DK Frederiksberg C, Denmark. 4 National Seed Bank, Australian National Botanic Gardens, Parks Australia, Acton, ACT, Australia. 5 Centre for Australian National Biodiversity Research, CSIRO, Acton, ACT, Australia. 6 School of Ecology and Environment Studies, Nalanda University, Rajgir, India. 7 Department of Genetics, Ecology and Evolution, Federal University of Minas Gerais, Belo Horizonte, Brazil. 8 Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan, China. 9 Millennium Seed Bank, Royal Botanic Gardens Kew, Wakehurst, UK. 10 USDA Agricultural Research Service, Eastern Oregon Agricultural Research Center, Burns, OR, US

We present SeedArc, a global initiative to mobilize seed germination data that has been produced by decades of studies on seed ecology and conservation. Seed germination is a key stage in a plant's life that is controlled by numerous environmental conditions. Yet, significant amounts of data on germination-environment relationships remain inaccessible and many plant lineages and biomes are underrepresented. Since available information is scattered into a multitude of sources and germination metrics, there is a need for a unified open-science database to archive and share primary seed germination data. To meet this need and to stimulate continental and global evolutionary research, we introduce SeedArc, a global archive of primary seed germination data (www.unioviedo.es/seedarc/; https://doi.org/10.1111/ nph.19143). We expect SeedArc to strengthen international collaborative projects in plant science, connecting the seed research community with other data synthesis initiatives. In this session, we demonstrate SeedArc's potential to address major research questions in seed ecology and describe the process for submitting your germination data and participating in collaboration. Most importantly, we extend an open invitation to all holders of seed germination data to submit data and collaborate on SeedArc research projects exploring seed ecology and evolution.
<figure><img src="image_88.png" /></figure>

# S.33 MECHANISMS AND CONSEQUENCES OF DISPERSAL IN LAND PLANTS: TOWARDS A MORE UNIVERSAL UNDERSTANDING. SESSION 2

## S.33.1 Drift in the tropics: Phylogenetics and biogeographical patterns in Combretaceae

Olivier Maurin¹, Artemis Anest²,³,⁴, Félix Forest¹, Ian Turner⁵,⁶, Russell L. Barrett⁷, Robyn C. Cowan¹, Lijia Wang⁸, Kyle W. Tomlinson²,⁹, Tristan Charles-Dominique⁴,¹⁰

1 Royal Botanic Gardens, Kew, Richmond, Surrey, UK. 2 Center for Integrative Conservation, Yunnan Key Laboratory for Conservation of Tropical Rainforests and Asian Elephants, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla, Menglun, Yunnan, China. 3 University of Chinese Academy of Sciences, Beijing, China. 4 AMAP, Univ Montpellier, CIRAD, CNRS, INRAE, IRD, Montpellier, France. 5 Singapore Botanic Gardens, National Parks Board, Singapore, Singapore. 6 Singapore Botanical Liaison Officer, Royal Botanic Gardens, Kew, Richmond, Surrey, UK. 7 National Herbarium of New South Wales, Australian Botanic Garden, Mount Annan, New South Wales, Australia. 8 University of Michigan School of Public Health, Ann Arbor, Michigan, USA. 9 Center of Conservation Biology, Core Botanical Gardens, Chinese Academy of Sciences, Mengla, Menglun, Yunnan, China. 10 Centre National de la Recherche Scientifique (CNRS), Sorbonne University, Paris, France

Certain plant clades are distributed across every continent, occupying various contrasting biomes. Within these globally distributed clades, the ecologically dominant Combretaceae family shows intercontinental disjunctions. Fruit morphology in Combretaceae suggests various seed dispersal modes, potentially explaining the observed geographical patterns. To enhance our understanding of the mechanisms that have facilitated such distribution, this study explores the dispersal strategies of the Combretaceae family, the acquisition of new modes throughout their evolutionary history, and their impact on present and past plant biogeography. The study integrates data on natural distribution, biomes, and dispersal types into comparative phylogenetic analyses. Estimates of the ancestral distribution of Combretaceae suggest a Gondwanan origin for this

family, with minimal transitions between different biomes. Our findings indicate that drift fruits, dispersing seeds by water, played a critical role in colonising new continental lands. Conversely, biome shifts appear unaffected by dispersal strategies. The study highlights a dispersal mode paradox, wherein specialised modes of seed dispersal can facilitate intercontinental dispersal but limit colonisation of new biomes and strengthen biome conservatism. Our study emphasises the implications for species preservation in changing environments and the importance of understanding the relationship between plant distribution and dispersal modes for effective conservation efforts.

## S.33.2 Unraveling the evolutionary history of island floras on the world's most isolated archipelagos.

Ángela Aguado-Lara¹,²*, Isabel Sanmartín², Johannes Le Roux³, Carlos García-Verdugo⁴, Sonia Molino⁵,⁶, Bettine Jansen van Vuuren⁷, Mario Mairal¹.

1 Universidad Complutense de Madrid, Madrid, Spain. 2 Real Jardín Botánico (CSIC), Madrid, Spain. 3 Macquarie University, Sydney, Australia. 4 Universidad de Granada, Granada, Spain. 5 Universidad Autónoma de Madrid, Madrid, Spain 6 Universidad Europea de Madrid, Madrid, Spain. 7 University of Johannesburg, Johannesburg, South Africa.

The sub-Antarctic archipelagos are some of the most remote and semi-pristine insular habitats in the world. The diverse array of ages of these archipelagos, combined with the large distances between them and to the nearest landmasses, create an exciting area for exploring austral biogeography. This includes phenomena such as extreme long-distance dispersal and colonization, facilitated by marine currents across the Southern Hemisphere. Understanding these dynamics requires reconstructing the patterns and processes explaining biodiversity assembly in the sub-Antarctic archipelagos involved in the sub-Antarctic region. To better understand the biogeography of the sub-Ant-
<figure><img src="image_89.png" /></figure>

arctic region we examined four vascular plant species exhibiting different modes of dispersal which are widespread across the region and nearby austral continents, along with three species restricted to a very remote sub-Antarctic province: the Southern Indian Ocean Biogeographic Province (SIOBP). We generated and compiled nuclear and plastid DNA sequence data for each species and estimated phylogenetic relationships, divergence times, inter-island migration rates and island carrying capacities. Our findings reveal that the nearby continental landmasses (South America, Australia or New Zealand) acted as source areas for the dispersal of these widespread vascular plant species to the sub-Antarctic archipelagos. This connection persisted for millions of years, spanning from the Miocene to the present day, and may have been driven by the eastward-moving winds and water currents, supporting long-distance dispersal as a key biogeographic process in this region. In addition, the isolation of the sub-Antarctic archipelagos played a role in the origin of endemics restricted to remote islands. Overall, our study suggests that the sub-Antarctic islands have served as important biodiversity refuges from the Miocene to recent times, emphasizing the need to establish priority conservation plans for these habitats, particularly in light of rapid climate change throughout this region.

### S.33.3 Patterns of sex expression and sex ratio with focus on insular bryophytes

Anabela Martins¹, Jairo Patiño², Ana Losada-Lima³, Manuela Sim-Sim¹,⁴

1 cE3c - Centre for Ecology, Evolution and Environmental Changes & CHANGE - Global Change and Sustainability Institute / MUHNAC - Museu Nacional de História Natural e da Ciência, Universidade de Lisboa, Lisboa, Portugal. 2 Island Ecology and Evolution Research Group. Instituto de Productos Naturales y Agrobiología (CSIC). La Laguna, Tenerife, Canary Islands, Spain. 3 Departamento de Botánica, Ecología y Fisiología Vegetal, Universidad de La Laguna, Tenerife, Spain. 4 cE3c - Centre for Ecology, Evolution and Environmental Changes & CHANGE - Global Change and Sustainability Institute, Departamento de Biología Vegetal, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, Lisboa, Portugal

Oceanic islands host remarkable biodiversity richness and represent important hotspots. Laurel forests are the relicts of an evergreen humid forest community, which occurred across the Mediterranean basin about

20 Mya. Since the onset of the Mediterranean climate, laurel forests only persisted in Macaronesia, where the oceanic environment buffered the climatic oscillations of the Pleistocene, and where these forest ecosystems are closely associated with the elevational belt of orographic cloud formation. The Macaronesian laurel forest harbors an exceptionally high diversity of bryophytes, with approximately 33 threatened endemic bryophyte species, the majority of which are dioecious. Dioecious bryophyte species often face challenges in sexual reproduction. Traditionally, asexual reproduction is thought to help maintain well-adapted phenotypes in stable environments, while sexual reproduction enhances genetic variability, increasing the population's chances of survival in new colonizing areas or during periods of environmental change. In contrast to flowering plants, most dioecious bryophytes exhibit a female bias among sex-expressing individuals. However, in a few cases, sex ratios have been found to be evenly balanced or that there is a male bias, as demonstrated in population studies of leafy liverworts. A significantly broader taxonomic range of moss species has been examined, but there is a noticeable lack of data for leafy liverworts. This research will be the first to investigate patterns of sex expression in Macaronesian laurel forest endemic bryophytes. Through the integration of data obtained from fieldwork conducted on the islands of Madeira and Tenerife, along with herbarium material (AZU, C, E, LD, LISU, E, TFC, VAL), we aim to examine the patterns of sex expression and sex ratio in four laurel forest bryophytes (Exsertotheca intermedia [endemic], Frullania polysticta [endemic], Frullania teneriffae, Porella canariensis), exploring their implications for effective sexual reproduction across their entire range.

### S.33.4 The ultimate dispersers? How dispersal has shaped global patterns of fern biodiversity

Emily B. Sessa¹, Weston L. Testo², Cody C. Howard³

1 William & Lynda Steere Herbarium, New York Botanical Garden, Bronx, NY, USA. 2 Department of Plant Biology, University of Vermont, Burlington, VT, USA. 3 Department of Plant Biology, Ecology, and Evolution, Oklahoma State University, Stillwater, OK, USA

Fern life cycles are characterized in part by their reliance on spores as the primary unit of dispersal. Unlike seeds, which may be a variety of sizes and often rely on animals to assist with dispersal, fern spores
<figure><img src="image_90.png" /></figure>

are almost uniformly microscopic and wind dis-
persed, and as a result, these plants have long been
recognized as being exceptional long-distance dis-
persers. We explore the consequences of this dis-
persal ability for fern distribution and natural histo-
ry, focusing on recent studies of globally dispersed
groups (including genera such as Asplenium and

*Dryopteris*, and families like *Blechnaceae*) and how their evolutionary history has been shaped by long distance dispersal. We also discuss the implications for various biogeographical patterns as they apply to ferns, such as island biogeography, classical examples of disjunct distributions, and fern species' ability to colonize of newly available habitats

# S.34 FLORAL SPECIALIZATION AND THE GENERALISED NATURE OF POLLINATION INTERACTIONS. SESSION 2

## S.34.1 The role of pollinator generalization in co-flowering community assembly

Gerardo Arceo-Gomez¹

1 East Tennessee State University, Tennessee, USA

Pollinators can play a key role in the assembly of plant
communities. Due their ability to carry and disperse
pollen among flowers they are central in mediating
facilitative and competitive plant-plant interactions.
These interactions can be direct (via pollen interac-
tions on the stigma) or indirect (by altering pollinator
visitation). However, their underlying complexity and
their relative role in mediating the assembly of high-
ly diverse plant communities is not fully known. To this
end, we collected data on floral traits, phenology and
pollinator visitation for over 40 plant species at the
serpentine plant communities in California and used
network tools to uncover novel patterns of co-flower-
ing and trait structure to help elucidate the role of pol-
linator generalization in co-flowering assembly. Results
show plant species are differentially assembled along
multiple trait-axes (flower color and flowering time) de-
pending on the efficiency and specialization level of the
main pollinator groups. The assembly of plant species
along multiple axes of trait differentiation limits pollina-
tor sharing (competition) and contributes to the main-
tenance of diverse co-flowering communities. We fur-
ther collected over 1000 flower visiting insects and plant
styles and characterized pollen loads on insect bodies
and flower stigmas. We reveal the importance of di-
rect pollen-pollen interactions in community assembly,
not only of those that occur on the stigma, but also of
those that take place on pollinators bodies during pol-

len co-transport. Specifically, pollen co-transport net-
works revealed differences in the number and identity
of competitors that pollen species encounter on insect
bodies across pollinator taxa. Pollen co-transport net-
works were modular, indicating some pollen species in-
teract more often on the bodies of pollinators than oth-
ers. Differences in pollinator generalization can lead to
vastly different competitive landscapes during pollen
transport and deposition, with important consequenc-
es for plant community assembly.

## S.34.2 Unraveling urbanization: a tropical perspective on pollinators, plants, and interactions

Victor H. D. Silva¹, Ingrid N. Gomes¹, Pietro K. Maruyama¹

1 Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil

Urbanization is a major threat to pollination, due
to the transformation of natural landscapes into
urban matrices. Previous studies indicate that
highly urbanized areas have a higher negative
effect on pollinators and their interactions with
flowers. However, most of this information comes
from non-tropical regions, emphasizing the need
for studies in the tropics. Here, we evaluated the
impact of urbanization on the richness of polli-
nators, plants, and interactions in a large tropical
metropolis, across twelve sampling points with
varying levels of impervious surface. Over a one-
year period, we conducted monthly visits to each
of these points, sampling over 1 km transects.
<figure><img src="image_91.png" /></figure>

During these visits, we observed and recorded all flowering plants, as well as interactions with pollinators. Throughout the study, we recorded 7,603 plants-pollinators interactions, involving 234 plant and 217 pollinator species. Urbanization did not influence pollinator ($pseudoR^2$= 0.154; $p$=0.151) and interaction richness; ($pseudoR^2$= 0.083; $p$=0.311), but it negatively influenced plant richness ($pseudoR^2$= 0.357; $p$ = 0.0129). Analyzing seasonal variation, we observed that plant, pollinator, and interaction richness were significantly higher in the rainy season compared to the dry season ($t$ = 3.69, $W$ = 125, $W$ = 127; $p$<0.001, respectively). The plant species most frequently visited included *Sphagneticola trilobata*, *Tridax procumbens*, *Leucanthemum vulgare*, and *Psychotria carthagenensis*. During the rainy season, *Sphagneticola trilobata* (a native species) emerged as the predominant resource for pollinators, while in the dry season, *Tridax procumbens* (non-native) took precedence. With the exception of *Psychotria carthagenensis* (Rubiaceae), all other frequently visited flowers belong to Asteraceae. These findings offer crucial insights into the impact of urbanization on biodiversity and ecosystem dynamics, with important implications for biodiversity conservation in tropical cities. Additionally, they underscore the substantial potential of Asteraceae plants and non-native species in supporting pollinators in urban areas.

### S.34.3 Do pollinator shifts or adaptive wandering explain floral divergence in generalist Dimorphotheca daisies?

Arjan Engelen¹, Allan G. Ellis¹

1 Stellenbosch University, Western Cape, South Africa

While pollinator-mediated selection undoubtedly contributes to diversification of plants with specialised pollination phenotypes, its importance in generalist plants is less clear. The adaptive wandering model of floral diversification posits that floral divergence in generalists, occurring in response to selection across quantitative pollinator mosaics, does not result in the trade-off in pollinator use (i.e., ecological specialisation) that is the hallmark of the alternate, pollinator-shift model. Here we test the importance of these alternate mechanisms in generalizing the striking spatially structured floral variation that characterises the annual *Dimorphotheca* daisy complex in South Africa.

