# S.167 LEGUME SYSTEMATICS: FROM COLLABORATIVE NETWORKS TO GENOME SEQUENCING

## S.167.1 Evolution of legumes: from introgression to whole genome duplication

Le Min Choo¹,², Rowan Schley¹

1 Department of Geography, University of Exeter, Exeter, UK. 2 Singapore Botanic Gardens, National Parks Board, Singapore

Introgression and duplication are two of the most dramatic phenomena that a genome may encounter, each with myriad outcomes, ranging from increased evolutionary potential through to a heightened likelihood of extinction. Both processes have greatly influenced the evolution of plant diversity, particularly within the legume family which holds more than 20,000 species. Among the most species-rich legume communities occur in tropical rainforests, which are emblematic for their diversity. Accordingly, here we explore the influence of introgression and genome duplication on legume tree genera found in tropical rainforests, spanning the tropics. *Sindora*, from the Detarioideae subfamily, comprises 20 species found in palaeotropical rainforests, and has experienced whole genome duplication in its history. As such, phylogenetic reconstruction is challenging within the group due to the high abundance of paralogues. *Sindora* is known to hybridise extensively, forming 'syngameons' of many interfertile species, and so here we investigate whether introgression has contributed to the present day ability of the genus to form interspecific hybrids using a bespoke phylogenetic pipeline that aims to account for paralogy. The extent of the syngameon and the nature of hybridisation are also studied using present day hybrids. *Inga*, from the Caesalpinioideae subfamily, comprises >300 species and typifies the rapid rainforest tree radiations that gave rise to most Neotropical tree diversity. Introgression can catalyse such events of rapid diversification, and so here we assess whether hybridisation catalysed rapid diversification in *Inga*. Firstly, we ascertain the extent of introgression across *Inga*, following which we test whether introgression of loci underlying chemical defences against herbivory

facilitated rapid speciation, given the ecological advantage of novel defences in resisting the relentless insect herbivory experienced by rainforest trees. Overall, we aim to assess whether transfer of adaptive variation through introgression helped generate the unmatched diversity of rainforest trees.

## S.167.2 The future of legume systematics: Embracing collaborative tools to enhance taxonomic, geographic and evolutionary knowledge

M. Marianne le Roux¹,², Domingos Cardoso³,⁴, Juliana Gastaldello Rando⁵, Anne Bruneau⁶

1 South African National Biodiversity Institute, Pretoria, South Africa. 2 University of Johannesburg, Johannesburg, South Africa. 3 Rio de Janeiro Botanical Garden, Rio de Janeiro, Brazil. 4 Federal University of Bahia, Salvador, Brazil. 5 Federal University of Western Bahia, Bahia, Brazil. 6 Université de Montréal, Montreal, Canada

The legume systematics community's long history of research and collaboration has greatly increased our understanding of the classification, biogeography and evolutionary history of the family. This is evident in the many activities and publications that have seen the light since 1978, when the first International Legume Conference was held at the Royal Botanic Gardens, Kew. In 2010, the Legume Phylogeny Working Group (LPWG) was founded, and has since published five papers under this name, including a new phylogeny-based subfamily classification and a revised higher-level classification for subfamily Caesalpinioideae. In 2020 the LPWG established five working groups to promote collaboration on specific aspects of legumes. The Taxonomy Working Group was created to work on a list of names that would feed into and support the underlay of the other four working groups. This has led to the publication of
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four versions of updated legume taxon names from which out of the 64 351 specific names ever published in the family we now accept 22 623 species and 797 genera. The Legume Taxonomy Working Group is now acknowledged as an active legume partner and contributor to the international World Flora Online project and Global Biodiversity Information Facility (GBIF) and serves as the authoritative source for legume names in both portals. A collaborative online platform, Rhakhis, was chosen for the community to continue updating names. Working with GBIF, the legume community developed a portal focused on the family, and this led to the launch of the first version of GBIF's phylogeny viewer based on a phylogenomic analysis of Caesalpinioideae. Other international collaborations to pursue phylogenomic analyses and data sharing, including morphological and geographical data, have been encouraged to facilitate monographic, phylogenetic, macro-evolutionary and biogeographical analyses of this ecologically important family.

**S.167.3 Resolve tribal relationships of Fabaceae using nuclear, plastid and mitochondrial genomic data**

Rong Zhang¹, Ting-Shuang Yi¹, Jin Jian-Jun¹,², Yin-Huang Wang¹,³, De-Zhu Li¹, Joseph Charboneau⁴, Xing-Xing Shen⁵, Stephen A. Smith⁴, Domingos Cardoso⁶, Luciano P. de Queiroz⁷, Anne Bruneau⁸

1 Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China. 2 Present Address: Columbia University New York, USA. 3 Present Address: Chongqing Normal University, Chongqing, China. 4 University of Michigan, Ann Arbor, USA. 5 Zhejiang University, Hangzhou, China. 6 Rio de Janeiro Botanical Garden, Rio de Janeiro, Brazil. 7 Universidade Estadual de Feira de Santana, Feira de Santana, Brazil. 8 Université de Montréal, Montréal, Canada

Tree of Life lies at the heart of almost all studies in evolutionary biology. However, it is challenging to resolve recalcitrant phylogenetic relationships caused by rapid evolutionary radiations, hybridizations, and whole genome duplications. Despite the ecological and economical importance of Fabaceae (Leguminosae), phylogenetic relationships among some subfamilial and tribal lineages remain poorly resolved. We reconstructed the backbone phylogeny of Fabaceae using plastomes, mitochondrial genes

and transcriptome data of 909 species across 486 genera, representing all six subfamilies and almost all tribal lineages. Our results strongly supported the monophyly of the six subfamilies and 64 tribal clades, and clarified many long-controversial relationships. However, 14 sets of nodes remained unresolved due to conflicts among trees based on plastid and nuclear data, or different analysis methods. Through gene-tree quartet analyses evaluating alternative topologies and consistently strong support in ML and Astral tree both on the nuclear data sets, we resolved 18 of these recalcitrant nodes. Using relative quantification analyses, our study identified incomplete lineage sorting as the main source of conflict for deeper nodes (e.g., the root of Fabaceae), gene flow is the main source for shallower nodes (e.g., Griffonia, ADA clade, Wisterieae), both are involved conflicts for some nodes (e.g., core Papilionoideae, Erythrophleeae, Tachigalieae, Moldenhawereae). The study suggested when ILS and gene flow coexist to a considerable extent, resolving contentious relationships becomes exceedingly difficult. Our study provides a robust phylogenomic framework for exploring legume evolution and supplies a typical case to apply multiple types of molecular data and analysis methods to resolve recalcitrant phylogenetic relationships and explore the underlying conflict.

**S.167.4 Phylogenomics understanding of the evolution and adaptation of RNS in Legumes**

Zijian Liu¹, Xiaoxiao Zhao¹, Wenfei Xian¹, Jeffrey J. Doyle², James Euan³, Shifeng Cheng¹

1 Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Dapeng District, Shenzhen, Guangdong, China. E-mail: chengshifeng@caas.cn. 2 School of Integrative Plant Science, Sections of Plant Biology and Plant Breeding & Genetics, Cornell University, Ithaca, NY, USA. Email: jjd5@cornell.edu. 3 The James Hutton Institute, Invergowrie, Dundee. Charity No. SC041796. Email: Euan.James@hutton.ac.uk

Leguminosae is the third largest angiosperm family in species richness, spreading and adapting globally in a wide range of ecology. Many legumes are economically important food crops offering highly nutritious sources of protein and micronutrients for human beings and animals, which can partly attributed to its successful establishment of the nitrogen-fixing root-nodule sym-
<figure><img src="image_391.png" /></figure>

biosis (RNS) with the mutualistic nitrogen-fixing bacteri. >95% of RNS are restricted in Legumes, however, how RNS in Leguminosae rised, stabilized, and widely adapted remains an unsolved mystery. With the development of genomic sequencing technologies and bioinformatics, the ability to "sequence everything" for an entire genus or even for an entire family, is becoming a reality. We thus initiated the "Legume Genome Sequencing Consortium" and started the Legume Nodulation and NFNC Phylogenomics v2.0 Project. 32 representative new legume species, carefully selected from all the six sub-families (Cercidoideae, Detarioideae, Duparqueo-tioideae, Dialioideae, Caesalpinioideae, and Papilion-oideae) as the diverse 'comparison pairs' of nodulating plants together with their closest non-nodulating relatives, are sequenced (PacBio + HiC), assembled and annotated into high-quality chromosome-level genome references. In addition, a matched phylo-pan-transcriptome from these species was constructed. This effort opens new ways to explore diversity of RNS in a broad comparative context (phylogenomics), to gain a deeper and more holistic view of the evolution in protein-coding genes, conserved non-coding elements, and differential gene expression for RNS, to utlinate re-veal the molecular evolutionary mechanism of RNS ei-ther by divergence from a common ancestor over 100 Mya or by convergence following independent origins over that same time period. This study also provides great opportunity to understand the diversification and adaptation of Legumes.

### S.167.5 Parallel domestication of New and Old World lupins: chromosome-level assemblies of Lupinus mutabilis and a crop wild relative

Romulo Segovia-Ugarte¹, Marco Todesco¹, Loren H. Rieseberg¹, Bruno Nevado², Colin E. Hughes³, Sean W. Graham¹

1 Department of Botany, University of British Columbia, Vancouver, Canada. 2 Centre for Ecology, Evolution and Environmental Changes, University of Lisbon, Lisbon, Portugal. 3 Department of Systematic and Evolutionary Botany, University of Zurich, Zurich, Switzerland

The Andes are the domestication centre of many important and minor crops, but the list of Andean crops with available high-quality genome assemblies is

biased towards crops with worldwide distribution, neglecting those cultivated locally that have significant agricultural and cultural value. *Lupinus mutabilis* (Fabaceae) is the only lupin crop domesticated in the New World, where it is cultivated for its high protein content and soil enrichment properties. There are high-quality genome assemblies and pan-genome data available for Old World lupin crops (i.e., *Lupinus albus* and *L. angustifolius*). However, there is no reference genome for *L. mutabilis*. We assembled the genomes of *L. mutabilis* and *L. piurensis*, its crop wild relative, using PacBio HiFi and chromosome conformation capture (Hi-C) reads into 828 Mb and 795 Mb assemblies, respectively. In each case we assembled 24 chromosomes (594 Mb and 591 Mb). In total we annotated 34,176 and 33,681 protein-coding genes in *L. mutabilis* and *L. piurensis*, respectively. BUSCO scores indicate 98% completeness for both annotations. Transposable element annotation reveals that 67% and 69% of the *L. mutabilis* and *L. piurensis* genomes are repetitive, with Copia the most abundant class. Synteny analysis between *L. mutabilis* and *L. albus* demonstrates that *L. mutabilis* chromosome 4, the largest super scaffold, represents a merger of *L. albus* chromosomes 4 and 24, resulting in a karyotype with one fewer chromosome total, which is characteristic of the western New World lupins. We are reconstructing the evolutionary history of the two Andean lupins in comparison to the predicted ancestral legume karyotype. We surveyed for genes under positive selection, and document gene-family evolution and the effects of parallel domestication in the New World and Old World lupin crops.

### S.167.6 Why are some lineages more successful than others? Comparing distribution patterns across subfamilies in Leguminosae

Jens J. Ringelberg¹, Moabe F. Fernandes², Anne Bruneau³, R. Toby Pennington²,⁴, Colin E. Hughes⁵, Juliana Gastaldello Rando⁶, Edeline Gagnon⁷, Joseph T. Miller⁸, Dario I. Ojeda⁹, Manuel de la Estrella¹⁰

1 University of Edinburgh, Edinburgh, UK. 2 University of Exeter, Exeter, UK. 3 Université de Montréal, Montréal, Canada. 4 Royal Botanic Garden Edinburgh, Edinburgh, UK. 5 University of Zurich, Zurich, Switzerland. 6 Universidade Federal do Oeste da Bahia, Barreiras, Brazil. 7 University of Guelph, Guelph,
<figure><img src="image_392.png" /></figure>

Canada. 8 Global Biodiversity Information Facility, Copenhagen, Denmark. 9 Norwegian Institute of Bioeconomy Research, Ås, Norway. 10 Universidad de Córdoba, Córdoba, Spain

One of the central aims in evolutionary biology is
explaining why some lineages are more evolution-
arily successful than others, for example in terms
of species richness, ecological versatility, and geo-
graphic distribution. Direct comparisons of lineag-
es are complicated by the impacts of confounding
factors such as differences in age and evolutionary
relatedness. The six subfamilies of the legume fam-
ily provide an ideal natural experiment to determine
why some lineages are more successful than others.
Since the origin of Leguminosae is a phylogenomic
tangle which gave rise to all subfamilies within a very
short time frame, the subfamilies are nearly identi-
cal in terms of age and relatedness. Notwithstanding
these similar starting conditions, this natural exper-
iment yielded vastly different results, ranging from
subfamily Duparquetioideae, consisting of a single
African species, to subfamilies Caesalpinioideae
and Papilionoideae, which have respectively circa
4,700 and over 14,000 species across the globe. The

other three subfamilies, Cercidoideae, Detarioideae,
Dialioideae, are also widely distributed but harbour
lower numbers of species. Here we assemble a le-
gume-wide quality-controlled occurrence dataset to
compare distribution patterns across the six subfam-
ilies. We quantify differences between subfamilies in
areas of species richness, ecological versatility, and
geographic extents, to investigate links between rich-
ness and distribution. Furthermore, by combining this
new geographic dataset with existing legume-wide
and subfamily-specific phylogenies, we compare
rates of long-distance dispersal and niche shifting
through time to examine the roles of evolvability and
phylogenetic conservatism in generating species
richness. Our analyses reveal important differences
between subfamilies in terms of their main centres
of diversity, ecological versatility, and biogeographi-
cal evolvability through time. Notably, invasion of the
temperate zone in Papilionoideae played a key role
in generating its extreme species richness, whereas
other subfamilies are largely constrained by tropical
niche conservatism. These results shed new light on
why some lineages are more successful than others.

S.168 SYSTEMATICS, BIOGEOGRAPHY,
ADAPTATION AND UTILIZATION OF THE GRAPE
FAMILY VITACEAE

S.168.1 The origin,
diversification, and
adaptation of the grape
family Vitaceae

Limin Lu¹,², Yichen You¹,²,³, Jinren Yu¹,²,³,
Zelong Nie⁴, Russell L. Barrett⁵, Jun Wen⁶,
Zhiduan Chen¹,²

1 Institute of Botany, Chinese Academy of Sciences, Beijing, China. 2 China National Botanical Garden, Beijing, China. 3 University of Chinese Academy of Sciences, Beijing, China. 4 Jishou University, Jishou, China. 5 Australian Botanic Garden, New South Wales, Australia. 6 National Museum of Natural History, Smithsonian Institution, Washington, USA

Faced with environmental changes, plants may either
"move" to track their ancestral habitats or "evolve" and
adapt to new habitats. Vitaceae, the grape family, has

evolved diverse adaptive traits facilitating a global ex-
pansion in wide-ranging habitats. This evolutionary
history makes Vitaceae ideal for investigating for any
preference between "move" or "evolve" strategies and
their underlying mechanisms. We inferred patterns of
biogeographic diversification and trait evolution in Vi-
taceae based on a robust phylogeny with dense sam-
pling including 495 species (~52% of Vitaceae species).
Vitaceae most likely originated from Asia, the diversity
center of extant genera and the major source of dis-
persals. India and Australasia are recognized as the
biggest sinks and dispersals to the two regions might
be largely facilitated by land connection. At least four
shifts in diversification rates have been detected re-
spectively in *Ampelocissus*, *Cyphostemma*, *Tetrastig-
ma*, and *Vitis*, which have either evolved key innovations
or showed adaptation to specific habitats. In particu-
lar, we find that the Oligocene-Miocene boundary is
a turning point in shifting strategies, before which Vi-
taceae lineages preferred to "move", and after which,
to "evolve". The increase of niche opportunities mainly
<figure><img src="image_393.png" /></figure>

contributed to the frequent employment of the “evolve”
strategy after the Oligocene-Miocene boundary, while
putative key trait innovations also played a role.

S.168.2 Genomic adaptation to aridity in Cyphostemma (Vitaceae)

Jin-Ren Yu¹,²,³, Ju Zhou¹,², Rindra M.

Ranaivosona¹,²,³, Romer N. Rabarijaona¹,²,³,

Zhi-Duan Chen¹,²,⁴, Li-Min Lu¹,²

1 Institute of Botany, Chinese Academy of Sciences, Beijing, China. 2 China National Botanical Garden, Beijing, China. 3 University of Chinese Academy of Sciences, Beijing, China. 4 Sino-Africa Joint Research Center, Chinese Academy of Sciences, Wuhan, China

Cyphostemma, the second largest genus (ca. 200 species) of the grape family, mainly distributes in continental Africa and Madagascar. Species of the genus have colonized various habitats including tropical rainforests, savanna, and deserts. The genus exhibits extraordinary diversity in morphological innovations, of which the development of succulent stem is the most striking. Previous studies revealed a disparity in genome size, which has been suggested to be related to the innovation of succulence. However, it remains unclear how the genome size diversity in Cyphostemma is generated, and how it contributes to the adaptation to various habitats. We investigate this problem from both the genomic and ecological aspects. We assembled genomes of two representative species of Cyphostemma, *C. dehongense* from the Asian rainforest and *C. currorii* from the African desert. We find that genomes of Cyphostemma are mainly inflated by the insertion of long terminal repeat retrotransposons (LTR-RT), especially the explosive insertions of several families in the Ale lineage. According to the dynamics of LTR-RT, these families show a preference for inserting into introns since the Oligocene-Miocene Boundary, when Vitaceae began to diversify rapidly. The intron-preferring LTR-RT families have a higher death rate, and the genes with longer introns are under more relaxed selection, suggesting the co-evolution of LTR-RTs and their host genome. To further investigate the relationships between LTR-RT amplification and environmental conditions, we collected genomic repeat profiles, genome size, and habitat climatic factors for more than 100 taxa in Cyphostemma. Analyses based on a linear model reveal that genome expansion is generally associated with adaptation to aridity but is limited by precipitation during the growing season.

Moreover, we find that succulent species from arid
regions of continental Africa and Madagascar exhib-
it different patterns of LTR-RT amplification, indicating
different mechanisms for the origin of succulence.

S.168.3 Unraveling
evolutionary diversification
dynamics of North American
grapes across different
biomes

Alicia Talavera¹,², Richard Hodel¹,³,
Elizabeth Zimmer¹, Jun Wen¹

1 Department of Botany, National Museum of Natural History, Smithsonian Institution, Washington. DC, USA. 2 Departamento de Botánica y Fisiología Vegetal, Universidad de Málaga, Málaga, Spain. 3 Data Science Lab, Office of the Chief Information Officer, Smithsonian Institution, Washington, DC, USA.

Understanding the origin and distribution of species is a focal point of evolutionary biology. In North America, the current distribution and diversity of species have been substantially influenced by environmental changes during climatic oscillations, as well as historic and contemporary geographic barriers. Phylogeographic studies have revealed conflicting patterns among co-distributed taxa with distinct ecological requirements, suggesting dispersal ability and other life-history traits are crucial for shaping the diversity and distribution of the region's biota. However, a complete understanding of North America's diversification dynamics is hampered by a limited number of comprehensive genome-scale phylogeographic studies that integrate phenotypic and environmental niche evidence. This is crucial for revealing intricate evolutionary mechanisms driving diversification, such as introgression/hybridization, ecological adaptation, and ecophenotypic plasticity, which might be overlooked through less integrative approaches. The grape genus Vitis, especially subgen. Vitis, serves as an ideal model for studying North America's biogeography and phylogeography. Recent progress in genomics, phenomics, and specimen digitization offers an unprecedented opportunity to explore the convoluted evolutionary history of two challenging species complexes: Vitis aestivalis and Vitis cinerea. These taxa have extensively overlapping distributions throughout several biomes in North- and Mesoamerica. In this study, we sampled both species complexes across their distributional range. We employed several phylogenomic inference approaches to identify relationships within
<figure><img src="image_394.png" /></figure>

these complexes and hybridization events. Complementary population structure analyses were used to assess genetic diversity within and among sampling locations. Environmental variables were used to evaluate ecological differentiation as a driver of diversification, and phenotypic differences in leaf and stem characters were quantified to determine the extent of the influence of ecophenotypic plasticity. Our results support the recognition of several species in both the *V. aestivalis* and the *V. cinerea* complexes. Geographic isolation was important for species differentiation, and key phenotypic differences were detected among different ecological zones.