First we show that the system is underlain by a weak spatial pollinator mosaic because all morphotypes interact strongly with a core group of widespread visitors, with use of these pollinators tracking their relative availability in communities, both within and across morphotype ranges. However, each morphotype is associated with some unique and effective pollinating species that are distributed in a largely qualitative spatial mosaic corresponding to floral morphotype distributions. Field experiments with model inflorescences suggest that, while the core pollinators are not exerting selection, the unique species in the assemblage are selecting on divergent trait phenotypes. Although evidence of trade-offs in pollinator use is limited, divergent floral phenotypes do produce weak but significant pollinator partitioning, and thus ethological isolation, in sympatry. While these results largely align with the adaptive wandering model of floral divergence, the fact that spatially discrete floral variation is underlain by largely qualitative gradients in the availability of a small subset of effective pollinators exerting selection on divergent floral traits is akin to the pollinator-shift model. Against the background of a core widespread generalist pollinator community, our results suggest that relative fitness gains associated with traits improving attraction and effective use of additional range-restricted pollinator species may explain the evolution of floral diversity in *Dimorphotheca*.

### S.34.4 A mosaic of local pollinator assemblages underlies floral trait divergence in a pollination-generalized plant.

Felipe Torres-Vanegas¹, Vanda Temesvári¹, Magne Friberg¹, Øystein H. Opedal¹

1 Department of Biology, Biodiversity Unit, Lund University. Lund, Sweden. SE.

Pollinators are a fundamental driver of the diversification of floral traits. Indeed, the astonishing diversity of floral traits represent, in part, an adaptation to the behavioural preferences and functional characteristics of the associated pollinators. The composition of the local pollinator assemblage is expected to differ
<figure><img src="image_92.png" /></figure>

across the distribution range of a particular plant species, with the potential to generate a geographic mosaic of divergent patterns of phenotypic selection that can represent an avenue of adaptation to the local pollinator assemblage. Thus, studies that relate geographical variation in floral traits to the distinct regimes of phenotypic selection exerted by the local pollinator assemblage are fundamental to understand the interplay between microevolution and macroevolution. This is particularly relevant for generalized plant-pollinator interactions, where a particular plant interacts with a broad range of pollinators. Here, we evaluated whether geographical variation in the composition of the local pollinator assemblage can establish distinct regimes of phenotypic selection and underlie the patterns of divergence in floral traits of *Viscaria vulgaris*. We observed a taxonomically and functionally broad range of diurnal and nocturnal pollinators that primarily included bumblebees, honeybees, solitary bees, syrphid flies, butterflies, and moths. We detected geographical variation in the composition of the local pollinator assemblage and the strength and direction of phenotypic selection on particular floral traits. Despite generalized plant-pollinator interactions, we detected that the observed variation in floral traits was associated with divergence in the composition of the local pollinator assemblage. We argue that generalized plant-pollinator interactions can exert phenotypic selection on floral traits and that distinct local pollinator assemblages can underlie the patterns of divergence in floral traits. Together, these outcomes indicate that specialized plant-pollinator interactions are not required to generate and maintain the diversity of particular floral traits.

flowering. Consequently, plants have evolved strategies to minimize heterospecific pollen transfer (HPT) and ensure efficient pollination. While floral trait diversity has been well-studied for its role in generating specialized interactions, the role of pollinators in maintaining this diversity is often overlooked. We investigated the foraging behavior of native pollinators in highly diverse, annually flowering, herbaceous communities of the Sky-islands of northern Western Ghats. We assessed floral abundance using a transect method across the flowering season to understand: i) the extent of flowering synchrony with respect to floral color and shape, and ii) shifts in plant-pollinator interactions as flowering phenology changed. Additionally, we examined the degree of floral constancy displayed by the most dominant pollinator group—bees—to assess the effect of floral traits and their abundance within a patch in a heterogeneous community. Despite the high species diversity in these seasonal communities, we found that the community as a whole exhibits higher asynchrony in its flowering phenology, potentially reducing competition for shared pollinators. Interestingly, while generalized interactions dominate at the landscape-level, our study revealed that pollinators display high floral constancy towards the most abundant species within a patch. This observation was further supported by the dominance of uni-floral pollen in their loads, highlighting the critical role of individual foragers in preventing HPT and maintaining floral diversity. Our findings unveil the paradox between floral specialization and generalized interactions by showing the presence of low HPT and specialized response of individual pollinators to locally abundant species in a hyper-diverse community with generalised network

## S.34.5 Unveiling the paradox: specialized individuals shape generalised networks in seasonal herbaceous communities of Indian Sky-islands

Sukhraj Kaur¹, Vinita Gowda¹

1 Indian Institute of Science Education and Research, Bhopal, India

Floral traits are recognised to play a key role in attracting pollinators, and the competition for limited pollinator resources can negatively impact a plant's reproductive fitness, especially in communities with synchronous

## S.34.6 Pollination of Dipcadi saxorum Blatt. (Asparagaceae: Scilloideae)

Suchandra R. Dutta¹, Hensal S. Rodrigues¹, Kiran G. Chakral¹

1 Department of Botany, R. D. & S. H. National College & S. W. A. Science College, Bandra (West), Mumbai, India

Dipcadi Medik. (Asparagaceae: Scilloideae), comprises of 44 species distributed in Africa, S. Europe up to Indian Subcontinent. In India, the genus is represented by 13 species and 2 varieties. The genus is characterised by bulbous geophyte plants, bearing long linear flat or channelled leaves, white-green-brick red tubular-campanulate flowers, emitting sweet-foul odour,
<figure><img src="image_93.png" /></figure>

some odourless. Initially, pollination in Hyacinthaceae (now Scilloideae) was known by bees, birds, butterflies, moths and rodents. Studies of pollination biology on members from Tribe Dipcadieae Speta ex J.C. Manning & Goldblatt, had not been undertaken until Manning et al. brought forth the pollination of *Dipcadi brevifolium* (Thunb.) Fourc. by owlet moth *Cornutiplusia circumflexa* (L., 1767) Syn: *Syngrapha circumflexa* (L) (Noctuidae) from Africa. Scientific study on Pollination of Indian Dipcadi is carried out by the in Mumbai suburban districts, Maharashtra, India during the flowering season from

June-August for a period of 2 years from 2020 to 2022.
Our work deals with pollination of *Dipcadi saxorum*. The flowers are nocturnal, producing scent and nectar at nightfall which extends up till morning attracting many visitors. The study concludes that the species is self-incompatible, and that the foul-acrid odour it produces is majorly because of aldehydes and esters. Settling moths, *Heliothus peltigera*, *Chrysodexis acuta* and hovering moths, *Macroglossum stellatarum* and *Cephanodes hylas* are the pollinators of *D. saxorum*.

**S.35 METAL HYPERACCUMULATORS: NEW ADVANCES ON A BOTANICAL CURIOSITY.**
**SESSION 2**

**S.35.1 Tomographic adventures in the selenium-enriched phloem sinks of a hyperaccumulator**

Maggie-Anne Harvey¹, Jeroen van der Woude¹, Ai Lin¹, Abel Muller¹, Cyril Stevens¹, Mirko Salinitro², Dennis Brueckner³, Hugh H. Harris⁴, Kathryn M. Spiers³, James H. Lovett⁴, Antony van der Ent¹

1 Wageningen University and Research, Wageningen, The Netherlands. 2 University of Bologna, Bologna, Italy. 3 Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany. 4 The University of Adelaide, Adelaide, Australia.

Selenium (Se) is an element beneficial but not essential to plant nutrition, and soils enriched with Se are typically hostile to plant growth and induce toxicity. Uniquely adapted hyperaccumulator plants are capable of growing on these enriched soils and accumulating rather extreme concentrations of Se in their tissues, without exhibiting serious toxicity. In Australia, the legume *Neptunia amplexicaulis* was found growing on seleniferous soils with >4400 µg Se g⁻¹ in its tissues, and under controlled conditions this plant is capable of concentrating up to 13,600 µg Se g⁻¹ in the youngest leaves. This plant has been shown to retain and circulate its accumulated Se, concentrating Se in phloem sinks such as developing tissues and recycling Se as these phloem sinks become phloem sources. While Se is generally metabolized into a non-toxic organic

form (selenocystathioinine), concentrating excessive Se can be a metabolically costly and risky process. Recently we have been able to use cutting edge X-Ray Fluorescence Tomographic Imaging at the German Synchrotron (DESY) to examine the tissue distribution of Se in phloem sinks. These tissues, such as the youngest developing leaves, apical meristem and root tips, would have been difficult to hand section without damaging the tissues or losing any phloem-bound Se. It was found that Se is present in the epidermis of the youngest leaves – unlike maturing leaves, less Se could be found in the vascular tissues of these developing organs. In contrast, the root tips showed ubiquitous Se concentrations in the ground meristem, with a gradual concentration of Se in vascular tissues as they begin to develop. Analysis of this distribution can reveal interesting implications about the mechanisms of Se cycling in this plant, as well as the evolutionary advantages of Se hyperaccumulation.

**S.35.2 Screening for metal hyperaccumulators - An analysis of herbarium material from Angola**

Tom Morgenstern¹

1 Institute of Botany, Faculty of Botany, Technical University Dresden, Germany

For the research on hyperaccumulator plants and their uses in phytomining, phytoremediation of soils and many other applications one crucial part is the iden-
<figure><img src="image_94.png" /></figure>

tification of species which are able to hyperaccumulate metals. To date there is some research done in the Democratic Republic of the Kongo but only a few on the native plants of its neighboring state, the Republic of Angola. In this context herbaria offer a broad spectrum of native plants that can be scanned for hyperaccumulators. The Herbarium Dresdense contains a big collection of over 3000 specimens from Angola. In cooperation with the Herbarium Hamburgense, for a master's project these were scanned for hyperaccumulating species from different regions of Angola, mainly from the north-western Province of Uíge as well as from the south of the country. The Global Hyperaccumulators Database was used to pre-select genera and families from which Hyperaccumulating species are known. Small samples of each specimen were analyzed for the concentration of several metals using ICP-OES. Over 130 different species, some with several samples from different locations and organs of the plants were tested to examine the influence of these factors on the metal concentrations. We were able to identify several new aluminum hyperaccumulators containing up to 40.000 mg/kg of its leaf dry mass, one new Mangan hyperac cumulator and discover a rather interesting genus for further searches on new hyperaccumulators.

### S.35.3 Bioaccumulation of potentially toxic elements in exotic and native woody species growing around a copper mine in Mufulira, Zambia

Charles Mulenga¹*, Darius Phiri², Daigard Ricardo Ortega-Rodríguez³,⁴, Martina Meincken⁵

1 Department of Biomaterials Science and Technology, Copperbelt University, Kitwe, Zambia. 2 Department of Plant and Environmental Sciences, Copperbelt University, Kitwe, Zambia. 3 Universidade de São Paulo, Escola Superior de Agricultura Luiz de Queiroz, Departamento de Ciências Florestais, Piracicaba, São Paulo, Brazil. 4 DendroOlavide-Dept. Sistemas Físicos, Químicos y Naturales, Universidad Pablo de Olavide, Sevilla, Spain. 5 Department of Forest and Wood Science, Stellenbosch University, Stellenbosch, South Africa

There is an international debate on the potential of exotic or native species in phytoextracting potentially toxic elements (PTEs) from contaminated soil to contain the impact of mining activities on forest ecosystems.

Exotic and indigenous trees have been evaluated as greenbelts to trap dust and thereby limit the dispersion PTEs. This study compares the potential of native (*Brachystegia longifolia*) and exotic (*Eucalyptus camaldulensis* and *E. grandis*) tree species growing around a copper mine in Zambia to accumulate PTEs and evaluate their ability to biomonitor metal pollution. Tree bark and leaves were collected from trees growing at the same site downwind from a copper-leaching plant. Topsoil was collected one metre from each sampled tree trunk. Portable X-ray fluorescence was used to analyse the elemental composition and concentration of heavy metals in biomass and soil samples. Pollution indices were used to establish the status and degree of soil contamination, while the bioaccumulation factor (BF) determined the ability of studied species to accumulate PTEs. PTEs (Cd, Fe, Cu, Pb, Ni, Mn, and Zn) were detected across biomass and soil samples, with a significant variation between species and plant parts. The pollution indices established that the soil at the study site is highly contaminated with Cu. The concentration of the studied heavy metals varied across species following the order *E. grandis* > *B. longifolia* > *E. camaldulensis* in both tree bark and leaves. The BF indicated that Zn, Mn, and Cd accumulates in all the studied species. Furthermore, a higher concentration of Cu was detected in *B. longifolia* bark, suggesting that this tree species has potential for biomonitoring Cu pollution attributed industrial activities. This study presents new insights in managing polluted environments through biomonitoring and bioaccumulation of PTEs, because these findings can guide policy on species selection when establishing greenbelts.

### S.35.4 Mediterranean native plants for the restoration of abandoned mining sites: where we stand and how we move forward

Maria E. Boi¹, Mauro Fois¹, Lina Podda¹, Marco Sarigu¹, Marco Porceddu¹, Gianluigi Bacchetta¹

1 Centre for the Conservation of Biodiversity, Sardinian Germplasm Bank (BG-SAR), Department of Life and Environmental Sciences, University of Cagliari, Cagliari, Italy

The Mediterranean Basin experienced a long history of mining exploitation that has left important consequences on environment, human health and biodi-
<figure><img src="image_95.png" /></figure>

versity. In this frame, phytoremediation can help in the
restoration of these environments, especially adopting
a multidisciplinary approach. A recent review (Boi et al,
2023) has discussed the main issues of metal(loid)s pol-
lution related to mine exploitation in the Mediterranean.
A list of native plants suitable for phytoremediation
and their related applications were given, categorizing
them into four groups basing on their biological forms,
vegetation types, and ecology: (1) hydro/hygrophilous
vegetation, (2) annual and perennial meadows, (3)
garrigues and maquis, (4) high maquis and woods. The
study allowed to highlight that mine environments are
often rich in taxa of phytogeographic and conservation
interest which must be taken in account for the remedi-
ation of abandoned mines. However, physiological and
ecological studies were deserved for a little amount of
Mediterranean plants and thus further efforts are nec-
essary to increase knowledge and detect other suitable
species. Acknowledgements: We acknowledge finan-

cial support under the National Recovery and Resil-
ience Plan (NRRP), Mission 4 Component 2 Investment
1.5 - Call for tender No.3277 published on December 30,
2021 by the Italian Ministry of University and Research
(MUR) funded by the European Union - NextGenera-
tionEU. Project Code ECS0000038 - Project Title eINS
Ecosystem of Innovation for Next Generation Sardinia -
CUP F53C22000430001- Grant Assignment Decree No.
1056 adopted on June 23, 2022 by the Italian Ministry of
Ministry of University and Research (MUR), and the Mu-
nicipality of Iglesias in the frame of "Progetto di conser-
vazione e traslocazione di Limonium merxmuelleri nella
Valle del Rio San Giorgio".

**References:** Boi ME et al (2023) Using mediterranean native plants for the phytoremediation of mining sites: an overview of the past and present, and perspectives for the future. Plants 12, 3823.

S.36 CULTIVATED PLANT TAXONOMY:
BRIDGING THE DIVIDE

S.36.1 Weeping willow
mystery solved: genome-
wide DArTseq genotyping
reveals origin of Salix
babylonica hybrids

Radim J. Vašut¹, Jana Geržová¹, Veronika Ciesaríková¹, Veronika Chodilová¹, Lucie Kobrlová¹, Zdeněk Špíšek¹, Jan Weger²

1 Palacky University Olomouc, Olomouc, Czechia.
2 Silva Tarouca Research Institute for Landscape and
Ornamental Gardening, Průhonice, Czechia

Weeping willow (Salix babylonica L.) and twisted wil-
low (S. matsudana Koidz.) are prominent ornamen-
tal willows in temperate regions of the world. In ad-
dition, their importance has been growing in recent
years due to their use as a promising crop for bio-
mass production. Despite their distinctive morphol-
ogies, the taxonomy of this ornamental group, which
also includes cultivars of presumed hybrid origin,
has been characterised by uncertainty and con-
fusion. Previous taxonomic concepts lacked exact
(genetic) data and relied instead on morphological
surveys. The considerable intraspecific phenotypic

variation in willows and the lack of knowledge of the
genetic relationships of taxa within *S. babylonica/S.
matsudana* group have prevented experts from
reaching a consensus on the number of accepted
taxa of weeping and twisted willows. In our study, we
used genome-wide genotyping using DArTseq (Di-
versity Arrays Technology) to analyse all major culti-
vars of *S. babylonica/S. matsudana* and their puta-
tive hybrids. By examining thousands of binary SNPs
for each chromosome pair, complemented by data
on genome size variation (estimated by flow cytom-
etry), we gained new insights into the delimitation of
taxa and cultivars within the *S. babylonica/S. matsu-
dana* group. The results clearly show their close ge-
netic relationship. We identified three major and four
minor hybrid groups, and elucidated the origin of
over 20 cultivars, including all the most widely grown
cultivars. By providing a solid basis for understand-
ing the genetic relationships between taxa within
the *S. babylonica* group, and by providing compel-
ling evidence for the origin of *S. babylonica* hybrids,
we propose nomenclatural changes. These pro-
posed changes aim to provide a stable taxonomic
concept for this immensely popular, widely cultivat-
ed and aesthetically pleasing group of ornamental
trees, potentially resolving uncertainties and errors
that have persisted for nearly two centuries.
<figure><img src="image_96.png" /></figure>

## S.36.2 The study of formerly cultivated plants, part of our historical, cultural and biological heritage

Daniel Guillot Ortiz¹

¹ Independent researcher, Valencia, Spain

The data obtained by the author during the period 2001-2023 as a result of the study of the Spanish ornamental flora, at present and during the 19th century and the first half of the 20th, are presented. The results have been used in the publication in scientific journals of numerous articles (more than four hundred) together with several monographs. The author have also participated in the creation, together with other botanist, of Bouteloua, a Spanish journal dedicated to the study of ornamental flora from the scientific perspective, the first volume of which was published in 2006. The results of this line of work have increased knowledge regarding the number of species and cultivars cultivated and/or commercialised in Spain. Over the course of this study numerous cultivars not mentioned in recent botanical or horticultural literature, and present in the Spanish alien flora, were discovered.

## S.36.3 A comprehensive phylogenomic analysis bridges micro and macro-evolutionary scales to explain the evolutionary history of Cannabis

Manica Balant¹,², Daniel Vitales¹, Zoltán Barina³, Branko Dolinar⁴, Lin Fu⁵, Tiangang Gao⁶,⁷, Teresa Garnatje¹,⁸, Airy Gras¹,², Muhammad Q. Hayat⁹, Marine Oganesian⁰, Jaume Pellicer¹,¹¹, Alireza S. Salami¹²,¹³, Alexey P. Seregin¹⁴, Nina Stepanyan-Gandilyan¹⁰, Nusrat Sultana¹⁵, Shagdar Tsooj¹⁶, Magsar Urgamal¹⁶, Joan Vallès²,¹⁷, Zhiqiang Wang¹⁸, Lisa Pokorny¹,¹⁹

¹ Institut Botànic de Barcelona (CSIC-CMCNB), Barcelona, Spain. ² Universitat de Barcelona, Barcelona, Catalonia, Spain. ³ WWF, Budapest, Hungary. ⁴ Botanical Society of Slovenia, Ljubljana, Slovenia. ⁵

South China Botanical Garden (CAS), Guangzhou, P. R. China. ⁶ Chinese Academy of Sciences, Beijing, China. ⁷ University of Chinese Academy of Sciences, Beijing, China. ⁸ Jardí Botànic Marimurtra-Fundació Carl Faust, Blanes, Spain. ⁹ National University of Sciences and Technology (NUST), Islamabad, Pakistan. ¹⁰ Institute of Botany (NAS RA), Yerevan, Armenia. ¹¹ Royal Botanic Gardens, Kew, Richmond, UK. ¹² Industrial and Medical Cannabis Research Institute, Tehran, Iran. ¹³ University of Tehran, Tehran, Iran. ¹⁴ Lomonosov Moscow State University, Moscow, Russia. ¹⁵ Jagannath University, Dhaka, Bangladesh. ¹⁶ Botanic Garden and Research Institute (MAS), Ulaanbaatar, Mongolia. ¹⁷ Institut d'Estudis Catalans, Barcelona, Spain. ¹⁸ Chengdu University, Chengdu, China. ¹⁹ Real Jardín Botánico (CSIC), Madrid, Spain

Cannabis sativa L. has been used by humans for millennia, providing important and versatile services in traditional medicine, for ritual purposes and to produce food and fibre. The process of domestication and subsequent dispersal by humans has resulted in the development of various landraces and cultivars, exhibiting a wide array of morphological features, phytochemical aspects, and an intricate genetic structure marked by numerous hybridisation events. Unravelling the origin of this plant and determining the existence of genuinely wild populations pose considerable challenges due to its complex nature. In this study, we relied on the Angiosperms353 probe set to investigate the genetic origins of over 90 accessions, encompassing a spectrum of wild/feral and landrace representatives. Phylogenomic and population genomics workflows were implemented to analyse the data generated. Both workflows consistently showed East Mongolian accessions as a distinct group, robustly supported as sister to all other sampled accessions. The remaining accessions segregate into two main clades, aligning with the geographical distribution and the classification proposed in earlier studies for the putative subspecies *Cannabis sativa* subsp. indica and *Cannabis sativa* subsp. sativa. The first clade encompasses accessions from China to Iran and southwards into the Indian subcontinent, with a dispersal to West Africa. These latter Indian and African accessions consist of landraces characterized by high Δ9-tetrahydrocannabinol (THC) content. The second clade is itself divided into two subclades. A Caucasian-Mediterranean clade and a mostly Turanian-Russian clade that also includes some Eastern European accessions. These distribution patterns are consistent with known routes of trade and migrations of human populations, in agreement with previous studies.
<figure><img src="image_97.png" /></figure>

### S.36.4 Classifying and naming in the network of cultivated plants

Valéry Malécot¹

1 Institut de Recherches en Horticulture et Semences, L'Institut Agro Rennes-Angers, Angers, France

Like done in any plant group, classification and naming are key process to spoke about cultivated plants. The same tools as those used for wild taxa can be used to reconstruct history of cultivated plants, but tools coming from genetics (whether population genetics, genetic diversity or heredity) may also be used to sort out genealogical links. Most of the questions raised by cultivated plants are on the bridge between genetics and phylogeny and have consequences on classificatory and naming process. Using various genera (such as Rosa, Cytisus...), we show how adding cultivated taxa in taxonomic studies can enlarge some aspects of taxonomic studies. The taxonomic and breeding histories need to be assessed before trying to classify and name some groups in these genera. Various techniques and process can be used to sort out these taxonomic and breeding histories, from purely historical ones to molecular ones. When such histories are reconstructed, in between inter-individual genealogies and inter-specific phylogenies, classification process can be applied in order to delimit pertinent taxa and may question taxa proposed while using wild plants only. In the same way, naming these taxa may need the exact same rules as for wild taxa, but may also rely on rules from the cultivated plant Code, or even other rules. Such situations, intermediate between wild and cultivated, requiring approaches from different disciplines, are common, may be particularly relevant when questioning the naturalness of some individual plants.

### S.36.5 Has taxonomy gone to pot? Why the needs and expectations of amateur gardeners and professional botanists lead to opposing desires

Alastair Culham¹

1 Herbarium School of Biological Sciences University of Reading, Reading, U.K.

Plant taxonomy is a scientific discipline which output is used directly by a broad range of other parties. Horticultural taxonomy has a particular impact on gardeners and the horticultural trade. This has led to tension between those who dislike any change and those who update names based on new evidence of relationships. The past three decades have seen major changes in plant names and the way plants can be identified due to the development of DNA based data. Through a series of examples, the impacts of nomenclatural changes, reactions to decisions made and the lessons learned from those experiences are examined.

### S.36.6 Integrative taxonomy, insights for wild and cultivated tomatoes classification

Iris Edith Peralta¹

1 Universidad Nacional de Cuyo y Consejo Nacional de Investigaciones Científicas y Técnicas, Mendoza, Argentina.

Cultivated tomato, Solanum lycopersicum L., is a leading vegetable crop worldwide. It is also a model organism for genetics and evolutionary research. Using integrative taxonomy, a predictive classification of tomatoes and closely related groups was proposed, as a framework for further studies (Peralta et al. 2008). Wild tomato species are distributed in Colombia, Ecuador, the Galapagos Islands, Perú, Bolivia and Chile in different ecosystems. The cultivated tomato belongs to a very recently derived group and was domesticated from its wild progenitor, S. pimpinellifolium. Domestication process initiated in South America and Mesoamerica, continued in Europe when cultivated tomatoes were introduced in the sixteenth century in Italy, and later its expansion throughout the world as the species was incorporated into consumption. In the 20th century, modern breeding and the emergence of seed companies increased the development of new varieties, and more recently, commercial hybrids mainly with resistance to abiotic factors, pests and diseases, and quality traits. This long process has generated an enormous cultivar diversity due to their vegetative and reproductive growth, and fruit shapes, sizes, colors, and quality characteristics. An attempt has been made to apply a nomenclatural system based on the International Code of Botanical Nomenclature to tomatoes, but it is so complex that it has not
<figure><img src="image_98.png" /></figure>

been used. Taking into account a comprehensive
taxonomy approach, a classification based on the
International Code of Nomenclature for Cultivated
Plants is proposed, which is simple and easy to ap-
ply to describe new tomato cultivars, and potentially
useful for harmonizing names and classification of
breeding collections among genebanks. Classifica-
tion criteria must contribute to understand and use
the diversity among wild and closely related do-
mesticated species.

References: Peralta I.E., Spooner D.M., S. Knapp. 2008. Taxonomy of Wild Tomatoes and their Relatives (Solanum sect. Lycopersicoides, sect. Juglandifolia, sect. Lycopersicon; Solanaceae) Systematic Botany Monographs Vol. 84
<figure><img src="image_99.png" /></figure>

Symposia Session 3

**S.37 PLANT RESPONSES TO STRESSES AND CLIMATE CHANGE. SESSION 1**

**S.37.2 Extreme fire severity and fire frequency: evidence for state shift and generation of fire feedback loops of a mesic forest**

Thomsen. A¹, Keith. D¹, Lemmon. J², Allen.
V², Ooi. M¹

1 Centre for Ecosystem Science, School of Biologi-
cal, Earth and Environmental Sciences, University of
New South Wales, UNSW Sydney NSW, Australia. 2
Office of Climate Change, Energy, the Environment
and Water, Wollongong, NSW

Shifts in the fire regime caused by climatic and anthro-
pogenic influences mean that we are currently experi-
encing more frequent and severe fires at larger scales.
Fire is a dominant disturbance in Eucalyptus forests
in temperate Australia. Wet sclerophyll forests are of-
ten bordered by dry sclerophyll and/or rainforests and
changes to the fire regime can shift these borders by al-
lowing drier sclerophyll or rainforest species to disperse
into current wet sclerophyll forests. The Black summer
fires of 2019-20 saw around 50% of the threatened Up-
land Basalt Eucalypt forests of the Sydney Basin Biore-
gion ecological community burnt, with some areas ex-
periencing extreme severity fire. Much of this ecological
community is often disturbed and fragmented, making
it potentially less resilient to disturbance changes, such
as extreme fire events. To assess how these wet sclero-
phyll forests respond to different fire severities, we sur-
veyed areas impacted by the 2019-20 Black Summer
fires one-year later. We quantified the mortality rates of
dominant tree species including Acacia melanoxylon,
Eucalyptus blaxlandii, Eucalyptus cypellocarpa and Eu-
calyptus radiata, and compared how the dominance
of different functional groups varied across the fire
severity gradient. We have uncovered evidence that
rainforest trees within wet sclerophyll forests may be
highly susceptible to extreme fire events especially and
at sites previously subjected to frequent fires. This study
highlights the importance of understanding how sever-

ity of fire impacts wet sclerophyll forests and can inform
management decision on conserving threatened eco-
logical communities.

**S.37.3 Improving salinity tolerance in rice: lessons from Oryza coarctata**

Ping Yun¹,², Lana Shabala¹,³, Meixue
Zhou¹, Zhonghua Chen⁴, Gayatri
Venkataraman⁵, Sergey Shabala²,³

1 Tasmanian Institute of Agriculture, University of Tasmania, Hobart, Australia. 2 School of Biological Science, University of Western Australia, Perth, Australia. 3 International Research Centre for Environmental Membrane Biology, Foshan University, Foshan, China. 4 School of Science, Western Sydney University, Penrith, Australia. 5 Plant Molecular Biology laboratory, M. S. Swaminathan Research Foundation, Chennai, India

Oryza coarctata is the only halophytic relative in the
genus Oryza that is tolerant to high salinity levels. In this
work, we investigated key traits that confer this excep-
tional salt tolerance in O. coarctata, comparing it with
responses of cultivated Oryza sativa species. We show
that O. coarctata is utilizing Na⁺ for stomata operation
and, as result, is capable to maintain unaltered stomatal
related traits (size, density, and opening speed). These
plants had also enhanced RuBisCO carboxylation and
RuBP regeneration in the Calvin cycle. Taken together,
these features allowed O. coarctata maintain relative
steady CO₂ assimilation under salt conditions. Upon ex-
posure to salinity, wild rice also quickly increased xylem
Na⁺ loading for osmotic adjustment but maintained
non-toxic level of stable shoot Na⁺ concentration by
increased activity of HKT1;5 (essential for xylem Na⁺ un-
loading) and a Na⁺/H⁺ exchanger NHX (for sequestering
Na⁺ and K⁺ into root vacuoles). In contrast, cultivated
rice prevented Na⁺ uptake and transport to the shoot at
the beginning of salt treatment but failed to maintain it
in a long-term. We also show that wild rice limits pas-
sive Na⁺ entry into root cells while cultivated rice relies
<figure><img src="image_100.png" /></figure>

heavily on SOS1-mediating Na+ exclusion, with major penalties imposed by the existence of the “futile cycle” at the plasma membrane. Other traits that contributed to superior salinity tolerance in *O. coarctata* included more efficient ROS signalling; desensitization of ROS-inducible cation channels and more superior K+ retention in the cytosol; more effective control of cytosolic Ca²⁺ homeostasis by Ca²⁺-ATPase and CAX-mediated Ca²⁺ efflux systems; and ability to prevent salt-induced downregulation of *RBOH* expression that may affect operation of the “ROS-Ca²⁺ hub” and signalling cascades under salinity.