**S.168.4 Genomic insights into adaptative evolution and climate change vulnerability of the grape genus (Vitis L.)**

Zhiyao Ma¹, Xiaodong Xu¹, Wenjing Peng¹,²,
Hua Xiao¹, Zhongjie Liu¹, Wenwen Liu¹, Fan
Zhang¹, Tianhao Zhang¹, Zhuyifu Chen¹,
Yuanyuan Luo¹, Yanling Peng¹, Jun Wen³,
Brandon S. Gaut⁴, Yongfeng Zhou¹

¹National Key Laboratory of Tropical Crop Breeding, Guangdong Laboratory of Lingnan Modern Agriculture, Key Laboratory of Synthetic Biology, Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, China. ²College of Agriculture, Guangxi University, Nanning, China. ³Department of Botany, Smithsonian Institution, National Museum of Natural History, Washington, DC USA. ⁴Department of Ecology and Evolutionary Biology, University of California, Irvine, CA, USA

Wild grapes show a high level of species diversity and have been widely recognized as important germplasm resources of wine, table grapes, and resistance to biotic and abiotic stresses for the grape industry. However, the local adaptation and drivers of diversification of genus *Vitis* L remain poorly understood. Hybridization and introgression events of *Vitis* are still lacking a comprehensive knowledge regarding their frequency, pattern, and adaptive landscape, particularly at the genome-wide scale. The reticulated evolutions, local adaptations and responses to future climate change of several widely distributed wild grape in East Asia are investigated based on population genomic data. Species network analyses identified several hybridization events within East Asian *Vitis*. These interspecific hybridization events

may have caused the topological discordances and relatively low support detected in our analyses. Ecological niche modeling shows that most of the diversification of East Asian *Vitis* species is driven by temperature and precipitation environmental variables. Sympatric parallel diversifications of major clades also may have facilitated the rich diversity in East Asian *Vitis*. Our results emphasize the significance of comprehensive genomic data, including single-nucleotide polymorphisms (SNPs), insertion/deletions and structural variations (SVs), to predict environmental adaption of wild grapes in the context of rapid climate change.

**S.168.5 Population genetics analyses reveal the genetic basis of important agronomic traits and facilitate grape breeding**

Hua Xiao¹, Zhongjie Liu¹, Nan Wang¹,
Ying Su¹, Qiming Long¹, Yanling Peng¹,
Lingfei Shangguan², Fan Zhang¹, Shuo
Cao¹, Xu Wang¹, Mengqin Ge², Hui Xue¹,
Zhiyao Ma¹, Wenwen Liu¹, Xiaodong
Xu¹, Chaochao Li¹,⁸, Xuejing Cao¹, Bilal
Ahmad¹, Xiangnian Su¹, Yuting Liu¹,
Guizhou Huang¹, Mengrui Du¹, Zhenya
Liu¹, Yu Gan¹, Lei Sun⁴, Xiucai Fan⁴, Chuan
Zhang³, Haixia Zhong³, Xiangpeng Leng⁵,
Yanhua Ren², Tianyu Dong², Dan Pei²,
Xinyu Wu³, Zhongxin Jin¹,⁸, Yiwen Wang¹,
Chonghuai Liu⁴, Jinfeng Chen⁶, Brandon
S. Gaut⁷, Sanwen Huang¹,⁸, Summaira
Riaz⁹, Andrew M. Walker⁹, Jinggui Fang²,⁵,
and Yongfeng Zhou¹, ⁸, *Hua Xiao¹, Zhongjie
Liu¹, Nan Wang¹, Ying Su¹, Qiming Long¹,
Yanling Peng¹, Lingfei Shangguan², Fan
Zhang¹, Shuo Cao¹, Xu Wang¹, Mengqin
Ge², Hui Xue¹, Zhiyao Ma¹, Wenwen Liu¹,
Xiaodong Xu¹, Chaochao Li¹,⁸, Xuejing
Cao¹, Bilal Ahmad¹, Xiangnian Su¹, Yuting
Liu¹, Guizhou Huang¹, Mengrui Du¹, Zhenya
Liu¹, Yu Gan¹, Lei Sun⁴, Xiucai Fan⁴, Chuan
Zhang³, Haixia Zhong³, Xiangpeng Leng⁵,
Yanhua Ren², Tianyu Dong², Dan Pei²,
<figure><img src="image_395.png" /></figure>

Xinyu Wu³, Zhongxin Jin¹,⁸, Yiwen Wang¹,
Chonghuai Liu⁴, Jinfeng Chen⁶, Brandon
S. Gaut⁷, Sanwen Huang¹,⁸, Summaira
Riaz⁹, Andrew M. Walker⁹, Jinggui Fang²,⁵,
Yongfeng Zhou¹,⁸,*

¹National Key Laboratory of Tropical Crop Breeding, Guangdong Laboratory of Lingnan Modern Agriculture, Key Laboratory of Synthetic Biology, Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, China. ²College of Horticulture, Nanjing Agricultural University, Nanjing, China. ³Institute of Horticultural Crops, Xinjiang Academy of Agricultural Sciences, Urumqi, China. ⁴Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou, China. ⁵College of Horticulture, Qingdao Agricultural University, Qingdao, China. ⁶State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, China. ⁷Department of Ecology and Evolutionary Biology, University of California, Irvine, CA, USA. ⁸National Key Laboratory of Tropical Crop Breeding, Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, China. ⁹Department of Viticulture and Enology, University of California, Davis, CA, USA

Grapevine is among the first domesticated crops. However, its breeding is impeded by the investigations of the genetic basis of important agronomic traits. Based on population genetics combining multiple new analytic methods, such as pan-genome and machine learning, we identified many important trait-related quantitative trait loci (QTLs), and revealed the importance of introgression in shaping the current texture of grapes. We constructed a variant map by interrogating resequencing data from grapevine cultivars based on pan-genome including 18 newly generated telomere-to-telomere gap-free haplotype-resolved assemblies and performed genome-wide association studies (GWAS) related to 29 traits. 148 significantly related QTLs were identified, and 8.1% QTLs of 12 traits were identified as significant contributions by structure variations (SVs). We also found some QTLs were located in regions of divergent selection between wine and table grapes. Given the previous studies showing genomic signals of introgression from wild to wine grapes in Europe, based on machine learning-based population genetic analyses, we predicted the time, mode, genomic pattern, and biological effects of these introgression events, and found that continuous gene flow between European wild grapes (EU) and cultivated grapes over the past ~2000 years, especially from EU to wine grapes, suggesting EU wild grapes were an important resource for improving the flavor of wine grapes. We further found that introgression

may be a historically important process for improving clonal systems like domesticated grapes. Despite the potential benefits of introgression in grape improvement, the introgressed fragments introduced a higher deleterious burden, and most of these deleterious variants were in a heterozygous state in grapes. In general, our study facilitates the breeding of superior cultivars via genomic selection of multiple traits and the understanding of beneficial and harmful effects of introgression is critical for genomic breeding of grapevine to take advantage of wild resources.

S.168.6 Cenozoic seeds of
Vitaceae reveal a deep
history of extinction and
dispersal in the Neotropics

Fabiany Herrera¹, Monica R. Carvalho²,
Gregory W. Stull³,⁴, Carlos Jaramillo⁵,
Steven R. Manchester⁶

¹ Field Museum of Natural History, Chicago, USA. ² Museum of Paleontology and Department of Earth and Environmental, University of Michigan, Ann Arbor, USA. ³ Department of Botany Smithsonian Institution, Washington, USA. ⁴ National Identification Services, USDA-APHIS-PPQ, Beltsville, USA. ⁵ Smithsonian Tropical Research Institute, Panama. ⁶ Florida Museum of Natural History, University of Florida, Gainesville, USA.

The remarkably diverse plant communities of the Neotropics are the result of diversification driven by multiple biotic (e.g., speciation, extinction, and dispersal) and abiotic (e.g., climatic fluctuations and tectonics) processes. However, in the absence of a well-preserved, thoroughly sampled, and critically assessed fossil record, the associated processes of dispersal and extinction are poorly understood. We report an exceptional case documenting patterns of extinction in the grape family (Vitaceae Juss.) based on fossil seeds discovered in four Neotropical paleo-floras dated between 60 and 19 Mya. These include a new species that provides the earliest evidence of Vitaceae in the Western Hemisphere. Eight additional species reveal the former presence of major clades of the family currently absent from the Neotropics and previously unknown dispersal events. Our results indicate that regional extinction and dispersal have substantially impacted the evolutionary history of Vitaceae. They also suggest that while the Neotropics have been dynamic centers of diversification through the Cenozoic, extant Neotropical bo-
# S.169 HERBARIOMICS, AN INVALUABLE RESOURCE FOR PLANT RESEARCH

## S.169.1 Herbariomics unveils endangered chemistry across the world's flora

Luiz L. Saldanha¹, Rodrigo Cámara-Leret¹

1 University of Zurich, Switzerland

Plants are essential for human health. However, little is known about how much of their chemical diversity is threatened with extinction. Here, we characterised the phytochemical space of endangered plant species and quantify, for the first time, how much chemodiversity may vanish globally with the loss of threatened plants. We find that 29% of the 1,463 species with chemical studies contain unique compounds. For the other 23,689 chemically neglected species, we developed a metabolomics workflow to accelerate their chemical study. Essentially, we test to what degree the diversity of alkaloids, terpenoids and flavonoids in living plants can be detected on herbarium specimens of the same species covering diverse taxa. Overall, our herbariomics approach to study chemodiversity brings a new dimension to conservation planning (beyond the currently used metrics of biological, phylogenetic and functional diversity) by highlighting priority species for future research (e.g. discovery of natural products) and imminent conservation action (i.e., threatened species with unique chemistry).

## S.169.2 Making the most of it: what can be done with poor data from bad specimens?

Catherine Kidner¹, Thibauld Michel¹

1 Institute of Molecular Plant Sciences, University of Edinburgh, Royal Botanic Gardens, Edinburgh.

Herbarium data can be of varying quality, some if surprisingly good, some is terrible. How do you decide if a sample is worth working with and what can you get from samples which will not yield up good

sequence? We use examples from our own work to explore the limits of herbarium genetics and present an outline of approaches which can maximise useful information from samples which would never pass quality control in a commercial company.

## S.169.3 Integrative species delimitation of Xanthium sect Acanthoxanthium based on herbarium specimens and type material

Manzo, Eleonora¹, Tomasello, Salvatore¹, Justus Müller-Kiefer ¹, Sebastian Nillies ¹, Diego G. Gutiérrez ²

1. Georg-August University Göttingen, Germany
2. Argentine Museum of Natural Sciences (MACN), Buenos Aires, Argentina

Xanthium is a peculiar genus of the Asteraceae family, identifiable by its spiny female capitula (burs) and wind-pollination. Features of the burs are the primary morphological characteristics used for identifying taxa within the genus. The high degree of their morphological variability has led to inconsistent taxonomic treatment in the past, resulting in numerous names given by different authors. The genus can be divided into two sections: sect.Xanthium, which includes all taxa with unarmed stems; and sect. Acantoxanthium comprising plants with stems with trifurcate spines. The taxa of sect. Acanthoxanthium are native of South America, but due to human-mediated dispersal, some of its lineages have now reached a cosmopolitan distribution. Apart from the widespread X. spinosum, several other taxa have been described within this section. The aim of the present study is to determine species boundaries in Xanthium sect. Acanthoxanthium. The study employed phylogenomics, including target enrichment of hundreds of nuclear loci and complete plastomes, and geometric morphometrics, alongside coalescent-based species delimitation approaches. To ensure a comprehensive range of morphological
<figure><img src="image_396.png" /></figure>

variations in the section, including types and original material for some taxa, we surveyed key herbaria such as P, B, WU, and BA. Analysis of herbarium specimens that are nearly 200 years old, comprising type material, along with modern extraction techniques and integrative taxonomy analyses, have enabled us to identify distinct evolutionary lineages within the section. This further confirms the existence of two previously acknowledged species, X. spinosum and X. ambrosioides (Tomasello 2018). Two further independent lineages are inferred as species, the neglected X. argentum, initially described by Widder (1923) based on a single herbarium voucher from the province of &quot;Nubles&quot; in Chile and nowadays probably extremely rare; and X. catharticum, differentiated from X. spinosum, is its vicariant in the high-altitude environments of the Andes.

## S.169.4 Systematics in the age of herbariomics: a tale of sweet potatoes

Pablo Muñoz-Rodríguez¹,², John R.I. Wood³, Tom Wells²,³, Bethany R.M. Williams², Tom Carruthers⁴, Robert W. Scotland²

1 Universidad Complutense de Madrid, Madrid, Spain. 2 University of Oxford, Oxford, UK. 3 Royal Botanic Gardens, Kew, London, UK. 4 University of Michigan, Ann Arbor, Michigan, United States.

Sweet potato (Ipomoea batatas (L.) Lam.) is a top ten food crop worldwide. Cultivated in all tropical and subtropical regions of the world, it is a hexaploid species with a complex evolutionary history. Dozens of studies have tried to untangle its origin since the 19th century, but they accumulated as much useful information as error. This was due to a poor understanding of the relationship between sweet potato and its wild relatives, poorly defined species boundaries, and lack of robust phylogenies. In summary, the lack of a good taxonomic knowledge has hindered progress in sweet potato research. Herbarium specimens are an extraordinary resource for molecular data. However, the optimal use of these data requires a decisive, parallel taxonomic effort. We have dedicated the last ten years to investigating the origin and evolution of sweet potato in the context of a taxonomic monograph of Ipomoea. During this time, we have studied ca. 30,000 herbarium specimens with molecular sequence data from ca. 3,000 specimens – Sanger sequencing, HybSeq and whole genome sequencing. This enabled important results pertaining to sweet

potato, including species delimitation, the identification of sweet potato closest diploid and tetraploid wild relatives, or their differentiation from modern hybrids. In this talk, we discuss the role and importance of herbarium specimens and herbariomics in our work, as part of a taxonomic monograph and with a focus on sweet potato. We discuss our previous results and ongoing work on the origin, evolution and domestication of this important crop, and will highlight the importance of integrating genomics and taxonomy using herbarium specimens to accelerate tropical plant diversity studies.

## S.169.5 Herbariomics for plant conservation: the study case of the presumed extinct Armeria arcuata

Giulia Albani Rocchetti¹, Thomas Abeli¹, Ana Isabel D. Correia², David Draper², Eric Gouda³, Isabel Marques², Lorenzo Peruzzi⁴, Fred Stauffer⁵, Manuel Tiburtini⁴, Gonzalo Nieto Feliner⁶, Salvatore Tomasello⁷

1 Roma Tre University, Rome, Italy. 2 University of Lisbon, Lisbon, Portugal. 3 Utrecht University, Utrecht, The Netherlands. 4 University of Pisa, Pisa, Italy. 5 Conservatoire et jardin botaniques, Chambésky, Switzerland. 6 Real Jardín Botánico, CSIC, Madrid, Spain. 7 Georg-August-University, Göttingen, Germany

Herbaria play fundamental roles in many fields, including plant taxonomy, systematics, and conservation biology. In this framework, Herbariomics offers invaluable-and sometimes unique-opportunities to obtain data that can inform species' conservation. While investigating extinct plant species in Europe, we found at the Utrecht Botanical Garden a living specimen labelled as *Armeria arcuata* Welw. ex Boiss. & Reut., a species endemic to Portugal assessed as extinct (EX). After many years in cultivation, the specimens' morphological characteristics changed making a correct identification difficult. To unequivocally identify the accession of "A. arcuata" from Utrecht, we compared its DNA with herbarium and fresh material from type material and other related *Armeria* species. We collected and analysed samples of *A. arcuata* from the living specimen at Utrecht and three ancient herbarium specimens (including type materials, ca. 1848; G, LISU herbaria), *A. neglecta*, also extinct, from the type specimen (1882; COI), *A. caespitosa* from recent herbarium specimens (2003-2009; JBAG, HSS), *A. pinifolia* (1970; GOET), *A. pungens* (1997; GOET), and *A. curvifolia* (living;
<figure><img src="image_397.png" /></figure>

Teplice Botanic Garden). We applied DNA extraction techniques suitable for old herbarium material, along with adaptation of the normal double-stranded library preparation protocols. Genome skimming was performed on a lane of a NovaSeq sequencer, which allowed us to gather about four Gb of sequence information per sample. After quality trimming of the received reads, we performed reference-based assembly of plastomes and of the nrDNA region (about 12,000 bp long) and inferred phylogenetic tree and networks. Results from the phylogenetic analyses indicate that the plants cultivated in Utrecht are not actually *A. arcuata*. Moreover, network analyses suggest a hybrid origin of these cultivated plants.

## S.169.6 Explainable AI powered by shape analysis to capture the phenotypic variation of plants in the field and in herbaria

Ladislav Hodač¹, Kevin Karbstein¹, Lara Kösters¹, Michael Rzanny¹, Hans C. Wittich², David Šubrt³, Patrick Mäder²,⁴,⁵, Jana Wäldchen¹,⁴

1 Department of Biochemical Integration, Max Planck Institute for Biogeochemistry, Jena, Germany. 2 Data-intensive Systems and Visualization Group, Technical University Ilmenau, Ilmenau, Germany. 3 Faculty of Science, Jan Evangelista Purkyně University, Ústí nad Labem, Czech Republic. 4 German Centre for Integrative Biodiversity Research - iDiv, Halle-Jena-Leipzig, Leipzig, Germany. 5 Faculty

of Biological Sciences, Friedrich Schiller University Jena, Jena, Germany

Plant leaves vary not only between species, but also within populations of the same species. A persistent challenge in botany is to accurately document this variation directly in a plant's natural environment. In this study, we investigate the phenotypic variation of buttercup leaves in natural plant populations using an easy-to-use smartphone app called Flora Capture, designed for citizen scientists. We exploit the strengths of machine learning and geometric morphometrics to address three key questions: (1) Can smartphone images effectively capture leaf variation comparable to traditional herbarium methods? (2) Do image differences detected by computer vision correspond to actual variation in leaf shape? (3) Can smartphone images of leaves achieve detailed resolution for identifying plants at high taxonomic levels? Our approach incorporates several deep learning techniques, including Grad-CAM for interpretable AI and EfficientNet for feature extraction from images. We analyze the same leaves using computer vision and quantitative morphology - by extracting their shapes using landmark geometric morphometrics. Through dimension reduction and covariation analysis, we discover common patterns in both the image features and the corresponding biological shapes. Using machine learning algorithms such as Random Forest and Support Vector Machine, we evaluate the suitability of leaf images for classifying plant populations at high taxonomic levels. The goal is to understand the decision-making processes of computer vision in biological shape classification. This work represents a step towards reliable and reproducible phenotyping of plants in their natural habitats.