## S.37.4 Tracing the resilience of African tree ferns: insights from the African humid period and beyond

Mwihaki J. Karichu¹,²,³, Boniface K.

Ngarega⁴, Jefwa JM⁵, Bette A. Loiselle⁶,

Emily B. Sessa²

¹The Lewis B. and Dorothy Cullman Program for Molecular Systematics, New York Botanical Garden, Bronx, NY. ²New York Botanical Garden, Bronx, NY. ³Graduate Center, City University of New York, New York, NY, USA. ⁴Department of Plant Biology, Ecology, and Evolution, Oklahoma State University, Stillwater, OK, USA. ⁵Pwani University, P.O Box 195, Kilifi, Kenya. ⁶Department of Wildlife Ecology and Conservation and Center for Latin American Studies, University of Florida

Tropical forests in Africa are rapidly declining due to increasing population, recurrent wildfires, logging, land use changes, agricultural intensification, and other socioeconomic factors. This study utilized maximum entropy modeling, paleoclimatic data, and future climate scenarios to evaluate the historical presence of tree ferns in tropical and Saharan Africa during the African Humid Period (AHP; ca. 14,500-5,000 years ago) and project the impacts of climate change on their future distribution and implications for African forests. Although precipitation increased substantially during the AHP, the distribution was variable and insufficient to support all tropical plant species. We found most African tree fern species persisted in refuge areas that survived the late Pleistocene extinction. These refugia provided a haven for tree ferns, enabling them to survive despite challenging and fluctuating conditions. This highlights the remarkable resilience and adaptability of tree ferns and the critical importance of refugial areas in safeguarding populations during climatic upheaval. Our study further shows tree fern species

had differential responses to climate change, with some experiencing minimal range contractions of 5.9% up to over 57% range expansion. Preserving these refugia not only protects tree ferns but also conserves overall forest biodiversity and ecosystem functioning. This knowledge is vital for targeted conservation to promote sustainable forest management and mitigate threats from climate change and human activities in African closed wet forests.

## S.37.5 Learning from the adaptive responses of wild emmer wheat to 28 years of global warming

Yong-Bi Fu¹

¹Plant Gene Resources of Canada, AAFC Saskatoon Research and Development Centre

Global warming has been documented to threaten wild plants with strong selection pressures, but how plant populations respond genetically to the threats remains poorly understood. We characterized the genetic responses of 10 wild emmer wheat (Triticum dicoccoides Koern.; WEW) populations in Israel, sampling them in 1980 and again in 2008, through an exome capture analysis. It was found that these WEW populations were under elevated selection, displayed reduced diversity and temporal divergence, and carried increased mutational burdens forward. However, some populations still showed the ability to acquire beneficial alleles via selection or de novo mutation for future adaptation. Grouping populations with mean annual rainfall and temperature revealed significant differences in most of the 14 genetic estimates in either sampling year or over the 28 years. The patterns of genetic response to rainfall and temperature varied and were complex. In general, temperature groups displayed more temporal differences in genetic response than rainfall groups. The highest temperature group had more deleterious single nucleotide polymorphisms (dSNPs), higher nucleotide diversity, fewer selective sweeps, lower differentiation, and lower mutational burden. The least rainfall group had more dSNPs, higher nucleotide diversity, lower differentiation, and higher mutational burden. These characterized genetic responses are significant, allowing not only a better understanding of evolutionary changes in the threatened populations but also for realistic modeling of plant population adaptability and vulnerability to global warming.
<figure><img src="image_101.png" /></figure>

## S.37.6 Phenological prints of climate change on Mediterranean flora: a retrospective study using herbarium specimens

Andros Solakis Tena¹, Federico Casimiro-Soriguer¹ & Noelia Hidalgo Triana¹

1 Universidad de Málaga, Málaga, Spain

Plant phenology is linked to environmental variables. However, in the last century, the global temperature (T) has increased by 1.1 °C, which has caused severe climatic changes that have affected the phenology of Mediterranean plants. In our study, conducted across the western Mediterranean area of the Baetic Range, we used data from herbariums to assess the phenological trends of 98 dominant species from Habitats of Community Interest (Group 4, 5, and 9), along with endemic plants and the overall phenology of the bioclimatic thermotypes present in the area, all over the past 200 years. Significant changes and trends in the day of

year (DOY) were detected in the studied phenophases of 71 species, whereas 27 showed no changes. In response to the time variable, seven species displayed significant shifts in the preflowering (FBF), 17 in the flowering (F), one in the fruiting (FS), and 10 in the growth (DVG) phenophase. In response to long-term climatic variables, 16 species showed no changes in FBF, whereas 22 did so in at least one. In F, 23 showed no changes whereas 49 did; in FS, 7 did not, whereas 13 did; and in DVG, 21 did not, whereas 29 did. Regarding overall phenology DOY with time, the thermo-Mediterranean (TM) thermotype showed no changes in the FBF and FS phenophases, whereas advances appeared in F and DVG. In the meso-supra-Mediterranean (MSM) and oro-cryoro-Mediterranean (OCM) thermotypes, all phenophases showed advancing trends. The DOY against climatic variables in the TM thermotypes showed advances in FBF and F, while FS showed no changes, and DVG advanced when P decreased. In the MSM and OCM thermotypes, all phenophases showed trends of advance. Tendencies were stronger in the orophilic than thermophilic group. Those results show the influence of climate change in the phenology of western Mediterranean flora.

## S.38 DISSECTING THE EVOLUTION OF SEDGES (CYPERACEAE): PERSPECTIVES FROM MULTIPLE LINEAGES

### S.38.1 The evolution and diversity of the genus Bulbostylis (Abildgaardieae, Cyperaceae)

Jeremie Morell¹², Fitiavana Rasaminirina³,⁶, Juliene Maciel-Silva¹,⁴,⁵, Vonjison Rakotoarimanana³, Vincent Savolainen², André S. B. Gil⁵ & Isabel Larridon¹,⁷

1 Royal Botanic Gardens, Kew, Richmond, UK. 2 Imperial College London, London, UK. 3 University of Antananarivo, Antananarivo, Madagascar. 4 Federal Rural University of Amazon, Belém, Brazil. 5 Museu Paraense Emílio Goeldi, Belém, Brazil. 6 Kew Madagascar Conservation Centre, Antananarivo, Madagascar. 7 Ghent University, Department of Biology, Systematic and Evolutionary Botany Lab, Gent, Belgium

The genus *Bulbostylis* (hairsedges) includes 227 species worldwide. The taxonomic history of this genus is quite complex, and limited DNA data is available. This lack of data means that species delimitation and evolutionary relationships are unclear, and thus the study of character evolution and biogeographical history of *Bulbostylis* is hampered. Thus, three PhD projects are currently devoted to this genus, working on three centres of diversity of *Bulbostylis*: Africa, Brazil and Madagascar. These different regions respectively include 127, 52 and 25 accepted species of *Bulbostylis*, representing around 90% of the total diversity of the genus. Consequently, the aim of these projects is to gain a better understanding of the diversity, evolution, and conservation of *Bulbostylis* on a large and local scale. To this end, a joint project is underway to reconstruct the evolutionary history of the genus as well as its biogeographical history using a phylogenomic approach. Secondly, integrative taxonomic studies at local
<figure><img src="image_102.png" /></figure>

scale are ongoing to clarify species delimitation based on phylogenomic data with extensive sampling of species complexes, (micro)morphological and anatomical data; and distribution data. Preliminary results for these regions include: thirteen herbaria visited, ten fieldtrips undertaken, discovery of new species to science (e.g. *Bulbostylis itremoensis* from Madagascar), new herbarium specimens and DNA samples of Africa, Neotropical and Madagascan species, discovery of new anatomical characters for species delimitation; 212 newly extracted DNA samples sequenced with the Angiosperms353 targeted sequencing probes. Our study generates a large, accurate dataset. The future integration of this information may contribute to demonstrate patterns of diversity that allow the identification of important areas for conservation studies and provide new insights on ecological and evolutionary understanding in areas of high biodiversity.

## S.38.2 Towards the consolidation of the Southeast Asian sedge knowledge

José Ignacio Márquez-Corro¹, Pedro Jiménez-Mejías², Millard Uy³, David A. Simpson¹ & Isabel Larridon¹

1 Royal Botanic Gardens, Kew, Richmond, UK. 2 Universidad Pablo de Olavide, Seville, Spain. 3 Independent researcher, Cebu, Philippines

Studies on global biodiversity are detecting common gaps in the taxonomic knowledge of tropical areas. While some of these areas are not necessarily considered hotspots, these are still incredibly diverse, and the knowledge gap extends widely to non-woody plants as they tend to be out of focus in regions dominated by tropical forests, which may underestimate diversity levels. For Cyperaceae, the third most biodiverse monocot family and among the top ten in angiosperms, Southeast Asia is the perfect example of such a situation. Despite having diversified worldwide into more than 5000 species with a particular preference for temperate areas in the largest genus *Carex* (40% of the family), there are over 600 taxa native to Southeast Asia. Nevertheless, the lack of historical collections, digitization of herbaria, and lack of support in Cyperaceae expertise for local research groups have turned the region into a darkspot for sedges. To protect current sedge diversity, understand their natural history,

and predict their future, we must put forward a set of international collaborations with in-country researchers to improve the available information for this economically and ecologically important family. We hope to start a much-needed new era of collaborations in Southeast Asian sedge-focused projects, aiming to generate taxonomic, geographical, morphological, molecular, genomic, biochemical, and ecological data, accompanied by photographic records of taxa and sets of data that will be publicly available. Objectives such as training resident taxonomists, digitization of local herbaria, or providing international long-term financial support are some of the crucial steps towards the establishment of sedge research lines in Southeast Asia.

## S.38.3 Progress on the phylogeny and evolution of Fuireneae s.l. (Cyperaceae)

Cassandra D. V. Bradshaw¹, Étienne Léveillé-Bourret², Isabel Larridon³,⁴, Robert F. C. Naczi⁵, Jeremy Bruhl⁶, Pedro Jiménez-Mejías⁷, Karen Wilson⁸, Julian R. Starr¹

1 University of Ottawa, Ottawa, Canada. 2 University of Montréal, Montréal, Canada. 3 Royal Botanic Gardens, Kew, Richmond, UK. 4 Ghent University, Ghent, Belgium. 5 New York Botanical Garden, New York, United States of America. 6 University of New England, Armidale, Australia. 7 Pablo de Olavide University, Seville, Spain. 8 Royal Botanic Gardens and Domain Trust, Sydney, Australia

Considerable changes to the classification of the large (ca. 5500 spp.), cosmopolitan monocot family Cyperaceae (“sedges”) have been made over the past 20 years. This is largely due to the reconstruction of relationships using molecular data. Nonetheless, many groups remain poorly studied, such as tribe Fuireneae s.l., a collection of ca. 153 spp. common to humid and wet habitats worldwide. The group possesses numerous annuals and perennials, and holds ecological and economic importance, often dominating their natural habitats and acting as noxious crop weeds worldwide. Though a recent molecular study found support to divide Fuireneae s.l. into four tribes and six genera, only 30% of the tribe’s diversity was sampled, sampling was poor where the group is most diverse (Asia), species level-complexes remain unresolved, and tribes could only be divided using impractical microscopic characters. Interestingly, this preliminary data suggested transitions from an annual to a perennial life history strategy,
<figure><img src="image_103.png" /></figure>

for which few examples are known in angiosperms, especially in monocots. Using a complete taxonomic sample of Fuireneae s.l., and molecular (Hyb-Seq), morphological and embryo characters, we are testing whether current tribal, generic and infrageneric classifications of Fuireneae s.l. are natural, and if some perennials in the grade are derived from annuals as suggested by preliminary data.

## S.38.4 The relevance of expert curation in taxonomically complex groups: a comprehensive database of occurrences for the genus Carex

María Sanz-Arnal¹, Pablo García Moro¹, Carmen Benítez Benítez², Marina Coca-de-la-Iglesia³, Angélica Gallego-Narbón³, Consolación Barciela¹, Fabrizio Bartolucci⁴, Matthew Bradley⁵, Asunción Cano⁶, Antoine Derouaux⁷, Sabina Donadío⁸, Marcial Escudero², Gabriele Galasso⁹, Sebastian Gebauer¹⁰, Mariela Fabbroni¹¹, Kerry A. Ford¹², M. Socorro González-Elizondo¹³, David Hamon¹⁴, Matthias H. Hoffmann¹⁰, Xiao-Feng Jin¹⁵, Jacob Koopman¹⁶, Bangze Li¹⁷,¹⁸,¹⁹, Raúl Lois ³,²⁰, Yi-Fei Lu¹⁵,²¹, Modesto Luceño¹, Santiago Martín-Bravo¹, José Ignacio Márquez-Corro¹, Attila Mesterházy²², Mónica Míguez¹, Ana Morales Alonso¹, Muthama Muasya²³, Paulo Muñoz-Schüler²⁴, Robert F.C. Naczi²⁵, Nora Oleas²⁶, Luciana Pereira-Silva²⁷, Radomír Řepka²⁸, Anton A. Reznicek²⁹, Katie K. Sanbonmatsu⁵, Enmily Sánchez²⁶, Daniel Spalink⁵, Arne Strid³⁰, Pieter Vanormelingen³¹, Filip Verloovee³², Karen L. Wilson ³³, Shuren Zhang¹⁷,¹⁸, Pedro Jiménez-Mejías¹

1 Universidad Pablo de Olavide, Seville, Spain. 2 Universidad de Sevilla, Seville, Spain. 3 Universidad Autónoma de Madrid, Madrid, Spain. 4 Centro Ricerche Floristiche dell'Appennino, Barisciano, Italy. 5 Texas A&M University, College Station, Texas, USA. 6 Universidad Nacional Mayor de San Marcos,

Museo de Historia Natural y Facultad de Ciencias Biológicas, Lima, Perú. 7 Natagora asbl, Namur, Belgium. 8 Instituto de Botánica Darwinión (CONICET-ANCEFN), Buenos Aires, Argentina. 9 Museo di Storia Naturale di Milano, Milano, Italy. 10 Martin Luther University Halle-Wittenberg, Geobotany and Botanical Garden, Halle (Saale), Germany. 11 Universidad Nacional de Salta, Salta, Argentina. 12 Allan Herbarium (CHR), Manaaki Whenua, Landcare Research, Lincoln, New Zealand. 13 CIDIR, Instituto Politécnico Nacional, Durango, México. 14 Independent Researcher, Lavau-sur-Loire, France. 15 Zhejiang Agricultural and Forestry University, Lin'an, Zhejiang, China. 16 ul.Kochanowskiego, Choszczno, Poland. 17 Institute of Botany, Chinese Academy of Sciences, Beijing, China. 18 China National Botanical Garden, Beijing, China. 19 University of Chinese Academy of Sciences, Beijing, 100049, China. 20 Universidad de León, Campus de Vegazana s/n, 24007, León, Spain. 21 College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China. 22 Independent Researcher, Celldömölk, Hungary. 23 University of Cape Town, Cape Town, South Africa. 24 Herbario CONC, Universidad de Concepción, Concepción, Chile. 25 New York Botanical Garden, Bronx, NY 10458, USA. 26 Universidad Indoamérica, Machala y Sabanilla, Quito, Ecuador. 27 Universidade Federal de Santa Catarina, Campus Trindade, Santa Catarina, Brazil. 28 Mendelova univerzita v Brně, Brno, Jihomoravský, Czech Republic. 29 University of Michigan Herbarium, Michigan, USA. 30 Bakkevej 6, Ørbæk, Denmark. 31 Natuurpunt Studie vzw, Mechelen, Belgium. 32 Meise Botanic Garden, Meise, Belgium. 33 National Herbarium of New South Wales, New South Wales, Australia

Species occurrences are essential for research requiring spatially-based information. The Global Bio-diversity Information Facility (GBIF) is the primary and most popular source of occurrences. However, species identification and georeferencing data in GBIF is often affected by imprecision. Accordingly, a curation process is needed to prevent downstream errors, biases and/or inaccuracies derived from their use. Because of its high species diversity, taxonomic complexity, and wide distribution, occurrence data from Carex L. (Cyperaceae) are not exempt from these errors. In this context, we present an expert-curated global compilation of Carex occurrences mined from different sources (online databases, herbaria, literature, and fieldwork collections). Beyond basic quality-control procedures, the dataset was reviewed by experts, who curated both geographical inaccuracies and misidentifications. The final expert-curated database contains 397,239 records. During the curation, 16,604 occurrences were flagged as potentially erroneous, and 7,672 taxa names were changed. The resulting dataset has occurrences for 1,679 Carex species (c. 80% of all currently accepted species). Finally, we compared the differences between
<figure><img src="image_104.png" /></figure>

the raw and the expert-curated database. Our work highlights the importance of high-quality taxonomic and geographic data and provides a significantly enhanced *Carex* occurrence database that is made available for future research in the genus.

**S.38.5 Niche partitioning or divergence? Sister-lineage comparisons reveal complex patterns of niche evolution in Carex**

Daniel Spalink¹

1 Texas A&M University, College Station, Texas, USA

With over 2000 species, *Carex* (Cyperaceae) is a nearly cosmopolitan genus with centers of diversity in temperate zones of each hemisphere. *Carex* exhibits tremendous ecological diversity, occupying habitats from coasts to mountaintops, deserts to wetlands, tundra to the tropics, and savannas to forest under-stories. Divergence across these ecological gradients has long been hypothesized to be a key driver of diversification in *Carex*. This hypothesis has been supported by phylogenetic comparative analyses, which show strong relationships between rates of speciation and niche evolution across the genus. These analyses to date have been based almost exclusively on the estimations of species niches from occurrence records, which are known to contain biased, missing, and incorrect data. Potentially more pernicious problems, however, are that 1) species distributions are dynamic through time; and few species likely 2) occupy the full extent of their fundamental niches or 3) occur in all geographic areas that are ecologically suitable. Consequently, significant findings of niche evolution in *Carex* may be spurious. Here, we use emerging methods that avoid the assumption that present distributions represent the full occupancy of species potential ranges. For nearly 500 sister-species pairs, we perform the Niche Overlap Test, which calculates niche similarity between two species in their total occupied environmental space, and the Niche Divergence Test, which calculates niche similarity in the subset of environmental space that is shared and accessible to both species. These tests are designed to detect whether species occupy distinct niches or simply have access to different environments. We then test whether significant niche evolution between sister species is related to their time since divergence, trait divergence, latitude, and total range size.

**S.38.6 Tackling rapid radiations – update on the understanding of the megadiverse genus Cyperus (Cyperinae, Cyperaceae)**

Isabel Larridon¹, Tamara Villaverde², Luciana Pereira-Silva³, Martin Xanthos¹, Megan Griffiths¹, Nina Münder⁴, Abubakar Bello⁴,⁵,⁶, Alexandra N. Muellner-Riehl⁴,⁵, Muthama Muasya⁷

1 Royal Botanic Gardens, Kew, Richmond, UK. 2 Universidad Rey Juan Carlos, Madrid, Spain. 3 Universidade Federal de Santa Catarina, Florianópolis, Brazil. 4 Leipzig University, Leipzig, Germany. 5 German Centre for Integrative Biodiversity Research, Halle-Jena-Leipzig, Leipzig, Germany. 6 Umaru Musa Yaradua University, Katsina, Nigeria. 7 University of Cape Town, Cape Town, South Africa

*Cyperus* or flatsedges represent the second most species-rich genus of Cyperaceae with almost 1000 species distributed mainly in the tropics and subtropics. Historically, many segregate genera were split from *Cyperus* sensu stricto to accommodate the breadth of morphological, functional and ecological variation present in this group. Over the last 15 years, new insights into the evolution, diversity and distribution of this group were gained by integrating molecular phylogenetic data with in-depth morphological and anatomical studies. In the current phylogenomics-based classification, subtribe Cyperinae is monogeneric, placing the 13 segregate genera previously recognised in the morphology-based classification within a broadened circumscription of *Cyperus*. We also have strong evidence that a biodiverse clade of species using the C4 photosynthetic pathway is nested within a grade of C3 species. However, rapid diversification has hampered our understanding of the relationships within the C4 clade. Now, with the advent of phylogenomics, first inroads are made to study trait evolution in *Cyperus*. Yet, to build a complete picture of this megadiverse genus, a concerted effort will be needed to generate a species-level phylogenomic framework to investigate its evolutionary history, generate an infrageneric classification, and resolve species limits. We present the current state of knowledge and explore plans for integrating efforts towards purging the gaps.
# S.39 PLANT CONSERVATION GENETICS: FROM IN-SITU AND EX-SITU CONSERVATION TO REINTRODUCTIONS AND RESTORATIONS.
## SESSION 1

### S.39.1 Genomic-guided conservation actions to restore the most endangered conifer in the Mediterranean Basin

José Carlos del Valle¹, Montserrat Arista¹, Carmen Benítez Benítez¹, Pedro Luis Ortiz¹, Francisco J. Jiménez-López¹, Anass Terrab¹, Francisco Balao¹

1 Departamento de Biología Vegetal y Ecología, Universidad de Sevilla, Sevilla, Spain

Species with extremely small population sizes are critically endangered due to reduced genetic diversity, increased inbreeding, and the added threat of hybridization. Genomic tools significantly advance conservation by revealing genetic insights into endangered species, notably in monitoring frameworks. Sicilian fir is the most endangered conifer in Europe with only 30 adult trees spread across an 84-hectare area. Using 20,824 SNPs from RAD-seq employing the silver fir genome assembly and a custom 120 SNP-array, we evaluated genetic diversity, mating patterns, and effective population size in adult trees, 118 natural seedlings, and 2,064 nursery seedlings from past conservation actions. We assessed introgression from neighboring non-native fir plantations and established an intra-population assisted gene flow program selecting the most genetically dissimilar individuals and investigating the outcome through simulations. Genomic analysis unveiled significant genetic diversity among adult Sicilian firs, comparable to non-endangered Mediterranean firs with larger populations. However, the genetic diversity of the forthcoming generation declined due to high self-fertilization, leading to marked inbreeding (FIS = 0.38) and an alarmingly low effective population size (Ne = 6). Nursery seedling monitoring revealed similar selfing rates

but significant introgression (~50%) from non-native firs. Although intra-population assisted gene flow could help to mitigate genetic loss, it may not alleviate the species vulnerability to imminent environmental challenges, perpetuating the risk of an extinction vortex. Hence, investigating the impact of Sicilian fir population decline and selfing on inbreeding depression, along with exploring the potential of hybrids for genetic load alleviation and future adaptation, is crucial for effective conservation strategies. This study stands as a compelling model for guiding conservation strategies in similarly imperiled species characterized by extremely small populations.

### S.39.2 Plant-animal interactions, population genetics and local community engagement to conserve the Malagasy baobab Adansonia suarezensi

Onja Hariveloniaina Morilline
Razanamaro¹, Tonisoa
Ranaivoandriamanantena², Laura
Parducci³, Fabio Attorre³, Elisa Rondoni³,
Dario La Montagna ³, Alex Byrne⁴, Barbara
Gravendeel⁵, Ester Sales⁶, Richard
Dominique Randriatsitohaina⁷, Lalaina
Fenosoa Ramiliarisona⁷, Juan Viruel⁴,

1 Parc Botanique et Zoologique de Tsimbazaza Rue Fernand Kassanga, Tsimbazaza, Antananarivo. Madagascar. 2 Madagasikara Voakajy, Andraiso-ro BP 5181 Antananarivo, MG. 3 Department of Environmental Biology, Sapienza University of Rome, Piazzale Aldo Moro 5, Rome, Italy. 4 Royal Botanic Gardens, Kew, Richmond, UK. 5 Naturalis Biodiversity Center, Endless Forms, Leiden, Netherlands and Institute of Biology Leiden, Leiden University, Leiden,
Netherlands. 6 Agrarian and Environmental Sciences Department, Institute of Environmental Sciences, University of Zaragoza, High Polytechnic School, Huesca, Spain. 7 Département biologie et écologie végétales, Université d'Antananarivo, Madagascar

Adansonia suarezensis, one of the six endemic baobabs in Madagascar, is currently classified as Endangered. Its narrow distribution in the arid areas of N Madagascar is expected to decrease from 1200 km² in 2010 to 17 km² in 2050 due to climate change, and alarming predictions indicated a likely extinction between 2050 and 2080 without intervention or conservation efforts. Madagascar has lost 37% of its natural forest since 1973, and despite the implementation of conservation programs, deforestation rates have increased in the last decade, mainly because population growth and political instability in the country. Ex situ and in situ conservation actions to recover threatened species require extensive knowledge from diverse disciplines to better understand the reproduction biology, ecology and demography of the target species. *Adansonia suarezensis* belongs to a section named Brevitubae, which include dry-season flowering baobabs with white flowers carrying short-staminal tubes pollinated by fruit bats and lemurs. Recently, a lack of natural regeneration has been observed in natural populations of *A. suarezensis*. In our project, we investigate the life cycle of *A. suarezensis* to reveal factors influencing the natural regeneration of the species, and we designed a restoration plan based on population genomics data by using a whole genome Single Nucleotide Polymorphism (SNP) screening approach (MigSeq). We explore also new plant-animal interactions with fruits, seeds and flowers using camera traps and direct observations. The aim is to elucidate the genetic patterns, investigate the level of gene flow, and explore relationships between population size and genetic variation in the species. The analysis of genetic diversity and population structure allow us to identify priority populations for restoration. Our project demonstrates also the importance of local community engagement in conservation by building nurseries, growing thousands of seedlings of baobabs and other economically plants as part of the conservation plan.

### S.39.3 The value of record-and molecular-based conservation management to exceptional species: a case study using Amorphophallus titanum

Justin Tutt¹, Jeremie B. Fant², Kayri Havens², Olivia G. Murrell³,⁴, Zoe Diaz Martin⁵

1 University of Central Florida, Orlando, USA. 2 Chicago Botanic Garden, Glencoe, USA. 3 Manchester Metropolitan University, Manchester, UK. 4 North of England Zoological Society, Chester Zoo, Chester, UK. 5 Spelman College, Atlanta, USA

If ex situ collections are to effectively conserve species, practitioners should strive to maintain genetically robust ex situ populations by (1) attaining a comprehensive understanding of the current estimates of kinship and other relevant population genetic measures and (2) making informed breeding decisions. The zoological community accomplishes these goals by using a combination of a record-based approach, where extensive documentation of mating between individuals is recorded and used to generate pedigrees, and a molecular-based approach, where individuals are genotyped to reconstruct the pedigree or fill in pedigree gaps. This raises the questions: how well do botanic gardens manage populations of exceptional plant species held across collections, and what lessons on how to manage collections can we learn and implement from the zoological community? We explore these questions and evaluate the utility of the zoo model for conservation using the characteristic arum *Amorphophallus titanum*, an exceptional species native to the Indonesian island of Sumatra that has been cultivated for over 130 years. In this study, we used available accession information to create a pedigree and genomic sequencing to generate a SNP-based molecular dataset. We present our anticipated results that will compare relevant population genetic measures and pairwise kinship estimates and their subsequent recommendations of optimal breeding pairs using the record- and molecular-based datasets. We expect to observe differences between kinship coefficients calculated from each dataset and predict that the molecular-based approach will provide higher precision given notable incompleteness in accession records.
To our knowledge, this is the first study to compare the record- and molecular-based approaches of managing exceptional species in botanic gardens. This work will be an important first step in improving collections management (i.e., standardizing the recording of accession data) and will highlight genetics as an effective tool when managing poorly pedigreed exceptional species.

highly degraded mining site. Our studies suggest that vulnerability cannot be easily predicted for co-distributed species, implying the need to obtain species-specific estimates to assess the extinction risks and inform decision-makers for conservation efforts. Based on our studies, we discuss the main challenges in reconciling conservation with mining activities in unique and highly diverse environments.

## S.39.4 Conservation genetics of rare and endemic plants: challenges and advances

Carolina S. Carvalho¹, Bárbara S.S. Leal¹, Jacqueline S. Lima¹, Carlos Eduardo Batista¹, Rafael Assis¹, Mauricio Watanabe¹, Cecílio Caldeira¹, Valéria Tavares¹

1 Instituto Tecnológico Vale, Belém, Brazil

Endemic species with limited geographic ranges are more prone to the negative effects of genetic drift and their consequences, especially when their habitat overlaps with economically valuable areas. This is the case of endemic plants associated with the Eastern Amazon cangas, a unique ecosystem limited by high iron concentration in the soils, which faces substantial threats from some of the world's largest iron ore mines. The conservation challenges in this ecosystem arise because just as plants adapted to iron-rich soils exhibit non-random distribution, often clustering in specific microclimatic and edaphic conditions; iron mining activities cannot also be haphazard due to variations in iron ore presence and quality. In response to this conservation dilemma, we have conducted comprehensive studies, integrating ecology and conservation and landscape genomics. By investigating the neutral genetic variation, we have found a scenario of conservation concern for three highly restricted endemic species, with simulations indicating a worsening of such scenario in a few decades if population sizes further decline. Conversely, certain species have demonstrated remarkable resilience, with genetic diversity and gene flow patterns remaining unaffected by mining-induced habitat loss. The use of genotype-phenotype-environment associations has offered further insights into adaptive genomic variation for restoration purposes. For example, our findings indicate that whereas local provenances were found optimal to restore a moderately disturbed site, a mixture of genotypes seems to be the most promising strategy for the recovery of

## S.39.5 Intraspecific genome size variation, cryptic speciation and outbreeding depression in Penstemon pachyphyllus

Rafael Urbina-Casanova¹,², Maria Giro¹,², Jeremy Foster¹,², Bryce Richardson³, Scott Jensen³, Jeremie B. Fant¹,² & Andrea T. Kramer¹,².