## S.170 FROM DEVELOPMENTAL SWITCHES TO BROAD EVOLUTIONARY TRAJECTORIES: UNDERSTANDING MECHANISMS OF SEX DETERMINATION

### S.170.1 Evolutionary developmental biology of sex in the Salicaceae

Quentin C. Cronk¹, MaKenzie Drowns², C.J. Tsai²

1 Department of Botany, University of British Columbia, Vancouver, Canada. 2 Warnell School of Forestry and Natural Resources, University of Georgia, Athens, USA
<figure><img src="image_398.png" /></figure>

The Salicaceae, and especially *Salix* and *Populus*, are becoming a leading model for the "clade biology" of sex determination, as dioecious, monoecious and polygamous forms are common. In angiosperms, dicliny (the production of single sex flowers), evolves through one of two contrasting evolutionary developmental mechanisms: organ deletion or organ abortion. Organ deletion, under which sex organs never form, may involve MADS-box genes, as these genes are required to produce floral organs of a given identity. Organ abortion, under which sex organs form but during development become vestigial or non-functional, are less likely to involve MADS-box genes, and more likely to involve late-stage developmental modifiers of organ development. Phylogenetic studies strongly suggest that there are necessary pre-adaptations for plant lineages to adopt either the deletion or abortion pathway. *Populus* and *Salix* are both examples of the deletion pathway, in which no trace of stamens is ever seen in female flowers, and no trace of carpels in male flowers. In poplar we show that the B-class MADS-box genes *PISTILLATA* (*PI*) and *APETALA3* (*AP3*) are strongly down-regulated in early female flower development. This is expected as these genes are required for stamen development under the canonical ABC model. A powerful way of dissecting the role of the MADS-box developmental module in dioecious species is provided by CRISPR/Cas9 gene editing. Recent developments in efficient poplar transformation coupled with engineered hyper-precocious flowering have facilitated novel experiments in this area. Preliminary experiments on MADS-box genes in poplar show the potential for functional dissection of pathways to dioecy. The development of CRISPR/Cas9 technology in other systems could greatly accelerate understanding of the evolutionary developmental genetics of dioecy in diverse lineages.

## S.170.1 The transition from dioecy to monoicy in liverworts is a case of convergent evolution

Giacomo Potente¹, Yasui Yukiko², Rachel N. Walstead³, Jane Grimwood³, Jeremy Schmutz³, Jim Leebens-Mack⁴, Eita Shimokawa², Tomoha Tanaka², Yuka Umeya², Shogo Kawamura², Katsuyuki T. Yamato⁵, Katsushi Yamaguchi⁶,

Shuji Shigenobu⁶, Masaki Shimamura⁷, Takayuki Kohchi² and Péter Szövényi¹

¹Dept. of Systematic and Evolutionary Botany, University of Zurich, Zurich, Switzerland ²Kyoto University, Kyoto, Japan ³HudsonAlpha Institute for Bio-technology, Huntsville, AL ⁴University of Georgia, Athens, GA ⁵Kindai University, B.O.S.T., Kinokawa, Japan ⁶Trans-Scale Biology Center, National Institute for Basic Biology, Okazaki, Japan ⁷Hiroshima University, Hiroshima, Japan

Sex chromosomes have evolved independently in many groups of plants and animals. Genetic sex determination ensures obligate outcrossing that may confer advantages against selfing. In liverworts, sex is expressed in the haploid phase and genetically determined by the presence of a single U (female) or V (male) sex chromosome. The UV sex determination is ancestral in liverworts and represents the oldest sex chromosome system ever described (430 million years old). Although the transition to monoicy occurred repeatedly in liverworts, its genomic basis has been investigated only in one species (Ricciocarpos natans). Here we report on the chromosome-scale genome assembly of the monoicous liverwort Marchantia quadrata, which diverged from the dioicous M. polymorpha 125 million years ago. We show that the M. quadrata genome consists of eight large and one 9th small chromosome (1/10th of the size of the large eight chromosomes). The eight large chromosomes are collinear with the eight autosomes of M. polymorphashowing many intra- but few inter-chromosomal rearrangements. By contrast, the small 9thchromosome contains genes homologous to the M. polymorpha V chromosome. Many genes homologous to U-chromosome genes of M. polymorpha are absent from the M. quadrata genome or are relocated to the autosomes, including the major sex determining gene These results imply that the transition from dioecy to monoicy was achieved by the translocation of few crucial U-genes to the autosomes and the loss of the U chromosome, while a larger proportion of V-linked genes was retained on a segment of the V chromosome as chromosome 9. Our findings are remarkably similar to the recent observation made on R. natans, raising the possibility that genomic changes underlying the transition from dioecy to monoecy in liverworts may be highly predictable.
<figure><img src="image_399.png" /></figure>

### S.170.3 Sex and/or asex: insights from brown seaweeds

Susana Coelho¹

1 Max Planck Institute for Biology Tübingen, Germany

In many eukaryotes, male and female sexes are determined during the (haploid) gametophyte stage of the life cycle, by the presence of a female (U) or male (V) sex chromosomes. The rise, evolution and demise of U/V systems have remained enigmatic to date. I present how we analyzing genomes spanning the entire brown algal phylogeny to decipher their sex- (and asex) determination evolutionary history.

### S.170.4 Analysis of sex determination system in Cannabis sativa

Ainhoa Riera-Begue¹, Matteo Toscani¹,

Jiaqi Shi¹, Susanne Schilling¹, Rainer

Melzer¹

1 School of Biology and Environmental Science, University College Dublin, Dublin, Ireland

Cannabis sativa is mainly a dioecious plant, which means that female and male flowers develop on separate individuals. This crop presents high sexual dimorphism, as we can see biochemical, vegetative and reproductive differences between the two sexes. C. sativa has a XY sex determination system: female have two X chromosomes and male plants carry an X and a Y chromosome; but the sexual system of this crop is much more complex, as the sex is not only determined genetically, but also by environmental and hormonal factors. While the developmental processes of male and female Cannabis sativa flowers clearly involves the orchestrated activation of numerous genes, it is conceivable that one or two key genes act as molecular switches for sex determination. The aim of our research is to understand how genes, phytohormones and environmental conditions interrelate and contribute to sex determination in hemp. Utilising transcriptomics analysis, we have pinpointed potential candidate genes on the sex chromosomes apparently associated with both female and male functional development in Cannabis sativa. Among those candidates are transcription factors with homologs know to be involved in reproductive development and sex determination.

We have also developed a PCR-based method to determine sex genotype and establish genotype-phenotype relationships. Our results indicate that sex determination is established very early during Cannabis development by a network of gene regulatory proteins, and that the genetic basis of sex determination is robust across different cultivars and chemotypes.

### S.170.5 The dynamic evolution of heteromorphic sex chromosomes in Cannabaceae

Sarah B. Carey¹

1 Hudson Alpha Institute for Biotechnology, Huntsville, USA

The Cannabaceae family has a deep history regarding their dioecious flowers and sex chromosomes. One century ago, the XY pair that controls the development of the sexes was identified in the hop *Humulus lupulus* var. *lupulus*, owing to the cytologically smaller Y chromosome relative to the X. Curiously, examinations across other species in *Humulus* and in the sister genera *Cannabis* uncovered varying cytological differences. In *Cannabis*, the Y chromosome is bigger than the X, while other varieties of *Humulus* remain homomorphic or have evolved a multiple sex chromosome system (e.g., XXYY). Despite these early discoveries, we know little about the Cannabaceae sex chromosomes at the molecular level. This is largely due to the complexities of assembling XY pairs in genome references. Here we use a combination of Illumina, PacBio HiFi, and Dovetail Omni-C data to assemble fully-phased genomes for XY males of *Cannabis* and *Humulus*. Using these assemblies, we first show that the XY pairs match the known cytology. We next use these assemblies to explore the dynamic patterns of gene gain and loss, as well as structural changes that have shaped the XY pairs. Ultimately these comparative analyses across Cannabaceae will uncover genes that control the development of the economically valuable females of hop and hemp.

### S.170.6 Long read sequencing to explore the repeated evolution of sex chromosomes in plants
<figure><img src="image_400.png" /></figure>

Alex Harkess¹

1 Hudson Alpha Institute for Biotechnology, Huntsville, USA

Sex chromosomes have evolved hundreds, if not thousands, of independent times across flowering plants. However, only a small number of master sex determination genes have been identified, largely due to a lack of high quality genomic resources for complex plant genomes that contain sex chromosomes. We have built computational pipelines for 1) assessing if a species has XY or ZW sex chromosomes,

2) assembling highly contiguous and fully-phased sex chromosome assemblies in heterogametic sexes, 3) delimiting the non-recombining sex determination region on these sex chromosomes, and 4) exploring the evolution of these regions in diverse plant species to identify putative sex determination genes in non-model plants. We use these computational tools to explore the repeated evolution of sex chromosomes in flowering plants, which is uncovering a hidden world of sex chromosome variation within and among species.

# S.171 PRESERVATION AND PROMOTION OF TRADITIONAL ECOLOGICAL KNOWLEDGE IN THE ERA OF OPEN SCIENCE: BENEFITS AND CHALLENGES

## S.171.1 The legitimacy of empirical sciences under the oversight of an Ethical Committee? Defending a critical ethnobotany

Adolfo Calvo Redondo¹

1 Centre de Recherche et de Documentation sur les Amériques, Université Sorbonne Nouvelle, Paris, France

In my ethnobotanical research in 2018 and my recent sociology-of-science fieldwork in Jujuy (Argentina), I address ethical challenges and question the need for expert committees in empirical sciences. I analyze how certain institutional mechanisms force a choice between the privatization of knowledge or its anonymization in the name of scientific universality. In Argentina, despite the 2023 Open Science Law (Law 27738), field research remains trapped in neo-developmentalism and patrimonialistic dynamics, especially when accessing biocultural information in an indigenous context. This reflects a dominance relationship of Jujuy institutions over scientists and their civil informants. The mistrust between academic and indigenist institutions is manifested in police controls and specific protocols for working with *Pueblos Originarios* (PO). Instead of addressing social exclusion from a Human Rights perspective, a contractual

practice is prescribed. Consent is seen as the legal solution to dialogue between "different cultures," leading to clientelistic practices among local authorities and researchers, namely by controlling field access. The patrimonialistic regime may promote these pre-research contracts to appropriate knowledge in favor to PO interests. Paradoxically, anti-colonial resistance is expressed through an anti-extractive discourse. In practice, then, control over science is transcended through underground dialogues, driven by a "community resistance" ethic that shapes the identity of both native populations and field scientists advocating empirical, inclusive, and critical methodologies within a "decolonial" framework. Critical ethnobotany is based on a "dialogue of knowledge" from a position of subalternity, promoting "interdisciplinarity" and "institutional coordination" to subvert the hierarchical and authoritarian structures of the neoliberal academic playground. However, this decolonial framework can perpetuate conservative dynamics if it does not overcome mistrust and secrecy in the transmission and production of knowledge. On a material level, effectively challenging the patrimonialistic regime requires expanding social justice policies by the State, ensuring academic autonomy in designing collaborative projects directly with the public.

## S.171.2 The language of land and life: importing data
<figure><img src="image_401.png" /></figure>

## sovereignty lessons and tools from linguistics into ethnobiology

Alex C. McAlvay¹, Stefanie Ramos Bierge¹, Tutupika Carrillo de La Cruz²

1 The New York Botanical Garden, New York, U.S.A.

2 Universidad Autónoma de Nayarit, Tepic, Mexico

Ecological knowledge and Indigenous languages are tightly interwoven and have been eroded in tandem in many parts of the world. To bolster or revitalize this cultural heritage, many Indigenous Peoples have worked with linguists and/or ethnobiologists, either from within or outside of their communities. Despite an increase in interdisciplinary projects at the intersection of these fields, there has been limited exchange of best practices, ideas, and resources related to ethical and effective work with communities. One area where linguists and ethnobiologists have innovated in parallel is data sovereignty. While linguists tend to deposit data in large centralized archives with different levels of access depending on community preferences, equivalent archives and graded-access capabilities for ethnobiology are much less common. At the same time, some scholars have been experimenting with Traditional Knowledge Labels and Biocultural Labels as a mechanism to ensure that community preferences for information use accompany the data. Linguistics and ethnobiology share significant overlaps not only related to the classification and encoding of biological knowledge, but also in the need for methodologies that foster ethical engagement with local communities. Cross-communication between the two disciplines on data sovereignty, community engagement, and other areas may lead to mutual benefit. Here we outline approaches taken to address these issues by both fields, examine opportunities for mutual enrichment, and share experiences from our project with Wixárika (Uto-Aztecan) communities in West-Central Mexico.

### S.171.3 The centers of origin as spaces for dialogue of knowledge

Alberto Betancourt Posada¹, Efraín Cruz Marín¹

1 Department of Intercultural Development and Management, Faculty of Philosophy and Letters, Mexico City, Mexico. 2 Master Degree in Environmental Education, Autonomous University of Mexico City, Mexico city, Mexico

Capitalism and colonization systematically persecuted and destroyed the knowledge of indigenous peoples. Currently, numerous argue the need for a new paradigm based on the dialogue of knowledge, which reverses positivist reification, vindicates traditional knowledge, decolonizes knowledge and establishes epistemic justice. In this article we maintain that since Nikolai Vavílov postulated the theory of the centers of origin of cultivated plants, these have become an important space for dialogue of knowledge between various scientific disciplines and the traditional knowledge of numerous cultures on topics such as the conservation of wild and domesticated biodiversity. The beginning of this dialogue sparked a scientific revolution, transformed the organization of knowledge and contributed significantly to the birth of multiple ethnosciences. In this text we ask ourselves: what importance does it have for the history of science that the centers of origin have become a space for dialogue between scientific disciplines and traditional knowledge? And what benefits has this dialogue brought to the various scientific fields? What are the main threats to the continuity of the centers of origin? And finally, how could they protect themselves? To answer these questions we follow bibliographically in the footsteps of Nikolai Vavílov, we integrate a large corpus of rom very diverse disciplines who recognize contributions of traditional knowledge; the factors that threaten them and the proposals to protect them. In our conclusions we agree with Casas and Parra (2016) who point out that farmers in the centers of origin constitute an evolutionary force, since of the total of 7000 species and hundreds of thousands of varieties of current agricultural and horticultural systems: "it is the result of evolutionary processes shaped by domestication, faster and more dynamic than those that occur in nature" (Casas and Parra, 2016).

### S.171.4 Historical documentation in the inventory of traditional knowledge associated with biodiversity: a tool for the recovery of neglec
<figure><img src="image_402.png" /></figure>

J. Esteban Hernández-Bermejo¹

1 Department of Forestry Engineering, Córdoba University, Córdoba, Spain

The author has participated in several initiatives in Spain and Latin America related to the inventory of knowledge associated with bio- and agro-diversity, always working with historical documentation, not only with that contained in textual sources but also from graphic and archaeobotanical sources. These include its participation in the IECTB and IECTBA (Spanish inventories of traditional knowledge associated with bio and agrodiversity), the FAFA (Agricultural and Forest Flora of al-Andalus), the CultIVA-CYTED network for the recovery of NUS (neglected and underutilized species) or various projects for the recovery and management of cultural landscapes in archaeological sites such as the Alhambra and Generalife or Medina Azahara in Andalusia (Spain) or the Jesuit reductions in the missionary-Guarani area. This experience demonstrates the interest of retrospective research in Historical Ethno-botany and its usefulness in recovering forgotten crops, phytotherapeutic knowledge, forms of agro-food consumption and forest uses that are once again of interest today in the fight against hunger, poverty and especially as elements of adaptation and even mitigation of climate change. They also make it possible to document the traditional value of many of these crops within a historical process of germplasm transfer between cultures, countries, and continents, turning apparent innovations subject to new codes of germplasm exchange and associated knowledge into archaeophytes, requiring a new analysis of the rights and sovereignty of peoples over part of their genetic and ethnobiological heritage. Some examples are examined such as the case of the argan tree [Argania spinosa (L.) Skeels] and its current potential cultivation in countries outside its natural range.

**S.171.5 Useful Plants and Fungi of Colombia: traditional knowledge for conservation, and conservation of traditional knowledge**

Mauricio Diazgranados¹,², Laura Kor²,³, William Milliken², Tiziana Ulian²,⁴

1 New York Botanical Garden, Bronx, NY, USA. 2 Royal Botanical Garden, Kew, Wellcome Trust Millennium Building, Wakehurst, Ardingly, West Sussex, UK. 3 King's College London, Bush House NE, London, UK. 4 University of Torino, Turin, Italy

Colombia is one of the world's most biodiverse and ethnically rich countries. As the nation continues resolving its socio-economic challenges, biodiversity, and the traditional ecological knowledge (TEK) associated with it are under increased threat by climate change, deforestation, habitat loss, acculturation, and other factors. The Useful Plants and Fungi of Colombia project aimed to develop pathways to enhance nature's contribution to people in Colombia by increasing, consolidating, and making accessible the knowledge of its useful plants and fungi for the benefit of society. The project produced catalogs of useful species, online portals, and a platform to facilitate sustainable value chain markets, alongside a wide range of publications, including books and scientific articles. A core research question was how the knowledge of useful plants could inform national conservation priorities and sustainable utilization. We applied the Important Plant Areas (IPA) approach to identify key sites for in-situ conservation, with 980 sites meeting IPA thresholds. We found 46 sites to be of high priority and highlighted 10 top-priority sites for further investigation and conservation actions. We also found that complementing scientific and traditional knowledge can better inform the conservation and support the sustainable use of plants in Colombia. We investigated whether citizen science could help mitigate the erosion of traditional ecological knowledge on useful plants through the use of digital surveys involving local citizen scientists. The trial in three pilot areas was successful, and we recommend building and strengthening the engagement of citizen scientists. Lastly, we carried out gap analyses at the spatial and taxonomic levels, to identify areas lacking floral inventories or ethnobotanical surveys, as well as underrepresented taxa. Overall, the project enhanced the baseline knowledge of useful plants and fungi, and the associated TEK, providing crucial information to support Colombia's transformation towards a greener, more sustainable economy whilst protecting its biodiversity.

**S.171.6 IECTB and CONECTe: two copyleft Spanish initiatives to document and**
<figure><img src="image_403.png" /></figure>

protect traditional plant
knowledge

Manuel Pardo-de-Santayana<fnref n="1" />,², Laura Aceituno-Mata³, Petra Benyei⁴, Airy Gras⁵, Almudena Lázaro³, María Molina³, Javier Tardío³, Victoria Reyes-García⁶,⁷,⁸

1 Departamento de Biología (Botánica), Universidad Autónoma de Madrid, Spain. 2 Centro de Investigación en Biodiversidad y Cambio Global (UAM), Universidad Autónoma de Madrid, Spain. 3 Instituto Madrileño de Investigación y Desarrollo Rural, Agrario y Alimentario, Madrid, Spain. 4 Dpto. de Economía y Geografía Aplicadas, Instituto de Economía, Geografía y Demografía, Centro de Ciencias Humanas y Sociales, CSIC, Madrid, Spain. 5 Departament de Biologia, Sanitat i Medi Ambient, Universitat de Barcelona, Spain. 6 Institució Catalana de Recerca i Estudis Avançats, Barcelona, Spain. 7 Institut de Ciència i Tecnologia Ambientals, Universitat Autònoma de Barcelona, Bellatera, Barcelona, Spain. 8 Departament d'Antropologia Social i Cultural, Universitat Autònoma de Barcelona, Bellatera, Barcelona, Spain

In the current global change framework, the erosion
of traditional ecological knowledge is pronounced,
remarkably in industrialized countries like Spain. Eth-
nobotanical studies conducted in the country re-
veal that this erosion varies across territories and
knowledge domains, with health or food exhibiting
a less profound decline. The overall erosion trend
is attributed to factors such as the abandonment

of traditional agricultural practices and shepherd-
ing, changes in land use, the diminished necessity
of wild plants for subsistence, reduced time spent
in the field, or negative associations with certain
species deemed famine food. Conversely, there are
multiple motivations that promote wild plants use.
Some view gathering as a recreational and com-
munity-building activity, while others appreciate the
unique flavors of wild plants that cannot be replicat-
ed by cultivated or commercial varieties. Addition-
ally, the medicinal role of food, particularly in uses
closely aligned with medicine such as beverages
and condiments, remains highly valued. Recogniz-
ing the significance of traditional ecological knowl-
edge, two initiatives in Spain are actively promoting
its documentation, protection, and reintroduction
to society. Firstly, the Spanish Inventory of Tradition-
al Knowledge related to Biodiversity compiles and
disseminates information on the traditional use and
management of biodiversity. Six volumes have been
published since 2014 and two more are being edit-
ed, including monographs about 837 plant species.
Secondly, CONECT-e (www.conecte.es) is an online
platform enabling citizens to document knowledge
and uses of both wild and domesticated species,
and includes 3772 monographs. By employing open
data and copyleft licenses, these endevours contrib-
ute to the traditional knowledge preservation in the
public domain, ensuring the unrestricted exchange
and reproduction of this valuable knowledge.