1 Plant Biology and Conservation, Northwestern University, Chicago, IL, USA. 2 Negaunee Institute for Plant Conservation Science and Action, Chicago Botanic Garden, Glencoe, IL, USA. 3 Rocky Mountain Research Station, USDA Forest Service, USA.

Outbreeding depression is a main concern when mixing multiple source populations in restoration, as it can reduce seed quantity and quality in seed production fields and restoration sites. The main causes for outbreeding depression are unnoticed taxonomic issues and fixed chromosomal differences, but these aspects are rarely assessed when designing and implementing a restoration project. Penstemon pachyphyllus was identified as a restoration priority in the Great Basin, where the Great Basin Restoration Initiative took a regional admixture provenancing approach, combining as many as 18 source populations in pooled seed production fields, to generate large quantities of genetically diverse seeds for wide-scale restoration across this region. However, outbreeding depression was previously reported for this species when mixing populations from two different ecoregions. Because this species extends beyond the Great Basin, we assessed genome size variation across the whole species range to study potential causes of this pattern of outbreeding depression. As we found a geographically structured pattern of genome size variation, with southern populations showing a smaller genome size, we designed a controlled crosses experiment to test if genome size difference is the main driver of outbreeding depression. Moreover, we took a phylogenomic approach to assess species delimitation on
<figure><img src="image_105.png" /></figure>

our target species including all closely related taxa (i.e., the Coerulei-Gentianoides clade). We found that genome size difference is the main driver of outbreeding depression in this species, with crosses between populations with different genome sizes showing a significantly lower seed production in terms of number and weight, than crosses between populations in different geographic regions but with the same genome size. Species delimitation analyses are consistent with this incipient reproductive barrier.

**S.39.6 Using assisted gene flow to save a population from depression**

Léa Auclair¹ & Olivier Brisset¹, José Utgé²,
Nathalie Machon¹, Paul Verdu²

1 CESCO, MNHN, CRNS, Sorbonne-Université, Paris, France. 2 Eco-anthropology, CNRS, MNHN, Université Paris Cité, Paris, France

After the drastic decrease of population sizes of *Arenga grandiflora*'s population in the Parisian region in the late 20st century, a translocation program was implemented in 1999. A preliminary study revealed that the main cause of the population's extinction was the low genetic diversity and the associated low fitness of the last individuals due to inbreeding depression. To save this species from extinction in this region,

non-local and local plants were multiplied by *in vitro* culture and introduced in three sites of their natural area. The three translocated populations were funded with the same mixture of 450 individuals, 2/3 being clones of local plants and 1/3 of clones from non-local ones. To evaluate the outcome of the program, annual monitoring of plant's fitness was carried out over 24 years, coupled with genetic analyses based on 13 microsatellite markers. First results highlighted that the newly-created populations showed an increase in genetic diversity compared to the founders, and displayed lower consanguinity due to higher heterozygosity level. These levels of diversity appear to be fairly stable from year to year, which was encouraging for the program's success. Interestingly, fluctuation in fitness were observed between individuals depending on the proportion of genome inherited from each founder population. To study this model in greater detail and gain a better understanding of the complex genetic processes that condition the success of programs based on assisted gene flow, we are currently resequencing the entire genomes of individuals sampled between 1999 and the present day. By comparing simulations to actual genotypic data obtained from whole genome sequencing, it will be possible to assess whether natural selection phenomena linked to crossbreeding, such as heterosis or hybrid depression, have influenced the patterns of genetic diversity and how introduction of genotypes drove the genetic evolution of the population.

**S.40 XYLEM ANATOMY AS A CENTRAL HUB LINKING FLUID TRANSPORT PROCESSES FROM THE INDIVIDUAL PLANT LEVEL TO ECOSYSTEMS**

**S.40.1 The role of drought on the evolution of woodiness in flowering plants**

Frederic Lens¹,², Timo Conradi³, Renske E. Onstein¹,⁴, Helge Bruelheide⁴,⁵, Alex Zizka¹,⁶

1 Naturalis Biodiversity Center, Leiden, The Netherlands. 2 Institute of Biology Leiden, Leiden University, Leiden, The Netherlands. 3 Plant Ecology, Faculty of Biology, Chemistry and Earth Sciences, Bayreuth University. 4 German Centre for Integrative Biodiversity Research Halle-Jena-Leipzig, Leipzig, Germany. 5 Institute of Biology/Geobotany and Botanical

Garden, Martin Luther University Halle-Wittenberg, Halle (Saale), Germany. 6 Department of Biology, Philipps-University Marburg, Marburg, Germany

Wood formation is a fundamental evolutionary innovation that gave rise to countless woody species (trees and shrubs), offering crucial ecosystem services such as carbon storage and its associated climatic feedback. Most of these species only have woody ancestors, whereas other lineages have independently evolved woodiness from non-woody (herbaceous) ancestors throughout evolutionary history. However, what drives species to become woody and why have so many unrelated herbaceous lineages evolved to new woody spe-
<figure><img src="image_106.png" /></figure>

cies? These key questions have baffled scientists since Darwin, but scientific information has been too fragmented to develop hypotheses that could be rigorously tested. Therefore, we have compiled a unique woodiness database in flowering plants, showing that most woody species with herbaceous ancestors occur in regions with recurrent drought cycles. This has laid the foundation for a novel hypothesis, proposing drought as the major driver of most transitions towards woodiness. We are testing this concept on islands and continents (1) by inferring via modelling whether there is a positive correlation between woodiness and drought, and (2) by assessing via experiments whether woody species with herbaceous ancestors are consistently more drought-tolerant than their herbaceous relatives. By bridging evolutionary biology and ecophysiology, this work shows increasing evidence for a novel strategy that reveals how plants respond to drought stress in a world facing global change.

## S.40.2 Assessing drought resistance in a population of cabbage (Brassica oleracea) with contrasting levels of stem woodiness

Anai Pereira¹,², Frederic Lens¹,²

1. Naturalis Biodiversity Center, Leiden, The Netherlands. 2. Leiden University, Leiden, The Netherlands

As the world's human population continues to grow and droughts become more intense and frequent, monitoring drought-responsive anatomical and ecophysiological traits in crops is crucial to guarantee food security in the future. *Brassica oleracea* is a relevant crop species with cultivars mostly consumed as vegetables. In our research, we crossed a woody accession of *B. oleracea* native to the island of Jersey- commonly known as Jersey cabbage - with an herbaceous, rapid flowering accession TO1000 of the same species. Five F2 genotypes were selected based on their contrasting levels of stem woodiness, along with the two grandparent lines. To evaluate whether stem woodiness is involved in the mechanisms leading to drought tolerance in cabbage, we monitored a broad range of key drought-responsive anatomical and ecophysiological traits under well-watered conditions and during con-

trolled drought. Examples of these traits are stem woodiness, pit membrane thickness, leaf water potential, leaf turgor loss point, embolism resistance in stems and leaves, hydraulic safety margin, and stomatal conductance. We hypothesize that (1) the more woody genotypes are more drought tolerant, (2) the more drought-tolerant genotypes use a coordinated set of anatomical and ecophysiological traits in both stems and leaves to reach a certain level of drought tolerance, and (3) these trait syndromes may differ among the genotypes studied.

## S.40.3 The vulnerability to drought-induced embolism-vessel diameter link: what needs explaining, and possible ways forward

Patricia Rivera¹, Tommaso Anfodillo², Mark E. Olson³

1. Unidad de Recursos Naturales, Centro de Investigación Científica de Yucatán, A.C., Mérida, Mexico. 2. Department Territorio e Sistemi Agro-Forestali, University of Padova, Legnaro, Italy. 3. Instituto de Biología, Universidad Nacional Autónoma de México, Mexico

The escalating global tree mortality crisis, with millions of trees succumbing to drought annually, impacts biodiversity conservation. The vulnerability of xylem conduits to drought-induced embolism is a critical factor in determining tree survival during hydric stress. If narrower xylem conduits are less vulnerable to embolism formation, then this vulnerability-diameter link would explain numerous well-established patterns. These include wider xylem conduits in moist environments and narrower conduits in drier environments, the consistent production of conduits that are well below the developmental possible maximum diameter of 700 µm+, widening of the conduits from the tip to the base of the plant, in a way that keeps conductance optimal with height growth, the distribution of wide to narrow conduits in earlywood to latewood, and the differential leaf shedding between plants with wider and narrower conduits. All of these patterns require explanation, and if a vulnerability-diameter link is rejected, alternative explanations are required. As part of a possible explanation for these patterns, we provide data suggesting that the vulnerability-diameter link
could involve the association between interves-
sel pit membrane thickness, vessel wall thickness,
and vessel diameter. We show ways in which the
vulnerability-pit membrane-wall thickness-vessel
diameter associations could account for the link
as well as why it is not found in some situations.

S.40.4 Developmental
trajectories of the xylem
vessel elements from
single-cell sequencing data

Philip Shushkov¹

1 Department of Chemistry, Indiana University, Bloomington, USA

Changes in transcriptional profiles are at the
heart of development. Understanding transcrip-
tional dynamics at the single-cell level is a cen-
tral challenge in deciphering developmental pro-
cesses. Current sequencing technologies allow
the estimation of transcription profiles at unprec-
edented single-cell resolution but the inference
of developmental progressions of cells is ham-
pered by sampling sparsity, technological noise,
and loss of spatial and temporal information.
The xylem cell populations have distinct tran-
scriptional profiles and are ideally suited to re-
construct transitions in transcriptional programs
during histogenesis. I use noise-resilient diffusion
mapping-based approaches to infer and reca-
pitulate developmental trajectories from large
single-cell datasets. Our new methods allow us to
reconstruct trajectories that occur on substan-
tially different timescales in development and
infer waves of gene expression changes along
the developmental progressions. We analyze the
development of vessel elements and compare it
to the development of other vascular cell types,
including sieve elements and companion cells, in
different plant organs. We recover known regula-
tors of vascular development that follow expect-
ed developmental dynamics based on previous
studies and identify putative regulatory factors.
We identify waves of gene expression changes
of transcription factors and their downstream
targets, which provide substantial resolution of
transcriptional dynamics in vascular cell devel-
opment.

S.40.5 Root pressure in
tomatoes slightly repairs
daily embolism at well-
watered conditions, but
its role during drought is
inconsistent

Giovanni Bortolami¹, Kathy Steppe², Jan
Van den Bulcke³,⁴, Salma Balazadeh⁵,⁶, Iris
Frederiks¹, Toon Gheyle³,⁴, Pierre Kibleur³,⁴,
Max Larter¹,⁷, Liselotte de Ligne³,⁴, Ajaree
Thonglim¹, Seppe Top², Joris Van Acker³,⁴,
Rune Vanbeylen², Brechtje van Helden¹,
Frederic Lens¹,⁶

1 Naturalis Biodiversity Center, P.O. Box 9517, RA Leiden, The Netherlands. 2 Laboratory of Plant Ecol- ogy, Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium. 3 UGent-Woodlab, Department of Envi- ronment, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium. 4 UGent Centre for X-ray Tomography (UGCT), Ghent University, Ghent, Bel- gium. 5 Max-Planck-Institute of Molecular Plant Physiology, Potsdam, Germany. 6 Institute of Biolo- gy Leiden (IBL), Leiden University, Leiden, The Neth- erlands. 7 University of Bordeaux, INRAE, BIOGECO, Pessac, France.

Inside plants, water moves from the roots up to the
leaves due to a gradient in negative pressure (i.e.,
tension) generated in the leaf mesophyll cells via
transpiration. During intense drought periods, the
tension in the xylem sap increases, leading to gas
bubbles (embolisms) inside the water-conducting
cells (i.e., xylem vessels), thereby obstructing the
long-distance water flow. Tomato plants are known
to also move water through a positive pressure gen-
erated in the roots. We hypothesize that root pressure
could help tomato plants remove gas embolism af-
ter release from drought after re-watering. There-
fore, we used a gantry-based X-ray microCT imag-
ing system, the EMCT scanner at the UGCT (UGent
Center for X-ray Tomography, www.ugct.ugent.be),
to visualize plant re-hydration and possible embo-
lism repair by scanning the plants before and after
drought, and at different times after re-watering.
In total, we followed 25 control well-watered plants
and 36 drought-stressed plants (covering a wide
range of drought intensities). Our results show that
all well-watered plants generated root pressure and
partially repaired the few embolized vessels over-
night. The role of root pressure in plants subjected
<figure><img src="image_107.png" /></figure>

to drought is more ambiguous: embolism repair af-
ter re-watering was also observed, but only in the
drought-stressed plants that developed root pres-
sure after re-watering (30%), while no root pressure
was detected in 70% of the drought-stressed plants
after re-watering. These results show that root pres-
sure development is not an efficient mechanism of

tomato plants' recovery from drought but shows the
ability of this embolism-vulnerable species to re-
move the first embolism events generated during
the day at well-watered conditions. In conclusion,
we show that root pressure in tomatoes can remove
the limited daily embolism under well-watered con-
ditions and only occasionally after drought.

# S.41 APPLICATIONS OF SPECTRAL REFLECTANCE IN PLANT SYSTEMATICS AND EVOLUTION

S.41.1 The evolution of
species specific spectra
in a diverse genus of
Neotropical trees (Guatteria,
Annonaceae) using
herbarium specimen

Paola G. Santacruz Endara¹,², Carlos
Rodrigues-Vaz², Denis Bastianelli³, Laurent
Bonnal³, Roy R.H. Erkens⁴, José Eduardo
Meireles⁵, Lily Bennett², Thomas L.P.
Couvreur¹,²

1 Pontificia Universidad Católica del Ecuador, Qui-
to. Apartado postal: 17012184. 2 DIADE, Université de
Montpellier, CIRAD, IRD, Montpellier, France. 3 SELMET,
Univ Montpellier, CIRAD, INRAE, Institut Agro, Mont-
pellier, France. 4 Maastricht Science Programme,
Maastricht University, Maastricht, The Netherlands.
5 University of Maine, School of Biology and Ecolo-
gy. Maine, USA.

Leaf reflectance spectra estimated via near-infra-
red scanning (NIRs) are increasingly used to untan-
gle the intricate complexity of plant diversity and
evolution. The variation of these spectra across the
tree of life and the impact of the evolution of leaf
traits on spectral differentiation between species is
still being explored. In this study, we used a frame-
work that combines NIRs of herbarium dried leaves
to infer the diversity and evolution of species specific
spectra within the most diverse genus of Annonace-
ae in the Neotropics: *Guatteria*. We scanned herbar-
ium preserved leaves of 148 out of the ca. 180 known
*Guatteria* species. Spectra were cleaned, normal-
ized, and analyzed using different multivariate ap-

proaches. We tested to see if these data could be
used to identify species with similar spectra. We
then analyzed this dataset within a comparative
phylogenetic framework using a large robust spe-
cies-level dated phylogeny of *Guatteria*. We tested
for the phylogenetic signal of leaf spectra and in-
ferred diversification rates across the evolution of
this genus. We tested if changes in spectral diversity
when present coincided with shifts in diversification
rates within the genus. Our results shed light on the
levels of constraint in the different spectral regions,
reflecting the evolution of underlying *Guatteria*-spe-
cific leaf traits. Finally, our study underlines the im-
portance of herbarium specimens to understand
the evolution of diversity and leaf traits using inno-
vative techniques such as NIRs.