S.172 NATURAL HYBRIDIZATION AND SPECIES
CONSERVATION. SESSION 1

S.172.1 Hybridization in
Mexican Pinus - implications
for management and
conservation

M. Socorro González-Elizondo<fnref n="1" />, Martha González-Elizondo<fnref n="1" />, Norma L. Piedra Leandro², Jorge A. Tena-Flores<fnref n="1" />, Christian A. Wehenkel³, José J. Corral-Rivas⁴, Lluvia Flores-Rentería⁵

Durango, Mexico. 3 ISIMA, Universidad Juárez del Estado de Durango, Durango, Mexico. 4 FCF, Universidad Juárez del Estado de Durango, Durango, México. 5 Department of Biology, San Diego State University, San Diego, CA, U.S.A.

Hybridization is common in plants. Its outcomes are
wide and contrasting, from detrimental or causing
merging of species, to increasing their evolution-
ary and ecological resilience and even resulting in
hybrid speciation. In western and northern Mexico,
hybridization frequency among Pinus species rang-
es from occasional (e.g., isolated individuals of Pi-
nus chihuahuana × P. lumboltzii) to stable popula-
tions (e.g., of P. arizonica × P. cooperi). There is also a
wide range of fitness outcomes, from hybrid swarms

<footnote n="1">1 CIDIR, Instituto Politécnico Nacional, Durango,
México. 2 Jardín Etnobiológico Estatal de Durango,</footnote>
<figure><img src="image_404.png" /></figure>

composed by low fitness individuals (e.g., *P. arizonica* × *P. engelmannii*) to populations exhibiting hybrid vigor (e.g., *P. herrerae* × *P. luzmariae*). Some are recent (isolated F1s), and others are part of ancient introgressions (e.g., *P. cembroides* × *P. discolor*; *P. strobiformis* × *P. flexilis*); a syngameon encompassing pinyons has been recently recorded showing how sequential hybridization might have accelerated the speciation in this group. Gene flow among these species increases along ecological or geographical range edges and in disturbed communities. The high rate of hybridization in the zone may be owed to the facts that: a) Mexican pines are relatively recent (6 - 10.5 Ma), allowing for species to hybridize in absence of genetic incompatible systems, b) northwestern Mexico is a very rugged region offering multiple and diverse niches that favor diversification, and c) it is a boundary area for many species reaching there their northern or southern limits. Despite the evidence about their commonness, natural hybrids are not yet considered in forestry policies in Mexico. Combined names indicating hybridization or introgression are not allowed in inventories and monitoring reports nor in official paperwork. A shift of paradigm is needed to adjust forestry policies and management to the real situation in which genetically merged biological entities are common in forest ecosystems.

## S.172.2 Multiple glacial refugia and impacts of environmental variables on hybridization between *Betula* species of differing ploidy levels

Lu Liu¹, Feifei Wang¹, Junyi Ding¹, Yuangui Wei¹, Wenting Wang², Julia López-Delgado³, Nian Wang¹

1 College of Forestry, Shandong Agricultural University, Tai'an, China. 2 School of Mathematics and Computer Science, Northwest Minzu University, Lanzhou, China. 3 School of Biology, University of Leeds, Leeds, UK.

Natural hybridization frequently occurs in nature and plays a crucial role in evolution of biodiversity. Here we use the genus *Betula* to address the following questions: (1) where are their glacial refugia? (2) does hybridization occur between *Betula* species of various ploidy levels, (3) which environmen-

tal variables explain the spatial patterns of genetic admixture? We collected samples representing *B. costata* (2x), *B. dahurica* (8x), *B. ermanii* (4x) and *B. platyphylla* (2x) in northeast China and *B. ashburneri* (2x) and *B. utilis* (4x) from north to southwest China. We integrated microsatellite data of 2,609 individuals, species distribution models and spatial autoregressive models to address these issues. Our results revealed multiple glacial refugia for *Betula* species. Microsatellite data analysis showed frequent bidirectional genetic admixture among *Betula* species, irrespective of the ploidy difference. The amount of genetic admixture from either *B. costata* or *B. ermanii* to *B. platyphylla* is partially explained by their overlapped range and from *B. dahurica* to *B. platyphylla* is partially explained by precipitation of the coldest quarter (PCQ) and changes in PCQ. The amount of genetic admixture from *B. ashburneri* to *B. utilis* is partially explained by the mean temperature of the wettest quarter (MTWQ). Our results show asymmetrical gene flow from high to low ploidy levels and indicate that environemtal factors play a role in shaping the patterns of genetic admixture between *Betula* species.

## S.172.3 Optimal mating strategy in secondary sympatry may oppose reinforcement of premating isolation – A whole-genome study of mangroves

Sen Li¹, Miles Tracy¹, Ziwen He¹, Chung-I Wu¹, Suhua Shi¹

1 State Key Laboratory of Biocontrol, Guangdong Provincial Key Laboratory of Plant Resources, School of Life Sciences, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Sun Yat-sen University, Guangzhou, China

Reinforcement, the process by which premating isolation is strengthened by selection against the investment in maladaptive hybrids, is a key prediction of the evolutionary theory. Exceptions may therefore shed light on this controversial topic. Two mangrove species (*Bruguiera gymnorhiza* and *B. sexangula*) on the Indo-Malayan coasts come into secondary contact on the Hainan island, China, but rarely elsewhere. Strikingly, hybrids account for 30% of the individuals in sympatry and exceed the number from the parental species, *B. gymnorhiza*. Further-
more, while the two species in allopatry have dif-
ferent flowering times, they both flower year-round
in Hainan. Counter-reinforcement thus appears to
be at work in *Bruguiera*. To analyze the unusual hy-
bridization, we sequenced and *de novo* assembled
the high-quality genomes of the two species, fol-
lowed by the re-sequencing of 63 individuals from
allopatric and sympatric populations. The genomic
analysis identifies both F1s and multiple generations
of backcrosses. The hybridization pattern suggests
weak postmating isolation and, likely, ecological
speciation. Based on these patterns, we develop a
theoretical model whereby the allele that does not
exercise mate choice can be advantageous. In this
model, the mating allele of the weaker species may
do better by mating with the stronger and more
common species. The case of *Bruguiera* thus shows
that the evolution of mating preference as well as
reinforcement is a function of genic fitness, rather
than the fitness of the species itself.

S.172.4 Genetic bases of the
flower color variation in a
natural hybrid zone

Pascaline Salvado¹, Valérie Hinoux-Delome¹, Joris Bertrand¹

1 Laboratoire Génome et Développement des Plan-
tes, Université de Perpignan Via Domitia, Perpignan,
France

The Angiosperm radiation can be characterized by
its outstanding flower color diversity, spanning the
entire color spectrum of human and pollinator vi-
sion. Moreover, flower color is an evolutionary labile
trait which can differ between sister species, among
populations or even among individuals at different
life-stages. In *Pedicularis comosa*, two subspecies/
morphs display distinct flower color, *Pedicularis co-
mosa comosa* has yellow flowers while *Pedicularis
comosa asparagoides* has pink flowers. In order to
better understand this species taxonomy, to get an
overview of the overall genetic diversity and differ-
entiation, and to unravel the possible evolutionary
origins and genetic bases behind both morpho-
types, we applied a population genomic approach.
The phylogenetic tree provided us keys to settle an
old debate on *Pedicularis comosa*'s classification,
supporting that the two "subspecies" are not recip-
rocally monophyletic, and that genetic structure is
rather consistent with geography. Cline analyses
and genetic structure results revealed the existence

of a hybrid zone between the two morphotypes,
suggesting a strong selection on certain parts of the
genome, therefore maintaining differences between
P.c. asparagoides and P.c. comosa. Finally, the in-
vestigation of potential outlier SNPs associated with
genetic structure as well as phenotypic and envi-
ronmental data allowed us to find genes involved in
color differentiation but also in light and cold stress-
es adaptation. To uncover the subtle color differen-
tiation from pink to yellow, we applied comparative
transcriptomic and metabolomic approaches. All
together, these results provided us key information
to draw recommendations for the conservation of
these species as a whole.

S.172.5 Can hybrids modify
the adaptive trajectories of
parental species in contact
zones?

Modesto Berbel¹, Ana García-Muñoz¹,
Andrés Ferreira-Rodriguez¹, Melissa
Moniz¹, José Carlos del Valle², Eduardo
Narbona³, Gonzalo Nieto Feliner⁴, A. Jesús
Muñoz-Pajares¹, Mohamed Abdelaziz¹.

1 Departamento de Genética, Universidad de Granada, Granada, Spain. 2 Departamento de Biología Vegetal y Ecología, Facultad de Biología, Universidad de Sevilla, Seville, Spain. 3 Departamento de Biología Molecular e Ingeniería Bioquímica, Universidad Pablo de Olavide, Seville, Spain. 4 Departamento de Biodiversidad y Conservación, Real Jardín Botánico, CSIC, Madrid, Spain

The species hybridization is a phenomenon which
consequences can go from the collapse and loss of
diversity to the rise of this diversity. Global change, are
promoting the secondary contact and hybridization
between diverging taxa with incomplete reproduc-
tive barriers. These hybrid zones have the potential to
shed light on evolutionary processes driving adapta-
tion and are particularly powerful natural systems for
studying the interaction between divergent genomes
to explore how the rise of phenotypic novelties can
modify the evolutionary trajectory. We have studied
a total of 480 plants belonging to six populations
and two transects of two *Erysimum* (Brassicaceae)
species presenting a contact zone in the Serra da
Estrela (Portugal). The plants were phenotyped using
25 traits and genotyped using more than 4000 SNPs
across their whole genomes. Plants also were tagged
during a complete session of reproduction and we
used different component to quantify their fitness. We analyzed the patterns of hybridization using genomic approach, identifying two hybrid zones in different stages of hybridization. When we explore the patterns of natural selection acting on the parental and hybrid plants, we found different traits related to plant size and reproductive investment driving the population evolution for the parental plants. However, the natural selective pressures acting on hybrids were variable depending on the evolutionary stage of the hybrid

zone. In the hybrid zone enriched with F1-hybrids we identified new adaptive trajectories acting on traits not selected in parental species. But the hybrid zone enriched by F2 and backcrosses, the adaptive trajectories were similar to those found at parental sites. Our results demonstrate that hybrids promote the rise of novelties in plant evolution but in absence of transgressive phenotypes the hybrid zone tends to become in parental secondary contact zones.

# S.173 ASSESSING AND CLASSIFYING THE POSITIVE AND NEGATIVE IMPACTS OF ALIEN PLANTS

## S.173.1 Employing the Environmental Impact Classification for Alien Taxa (EICAT) on coastal areas: the example of the Costa Brava (Spain)

Carlos Gómez-Bellver¹*, Neus Ibáñez¹, Neus Nualart¹, Arnau Guiu-Bosch¹, Jordi López-Pujol¹

1 Botanic Institute of Barcelona, CSIC-CMCNB, Barcelona, Spain. *carlosmiguel.gomez@ibb.csic.es

High human population density and habitat fragmentation, in a future climate change scenario, may lead to an accelerating trend of biological invasions, particularly in Mediterranean coastal areas. A paradigmatic example of this is represented by the Costa Brava, a coastal strip of ca. 250 km stretching from the French-Spanish border to Blanes (Girona Province, Spain). It harbours a great richness of native species, some of them endemisms, as well as diverse natural habitats. However, it is under enormous anthropogenic pressure due to passive urbanization, mass tourism, and the presence of numerous invasive plant species, mostly escaped from gardens. In order to minimize new introductions of alien plants and improve the management of invasive species present in the Costa Brava, an European Union LIFE Program project (LIFE MEDCLIFFS https://lifemedcliffs.org/es/) is underway. The initial phase required the design of around a

hundred transects of 0.5-1 km in length distributed throughout the coastline to monitor by trained volunteers the evolution of 33 invasive or potentially invasive plant species populations (i.e., using citizen science). Large amount of data was gathered as a result of our field trips derived from examining the areas for the transects, together with the first results provided by the volunteers [which included the area (m²) and the size (census) of the detected populations of alien species]. All this information let us to identify and categorize the main impacts of these 33 invasive (or potentially invasive) species on native communities according to the Environmental Impact Classification for Alien Taxa (EICAT) criteria of the IUCN (2020). To our knowledge this is the first time that this standard classification has been used in a LIFE project and, also, specifically for a coastal area. Future projects employing this assessment methodology will contribute to a more accurate understanding of these impacts.

## S.173.2 Positive and negative invasive plant species ecological impacts in Europe

Montserrat Vilà¹², Alejandro Trillo¹, Pilar Castro³, Belinda Gallardo⁴, Sven Bacher⁵

1 Estación Biológica de Doñana (CSIC), Sevilla, Spain. 2 Department of Plant Biology and Ecology, Universidad de Sevilla, Sevilla, Spain. 3 Department of Live Sciences, University of Alcalá, Alcalá
de Henares, Spain. 4 Applied and Restoration Ecology Group, Pyrenean Institute of Ecology (CSIC), Zaragoza, Spain. 5 Department of Biology, University of Fribourg, Fribourg, Switzerland

Despite that many invasive species threaten biodiversity and ecosystems, their effects are highly idiosyncratic and context dependent. We compiled a comprehensive database of 287 publications on 4622 field studies on the ecological impacts of 114 invasive alien plants in 30 European countries to analyse their direction and frequency. Impacts were categorized across levels of ecological organization, taxa and feeding mode. Furthermore, if the information was available, the impacts were also classified based on the feeding mode of the impacted species or communities. Forty-four percent of the studies found significant impacts with more significant decreases (26.8%) than increases (16.6%) on the impact response variables. Negative impacts (i.e., statistically significant decreases) were more frequent on species and communities than on ecosystem properties. The impacts on ecosystem properties were highly variable. The frequency of negative impacts to species and communities was higher on plants than on animals, but the frequency of finding positive impacts (i.e., statistically significant increases) was similar between taxa. Finally, the frequency of negative impacts was higher on primary producers, herbivores and parasites than on decomposers, pollinators and predators, while the frequency of positive impacts was generally similar among feeding modes. Our database provides the first information system on field studies of the ecological impacts of invasive plant species at the continental scale. This information can be of interest for academic, management and policy purposes.

### S.173.3 Not all impacts are negative: a bidirectional impact assessment of the alien tree Robinia pseudoacacia

Giovanni Vimercati¹, Leon Strotz, Sven Bacher¹

1 University of Fribourg, Fribourg, Switzerland

Alien species cause various environmental impacts worldwide. Negative impacts have received considerable attention, as impactful alien species, named invasive, are a major driver of biodiversity decline.

The IUCN Environmental Impact Classification for Alien Taxa (EICAT) has been developed to assess negative impacts on native biodiversity. Alien species can also have positive impacts, e.g., through habitat/food provisioning or dispersal facilitation. However, these impacts have never been systematically examined, mostly due to the absence of a standardized protocol to assess their magnitude and diversity. We have filled this gap by developing EICAT+, a framework which structurally resembles IUCN EICAT, can be applied to all alien taxa at different spatial and organizational scales, and uses five semi-quantitative scenarios to describe the magnitude of observed positive impacts of alien taxa on native biodiversity. In EICAT+, the highest levels of impact magnitude are assigned to alien taxa increasing the area of occupancy of native species through local population re-establishment or preventing local extinction. EICAT+ also describes the underlying impact mechanisms by which alien taxa facilitate native biodiversity. To illustrate the functionality and utility of the framework, we have applied EICAT+ to studies reporting positive impacts of the alien tree black locust (Robinia pseudoacacia) and used IUCN EICAT to assess its negative impacts. About one third of all impacts were positive. Most impacts, both negative and positive, involved changes in population size of native species and were caused by alterations to abiotic conditions. Species that benefit from the introduction of the black locust show a preference for nutrient-rich soils and higher light intensity, in contrast to species negatively affected. Our results align with the existing literature on the black locust and demonstrate how the combined use of IUCN EICAT and EICAT+ enables to obtain a comprehensive understanding of alien species impacts on native biodiversity.

### S.173.4 Assessing the Ecological Impacts of Invasive Alien Plants of France using adapted protocols of EICAT and EICAT+

Thomas de Solan¹, Aurélien Caillon¹, Guillaume Fried², Yohann Soubeyran³, Arnaud Albert⁴

1 Conservatoire Botanique National Sud Atlantique, Audenge, France. 2 Anses, Laboratoire de la Santé
<figure><img src="image_405.png" /></figure>

des Végétaux, Unité entomologie et botanique, Montferrier-sur-Lez, France. 3 French committee of the International Union for Conservation of Nature, Montpellier, UMR AMAP (Univ Montpellier, CIRAD, CNRS, INRAE, IRD), France. 4 French Biodiversity Agency, Research and Scientific Support Department, Nantes, France

The creation of prioritized checklists ranking alien taxa depending on their risk for biodiversity is an essential step for an effective management of invasive alien species (IAS). In France, such lists for alien plants are drawn up by different structures at regional and national scales using various risk assessment methods. This lack of homogenization in assessment methods limits the implementation of the French national strategy on IAS. To address this problem, we investigated if two recent frameworks, EICAT and EICAT+, have the potential to become standards for assessing the ecological impacts of alien plants in France. We first identified the needs and constraints associated with the use of these two protocols, and proposed several modifications. We then assessed with the modified EICAT/EICAT+ (i) the regional impacts of

100 alien plants naturalized in a French region, and (ii) the national impacts of the 230 plants identified as invasive in France. Modifying EICAT and EICAT+ is tricky, as too many changes would compromise the standardization feature of these methods and limit comparison with assessments from other regions. Firstly, we propose criteria to better define some concepts (e.g. local population), which were difficult to handle with plants in a continental context. Sourced impact data are required in both methods, but are either non-existent or fragmentary for most species at a regional scale, leading to a high number of "data deficient" species. Therefore, we include local knowledge from botanists and wildlife specialists, in addition to literature review. Finally, we propose a key for translating field observations into impact levels, because local knowledge is usually related to species behaviour and often cannot be translated into EICAT(+) impact. This work provides the most comprehensive assessment to date of the current ecological impacts in France of 280 alien plants on a local and/or national scale.

# S.174 PARADIGM SHIFTS IN FLOWER COLOR

## S.174.1 New insights into the impact of non-pollinator agents of selection in flower color over the past ten years

José Carlos del Valle¹, Justen B. Whittall², M. Luisa Buide³, Eduardo Narbona³

1 Departamento de Biología Vegetal y Ecología, Universidad de Sevilla, Seville, Spain. 2 Department of Biology, Santa Clara University, Santa Clara, California, United States of America. 3 Departamento de Biología Molecular e Ingeniería Bioquímica, Universidad Pablo de Olavide, Seville, Spain

Besides the predominant role of pollinators in floral evolution, it has been suggested that the selective pressures exerted by abiotic agents may also contribute to variability in flower color. In the past ten years, we used the shore campion (Silene littorea) as a model species to develop studies aimed at gaining a better understanding of modifications of flower color driven by these selection agents, mainly focusing on the effect of solar, and more specifically in the UV light. We used different approaches to unveil color variation in this species, such as com-

mon garden experiments, transcriptomic and biochemical studies. Our results have highlighted that flower color in S. littorea is influenced to some extent by sunlight and UV radiation. However, the specific regulation of anthocyanin production in petals, the main pigments responsible for the flower color in this species, may reduce drastic changes on that trait, presumably due to its importance for pollinators. This is manifested in the lower plasticity observed in anthocyanin biosynthesis in petals compared to vegetative tissues. Additionally, transcriptomic and biochemical analyses have evidenced that petal pigmentation is tissue-specific regulated, resulting even in the occurrence of individuals with white flowers lacking anthocyanins without hindering anthocyanin accumulation in other tissues. Pollinator attraction seems to be at least partially favored by the presence of petal anthocyanins, although white-flowered plants can overcome this limitation by utilizing their mixed mating system and reproducing through autonomous selfing. Finally, the variety of plants lacking anthocyanins in both vegetative and reproductive tissues suggest that anthocyanins in photosynthetic tissues are the targets of non-pollinator mediated selection. In conclusion, we suggest that environmental stressors may drive antho-
cyanin accumulation in the whole plant, although it
may be limited in petals, at least in S. littorea. Future
research will incorporate further transcriptomic and
biochemical analyses.

S.174.2 Does allopolyploidy
lead to transgressive red
pigments in Silene?

Andrea E. Berardi¹

1 Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, USA

Determining how and why reproductive traits evolve between and within species is key to understanding patterns of speciation. In plants, both genome duplication as well as selection on floral traits, such as color, have been major drivers of angiosperm diversification. *Silene* (Caryophyllaceae) is a largely white- and pink-flowering diploid genus distributed throughout temperate regions. However, at least two independent polyploidization events occurred in the genus's expansion into North America, and most extant North American species are at least tetraploids. Two novel floral pigmentation strategies arose in the North American polyploids – red flower color and UV bullseyes. Insights into how these two floral color traits arose as well as the revised phylogeny of North American *Silene* polyploids are made from target capture of herbarium specimens, gene silencing, and character mapping of floral pigments to answer the following questions: What is the most likely driver of floral color evolution in *Silene* - pollinator shifts, habitat specialization, sympatric competition, abiotic factors, or polyploidy?

S.174.3 The impacts of
global change on floral
pigmentation

Matthew H. Koski¹

1 Department of Biological Sciences, Clemson University, Clemson, SC, USA

Floral coloration is nortoriously important for medi-
ating plant-pollinator interactions though support
for abiotic selection on floral pigmentation has in-
creased over that last few decades. Pigments pro-
duced in the anthocyanin biosynthetic pathway
both give rise to floral color diversity and are critical
compounds for abiotic stress responses. As such,

rapid abiotic shifts associated with global change
have the potential to drive temporal change in flower
coloration. I discuss results from collections-based
studies exploring whether flower color has changed
over the last ~100 years across a diverse set of angio-
sperm taxa and whether such change is linked with
abiotic factors such as temperature and ozone (a
proxy for ultraviolet irradiance). I then present data
testing whether abiotic-induced flower color plas-
ticity has the potential to contribute to responses of
flower color to global change. Results show that taxa
that have experienced stronger temporal changes
in ozone and temperature have experienced great-
er change in floral pigmentation. The response to
abiotic change depended on whether anthers are
exposed to external environmental conditions or
concealed by petal tissue, which is in line with petal
pigmentation's role in protecting pollen from abiotic
stress. Plasticity studies revealed that UV exposure
induced a pigmentaiton response in one focal spe-
cies, but not in all populations. Thus, there is genetic
variation for pigmentation plasticity. Temperature
change induced a weaker pigmentation response
that was not population-specific. Together, results
show that floral pigmentation has responded to
global change, and plasticity is one mechanism by
which plants have elicited phenotypic responses.
This change in pigmentaiton has the potential to
impact plant-pollinator interactions, and plant re-
productive output.

S.174.4 Flower colour
communities across
elevation - a case study
on calcareous vs silicious
bedrock from the European
Alps

Svenja Wanke¹*, Mani Shrestha¹, Andreas
von Heßberg¹, Anke Jentsch¹

1 University of Bayreuth, Germany *svenja.wanke@uni-bayreuth.de

Flower colour plays an important role in structuring
plant communities and is under selection by pol-
linators. Recently, the significant discrepancy be-
tween flower colour vision by humans versus pol-
linators has been discussed, which must be taken
into account when considering the role of flower co-
lour in communication with pollinators. Since bed-
rock and soil conditions are known to affect plant
<figure><img src="image_406.png" /></figure>

community composition, and can also affect flower colour directly, studies on flower colour composition of a community should include bedrock as an environmental variable. Elevational gradients in high mountain ranges are particularly suitable to study patterns and drivers of plant community diversity, including plant functional trait distribution relevant for trophic interactions. We measured flower colour spectra within six highly diverse grasslands on both calcareous and silicious bedrock across an elevational gradient in the European Alps: Hohe Leite (500 m asl), Brunnenkopfalm (1500 m), Obergurgl (2500 m), Kaserstattalm Stubai (1850 m), Arnspitze (1700 m) and Furkapass (2400 m). We collected flowers or petals of 315 locally occurring, insect-pollinated plants and measured their light reflectance properly using ocean optics spectrophotometer. We used the 'bumblebee color vision model' to translate spectral data into "bee-colours" because bumblebees are representative insect pollinators from low to high elevation in the European Alps. Our primary results suggest that for humans 'white' and 'yellow' flowers increase with higher elevation, whereas for bees 'blue' and 'green' flowers increase. Our data shows that calcareous grasslands have a higher color diversity than siliceous grasslands based on human vision, but higher number of blue and blue-green flowers based on hymenopteran vision. Subalpine area shows a higher color diversity based on human vision, despite alpine regions harboring a higher number of plant species. Based on pollinator vision, more species with blue and bluegreen flowers occur in the alpine than in the subalpine region.

### S.174.5 On colour to rule them all: the evolutionary rise of the betalains

Samuel F. Brockington¹

1 Department of Plant Science, University of Cambridge

The evolution of betalains within the order Caryophyl- lales, was a singular evolutionary event, both in the evolution of a completely new biosynthetic pathway to make pigments, but also in its replacement of the anthocyanin pathway. It has had profound implica- tions for the ecology and evolution of Caryophyllales, from the evolution of floral colour to the structural arrangement of the genomes. In talk I outline how our knowledge of the evolutionary fate of the betalain pathway and anthocyanin pathways in Caryophyl-

lales has been transformed, through transcriptomic and genomic resources, coupled with synthetic biolo- gy approaches. I address the role of modulation of the interface between primary and secondary metabo- lism, the prevailing importance of gene duplications, and the evidence for convergent origins of betalain pigmentation and iterative loss of anthocyanins. I ex- plore how flavonoid pathways have more generally changed in the context of a shift to betalain pigmen- tation, and discuss the impact of this on the variety of floral colour hues within Caryophyllales. Given the increasingly strong support for convergent evolution of betalains, I revisit hypotheses for the evolution of betalains and consider new and emerging evidence for their functions in the context of floral adaptation, and the unique adaptational advantages they offer with respect to flower-pollinator interactions.

### S.174.6 Identification of TFs controlling pigment synthesis in the endoholoparasite Pilostyles boyacensis and its host Dalea cuatrecasasii

Estefania Elejalde-Baena¹, Favio Gonzalez², Juan F. Alzate³, Natalia Pabon-Mora¹

1 University of Antioquia, Medellin, Colombia. 2 Na- tional University of Colombia, Bogota, Colombia. 3 National Center of Genomic Sequencing, Medellin, Colombia

Chlorophylls, carotenoids, flavonoids, and beta- lains generate colors in plants. Anthocyanins, a type of flavonoids, are the pigments responsible for red, pink, and purple hues in vegetative and reproductive tissues of plants. The synthesis of these pigments is primarily regulated at the tran- scriptional level by R2R3-MYB transcription factors assigned to subgroup 6. These genes have exten- sively studied in model plants such as *Arabidopsis thaliana*, *Antirrhinum majus*, and *Petunia hybrida*. In this study, we focused on a system composed by the endoholoparasite *Pilostyles boyacensis* and its exclusive host *Dalea cuatrecasasii*, both exhib- iting purple perianth organs. Two R2R3-MYB homo- logs from subgroups 5 and 7 were identified in *P. boyacensis*, and four homologs from subgroup 6 were isolated from *Dalea cuatrecasasii*. We found that R2R3 MYB genes from subgroups 5 and 7 are
<figure><img src="image_407.png" /></figure>

actively expressed in the endophyte, flowers, and
fruits of *P. boyacensis*. Meanwhile, R2R3 MYB sub-
group 6 homologs are expressed in the in the *D.
cuatrecasasii* inflorescences, as expected. Inter-
estingly, R2R3 MYB S6 homologs were also found
expressed in the *P. boyacensis* endophyte. These
results suggest that anthocyanin synthesis in the
flowers and fruits of the holoparasite *P. boyacen-
sis* may be mediated by host-specific R2R3 MYB

S6 genes, which are known to be mobile at least,
from *Citrus* to *Cuscuta*. Alternatively, anthocyanins
in *P. boyacensis* can be endogenously produced
R2R3 MYB genes from subgroups 5 and 7. We dis-
cuss our results in the broader context of conver-
gent floral pigments and the resulting color hues
in parasites with significant reduction in their body
plan and complete dependence upon their hosts.

# S.175 PLANT RESPONSES TO STRESSES AND CLIMATE CHANGE. SESSION 8

S.175.1 How will leaf nutrients in C2 vs C3 salad crops fare under climate change?

Catherine A. Walsh¹, Marjorie R. Lundgren¹

1 Lancaster University, Lancaster, UK

Climate change is placing immense pressure on
agricultural systems to ensure sufficient production
for an ever-growing global population. Producers
must not only increase yields to satisfy these de-
mands, but also safeguard crop nutrition to pre-
vent deficiency-induced diseases and consequent
malnourishment. Some crop species experience
nutritive decline under ambient atmospheric CO2
concentrations in comparison to crops grown his-
torically when atmospheric CO2 levels were much
lower. Of course, elevated CO2 does not occur in
isolation and the problem is further exasperated
by other environmental factors such as high tem-
peratures or drought. The extent of nutritional de-
mise, however, may be determined by the mode
of photosynthesis employed by the crop. Currently,
the only commercial food crop to use the rare C2
type of photosynthesis is wild rocket (Diplotaxis te-
nuifolia), a nutritionally dense salad crop with rich
potential to provide both high nutrients and yield
under climate change. Unlike C3 species, plants us-
ing C2 photosynthesis divide the photorespiratory
pathway across mesophyll and bundle sheath cells,
which functions to effectively concentrate and re-
assimilate carbon released from photorespiration to
ultimately improve photosynthetic efficiency under
warm and dry conditions. However, how C2 physiol-
ogy influences leaf nutrients remains unknown. Here
we present the first study of intraspecific diversity for

C₂ photosynthesis across 15 cultivars of wild rocket. We investigate how mineral and phytonutrient profiles change across a range of growth temperatures and in response to elevated CO₂ concentration across these C₂ wild rocket cultivars as compared to closely related C₃ salad greens. Our results suggest that wild rocket will retain nutrients better than related C₃ salad crops under climate change. This work has significant commercial application for current and future agricultural systems.

S.175.2 The ultimate guide of mycorrhiza and abiotic stress polytolerance in woody plants

Lauri Laanisto¹

1 Biodiversity and Nature Tourism, Estonian University of Life Sciences, Tartu, Estonia

Mycorrhizal symbiosis is an ancient association
between plants and fungi, crucially important es-
pecially for woody plants, as they have to main-
tain aboveground structures. Our current under-
standings of how woody plants respond to abiotic
stress and how mycorrhiza mitigates this stress is
very limited and considers almost exclusively just
a single stress factor at a time. Functional diver-
sity of both woody plants and mycorrhizal fungi
interacting with them, variability of the strength
and composition of multiple stress conditions in
different regions of the world – all this makes it
difficult to predict the patterns of these interac-
tions from both the adaptational and mitigational
point of view. In this study we used top-down ap-
proach, where known interactions are partitioned
into functional and biogeographical groups, and then the stress tolerances and interactions are mapped into overlapping heatmaps to provide us large-scale patterns of these associations. We compiled a concordant dataset of 621 woody species stress polytolerance (including shade, drought, waterlogging, cold stress) and their known species-specific mycorrhizal interactions. We tested how stress polytolerance correlates with different mycorrhiza functional types. Our results confirmed contrasting patterns between single vs. multiple type, arbuscular vs. ectomy-corrhiza, and obligate vs. facultative mycorrhizal interaction. Functionally different symbionts form significantly polarizing abiotic stress mitigation patterns with woody species with different life forms, growth forms, and biogeographical origin. These results provide insight into both evolutionary and biogeographic patterns related to the development of plant-mycorrhiza interactions.

### S.175.3 miRNA profiling in maritime pine (Pinus pinaster Ait.) subjected to water stress using grafting as an experimental system

Lorenzo F. Manjarrez¹, Nuria de María¹, María Á. Guevara¹, María D. Vélez¹, José A. Cabezas¹, José A. Mancha¹, Paula Ramos¹, Alberto Pizarro², Miriam López-Hinojosa¹, Irene Cobo-Simón¹, María C. Díaz-Sala², María Teresa Cervera¹

1 Institute of Forest Science - National Institute for Agricultural and Food Research and Technology, Spanish National Research Council, Madrid, Spain.
2 University of Alcalá, Alcalá de Henares, Spain

Growing body of evidence has established miRNAs as a key regulation component of multiple biological processes across diverse species. Yet, studies focusing on miRNAs in forest species, particularly in gymnosperms like conifers, remain scarce. This scarcity limits our comprehension of how forest trees sense and respond to environmental stimuli to manage stress. Pinus pinaster Ait., a native conifer to the western Mediterranean, has held significant ecological and socioeconomic value for decades, emerging as a European model species due to its remarkable genetic variability, which has made it possible for this species to thrive in a wide variety of habitats,

ranging from Mediterranean areas to the Atlantic coast. All these features make P. pinaster exceptional for studying the response and adaptation of conifers to water stress. In this study, we explored miRNA profiles from sampled stems of four different graft constructs. These grafts combined four genotypes that showed contrasting response to drought in previous studies. We used Gall056 (drought-sensitive) and Oria6 (drought-tolerant) as scions grafted onto either RIS (drought-sensitive) or R18T (drought-tolerant) rootstocks. The rootstock genotypes are F1 siblings derived from the controlled cross Gall056 × Oria6, vegetatively propagated. Three biological replicates of each graft construct were grown under water deficit and control conditions. This study will likely reveal potential roles of some miRNA families regulating transcripts associated with the response and tolerance to water stress. We can envisage they might regulate the expression of genes involved in response to osmotic and oxidative stresses, signaling of growth regulators (abscisic acid or ethylene) and controlling compatible solute metabolism. Additionally, other miRNA families, including two novel families, are involved in regulation of defence response and signal transduction.

### S.175.4 The effects of rising temperatures on the foliar and reproductive traits of the snowbed specialist Salix herbacea L.

Estela Illa Bachs¹,², Iu Vilaseca Juan¹, Núria Llauradó Nolla¹, Núria Roca Pascual¹

1 Departament de Biologia Evolutiva, Ecologia i Ciències Ambientals. Facultat de Biologia, Universitat de Barcelona, Barcelona, Spain. 2 Institut de Recerca de la Biodiversitat, Universitat de Barcelona, Barcelona, Spain

Snowbeds are those environments in temperate high mountains that remain covered by snow until mid-summer, and where only a set of specialised species are able to thrive. Climate change represents a major threat for these ecosystems and species. The rising temperatures cause an earlier snowmelt, and snowbeds become then favourable environments for the more competitive species growing in the surrounding alpine grasslands. The way snowbed specialists respond to changing environmental conditions through their vegetative and reproductive performance may be crucial
<figure><img src="image_408.png" /></figure>

to face not only new abiotic stresses but also the higher competition exerted by grassland species. In 2017, we set four pairs of control and OTC (Open Top Chamber) plots in four Pyrenean study sites with presence of the dioecious dwarf shrub snowbed specialist *Salix herbacea* L., with the aim to induce a moderate increase of temperatures on this species. In year 2021, we collected from 3 to 5 *Salix herbacea*'s female ramets with mature fruits per plot, and we analysed a set of foliar and reproductive traits, among which leaf area, LDMC, SLA, viable seeds/ fruit, total seeds/fruit or proportion of viable seeds. Preliminary results showed a site dependent trend towards a higher leaf productivity inside OTC, where we usually observed larger leaves with higher SLA and lower LDMC. Moreover, we also observed an increase of soil N in many of the OTC plots. Concerning reproductive traits, we found a trend towards a lower reproductive performance of *Salix herbacea* inside the OTC, with lower seed production and seed viability. Overall, the results suggest that increased temperatures have a fertilising effect in snowbeds, that enhance leaf productivity of *Salix herbacea*. However, the lower reproductive performance inside the OTC suggests that *Salix herbacea* may be approaching to its thermal fertility limit, which may pose a threat to its colonizing ability.

**S.175.5 Comparisons of genetic diversity between congeneric plants elucidates vulnerability of species in the face of environmental change**

Jennifer M. Cruse-Sanders¹, Jennifer N. Boyd², Jill T. Anderson¹, Carol Baskauf³, Jessica Bryzski⁴, Annie Lindsay³, Jared Odell², Will Rogers¹

1 University of Georgia, Athens, Georgia, USA. 2 University of Tennessee at Chattanooga, Chattanooga, Tennessee, USA. 3 Austin Peay State University, Clarksville, Tennessee, USA. 4 Seton Hill University, Greensburg, Pennsylvania, USA

Environmental change, accelerated by anthropogenic activities, threatens many species and can be particularly challenging for rare plants with potentially limited capacity for migration and adaptation relative to more common species. Plant species differ dramatically in their prevalence in

natural landscapes with some species characterized as rare due to restricted geographic distribution, low local abundance or habitat specialization. Several questions arise from these observed patterns, including ecoevolutionary causes and consequences of rarity. Our metanalyses of population genetic diversity, fitness and functional traits in rare and common congeneric plant species compared 252 rare species and 267 common congeners. Patterns identified for rare species included reduced population genetic diversity and fertilization efficiency, inbreeding depression and depressed fitness among other traits. Our recent comparisons of phenotypic plasticity and genetic diversity between rare and common congeners collected from natural populations elucidate rarity and vulnerability of an endangered riparian plant, *Pityopsis ruthii*, compared to more common congener, *P. graminifolia*. *Pityopsis ruthii* (Asteraceae) is a narrowly endemic perennial herb that grows in rocky crevices within a short distance along the banks of two rivers in Tennessee, USA. *Pityopsis graminifolia* is widespread within wooded savannahs, open grasslands and roadsides across the southeastern USA. Genetic diversity across the range of both species was estimated at nine punitatively neutral microsatellite loci. Results indicated that *P. ruthii* exhibited reduced genetic diversity relative to its more common congener. These results were compared with other population genetic studies of field collected congeneric pairs of rare and common perennial herbaceous species in the southeastern USA and evaluated along with other measures of fitness. Overall, rare species exhibited lower levels of genetic diversity and limited capacity to adapt or migrate in response to environmental change, highlighting potentially significant vulnerability to future change.

**S.175.6 Regulation of metal micronutrient homeostasis and toxicity by glutathione and phytochelatins in Marchantia polymorpha**

Erika Bellini¹,², Carlo Sorce¹, Andrea Andreucci¹, Valentina Vitelli¹, Alessandro Saba¹,³, Mingai Li⁴, Claudio Varotto⁴, Luigi Sanità di Toppi¹
1 University of Pisa, Pisa, Italy. 2 Sapienza University of Rome, Roma, Italy. 3 Center for Instrument Sharing of the University of Pisa, Pisa, Italy. 4 Research and Innovation Centre, Fondazione E. Mach, Trento, Italy.