S.41.2 Classifying stone oaks
(Lithocarpus) in herbaria
with reflectance spectra

Barbara M. Neto-Bradley¹, J. Antonio
Guzmán Q.², Jeannine Cavender-Bares²,
David A. Coomes¹

1 Department of Plant Sciences, University of Cam-
bridge, UK. 2 Department of Ecology, Evolution and
Behavior, University of Minnesota, USA

Global biodiversity datasets contain key gaps, this
is particularly obvious when considering taxa in the
Global South. Natural History Collections have the
potential to help fill these gaps, however many re-
cords are unidentified, precluding them from use. A
need to non-destructively sample from these col-
lections further limits their potential. We evaluate the
added value gained by taking spectral reflectance
<figure><img src="image_108.png" /></figure>

measurements of leaf tissue from herbarium specimens. We use a previously undigitised set of 1539 herbarium specimens from the Herbarium at Kew to ask how well we can discriminate between species using spectral data. We also evaluate the feasibility of inferring traits from these herbarium derived spectra. Spectra can accurately be used to identify species, however further work is still needed to extend this application to trait estimates. We highlight that leaf reflectance spectra provide a novel and non-destructive way to add value to natural history collections and can be applied to fill systemic knowledge gaps.

### S.41.3 NIRS as a innovative tool for valorizing herbarium specimens: ongoing research at CIRAD

Samantha Bazan¹,², Morgane Dendoncker³, Elodie Baby¹,², Denis Bastianelli¹,², Jennifer Mersni¹,², Alexia Bennert³, Mia Svensk¹,², Simon Taugourdeau¹,², Laurent Bonnal¹,²

1 CIRAD, UMR SELMET, Montpellier, France. 2 SELMET, Univ Montpellier, CIRAD, INRAE, Institut Agro, Montpellier, France. 3 Université catholique de Louvain, Earth and Life Institute, Environmental Sciences, Croix du Sud, 2, box L7.05.24, Louvain-la-Neuve, Belgium

Near infrared spectroscopy (NIRS) is a promising tool for valorizing herbarium specimens as it allows the gathering information on plants without damaging valuable and historical samples. The CIRAD herbarium is currently testing various approaches. These research can be separated into three areas: Firstly, we are using the NIR spectrum as a trait, studying its covariation with some other more "classic" functional traits such as Specific Leaf Area and leaf thickness. We are working on a dataset of *Combretum* genus from West and Central Africa to assess relationships between functional traits and a rainfall gradient. We are also examining functional traits of a Sahelian species, *Balanites aegyptiaca*, in relation with water availability. Secondly, we are experimenting with NIRS as a tool for systematics discrimination. Assuming that intraspecific variations in the spectrum are smaller than interspecific variations, it may be possible to make identifications in herbarium within some genus, using appropriate calibration. A test was carried out on two *Pistacia* species and

their hybrid, with some specimens dating back 150 years and others collected recently. Using these two datasets, we evaluate the ability of NIR models built on older collections to discriminate recent samples or, in opposite, the possibility of discriminating older samples with recent collections. Finally, we are experimenting NIRS for the prediction of leaf chemical composition. As prediction of fodder chemical value is already possible from the use of NIRS on grounded vegetal material, we are building a large calibration set to test the feasibility of applying the same method non-destructively on complete herbarium specimens. We present an overview of the research underway in our laboratory, spanning these three areas with practical case studies and initial results.

### S.41.4 Leaf spectral properties and leaf traits of four alpine species of butterworts (Pinguicula L.)

Sara Natale¹, Marco Canella¹, Garima Singh¹, Mariano Battistuzzi¹, Silvia Moschin¹, Anna Pasinato¹, Valentina Boscariol¹, Barbara Baldan¹, Francesco Dal Grande¹, Nicoletta La Rocca¹, Alessandro Alboresi¹

1 University of Padua, Padua, Italy

Understanding the traits that influence the responses of species to climate change is a crucial aspect of predicting future community composition and ecosystem function. Leaf functional traits are informative data to understand adaptation and acclimatization of species to ecological niches. Moreover, different plant species could have unique spectral reflectance signatures that might provide information to recognize and discriminate plant species. We characterized leaf morpho-functional traits of four species of *Pinguicula* with different ecological niches, from dump meadows to rocky walls. Specifically, we studied *P. alpina*, *P. leptoceras*, *P. vulgaris* and *P. poldinii*. Sampling activities took place in the Dolomites, north-eastern side of European Alpine Arc. We measured the following environmental parameters: temperature, relative humidity, light intensity, light spectrum. As leaf functional traits we considered leaf dry matter content and specific leaf area. In order to characterize the photosynthetic adaptation in alpine butterworts, we quantified leaf pigment contents, and we measured photosynthesis *in-situ* and leaf reflectance (400-1000 nm). Also, we ob-
<figure><img src="image_109.png" /></figure>

served leaf surface features, trichomes and glands morphology via scanning electron microscopy (SEM). Despite a lower concentration of chlorophylls, *P. poldinii* and *P. alpina* showed a higher photosynthetic efficiency (e.g. yield of open photosystem II reaction centres in light) compared to the other two species. Since the different species showed different leaf reflectance spectra, our findings corroborate the ongoing hypothesis that leaf reflectance can be a useful tool to interpret data related to leaf traits, pigment composition and tissues organization. By providing a comprehensive scenario of leaf morpho-functional characteristics and leaf spectra signature, we show that the studied-species are coherently optimized to their prevailing environmental conditions.

**S.41.5 Plants show their true colors: leveraging leaf spectra to understand the diversity and evolution of plant form and function**

Jose Eduardo Meireles¹, Dawson White², Cornelius Nichodemus¹, Jeannine Cavender-Bares³

1 School of Biology and Ecology, University of Maine, USA. 2 Herbarium, Harvard University, USA. 3 Department of Ecology, Evolution, and Behavior, University of Minnesota, USA

Understanding the diversity and evolution of plant function is critical at a time of rapid global change. However, traditional methods for assessing plant function, including measuring their traits and physiological strategies, are too expensive and time-consuming to be done at scale. First, we introduce leaf spectra and how they can be used to rapidly measure plant functional traits and physiological attributes. We cover the basics of leaf optics and techniques used to measure leaves Second, we cover the prospects and challenges of assessing plant function using herbarium specimens instead of fresh leaves. We talk about the challenges imposed by the background mounting paper and glue on the measurements and how to mitigate them. Finally, we describe and exemplify how to model the evolution of leaf spectra and their underlying traits to understand evolutionary rates and convergent evolution.

## S.42 CURRENT RESEARCH IN BORAGINALES

**S.42.1 An updated phylogeny of Boraginales based on the Angiosperms353 probe set: a roadmap for understanding morphological evolution**

Maria-Anna Vasile¹*, Tim Böhnert¹, Julius Jeiter², Maximilian Weigend¹

1 Bonn Institute for Organismic Biology, Plant Bio-diversity Section, University of Bonn, Bonn, Germany. 2 Faculty of Biology, TUD Dresden University of Technology, Dresden, Germany * email: m.vasile@uni-bonn.de

Boraginales are a subcosmopolitan order with ca. 2,700 species recognized in 11 families. We analyzed relationships using the Angiosperms353 probe set. In addition to the Angiosperms353 nuclear gene

data from 109 Boraginales accessions available in PAFTOL, phylogenomic data from 161 samples were generated, collectively representing all Boraginales families and approximately 84% of genus diversity. The results are consistent with previous phylogenetic studies, with some novelties and overall better resolution. Well-resolved species trees are retrieved with consistently high support for both data sets used (exons and supercontigs) and both analyses implemented (concatenation and multi-locus species tree estimation approach). Among the most important findings, Lennoaceae are found nested in Ehretiaceae and we propose their inclusion in Ehretiaceae. The tribal relationships of the highly diverse Boraginaceae subfam. Cynoglossoideae are fully resolved for the first time. Residual ambiguities remain: Codonaceae are found as basal branch in Boraginales I (sister to [Wellstediaceae and Boraginaceae]) in the exonic data set, in agreement with previous studies, but they are retrieved as basal branch of Boraginales II in reconstructions based on exons and introns. Also, the sister relationship of Na-maceae and Hydrophyllaceae is not clear, probably
<figure><img src="image_110.png" /></figure>

due to limited sampling especially in *Nama*. Similarly, tribe Cynoglosseae was summarily sampled and intergeneric relationships are not at present fully resolved. Overall, the results highlight the effectiveness of the target enrichment probe set for understanding the evolutionary history of Boraginales and pave the way for resolving remaining taxonomic issues and a comprehensive study of the evolution of group-specific morphological traits. Our studies on the internal ovary architecture using scanning electron microscopy and micro-computed tomography reveal that changes in placentation and ovule number are key characters in the macroevolution and divergence of Boraginales families. Combining highly resolved phylogeny with critical morphological analyses promises a deep understanding of evolution trajectories in Boraginales.

## S.42.2 Unraveling the evolutionary history of Cynoglossinae (Boraginaceae): insights from molecular dating, biogeography, and diversification patterns

Zohreh Pourghorban¹, Yasaman Salmaki¹, Maximilian Weigend²

1 Department of Plant Science, University of Tehran, Tehran, Iran. 2 Bonn Institute of Organismic Biology, University of Bonn, Bonn, Germany

Cynoglossinae has radiated into a diverse assemblage comprising approximately 200 species, and its evolutionary history has not yet been comprehensively explored. Here, we examine the historical biogeography of Cynoglossinae, employing molecular dating and ancestral area reconstruction to understand how past climate changes and geographical patterns have provoked the diversification of Cynoglossinae. We inferred the evolutionary relationships of Cynoglossinae based on a dataset of 154 taxa, utilizing four plastid markers (*trnL-F*, *rps16*, *rpl16*, and *trnK-matK*) and one nuclear region (ITS) through PacBio sequencing. The monophyly of Cynoglossinae is strongly confirmed under Bayesian and maximum likelihood approaches. However, the nested position of all segregates confirms the broad concept of *Cynoglossum*, encompassing all generic segregates. Our biogeographic analysis results

suggest that this subtribe likely originated during the middle Miocene in a large area, including East Asia and the Irano-Turanian region. It underwent rapid initial diversification, subsequently and repeatedly expanding its range both westward to Europe (including the Mediterranean region) and Africa, and southward to Australia during the Middle-Miocene to Pleistocene, with a preferential radiation into northern temperate regions. The timing of the diversification of the species-rich clades corresponds with Miocene tectonic events and global climate changes, resulting in increased aridity across Eurasia. Diversification analyses demonstrated a relatively constant diversification rate for all Cynoglossinae. Our results highlight the complexity of processes shaping diversifications in Cynoglossinae, emphasizing a complex interaction among climatic modifications providing opportunities for diversification.

## S.42.3 An update on the phylogeny, classification, and biogeography of the popcorn flowers (subtribe Amsinckiinae, Boraginaceae)

Michael G. Simpson¹, C. Matt Guilliams²
Kristen Hasenstab-Lehman², Makenzie E. Mabry³

1 San Diego State University, San Diego, USA. 2 Santa Barbara Botanic Garden, Santa Barbara, USA 3 University of Florida, Gainesville, USA

Subtribe Amsinckiinae, colloquially known as the popcorn flowers, is currently recognized with 17 genera and approximately 302 species. The subtribe exhibits a remarkable diversity of form and is one of the most taxon-rich groups of family Boraginaceae. Several new species in the subtribe have been discovered in the past five years, with more yet to be described. Analyses of new chloroplast DNA and ddRadSeq sequence data resolve some past ambiguities and confirm generally strong support for the monophyly of the 17 genera and of a separate group of *Cryptantha* species termed the *Maritimae* clade. We report the status of resolving “backbone” interrelationships among these major clades, which are often poorly resolved. We summarize major evolutionary shifts in the Amsinckiinae in plant duration, leaf size, fruit morphology, and reproductive biol-
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ogy. Long-distance dispersals within the subtribe occurred a minimum of 18 times, each resulting in an American amphitropic disjunction. Based on sequence data and comparative morphology, we now expand membership of the *Maritimae* clade to include up to 7 North American and 12 South American species, the latter resulting from evolutionary divergence following a single long-distance dispersal event. Finally, we present evidence that the *Maritimae* clade is inclusive of a monotypic genus previously not confirmed as a member of the subtribe, and we evaluate whether the *Maritimae* clade might best be treated as a separate genus of the popcorn flowers.

### S.42.4 A systematic analysis of the family Heliotropiaceae Schard. in India based on morphology, anatomy and molecular data

Anna Ancy Antony Attullil¹, Etna Bivera¹

1 Department of Botany, St. Albert's College, Ernakulam, India

Systematic concepts of Boraginaceae have been highly problematic and controversial over decades. Traditional classifications subdivided the family Boraginaceae into five subfamilies. Recent molecular data-based results, supports the recognition of eleven families namely the Boraginaceae s.str., Codonaceae Weigend & Hilger, Coldeniaceae J.S.Mill. & Gottschling, Cordiaceae R.Br. ex Dumort., Ehretiaceae Mart., Heliotropiaceae Schard., Hoplestigmataceae Gilg, Hydrophyllaceae R.Br., Lennoaceae Solms, Namaceae Molinari, and Wellstediaceae Novák. under Boraginales. The family Heliotropiaceae Schard. comprises four genera, *Heliotropium* L., *Euploca* Nutt., *Ixorhea* Fenzl and *Myriopus* small. with about 450 species world wide. The members are distributed in the tropics and temperate regions of the world and occur mostly along the plains, coastal areas, dry arid zones. In India, the family is represented by two genera *Heliotropium* L. and *Euploca* Nutt. and with a high percentage of endemism. The members are annual or perennial herbs characterised by scorpioid cymes, epipetalous stamens, bicarpellary gynoecium, tetralocular ovary, terminal style, uniquely modified stigmatic head with anulus base and infertile apex, four seeded fruits.

The family is studied in India based on field and herbarium data and the characters were recorded. Entire morphological variation occurring within the family is considered. Diversity and adaptive value of foliar architecture and pollen morphology of species is analysed. Hypodromous, brochi-dodromous type I, II, III, craspidodromous and reticulodromous venation patterns are noted within the members. Kranz anatomy is restricted to the *Euploca* clade. *Euploca* is characterised by medium-sized pollen grains, except in *E. ovalifolia*. Pollen grains are prolate in *Heliotropium* and prolate spheroidal in *Euploca* with the absence of endocingulum and medial constriction. Phylogenetic analysis using combined sequence data from regions ITS, trnL-trnF, matK and rps16 were conducted. These approaches prove to be highly informative in delimiting members within the family.