As with all living organisms, photoautotrophs face challenges in regulating metal homeostasis. Plants have evolved mechanisms to control the uptake and accumulation of both essential and non-essential metals, including chelation and sequestration by ligands such as phytochelatins (PCn). These thiol-peptides are synthesized by the phytochelatin synthase enzyme (PCS). PCn production is a widespread strategy for detoxifying harmful metal(loid)s, such as cadmium (Cd), mercury (Hg), lead (Pb), and arsenic (As). However, recent studies suggest that PCn and PCS roles may extend beyond detoxification of harmful metal(loid) s, to include responses to homeostatic needs and toxicity of essential metals, including copper (Cu), zinc (Zn), and iron (Fe). Yet, PCS function in early plant lineages remains largely underexplored. This study focuses on the PCS of the model liverwort Marchantia polymorpha L., namely MpPCS. Using a recombinant MpPCS protein, we confirmed that the MpPCS gene encodes a 530 amino acids poly-

peptide with a molecular mass of ~57 kDa. In vitro activity assays with Zn, Cu, and Fe demonstrated that MpPCS catalyzes a transpeptidation reaction, with activation effectiveness in the order of Zn, Cu, and, to a lesser extent, Fe. Gene expression and in vivo function of PCS were investigated under conditions of starvation, metal excess and physiological concentrations of Fe, Cu, and Zn, with a range of different exposure times. Metal-induced impacts were monitored through photochemical efficiency analysis using a chlorophyll fluorometer, offering insights into light absorption, trapping, photochemistry, electron transport, and PSII structure. Our results reveal that MpPCS plays a crucial role in detoxifying metal excess and regulating the homeostatic control of metal micronutrients. Thus, thiol-peptides such as glutathione and PCn have proven their efficiency in managing metal needs and detoxifying their excess. It is worth noting that PCn production appears to be regulated by enzyme activity rather than gene expression levels. Furthermore, this study highlights a potential novel extracellular detoxifying role for PCn and glutathione in M. polymorpha.

**S.176 SPEEDING UP CONSERVATION IN POORLY KNOWN ORGANISM GROUPS – NEW APPROACHES TO BRYOPHYTE CONSERVATION. SESSION 2**

**S.176.1 A systematic review of the success of translocation experiments for bryophyte species.**

Izadora Kervin¹*, Jennifer Rowntree¹, Scott J. Davidson¹, Neil Bell²

1 School of Biological and Marine Sciences, Faculty of Science and Engineering, University of Plymouth, Devon, UK. 2 Royal Botanical Gardens Edinburgh, Edinburgh, Scotland. *izadora.kervin@plymouth.ac.uk

An estimated one-fifth of all botanical species are at risk of extinction with the most vulnerable of these species being bryophytes. This is due to them being

poikilohydric, causing bryophytes to be sensitive to changes in the environment. Translocation and re-introduction practices can be used to help reintroduce species into their native ranges, and ultimately help reduce the risk of extinction. However, currently there are no available guidelines specifically tailored towards the translocation and reintroduction of bryophyte species. We undertook a systematic review to advance our understanding of successful translocation and reintroduction projects. Using a standardised methodology, we systematically searched literature databases for articles on bryophyte translocations. The academic papers were filtered to 144 relevant articles (WoS = 138, EthOs = 4, ProQuest = 1, and Scorpus = 1), in which the subject and focus were assessed. From the remaining literature the estimated location of the translocation, distance transferred, reason for translocation, bryo-
<figure><img src="image_409.png" /></figure>

phyte species, duration of experiments and transplant method were extracted. The results show that most transplant experiments are for biomonitoring projects, with very little literature dealing with bryophyte transplants for conservation efforts. Most of these studies are short-term experiments with very limited long-term translocation projects. There were also disparities between bryophyte species utilised, for example, out of the 144 journal articles there was a total of 133 bryophyte species, of which 82.7% were moss species and only 17.3% were liverworts (4.7x more moss species compared to liverworts). This suggests that liverwort species are underrepresented within translocation and reintroduction experiments. This systematic review has highlighted a research gap in the frequency of academic papers focusing on conservation-based translocation experiments. To speed up bryophyte conservation we need to focus on increasing research efforts for translocations and reintroduction projects. Furthermore, we need to develop standardized guidelines for bryophyte reintroduction and translocations.

tralasia. 70% of threatened bryophytes are found in tropical forests, and the major threat is habitat loss. We hope this is a significant step towards extinction risk evaluation of bryophytes, but acknowledge its flaws. It is time-consuming, taxonomic knowledge may be lacking, publication is complex, information can already be out of date when it is published, species classed as Not Evaluated or Data Deficient due to taxonomic issues and/or poor recording are not adequately dealt with. New artificial intelligence techniques using morphological traits are being developed to indicate which species are pre-disposed to becoming threatened, tackling uncertainties and narrowing down the species to prioritize for IUCN Red List assessment while also speeding the process. However, collaboration between taxonomic specialists, ecologists and conservationists is paramount to fully estimate and understand the true number of bryophytes at risk, which and where they are, what threatens their survival and how to halt their extinction.

### S.176.2 Global assessments of bryophyte extinction risk

Neil Brummitt¹, Ana Claudia Araujo¹, Sinan Gürlek¹,², Kathryn Fowler¹, Giuseppe Signorino¹

1 Natural History Museum London, UK. 2 Imperial College London, UK

Bryophytes are fundamental ecological components of peatlands and tropical forests and provide important services such as carbon storage and nitrogen fixation. About 20% of species worldwide might be threatened with extinction (Brummitt et al. 2015), yet fewer than 3 % globally have been assessed under IUCN Red List criteria. Of those that have, two thirds are threatened with extinction (Vulnerable, Endangered or Critically Endangered) or Near Threatened, a further 10% classified as Data Deficient, but almost one third already need updating. These results reflect bias in the species selected for assessment; instead, we have randomly selected and assessed 1500 bryophyte species from around the world, and show that 215 are at risk of becoming extinct, 68 were classified as DD and 725 are Least Concern; the rest are poorly known and left as Not Evaluated. The greatest number is found in Africa, followed by the American continent, Asia, and Aus-

### S.176.3 Breaking the homogeneity of the eastern North American boreal forest: identifying bryophyte biodiversity hot spots for conservation

Nicole J. Fenton¹, Nils Ambec¹, Manon Carboni¹, Enrique Rodriguez Hernandez¹, Deanna Holt-Schmitt¹, Marion Noualhaguet¹, Xiangbo Yin¹

1 Université du Québec en Abitibi-Témiscamingue, Rouyn-Noranda, Canada

Conservation of bryophytes across boreal North America is complicated by the large spatial scale of the landscapes, the difficulty of access of many areas, and the relatively few bryologists studying this landscape. Consequently, the detailed habitat niche of many species are not identified, and many species are described as simply "boreal forest", which does not permit meaningful evaluation of the amount of habitat available for a given species, or even estimations of population sizes. Combining results from several large-scale field studies including nearly 1000 systematically sampled plots, we are able to determine
<figure><img src="image_410.png" /></figure>

patterns at different spatial scales from sites to
regions that illustrate the heterogeneity of bryo-
phyte species distribution patterns, but also of
community diversity. Individual species showed
clear patterns of frequency among ecosystem
types and across landscapes, linked to their spe-
cies traits and their ability to find the right micro-
habitat conditions with different combinations of
macrohabitat factors. While alpha diversity with-
in an ecosystem type varies with microsite rich-
ness as previously described, beta and gamma
diversity varied across landscapes within eco-
system types, in association with topographical
elements and regional gradients. Consequently,
describing site, landscape and regional patterns
in bryophyte communities allows us to refine the
existing concept that boreal bryophyte species
are inherently not rare because of the size of their
potential biogeographic niche. With these results
we are able to identify areas of diversity and high
interest for conservation.

S.176.4 About disturbed
landscapes and the unseen
diversity of a miniature
plant

Enrique Hernández-Rodríguez¹,²*, Adriel M. Sierra²,³,⁴,⁵, Marta Alonso-García²,³,⁴,⁵, Juan Carlos Villarreal²,³,⁴,⁵, Nicole J Fenton¹,²

1 Institut de recherche sur les forêts, Université du Québec en Abitibi-Témiscamingue, Rouyn-Noranda, QC, Canada. 2 Centre d'étude de la forêt, Québec, QC, Canada. 3 Département de Biologie, Université Laval, Québec, QC, Canada. 4 Institut de Biologie Intégrative et des Systèmes, Université Laval, Québec, QC, Canada. 5 Centre d'études nordiques, Université Laval, Québec, QC, Canada.
*enrique.hernandezrodriguez@uqat.ca

Habitat loss is the primary driver of species diversity
decline. One effect of habitat loss is changes in the
occurrence of new land cover (composition) and
the spatial pattern of different land cover (config-
uration) at different scales across the landscape.
Although the effects of changes in landscape
composition and configuration (LCC) on species
richness are increasingly well understood, little is
known about the response of genetic diversity to
such changes, particularly in non-vascular plants
such as mosses. In this study, we used a multiscale
approach to assess the effects of changes in LCC
on the population genetics of the dominant asexual

reproduction moss *Dicranum flagellare*. We sampled and SNP-genotyped 191 stems of *D. flagellare* from 12 old-growth boreal forest fragments in eastern Quebec, Canada. Our results indicate that genetic diversity (e.g., nucleotide diversity and allelic frequency) of *D. flagellare* is primarily negatively influenced by old forest reduction at large landscape scales (8-9 km radii; landscape composition). In addition, patch density (landscape configuration) of young forest areas reduces the genetic diversity of *D. flagellare* from old forest at small scales (2 km radius). We also found evidence of spatial genetic structure between and within populations depending on the old forest percentages (<40 % and >40 %) in the landscape. Our study shows that habitat loss at large scales and aggregation of recent disturbance areas at small scales affect the genetic diversity and spatial structure of non-vascular plants. Because of habitat loss at large scales, our results are useful for establishing logging thresholds that promote species conservation in landscapes with logging plans.

S.176.5 Bryophyte
communities in broad-
leaved forests in relation to
substrate and environment
along forest age
chronosequence

Evita Oľehnoviča¹, Anna Pastare-Skutele¹,
Ligita Liepiņa¹,², Anna Mežaka¹

1 Department of Biosystematics, Institute of Life Sciences and Technology, Daugavpils University, Daugavpils, Latvia. 2 University of Latvia, Faculty of Biology, Riga, Latvia

Broad-leaved forests are rich in bryophytes and pro-
vide microhabitats for many rare bryophyte species.
Forest transformation to agricultural lands in the
past and intensive forestry has led to fragmentation
of broad-leaved forests in Latvia. Previous studies in
broad-leaved forests mostly focused on bryophyte
communities in old-growth forest stands, but there
is still a lack of bryophyte studies in young and mid-
dle-aged forest stands. We studied bryophyte com-
munities in 30 forest stands dominated by European
ash (Fraxinus excelsior L.) and linden (Tilia cordata
Mill.) in Latvia. The aim of the study was to evalu-
ate the changes in bryophyte communities along
the forest age chronosequence from younger forest
<figure><img src="image_411.png" /></figure>

stands to middle aged and old-growth forest stands in relation to substrate and environmental variables that affect bryophyte communities in each forest age class. The results of the study can be applied to determine the best conservation practices to enhance bryophyte species diversity in broad-leaved forests of different ages.

**S.176.6 Spore germinability and longevity of four Sphagnum species along the gradient of altitude in the Changbai Mountains**

Zhao-Jun Bu¹², Jia-Qi Zhang¹,²

1 Key Laboratory of Geographical Processes and Ecological Security in Changbai Mountains, Ministry of Education, School of Geo-graphical Sciences, Northeast Normal University, Changchun, China. 2 State Environmental Protection Key Laboratory of Wetland Ecology and Vegetation Restoration, Institute for Peat and Mire Research, Northeast Normal University, Changchun, China.

Elevation can strongly influence the vitality and longevity of sexual propagules (seeds and spores) of mountain plants. Germination behavior of sexual propagules at a certain elevation can vary with

microhabitats. To date, little is known about how elevation and microhabitat differences in peatland ecosystems affect the germination and longevity of *Sphagnum* (peat mosses) spores. Here, we selected five peatlands in the Changbai Mountains along an elevation gradient (562-1472 m.a.s.l.) and conducted a three-year burial experiment with spores of four *Sphagnum* species. Our aim was to explore the effects of elevation and microhabitat on the germinability, dormancy percentage, and longevity of *Sphagnum* spores. We found that intermediate elevations were favorable for the germination of *Sphagnum* spores but unfavorable for dormancy maintenance and spore bank formation. Overall, *Sphagnum* spores had greater longevity in hollow microhabitats at low elevations and in hummock microhabitats at high elevations. Correlation analysis showed that spore germination of hollow species was more sensitive to temperature change than that of hummock species, with *S. squarrosum* demonstrating the clearest response to both temperature and precipitation changes. Our study suggests that elevation and habitat may interactively affect spore germinability and persistence of spore banks in peatlands. This highlights the importance of considering the difference of both elevation and microhabitat in vegetation restoration by using spore banks in degraded peatlands in the future.

**S.177 SYSTEMATICS, FLORISTICS, AND CONSERVATION: FACILITATING DATA INTEGRATION TO PROMOTE SOUND SCIENCE.**
**SESSION 3**

**S.177.1 Conservation**
**Challenges and Priorities for**
**Rafflesia (Rafflesiaceae) in**
**the Philippines**

Tobias, A.B.¹,², Malabrigo Jr., P.L.²,³,⁴, Maldia, L.S.J.²,⁵, Buot Jr., I.E.⁶, Chris Thorogood⁷,⁸

1 Graduate School, University of the Philippines Los Baños, Philippines. 2 Department of Forest Biological Sciences, University of the Philippines Los Baños, Philippines. 3 Museum of Natural History, University of

the Philippines Los Baños, Philippines. 4 Land Grant Management Office, University of the Philippines Los Baños, Philippines. 5 Makiling Center for Mountain Ecosystems, University of the Philippines Los Baños, Philippines. 6 Institute of Biological Sciences, College of Arts and Sciences, University of the Philippines Los Baños, Philippines. 7 University of Oxford Botanic Garden, Rose Lane, Oxford, UK. 8 University of Oxford Department of Biology, Oxford, UK

We present a synthesis of the taxonomy, distribution, and conservation status of *Rafflesia* in the Philippines. Through meticulous taxonomic review, field surveys, and data compilation, 16 *Rafflesia* species have been identified in the country, with the addition of a newly
<figure><img src="image_412.png" /></figure>

discovered species, *R. balatociana*. The IUCN Catego-
ries and Criteria assessment categorizes ten species
as Critically Endangered, five as Endangered, and one
as Vulnerable, emphasizing the urgent need for con-
servation action. The study highlights various threats
faced by *Rafflesia*, including deforestation driven by
commodities, shifting agriculture, urbanization, ille-
gal forest harvesting, and plantation establishment.
Alarmingly, half of the recorded *Rafflesia* sites are lo-
cated outside of designated protected areas, under-
scoring the necessity for targeted conservation effort.
Even within protected areas, some *Rafflesia* species
face habitat degradation and tree cover loss. To en-
sure their conservation, actions are recommended
to address the drivers of habitat loss, promote sus-
tainable land-use practices, and establish more pro-
tected areas. Effective monitoring and collaboration
among local communities, government agencies,
and conservation organizations will be vital for achiev-
ing positive conservation outcomes. Furthermore, we
identify strengthening legal frameworks and enforce-
ment, habitat restoration efforts, regular monitoring,
and raising awareness as essential steps towards the
recovery of *Rafflesia* in the Philippines.

S.177.2 Mulshi Watershade
Catchment, NWG, India:
Floristic Assessment, impact
analysis and Biodiversity
Conservation

Rani B. Bhagat¹

1 Department of Botany, PDEAs Baburaoji Gholap College, Sangvi Pune, Maharashtra, India

The Western Ghats (WG) are a UNESCO World
Heritage site in India and one of the world's rec-
ognized biodiversity “hot spots.” It is shrunken by
25% over the past decades, degrading unique &
valuable ecosystem. The physiography and cli-
mate of the NWG sustain tropical semi-evergreen
to moist deciduous forest vegetation, which has
enormous worldwide relevance for biodiversity
conservation. Research area is biogeographical-
ly located in the Sahyadri mountains and spans
multiple ecological zones such as the crestline of
the Western Ghats, hill ranges, and a gently slop-
ing section abutting the Deccan plateau, sup-
porting numerous plant communities. Due to high
rainfall, processes of soil formation are relatively
faster than low rainfall areas on plains, though

happening at geological time scale. All of these
distinct physical characteristics have resulted
in the formation of distinct habitats. Study area
is centrally located in NWG at 18 °25'N to 18 ° 41' N
and 73 °20' E to 73 °25' E. which covers 240 sq. km
in Pune district, Maharashtra. Extensive surveys
reported 1592 flowering species from 776 gen-
era and 162 families. Number of flowering spe-
cies from dicotyledones were 1185 and 337 from
monocotyledones, of which 150 are endemic spe-
cies and more than 50 IUCN categorized. Study
also documented 74 sacred groves with rich flo-
ra. In 2013, state government designated Tamhi-
ni as a wildlife sanctuary from Mulshi. Important
water catchments includes-Mulshi, Temghar,
Varasgaon and adjoining Pawana dam. Besides
a private planned cities like- Lavasa, Sahara and
Amby valley lies in the study area. Further, more
hill cities are proposed in various parts of Mulshi
threatening the biodiversity. Iconic' Western Ghat
is facing a threat due to forest loss, encroach-
ment & conversion. With this backdrop the doc-
umentation of floristic diversity, impact analysis
and conservation of diversity was carried out for
future ecosystem management.

S.177.3 Plagiarism in Botany
and Plant Classification:
Unearthing Ethical
Dilemmas in Scientific
Research

Orlando Alvarez¹,²,³

1 The SAR Group, LLC, Florida, USA. 2 Plant Biology Department, Herbarium, Michigan State University, Michigan, USA. 3 Herbarium, University of Florida, Florida, USA

Plagiarism in botany manifests itself in various
forms, ranging from word-for-word replication of
a text without due recognition to the appropria-
tion of experimental data and research discover-
ies; the latter materializes most of the time in new
combinations of nomenclature that eliminate
and also obscure any original hip. However, this
secondary result could also be an asset to use
in a court of law in a civil case for plagiarism or
copyright infringement, or both. The consequenc-
es of such unethical practices extend beyond ac-
ademic misconduct because they undermine the
foundations of scientific progress and thwart the
development of accurate plant classification sys-
<figure><img src="image_413.png" /></figure>

tems. It is no secret that plant classification within botany has played and will continue to play a fundamental role in expanding our understanding of the diverse plant kingdom, encompassing taxonomy, ecology, and evolutionary biology. However, in our haste to use phylogenetic data to validate, change, or generate new hypotheses about the evolutionary relationships within said plant kingdom, we are once again nullifying all scientific integrity with renewed acts of plagiarism; acts that unfortunately will be overlooked and little criticized and their perpetrators will feel covered, as happened in the past, under the false cloak provided by the international code of botanical nomenclature. The presentation explores specific instances of plagiarism within the large fern family Thelypteridaceae over the past 60 years, and its implications for the reliability of some taxonomic and genomic databases, as well as for regional floras; all of this hinders true progress in our understanding of the diversity and evolution of Thelypteridaceae. It also emphasizes the need for collective efforts within the scientific community to combat plagiarism in botany.

**S.177.4 Assessing curatorial practices and improving access to natural resource data**

Celia C. Aceae¹

1 School of Biological Sciences, University of Edinburgh, Edinburgh, UK

Herbaria represent vast repositories of data that are integral for conservation work and have the potential to contribute to research across biological disciplines. The value of herbaria and the data of the specimens within depends largely on the accessibility of this information and the accuracy of specimen identification, which in turn depends on curatorial work. Here, we assessed the current practices in curation and identification in a global survey for herbarium curators. The results were compared by the size of the responding herbaria, which were identified as small, medium, or large institutions based on the number of specimens held. This assessment found that there is a vast need for more global standards in herbarium curation. Curators of herbaria are limited by the resources they have available and the training in curation they have received. Curation practices are often varied within herbaria themselves and differ by size and geographical region of the herbarium. Curators each have different

approaches to continually changing taxonomic information, and these discrepancies cause further challenges for the researchers using herbarium specimens and data. Here, we present the survey results, an overview of current herbarium curation practices under different resource limitations and propose a recommended workflow for curators with varying resource restrictions. Three levels of curation standards were identified, and a checklist associated with each level was developed for use in future curatorial work.

**S.177.5 Advances in the systematics of genus Ternstroemia Mutis ex L.f. (Pentaphylacaceae s.l.): Towards a taxonomic monograph**

José R. Grande A.¹

1 Facultad de Ciencias, Universidad de Granada, Granada, Spain

Ternstroemia Mutis ex Lf. has been, until recently, a poorly known genus from tropics and subtropics of America, Africa, Australia, and Southern Asia, with one species reaching Korean peninsula and Japan. In the last years a wealth of contributions spanning from classic taxonomy and biogeography to phylogenetic systematics have shown the existence of two main clades. As a product of a revision of literature and a new combined analysis, including DNA, morphology, and anatomy, major transitions within the genus can be traced along with morphologic shifts in flowers, related with pollination syndrome, and for the most part following a geographic structure. As a consequence of the new, updated phylogeny, the Asian genus Anneslea Wall. is transferred to Ternstroemia, resulting in an expanded circumscription where tribe Ternstroemieae becomes monogeneric. Six monophyletic clades can be distinguished, with type species (Ternstroemia meridionalis Mutis ex Lf.) embedded within a pantropical super-clade, still to be solved. Pistil morphology, especially in the stigmas are correlated with corolla, stamen, calyx and bracteole morphology, and all of them to certain anatomic features in leaves. After the present revision Ternstroemia Mutis ex Lf. is a genus with ca. 141 species, 87-103 of which are present in the Neotropics, ca. 49 in Asia (or its adjacent islands, including Papuasia), and 4 in Africa. A taxonomic overview of the genus is presented, including major areas of
<figure><img src="image_414.png" /></figure>

distribution and endemism, number of species per
country, and species complexes. Future venues for
systematic research in the genus are suggested,
and a provisional species list is offered.
<figure><img src="image_415.png" /></figure>

# S.178 MOUNTAIN BIODIVERSITY AND EVOLUTION. SESSION 2

## S.178.1 Origin and assembly of the alpine flora in the Northern Hemisphere

Wen-Na Ding¹,², Richard H. Ree³, Michael R. May⁴, Alex Skeels⁵, Philipp Brun², Loïc Pellissier²,⁶, Yao-Wu Xing¹, Niklaus E. Zimmermann²

1 CAS Key Laboratory of Tropical Forest Ecolоgy, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla, Yunnan, China. 2 Swiss Federal Institute for Forest, Snow and Landscape Research WSL, Birmensdorf, Switzеerland. 3 Negaunee Integrative Research Cen ter, Field Museum, Chicago, IL, USA. 4 University Herbarium and Department of Integrative Biolo gy, University of California, Berkeley. 5 Research School of Biology, Australian National University, Canberra, Australia. 6 Department of Environ mental Systems Science, Ecosystems and Land scape Evolution, Institute of Terrestrial Ecosys tems, ETH Zurich, Zurich, Switzerland.

The alpine biota above the treeline of the high mountains harbors unique floristic characters adapted to stressful environments and provides critical ecosystem services to our society. Yet, the origin, assembly pattern, and biotic interchange across different mountain regions remain enigmatic. Here, we investigated the diversification and assembly history of alpine biotas in the North- ern Hemisphere aligning with paleoenvironmental changes. Our findings reveal that the alpine bio- diversity is disproportionally assembled by colоnization and in situ diversification. Alpine lineages initially diversified in the Tibet-Hengduan-Himalа- ya (THH) region as early as the Oligocene, playing a pivotal role as the primary source of alpine flora until it was overtaken by the arctic-boreal region by the Pliocene. The diversity of alpine plants in other major mountains of the Northern Hemisphere be- gan accumulating since the early to middle Miо- cene, but significantly diversified until climatically suitable habitats expanded towards the end of the Miocene. Our study highlights the significant role of the THH region as both the cradle and museum of cold-adapted plants, with a subsequent increase in the importance of the arctic-boreal region.

## S.178.2 Evolution and species diversification of seed plants in the Hengduan Mountains

Lianming Gao¹,⁴, Chaonan Fu¹,², Hongtao Li², Zhiqiong Mo¹,³, Mingshu Zhu¹,³, Tingshuang Yi², Dezhu Li²

1 CAS Key Laboratory for Plant Diversity and Bioge ography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan, China. 2 Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan, China. 3 University of Chinese Academy of Sciences, Beijing, China. 4 Lijiang For- est Biodiversity National Observation and Research Station, Kunming Institute of Botany, Chinese Acad emy of Sciences, Lijiang, Yunnan, China

The Hengduan Mountains, also named Mountains of Southwest China (MSC), is one of the global biodiversity hotspots with the most extensive- ly elevated surface on Earth, harboring around 10840 seed plant species from 1542 genera and 194 families. The origin and evolutionary history of the Hengduan Mountains flora remains unclear. We analyzed a time-calibrated phylogenetic tree of over 21000 seed plant species with 206 fossil calibrations based on 80 plastid genomic genes to investigate the evolutionary history and spe- cies diversification covering over 70% seed plant species from the Hengduan Mountains (7663 species from 1435 genera and 194 families). Our results showed that extant lineages (genera) in the Hengduan Mountains mainly immigrated from other floras and emerged after the early Oligocene. The rapid species diversification and accelerated diversification took place after the middle Miocene (ca. 15 Mya), especially in Plio- cene (5.3–2.6 Mya) and Pleistocene (2.6–0.1 Mya), and the regional endemic species had the simi- lar diversification pattern, which was likely driven jointly by mountain building, intensification of the Asian monsoon, Neogene climate cooling, and Quaternary climate fluctuations. The results of the molecular dating, species diversification, and ancestral area reconstruction analyses on nine representative genera of seed plants showed
that the early lineages mainly immigrated from other floristic regions, and accelerated from early Miocene (ca. 20 Mya), and peaked in ca. 5 Mya, mainly from the Saharo-Arabian realm. The *in situ* speciation with an increased rate of *in situ* speciation started from the later Miocene-early Pliocene to Pleistocene, likely triggered by Hengduan Mountains orogeny, the intensified Asian summer monsoon, and Quaternary climate fluctuations.

## S.178.3 Reconstructing the ecological and evolutionary assembly of the Alpine Flora

Sébastien Lavergne¹, the PhyloAlps consortium

Laboratoire d'Ecologie Alpine, CNRS - Université Grenoble Alpes - Université Savoie Mont-Blanc, Swiss Federal Research Institute, Jardin du Lautaret, Genoscope-Evry, Conservatoire Botanique National Alpin, Conservatoire Botanique National Méditerranée, Parc National des Ecrins, Parc National de la Vanoise, Parc National du Mercantour, Info-Flora, Institute of Biological Research Cluj-Napoca, Botanic Garden - Babeş-Bolyai University, Charles University, Institute of Botany Krakow, Institute of Botany Bratislava, Museum National d'Histoire Naturelle, Evolution et Diversité Biologique Toulouse, Universitat Autònoma de Barcelona, Universität Wien, Universität Innsbruck, Università di Pisa, British National Museum

Biodiversity genomics is an emerging field that leverages advanced sequencing technologies to analyze the genetic makeup of entire species assemblages in order to gain insights into their evolutionary history, adaptation, and ecological roles. I present the PhyloAlps project, which has now been running for over 10 years, and consisted in sampling and sequencing all vascular plant species of the Alps, a biodiversity hotspot harboring 20% of the European flora. This initiative has now expanded onto the flora of the arctic realm and other mountain ranges through the work of companion projects, thus providing an ever-growing, unique genomic resource aiming toward a comprehensive genomic characterization of the arctic-alpine flora. I finally present some use of these data to study the phylodynamics of alpine plant clades, the delimitation of novel species and the metabarcoding of sediment-cores time-series.

## S.178.4 Genetic diversity assessment of Juniperus seravschanica populations from Central Asia

Moldir Yermagambetova¹, Shyryn Almerekova¹, Orzimat Turginov², Ormon Sultangaziev³, Saule Abugalieva¹, Yerlan Turuspekov¹

1 Institute of Plant Biology and Biotechnology, Almaty, Kazakhstan. 2 Institute of Botany, Tashkent, Uzbekistan. 3 Fauna and Flora International, Bishkek, Kyrgyzstan

Juniperus seravschanica Kom. is a widespread and important tree species in Central Asia, and grows extensively in the mountain ranges of Oman to Uzbekistan, Kyrgyzstan, and Kazakhstan. It is crucial in forming shrub-forest massifs in mountainous regions, contributing to soil drainage and stabilization at medium to high altitudes. To gain a comprehensive understanding of the current status of *J. seravschanica* resources and develop effective conservation strategies, a thorough study of the species' genetic diversity and population structure was conducted. To assess genetic diversity and population structure leaf samples were collected from 15 *J. seravschanica* populations across Uzbekistan, Kyrgyzstan, and Kazakhstan. Utilizing 11 polymorphic simple sequence repeat (SSR) markers genetic diversity parameters such as the number of alleles, number of effective alleles, percentage of polymorphic loci, and Nei's genetic diversity index were evaluated. Nei's genetic diversity index for the *J. seravschanica* populations averaged 0.450, ranging from 0.407 to 0.566. The AMOVA indicated that 90.3% of the total genetic variation was distributed within populations, suggesting a high level of genetic diversity within each population. Gene flow calculated using alleles from all populations, was estimated at 4.654. Population structure analysis revealed weak clustering among the studied populations, confirming the AMOVA findings. The results of this study provide valuable insights into the genetic diversity and population structure of *J. seravschanica* in Central Asia. The high genetic diversity within populations suggests that the species has a strong capacity to adapt to environmental changes. These findings can be effectively utilized to develop conservation strategies for *J. seravschanica*, ensuring the long-term sustainability of this important tree species in Central Asia.
Acknowledgments: This work was supported by Grant No.AP09259027 of the Science Committee of the Ministry of Science and Higher Education of the Republic of Kazakhstan.

## S.178.5 The signature of mountain uplift on biogeography, trait evolution, and phylogenomic complexity of the Andean flora

Laura P. Lagomarsino¹, Ana M. Bedoya¹, Laura A. Frost²

¹Shirley C. Tucker Herbarium, Louisiana State University, USA. ²University of South Alabama, Mobile, USA

The Andean mountains of western South America are one of the World's most species-rich biodiversity hotspots and are home to almost 15% of all angiosperm species. Much of this diversity origi-

nated in concert with relatively recent mountain uplift and the resulting landscape changes, including high levels of habitat heterogeneity. In this talk, I discuss how Andean uplift has left its mark on the evolution of its flora using *Freziera* (Pentaphylaceae) as a primary example. *Freziera* is an Andean-centered clade of 75 species of trees and shrubs that diversified during the late stag- es of Northern Andean uplift in the last 12 million years. Subsequently, *Freziera* disperded into the Central Andes repeatedly. Across the cloud forest habitats that it can now be found, it displays an impressive diversity of leaf morphology, which we demonstrate is connected to local environmen- tal conditions (especially precipitation and sea- sonality). Finally, *Freziera's* high levels of phylog- enomic complexity can be attributed, in part, to high diversification rates and short times between speciation and introgression— both scenarios whose impacts were heightened by Andean up- lift. Throughout the talk, comparisons between *Freziera* and other Andean plant clades will high- light generalities—and idiosyncracies—of Andean plant evolution.

## S.179 PHYLOGEOGRAPHY

### S.179.1 The role of hybrid zones on the phylogeography of two willow species pairs (Salix L.) in the European Alps

Loïc Pittet¹, Natascha Wagner¹, Pia Marinček¹, Elvira Hörandl¹

¹Department of Systematic Botany, University of Göttingen, Germany

The geographic distribution of genetic lineages is highly impacted by climate fluctuations. Cold-adapted plant species have survived the last glaciations on nunataks within the glaciated area of the European Alps, unglaciated areas at the periphery of the ice sheet, in the marginal lowlands, or in other mountains of the European Alpine Systems that have been less glaciated. Following the last glacial period, cold-adapted species started to spread to newly available areas. The colonization is influenced by species-specific factors as well as

evolutionary processes like hybridization and is still on-going. Two closely related but previously isolated species that merge their marginal ranges during recolonization create a secondary contact zone where hybridization can occur and potentially block the recolonization. Using Restriction-Site-Associated DNA sequencing, morphometrics, species distribution modelling and fine-grained environmental data, we aim to identify potential glacial refugia and recolonization routes of two willow (Salix L.) sister species pairs that show vicariant distributions and secondary contacts in the European Alps. We aim to characterize the hybrid zones genetically, morphologically, and ecologically to test the hypothesis that hybrid zones influence biogeographical patterns of parent species. Different potential glacial refugia were found in the southern European mountain systems or alpine periphery and suggest recolonization of the Alps from both eastern and western marginal areas. Hybrid zones show patterns of introgression rather than hybrid speciation and extend over several hundred kilometers. A discrepancy between phenotype and genotype could have resulted from backcrossing and segregation of later generation
hybrids. Suitable environmental conditions were found for each species beyond their current distributions. Ecological relevees of secondary contact zones indicate high ecological niche similarity with parents but also niche expansion. No apparent reduction of fertility of hybrids was observed in the field. Altogether, our results support the hypothesis that hybrid zones can block the recolonization of parents.

## S.179.2 Phylogeographic insights to delimitate Mediterranean protected endemic species *L. cuspidatum* evolving in sympatry with close taxa

Cyllène Chatellier¹,², Baumel Alex³,
Christine Tollon², Sylvain Santoni², James
Molina¹& BouchaibKhadari¹,²,⁴*

1 Conservatoire Botanique National Méditerranéen de Porquerolles, Antenne Occitanie-Méditerranée-Occitanie, Montferrier-sur-Lez, France. 2 AGAP Institut, University of Montpellier, CIRAD, INRAE, Institut Agro, Montpellier, France. 3 IMBE, Institut Méditerranéen de Biodiversité et d'Ecologie marine et continentale, Aix Marseille University, Station Marine Endoume, ch. Batterie des Lions Marseille, France. 4 Conservatoire Botanique National Méditerranéen, UMR AGAP Institut, Montpellier, France

While the protection statute of threatened plants is designed at species level, delimitating plant species is a challenge for conservation biologists, especially for diversified taxa evolving in sympatry. Species delimitation exist at the interface of population genetic and phylogenetic analyses. Here, we aimed to delimitate species to question the species statue of a rare and protected taxa in the *Limonium confusum* group. *Limonium cuspidatum* (Delort) Erben has a restricted range geographical distribution from Narbonne to Marseille in south France. Morphological descriptors are not sufficient to clearly distinguish this taxon from three others taxa cited in the same environments in the south of France: *Limonium confusum* (Godr. & Gren.) Fourr. 1869; *Limonium legrandii* (Gaut. & Timb.-Lagr.) Erben, 1978; *Limonium densissimum* (Pignatti) Pignatti, 1971), all belonging to the *Limonium confusum* group. To understand the evolutionary history of these populations and taxa belonging to Mediterranean lineage of *Limonium* genus, we analysed a comprehensive sampling of

populations of *L. confusum* (23 from France, 8 from Spain) and 29 Mediterranean *Limonium* species by combining population genetic and phylogenetic approaches. First, using population genetic analysis of 31 populations of *L. confusum* with 12 Simple Sequence Repeat loci, we clearly identified three genetic groups. A first group includes Iberian-Provençal population, from the salty mud of the Bouches-du-Rhône (France) to the Tarragona and Ebro Delta region (Spain). The second group includes populations of the salt marshes located in the Languedoc coast between Leucate and Vendres (France). The third group includes three disjunct populations which however share similar ecological conditions. Second, we selected samples based on genetic and geographic origin and added representative species of Mediterranean Lineage of *Limonium* genus for phylogenetic analysis using plastid DNA variation. We expect to give insights for delimitating species of *L. confusum* and to define populations, which have to be protected in the future.

## S.179.3 Phylogeography of Magnolia Sect. Macrophylla in Mexico: insights into its origin, diversification and evolutionary history

Gerardo Hernández-Vera¹*, Marco Ibarra-Martínez¹,², José A. Vázquez-García¹,
Eduardo Ruiz-Sánchez¹, Marco Cerna²,
Miguel A. Muñiz-Castro¹, Arturo Sánchez-González³

1 Centro Universitario de Ciencias Biológicas y Agropecuarias, Instituto de Botánica, Universidad de Guadalajara. Zapopan. Jalisco, Mexico. 2 Carrera de Biotecnología de los Recursos Naturales, Universidad Politécnica Salesiana. Quito, Ecuador. 3 Centro de Investigaciones Biológicas, Universidad Autónoma del Estado de Hidalgo, Pachuca, Hidalgo, Mexico. \*gerardohvera@hotmail.com

In Mexico there are approximately 40 species of the genus *Magnolia* (Magnoliaceae), five of them included in the Sect. *Macrophylla*: *M. alejandrae*, *M. dealbata*, *M. nuevoleonensis*, *M. rzedowskiana* and *M. vovidesii*. Although most of these are threatened and some have sociocultural importance, its evolutionary history and the biogeographical events underlying their diversification and distribution are still unknown. In the present study, we analyse sequence data from four cpDNA regions (trnH-psbA, ORF350, trnK-matK and trnL-rpl32) of
<figure><img src="image_416.png" /></figure>

*Magnolia* species Sect. *Macrophylla* distributed in Mexico in order to: (1) infer their phylogenetic relationships, (2) assess its genetic diversity and population structure, (3) evaluate possible signatures of demographic expansion and (4) infer its temporal and geographical origin. Bayesian inference and maximum likelihood analyses corroborated the monophyly of Sect. Macrophylla. Interspecific relationships were not resolved and *M. alejandrae* and *M. nuevoleonensis* were recovered as a single monophyletic group. The highest genetic diversity was found in populations of *M. dealbata*, *M. alejandrae* and *M. nuevoleonensis*. Bayesian Skyline Plots and mismatch distribution analyses indicate that, overall, the studied populations are in a process of demographic expansion, particularly, the populations of *M. alejandrae* and *M. nuevoleonensis*. Analyses of genetic and phylogeographic structure indicate that geography has played a fundamental role in shaping the genetic structure of *Magnolia* Sect. *Macrophylla* in Mexico. Based on molecular clock analyses, initial diversification of Sect. *Macrophylla* is estimated to have occurred ca. 16.1 Ma, whose ancestral area probably included the Sierra Madre Oriental, Sierra Madre del Sur and Trans-Mexican Volcanic Belt. Given that the origin of the Section broadly coincides with the climatic changes of the middle Miocene and with the emergence of the Trans-Mexican Volcanic Belt, these events could have played a fundamental role in the diversification of *Magnolia* Sect. *Macrophylla* in Mexico.

### S.179.4 Phylogeographical patterns in Peruvian coastal desert Oxalis reveal two independent lineages

Leonardo Gaspar¹, Feng Song², Ning Fu², Lu Jin², Frank Guzmán³, Asunción Cano¹, Mónica Arakaki¹, Xue-Jun Ge²

1 División Botánica, Museo de Historia Natural - UNMSM, Lima, Perú. 2 Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, China. 3 Universidad Científica del Sur, Lima, Perú.

The genus *Oxalis* is a rather notable and constitutive plant group in Peruvian lomas formations, displaying conspicuous traits to cope with aridity. The flora of this seasonal ecosystem is highly endemic, much like its inhabiting *Oxalis* members, thus placing this genus as optimal for testing evolutionary and biogeographic hypotheses. While elucidating the diversity and distribu-

tion of *Oxalis* along the lomas extension (latitude from 6° to 18° S), a total of 11 species were identified, largely showing connections with western Andean slopes. Two main lineages were identified and further examined, corresponding to succulent herbs in section *Carnosae* (subg. *Oxalis*) and sub-shrubs in section *Thamnoxys* (subg. *Thamnoxys*). The phylogenetic reconstruction revealed differing evolutionary stories for each lineage, directly linked with alternative adaptive strategies and geographical colonizing patterns. This study also allowed filling information gaps related to understudied and endemic taxa, that, in addition to the non-monophyletic arrangement detected in some endemic species, exposes the need for a taxonomic update in the genus. Furthermore, a set of genomic resources, such as plastidial markers and nuclear SSRs, were generated for future phylogenetic and population genetic studies, mainly centered in the broadly distributed sect. *Carnosae*. Lastly, these results provide valuable insights in the understanding of the origin of plant diversity in Peruvian coastal lomas.