### S.42.5 Evolutionary trends in gynoecium and fruit evolution in Boraginales

Julius Jeiter¹*, Maria-Anna Vasile², Maximilian Weigend²

1 Chair of Botany, Faculty of Biology, TUD Dresden University of Technology, Dresden, Germany. 2 Bonn Institute for Organismic Biology, Plant Biodiversity Section, University of Bonn, Bonn, Germany *email: julius.jeiter@tu-dresden.de

Boraginales constitute a medium-sized order in the Lamiids with a chequered taxonomic history due to considerable diversity in flower and fruit morphology. In recent decades, molecular phylogenetics established Boraginales as a well-defined order with two major clades. Both comprise families characterised by either capsules or nutlets and capsules, schizocarps and/or drupes. A reduction in ovule number from many to four appears to have preceded the transition from capsular to indehiscent fruits. We address two main questions: Why did indehiscent fruits derived from a bicarpellate, syncarpous gynoecium evolve twice independently? What are the main factors driving fruit evolution in Boraginales? We combine scanning electron microscopy and X-ray micro-computed tomography in the context of recent phylogenies to understand gynoecium and fruit morphological evolution within Boraginales. We focus on three examples from Boraginales II. Our studies of gynoecium and
fruit morphology address patterns in ovule and seed number evolution. In Hydrophyllaceae we found multiple transitions from pluri-ovulate to four-seeded fruits, but reversals of this character are rare. We argue that the reduction of ovule number to four fixes an ontogenetic trajectory, ultimately leading to indehiscent, four-seeded fruits. Conversely, in holoparasitic *Pholisma arennarium*, parasitism has likely led to a departure from the "two carpel-four ovule paradigm" in Boraginales, resulting in a multiplication of carpels to increase seed number. Finally, in Heliotropiaceae, we found fruit architecture to be highly conserved

and divergent fruit morphology and function is realized by histological changes. Multiple transitions between schizocarps and drupes occurred even within closely related taxa within the family. These morphological and developmental studies underscore the multiple ontogenetic trajectories available for morphological diversification and functional convergence, even where particular character combinations are highly conserved. Traditional Boraginales systematics have emphasised mature fruit morphology – phylogenetic data clearly demonstrate that this often fails to reflect relationships correctly.

# S.43 GEOPHYTIC MONOCOTS: TAXONOMY AND THE EVOLUTION OF SUCCULENCE

## S.43.1 Delimiting species boundaries in Indian Pancratium (Amaryllidaceae): a cytotaxonomic approach

Shrirang R. Yadav¹

1 Shivaji University, Kolhapur, India

Pancratium species grow sporadically throughout the country and individual plants are usually sparsely distributed. The species are ephemeral bulbous perennials. The emergence of scape and flowering mostly depends on the pre-monsoon rain pattern. Following the first pre-monsoon showers, the species flower only once in a year for a short period (10-15 days). Therefore, catching the species in flowering is a difficult task. Consequently, the genus is taxonomically poorly studied. Moreover, the species have been distinguished based on overlapping quantitative characters. Protologues are very brief, and the type specimens are either not available or poorly preserved. Therefore, there is a need to revisit the taxonomy and develop standard taxonomic methods to circumscribe species unambiguously. Accordingly, attempts have been made over the last 20 years to study the Indian species (distributed across ca. 75 accessions from various localities in India) in the field as well as under cultivation. I recognize nine species in India and provide an account of diagnostic characters and standard terminology that can be used

to distinguish them. Additionally, chromosomal and crossability data are used to understand species limits. Most of the *Pancratium* species are diploid with 2n = 2x = 22 chromosomes but a few, viz. *P. triflorum* and *P. zeylanicum* exhibit triploid (2n = 3x = 33) cytotypes as well. Karyotypes are represented by m (metacentric), sm (submetacentric), st (subtelocentric) and t (acrocentric) chromosomes. About 60 interspecific crosses involving eight species were made. Interspecific crosses among the Indian species produced fully fertile hybrids indicating that they represent a single biological species. The differences in characters (fixed genetically) among the populations (of species) probably represent ecotypes adapted to local environmental conditions.

## S.43.2 Taxonomy of Indian Dipcadi: Does cytogenetics provide all answers?

Manoj M. Lekhak¹, Priya E. Shelke¹, Shrirang R. Yadav¹

1 Shivaji University, Kolhapur, India

Dipcadi Medik. (Asparagaceae) is a bulbous monocot with less than 50 species distributed in Africa, Europe, the Indian subcontinent and Madagascar. India has 13 species with a maximum concentration (11 species) in peninsular India. Cytogenetically the genus is well-known for its bimodal karyotype wherein two sets of chromosomes of contrasting sizes are found. Taxon-
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omy of the genus has always been convoluted on account of the uniformity of morphological features and their short flowering period. Moreover, these species make poor herbarium specimens because of their succulent nature and hence are underrepresented in Herbaria across the globe. Here, we studied the cytogenetics of 23 accessions distributed across 11 species to refine their taxonomy. The variation in diploid chromosome number (2n) indicated the presence of three chromosome counts, i.e., 2n = 12, 20 and 22 chromosomes. *D. concanense*, *D. goaense*, *D. janae-shrirangii*, *D. reidii*, *D. saxorum*, accessions collected from Ankai fort, Delhi, Gautala and Mhaismal had 2n = 12 chromosomes whereas *D. coimbatorensis*, *D. krishnadevarayae*, *D. montanum* var. *madrasicum*, *D. ursulae*, *D. ursulae* var. *longiracemosum*, accessions collected from Ajara, Badami, Belgaum, Dapoli, Dolkhamb, Halkarni, Rantale and Yercaud had 2n = 20 chromosomes. *D. erythraeum* had 2n = 22 chromosomes. The mean chromosome length (MCL) varied from 1.52 – 9.31 µm and the total length of the haploid complement (THL) was 24.76 – 48.91 µm. Both bimodal and trimodal karyotypes were observed. Trimodal karyotypes were confined to species with 2n = 20 and 22 chromosomes. Male meiosis was found to be normal. Our findings suggest that cytogenetic information, viz. chromosome number and morphology cannot fully discriminate between species and hence an integrated approach involving molecular cytogenetics or molecular phylogenetics should be applied to resolve the identity of species.

### S.43.3 Geophytes in the Anthropocene: an integrated approach to understanding geophyte responses to rapid change

Syd Ramdhani¹, Şerban Procheş², Sershen³, Angus Macdonald¹

1 School of Life Sciences, University of KwaZulu-Natal, Durban, South Africa. 2 Discipline of Geography, University of KwaZulu-Natal, Durban, South Africa. 3 Department of Biodiversity and Conservation Biology, University of Western Cape, Bellville, South Africa

Geophytes are generally plants with an underground perennation organ (e.g. bulb, corm, tuber, rhizome) that has a storage function and

leaves that normally die back annually. This life-form evolved to avoid or cope with adverse environmental conditions and associated stressors both spatially or temporally. A predisposition to stress has arguably resulted in positive and negative consequences, but offers an interesting perspective on responses to rapid global change stressors of the Anthropocene especially climate change. While research focused on geophytism has gained pace recently, more integrated approaches are required. Here we offer recommendations on an integrated approach of understanding geophyte responses to rapid change from evolutionary, ecological, and physiological perspectives for monocots. This approach should begin with a deeper understanding of the evolution of the geophytism (particularly lineages with high levels of geophytism) to determine whether the trait is basal/advanced, when it evolved and under what conditions the trait emerges and persists. Ecological and physiological studies on geophytes focus on distribution, richness, endemism, growth, phenology, reproduction and dormancy. We argue for the implementation of more *in situ* and greenhouse studies (simulated stress scenarios) as these can provide excellent opportunities to understand geophyte responses specifically to climate change at various levels, and potentially provide data for predictive models. Spatial modelling of geophytes under climate change scenarios has been initiated in various parts of the world, but more of such studies are needed. While we have focused on climate change as a primary Anthropocene stressor, we need to acknowledge other interlinked stressors such as land use change, overharvesting, overgrazing, disease, fragmentation, and ecosystem degradation that are increasing the extinction rate of geophytes globally. An integrated approach will allow for a better understanding of the risks posed by climate change and multiple stressors of the Anthropocene, as well as the potential conservation options.
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## S.43.4 Diversity and phylogeny of the rapidly expanding Asian genus Peliosanthes (Asparagaceae)

Maxim S. Nuraliev¹,², Anastasia A.

Krinitsina¹, Nikolay A. Vislobokov¹,², Leonid

V. Averyanov³, Khang Sinh Nguyen⁴,⁵,

Dmitry D. Sokoloff¹, Pavel A. Nikitin¹

Maxim S. Nuraliev¹,², Anastasia A. Krinitsina¹, Nikolay

A. Vislobokov¹,², Leonid V. Averyanov³,

Khang Sinh Nguyen⁴,⁵, Dmitry D. Sokoloff¹,

Pavel A. Nikitin¹

¹M.V. Lomonosov Moscow State University, Moscow, Russia. ²Joint Russian-Vietnamese Tropical Scientific and Technological Center, Hanoi, Vietnam. ³Komarov Botanical Institute, St. Petersburg, Russia. ⁴Institute of Ecology and Biological Resources, Hanoi, Vietnam. ⁵Graduate University of Science and Technology, Hanoi, Vietnam

Considered recently to contain only a few species, the genus *Peliosanthes* is currently under active discovery for its taxonomic diversity. Indeed, by the end of the 20th century, it was known to comprise about 15 species, whereas more than 60 formally accepted species are presently known. However, the more taxonomic novelties are published, the less clear the interspecific boundaries become. From a morphological point of view, *Peliosanthes* can hardly be subdivided into species groups. In the virtual absence of molecular phylogenetic data, this makes the ideas of species proximity and their discrimination from each other highly subjective and often ambiguous. We used high-throughput sequencing approaches to obtain sequences of the entire plastid genomes of 20 specimens representing 18 species of *Peliosanthes*. Our results indicate that plastome data allow a well-resolved phylogenetic reconstruction of the genus. In some cases, the reconstruction was in congruence with the morphological similarity between species. For example, *P. triandra* and *P. weberi*, representing the earlier recognized genus *Neolourya*, formed a clade. In other cases, in contrast, the phylogenetic pattern obtained has never been predicted. In particular, our results support the distinctness of *P. graminea*, which was considered to be very close to the widespread and variable *P. teta* but appeared to be distantly related to the latter. We show homoplastic evolution of several structural traits, including the inflorescence type (raceme vs. thyrse), the absence or

presence of a pedicel, the shape of the perianth (rotate vs. more or less cup-shaped), and the shape of the androecial corona (entire vs. lobed). We therefore argue that the significance of these characters for the systematics of *Peliosanthes* was overestimated, and we refute a number of earlier proposed interspecific relationships. Supported by the Russian Science Foundation, project 24-44-04001.

## S.43.5 New insights on generic circumscription in Hyacinthaceae based on comprehensive morphological, molecular and distributional data

Mario Martínez-Azorín, Manuel B. Crespo,
María Ángeles Alonso-Vargas

University of Alicante, Alicante, Spain

Generic circumscription and systematics in *Hyacinthaceae* (= *Asparagaceae* subfam. *Scilloideae*), have been controversial in recent decades, with contrasting treatments based on preliminary and partial studies focusing only on morphology and/or limited DNA sequence data. This created considerable instability at both generic level and generic relationships. Recent treatments in subfamilies *Ornithogaloideae* and *Urgineoideae* have recognized a single genus *Ornithogalum* or up to four genera in *Ornithogaloideae* or only two genera in *Urgineoideae*, with a very broadly conceived *Drimia*. These proposals have been accepted for the last years by influential repositories (such as POWO, etc) and represented synonymization of several traditionally accepted genera with very distinct morphology, such as *Galtonia*, *Neopatersonia* and several others into *Ornithogalum* s.l.; several groups traditionally included in *Ornithogalum* into *Albuca* s.l.: or *Schizobasis*, *Thuranthos*, *Litanthus*, etc in *Drimia* s.l. In this way, recognition of such very broad concepts compromise acceptance of most genera in flowering plants and in petaloid monocotyledons and it is our aim to provide an alternative solution based on comprehensive studies combining morphological, molecular and distributional data. Our latest phylogenetic analyses involving plastid and nuclear DNA regions and a selection of 40 morphological characters covers 293 samples of *Urgineoideae* (ca. 80% of its diversity) and yielded 31 well-defined clades or lineages, most corresponding to previously described genera, although some have required description or revised
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circumscription. Regarding subfamily *Ornithogaloideae*, our studies evidence the existence of 19 lineages that can be accepted at genus rank and are linked to a defined biogeography. As in other monocot families, a considerable degree of homoplasy was observed in morphology, especially in those groups with unspecialized flowers; nonetheless, consistent syndromes of traditional and novel characters are shown to support clade recognition at genus rank.

### S.43.6 Integration of genomic phylogeography, morphology and climatic niche to elucidate speciation within Phycella (Amaryllidaceae)

Nicolás García¹, Taryn Fuentes¹,², Ryan A. Folk³

1 Universidad de Chile, Santiago, Chile; email: ngarcia@uchile.cl. 2 Wildland Ecobenefit Conservancy (WEConserv) Foundation, Santiago, Chile. 3 Mississippi State University, Mississippi State, USA

*Phycella* (Amaryllidaceae) is a clade of approximately 13 species of lilies endemic to the Mediterranean-type ecosystem of Chile. An unclear taxonomy at the species level and conservation interest makes it a priority to clarify the number and limits of species which hinders addressing questions about speciation mechanisms in this key endemism area. To approach the evolutionary

history of *Phycella*, we sampled a total of 137 individuals from 47 populations, including all described taxa throughout the geographic range of the group, and sampled 893 nuclear genes (1135 exons) through hybrid capture. These data largely resolved the phylogeny of *Phycella* with high support and demonstrated substantial phylogenetic resolution at the population level. The combination of high-resolution molecular data with morphological and ecological data further yielded insight into speciation mechanisms, and near-complete plastomes were extracted from raw reads and assembled for all samples to compare with the nuclear framework and examine complex evolutionary processes. Using phylogenetic network and modeling approaches, we identified major cytonuclear discord, attributable to proximity-based gene flow among recently diverged species through the speciation process, largely involving cytoplasmic DNA. Analyses of niche overlap among species and nuclear clades suggest that the diversification of *Phycella* was associated with niche divergence, supporting a predominantly geographic mode of speciation in the group, likely driven by the mountainous landscape characteristic of diversity and endemism center of this clade in central Chile. Finally, we present a major integrative taxonomic proposal that divides *Phycella* into 18 species on the basis of molecular, morphological, and ecological data. Overall, our findings highlight the value of strong sampling of both populations/individuals and genetic loci for speciation studies, which was key to identifying both evolutionary processes and a confident taxonomic framework for contextualizing these processes.

## S.44 MECHANISMS UNDERLYING PLANT DIVERSITY PATTERNS IN THE EAST ASIAN–AUSTRALASIAN REGION

### S.44.1 What factors contribute to the biased latitudinal species diversity gradient in the East Asia–Southeast Asia–Australasian region?

Buntarou Kusumoto¹, Takayuki Shiono², Yasuhiro Kubota²

1 Faculty of Agriculture, Kyushu University, Japan. 2 Faculty of Science, University of the Ryukyus, Japan

The East Asia–Southeast Asia–Australasian region constitutes a continuum of plant diversity hotspots across a broad latitudinal gradient, contributing significantly to global plant diversity. The higher latitudes in the region, including East Asia and Australia, are the richest source of botanical information (i.e., plant species occurrence records) in the world, while the lower latitudes present a global cold spot in such records. This spatial inequality of sample completeness has made it difficult to scrutinize the