### S.179.5 Insights into the evolutionary and biogeographic history of the Carduus-Cirsium group in the African Sky-Islands

Lucía D. Moreyra¹, Jennifer R. Ackerfield², Carme Blanco-Gavaldà³, Christian Brochmann⁴, Juan A. Calleja⁵, Kazumi Fujikawa⁶, Mercè Galbany-Casals³, Abel Gizaw⁴, Cristina Roquet³, Alfonso Susanna¹

1 Botanic Institute of Barcelona, CSIC-Ajuntament de Barcelona, Spain. 2 Department of Biology, Colorado State University, U.S.A. 3 Systematics and Evolution of Vascular Plants - Associated Unit to CSIC by IBB, Autonomous University of Barcelona, Spain. 4 Natural History Museum, University of Oslo, Norway. 5 Autonomous University of Madrid, Spain. 6 Kochi Prefectural Makino Botanical Garden, Japan.

The Afromontane archipelago consists of isolated humid and temperate areas on the highest mountains in Tropical and South Africa. This biome hosts a unique flora characterized by a high proportion of endemic species. Despite its uniqueness, the Afromontane flora is understudied from an evolutionary and biogeographic point of view. Several afromontane endemics belonging to subtribe Carduinae (Asteraceae) have
<figure><img src="image_417.png" /></figure>

traditionally been ascribed to the *Carduus-Cirsium* group, based on superficially similar morphologies. To investigate their evolutionary origin, we applied the Hyb-Seq approach on >500 samples comprising all genera of subtribe Carduinae. We recovered sequences for >1000 conserved orthologous nuclear loci plus the complete plastome, and conducted phylogenetic inference using concatenated and coalescence-based approaches. The Afromontane species constituted three distinct evolutionary lineages, one of them not even belonging to the *Carduus-Cirsium* group. Given the evolutionary and morphological distinctiveness of these three lineages, we propose three new genera, all endemic to the Afromontane flora:

*Afrocarduus*, *Afrocirsium* and *Nuriaea*. Building on our highly resolved phylogeny, we conducted ancestral range estimation analyses. Our results indicate that these three endemic genera originated after two independent long-distance dispersals from the Mediterranean region. The first occurred during the Middle Miocene and resulted in *Nuriaea*, and the second occurred in the Late Miocene and gave rise to *Afrocarduus* and *Afrocirsium*. The current diversity within the three genera mainly results from speciation during the Pliocene and Pleistocene. The increased orogeny in Tropical East Africa during these periods, combined with the dramatic Pleistocene climate oscillations, likely fueled the diversification.

# S.180 MORPHOMETRICS

## S.180.1 A machine learning approach to morphometry of Armeria (Plumbaginaceae) in peninsular Italy

Manuel Tiburtini¹, Giovanni Astuti¹, Fabrizio Bartolucci², Liliana Bernardo³, Fabio Conti², Duilio Iamonico⁴, Mauro Iberite⁴, Simone Orsenigo⁵, Luca Sandroni¹, Lorenzo Peruzzi¹

1 University of Pisa, Italy. 2 University of Camerino, Italy. 3 University of Calabria, Rende, Italy. 4 Sapienza University of Rome, Italy. 5 University of Pavia, Italy

In the contemporary era of genome-wide phylogenomic analyses, the significance of morphology in Botany is crucial but often overlooked. Morphometry emerges as a method for scientifically and statistically handling morphological data, particularly in intricate groups like *Armeria* (Plumbaginaceae) in peninsular Italy, which has undergone multiple taxonomic revisions without yielding conclusive outcomes. In this study, we measured 27 features from 587 herbarium specimens across 34 populations spanning across the Apennines. The specimens encompass the *A. arenaria*, *A. denticulata*, *A. canescens*, and *A. macropoda* complexes for a total of 11 taxa, 10 of which putatively endemic to Italy. Since *A. gracilis* has been always traditionally related to the Balkan *A. canescens*, we also included the latter species for comparative purposes. To analyze the data, we employed two innovative machine learning approaches.

Dimensionality reduction was achieved using a novel non-linear technique called UMAP (Uniform Manifold Approximation and Projection). Additionally, Gaussian mixture models (GMM) were used to test alternative grouping hypotheses. The 3D - UMAP analysis along the Italian peninsula reveals six main groups, i.e., from northern to southern Italy: *A. arenariasubsp. praecox*, *A. arenaria subsp. marginata*, *A. denticulata* complex (*A. denticulata* and *A. saviana*), two groups within the *A. canescens* complex (including also *A. gracilis*, *A. garganica*, and two populations referred as 'A. macropoda'), and the *A. macropoda* complex (*A. aspromontana*, *A. brutia*, *A. macropoda* s.str., and a population from the Pollino Massif referred as 'A. gracilis'). Bayes factor calculations, fitting a supervised GMM, support the recognition of these six groups over other grouping hypotheses, constituting a comprehensive overview of our taxonomic exploration. Seed morphometric, karyological, and phylogenetic studies are ongoing and will be integrated into a conclusive taxonomic study.

## S.180.2 Morphology 2.0 - discovery through 3D imaging and big data of cacao and its relatives (Malvaceae: Byttnerioideae)

Katherine A. Wolcott¹, Edward L. Stanley², Osman A. Gutierrez³, Stefan Wuchty¹,⁴, Barbara A. Whitlock¹
<figure><img src="image_418.png" /></figure>

1 Department of Biology, University of Miami, Coral Gables, FL, USA. 2 Department of Natural History, Florida Museum of Natural History, Gainesville, FL, USA. 3 Subtropical Horticultural Research Station, USDA-ARS, Miami, FL, USA. 4 Department of Computer Science, University of Miami, Miami, FL, USA

We are beginning to realize the potential for discovery using big data and biodiversity archives. As 3-dimensional (3D) imaging with micro-computed tomography (micro-CT) and photogrammetry becomes more common, workflows to streamline processes and develop novel studies using large-scale digital imaging are needed, especially for botanical specimens in a vertebrate-focused field. We present a case study on the 3D pollination biology of *Theobroma cacao* and its relatives (Malvaceae: Byttnerioideae) that highlights discoveries possible through combining cutting-edge imaging modalities and big data, which we call Morphology 2.0. Despite the importance of chocolate, little is known about its specialized pollination (likely by micro-dipterans and/or -hymenopterans) and even less about other Byttnerioideae. First, we used micro-CT, 3D geometric morphometrics (GMM), and scanning electron microscopy to precisely quantify plant-pollinator geometry, functional size limits for pollinators, and floral reward structures in cacao. Then, we extended our methods to *Ayenia euphrasiifolia*, an endemic of endangered Florida pine rocklands. To gain baseline data on putative pollinators, we used a GoPro camera trap (modified for subjects <1cm), 3D imaging with micro-CT and DSLR camera photogrammetry, then 3D GMM analysis. After refining methods for cacao and *Ayenia*, we applied them to other Byttnerioideae (*Guazuma*, *Herrania*, *Byttneria*, *Commersonia*). Processing and analysis pipelines are open source and available on GitHub, CT datasets on MorphoSource, and 3D models for outreach on Sketchfab. By combining lab and field-based imaging technologies, we developed a novel tool for studying pollination biology of an important crop. Our 3D imaging also enabled high resolution comparative study of one of the nine major Malvaceae subclades, characterized by unusual floral morphology not captured in 2D. Moreover, we produced a large annotated imaging dataset (N>200) that will be repurposed to train computer vision models that automate 3D shape prediction and digitization of botanical specimens.

### S.180.3 Innovative multi-scale phenotyping approach on Amaranthus spp. leaves reveals contrasting differentiation traits at young stage

Dora Scarpin¹, Giacomo Este¹, Francesca D'Este¹, Francesco Boscutti¹, Marco Vuerich¹, Elisa Petrussa¹, Enrico Braidot¹

1 University of Udine, Udine, Italy

Imaging analysis can aid in phenotyping plant morphological traits at different scale levels, with options ranging from microscopy images to remote sensing acquisitions. Through this approach, it is possible to gain a minimally invasive understanding of physiological and structural trait diversity, as well as the relationship between plants and the environment. Much attention has already been paid towards the high phenotypic plasticity of the *Amaranthus* genus, as it includes highly invasive weed species harmful to summer crops. The identification of morpho-anatomical traits in amaranth leaves at early stage could be beneficial in gaining a greater understanding of the relation between their structural characters and their performance in the agroecosystem. In this work, we applied an innovative multi-scale phenotyping approach on single leaf morphometric traits of four *Amaranthus* species (*A. hybridus*, *A. palmeri*, *A. retroflexus* and *A. tuberculatus*) by processing digital images of intact leaves and confocal reflection microscopy acquisitions on adaxial surface imprints. The analysed morphometric variables belonged to three groups: macroscopic leaf traits, evapotranspiration-related traits and microscopic traits related to the leaf surface. The results evidenced that different investigation levels contribute to revealing species-specific characters even at a juvenile stage, which is crucial for plant development and the establishment of competition. In detail, some variables concerning macroscopic and evapotranspiration-related traits were those that best discriminated *A. tuberculatus* from the other three species. The approach proved to be an effective, reliable, and low-impact procedure for describing the functional characteristics of phylogenetically related species and quantifying traits associated with taxonomy and biodiversity studies.
<figure><img src="image_419.png" /></figure>

S.180.4 Morphometric
analysis of freshwater
estuary endemic Bidens
(Asteraceae)

Maurane Bourgouin¹,², Robert F. C. Naczi³,
Simon Pesant¹,², Étienne Léveillé-Bourret¹,²

1 Centre sur la biodiversité, University of Montreal,
Montreal, Québec, Canada. 2 Marie-Victorin Herbarium, University of Montreal, Montreal, Québec, Canada. 3 New York Botanical Garden, New York, USA

The unusual ecological conditions provided by
freshwater estuaries, namely, high rate of sedi-
mentation of nutrient rich minerals, twice-daily
tides, waves, and ice-scouring, have induced the
evolution of a highly specialized flora containing
several endemics. The origin of this flora appears
to be recent, dating at most to the last deglaci-
ation 10,000-12,000 years ago, which is probably
the cause of relatively minor morphological dif-
ferentiation of most of the endemic taxa. Unfor-
tunately, freshwater estuaries also are heavily ur-
banized and industrialized, contributing to habitat
degradation and loss. Endemic Bidens taxa are
an important component of the freshwater estu-
aries of Northeastern America and offer a great
insight into understanding the recent evolution of
the North American estuarine flora. Many of these
endemics are hypothesized to have originated
from hybridization and polyploidization (allo-
polyploidy). Unfortunately, taxonomic uncertain-
ties of many of the Bidens taxa of Northeastern
America are hampering conservation action. For
instance, unclear morphological limits between
species frequently result in misidentifications,
which can lead to over- or under-estimation of
the frequency of threatened taxa. We conduct-
ed a morphometric analysis to test species de-
lineation, explore the relevance of new traits for
identification, and test hybridization hypotheses.
Most of the dissimilarity between species is cor-
related to flower and fruit traits, while vegetative
characters show great lability that is largely un-
correlated with taxonomic boundaries except for
species with composed leaf. We have obtained
clear delineation between B. eatonii and B. in-
firma and between B. tripartita and B. comosa,
species for which taxonomic status until this day
were still uncertain. Even though more sampling
is needed to better assess the full range of phe-
notypic variation of each species, the recogni-

tion of *B. infirma*, an endemic of the St-Lawrence
estuary, Canada, would have a great impact on
conservation action.
**S.181** SYNANTHEROLOGY RELOADED:
RECENT ADVANCES AND THE FUTURE OF
EVOLUTIONARY STUDIES IN COMPOSITAE.
SESSION 1

**S.181.1 Dancing with Thistles**

Alfonso Susanna¹

1 Botanic Institute of Barcelona, Spain

Our research in tribe Cardueae was a challenge from the beginning for several reasons. Field collections faced many challenges due to the geographical distribution of thistles in complicated places. When we started investigating the group, our Botanic Institute in Barcelona was a very small research center with limited resources for studying such a large tribe. Obtaining an accurate classification was particularly challenging due to the thistle-like architecture of many genera, which is shared by many unrelated genera of the tribe. Development of molecular methods, from single-region DNA sequencing in the nineties to high-throughput sequencing in recent times, provided the tools for unraveling the true phylogeny of the tribe and allowed us to define a new classification. The new subtribal scheme in twelve subtribes was anticipated by morphological evidence, and molecular data confirmed that the system is robust. This presentation summarizes thirty years of wrestling with this fascinating group of plants.

**S.181.2** An updated subtribal classification of Compositae tribe Anthemideae based on extended phylogenetic reconstructions

Christoph Oberprieler¹, Nadine Benda¹,
Alisha Töpfer¹, Marco Dorfner¹, Miriam
Stock¹, Robert Vogt²

1 University of Regensburg, Regensburg, Germany. 2
Freie Universität Berlin, Berlin, Germany

An updated subtribal classification of the Compositae-Anthemideae is presented based on gene- and species-tree reconstructions considering sequence variation from the plastid (cpDNA *ndhF*, *trnL-trnF*) and nuclear genome (nrDNA ITS1-5.8S-ITS2, ETS; nDNA dual specificity phosphatase DSP, plus-3 domain-containing protein of the vernalization independence 5 complex VIP5, major facilitator superfamily protein of the Nitrate Transporter1/Peptide Transporter NPF3.1) for 113 of the 115 hitherto accepted genera of the tribe. As a result, six subtribes were newly circumscribed [i.e., Anthemidinae (including also Xylanthemum), Artemisiinae (including also Hulteniella, Lepidolopha, Opisthopappus, and Triadactylina), Glebionidinae (including also Otoglyphis, Endopappus, and Nivellea), Handeliinae (including Cancrinia, Polychrysum, and excluding Xylanthemum), Leucantheminae (including also Daveaua, Heteromera, and Otospermum), Leucanthemopsisinae (including Phalacrocarpum)]. Additionally, five unigeneric subtribes (i.e., Brocchiinae, Inulantherinae, Lepidophorinae, Lonadinae, Vogtiinae) were described as new to science due to their phylogenetically isolated positions, resulting in 18 subtribes accepted presently for Compositae-Anthemideae. A focal subtribe of a more detailed phylogenetic analysis is based on the W Mediterranean genus Vogtia Oberpr. & Sonboli that comprises two species, i.e., V. annua (L.) Oberpr. & Sonboli and V. microphylla (DC.) Oberpr. & Sonboli. To infer the phylogenetic position of this subtribe the complete chloroplast genomes of the two species and seven other representatives of additional subtribes of the Anthemideae were sequenced using long-range PCR and Oxford Nanopore technology (MinION) and subjected to maximum-likelihood tree reconstructions. The phylogenetic analysis confirmed the placement of Vogtiinae as sister to subtribes Anthemidinae+Mat-
<figure><img src="image_420.png" /></figure>

ricariinae. Results of additional analyses concern-
ing the phylogenetic relationships and the subtribal
status of the monogeneric orphan genera *Brocchia*,
*Lepidophorum*, and *Lonas* will be also presented.

S.181.3 Exploring the future
of Compositae: advances in
genomics, transcriptomics,
and functional genetics

Jennifer R. Mandel¹, Mauricio Bonifacino²,
Erika R. Moore¹, Paige A. Ellestad¹, Linda E.
Watson³, Carolina Siniscalchi¹, Ramhari
Thapa¹, Daniel S. Jones⁴, Reid Selby⁴,
Brannan Cliver⁴, Vandana Gurung⁴, Emily
Yaklich⁵, Jakub Baczyński⁵, John M. Burke⁵,
Ashley Crook⁶, Zachary Nimchuk⁶, Alex
Harkess⁷, Zachary Meharg⁷

1 Department of Biological Sciences, University of
Memphis, Memphis, USA. 2 Departamento de Bi-
ología Vegetal: Laboratorio de Botánica, Univer-
sidad de la República, Montevideo, Uruguay. 3
Department of Plant Biology, Oklahoma State Uni-
versity, Stillwater, USA. 4 Department of Biological
Sciences, Auburn University, Auburn, USA. 5 Plant
Biology Department, University of Georgia, Athens,
USA. 6 Department of Biology, University of North
Carolina, Chapel Hill, USA. 7 Hudson Alpha Institute
for Biotechnology, Huntsville, USA

Asteraceae or Compositae comprise 10% of all angiosperm species and include numerous species of economic, medicinal, and industrial value. Despite this, relatively few large-scale genomic resources exist for the family, especially given its size/diversity, which limits our ability to understand the evolution of this fascinating family and to link genes to phenotypes. Moreover, studies of genomic and evolutionary mechanisms that have driven the success of Asteraceae have been hampered by the lack of a well-resolved phylogeny. We have used a phylogenomic framework to fully resolve the Asteraceae backbone. This work supported a late Cretaceous origin followed by explosive diversifications that resulted in the family's 30,000+ extant species. Recently, our group has been working toward enhancing available resources for the family with genomic and transcriptomic resources for diverse species, including several key outgroups. We are also addressing the outstanding need for genome editing tools by working toward developing novel approaches in Asteraceae. These resources will be made freely available to enable

comparative and functional genomics, as well as
crop improvement efforts in the broader community.
This work provides an integrative framework for future
studies aimed at understanding the role of the mac-
roevolutionary patterns and processes that generat-
ed the enormous species diversity of Asteraceae.

S.181.4 Anatomy of
Asteraceae: methodological
advances for an old science

Makeli G. Lusa¹, Benoit F.P. Loeuille²

1 Department of Botany, Universidade Federal de Santa Catarina, Florianopolis, Brazil. 2 Royal Botanical Gardens Kew, Richmond, UK.

Plant anatomy used to be considered an ancient sci-
ence, or a simple tool for answering equally simple
questions. Thanks to recent advances in the molecular
era, the understanding of systematics of the large and
complex family Asteraceae has greatly advanced. In
this process, plant anatomy has re-emerged as an
important ally, enabling the investigation of complex
evolutionary questions, answering new and old tax-
onomic questions, and, jointly with plant molecules,
providing a wide range of data for the arsenal of new,
species-level information, in addition to helping de-
tect sources of secondary metabolites, through the
integration of histochemistry and phytochemistry. The
present work aims to address the most recent plant
anatomy techniques used in investigations in the
Asteraceae, focusing on methodological advances
in the following sectors: evolutionary studies of mor-
phoanatomical diversity; morphoanatomy to answer
complex taxonomic questions; integration of anato-
my and phytochemistry through histochemistry and
*in situ* hybridization; and new developments in investi-
gations using Transmission Electron Microscopy. Thus,
this proposal intends to show botanists that this an-
cient science persists and thrives, uniquely contribut-
ing to increasing our knowledge of plants.

S.181.5 The Compositae in the
Plant and Fungal Tree of Life

Benoit Loeuille¹, Oriane Hidalgo², Carolina M. Siniscalchi³, Olivier Maurin¹, Raquel Negrão¹, Jennifer Mandel⁴, William Baker¹

1 Royal Botanic Gardens, Kew, UK. 2. Institut Botànic de Barcelona, Spain. 3 Mississippi State University, USA. 4 University of Memphis, USA
<figure><img src="image_421.png" /></figure>

The Compositae (or Asteraceae) are one of the largest flowering plant families with ca. 1,630 genera and ca. 24,700 species. They are distributed worldwide and display an amazing morphological diversity. Despite the challenging size of the family, molecular phylogenies have recovered a fairly consistent tribe-level backbone, which resulted in the current classification with 16 subfamilies and 51 tribes. However, up to now, phylogenomic studies have included less than 16% of the generic diversity of the family. Here, we present a robust phylogeny including ca. 90% of the genera of Compositae and all subtribes but one, based on combined data from two different probe sets (Angiosperms353 and Compositae1061). The monophyly of the family is challenged by the position of Calyceraceae as sister group of Barnadesioideae in some of the inferences. The evolutionary relationships between the main lineages are mostly in agreement with previous phylogenomic studies, except for Hecastogleidoideae emerging as sister to the clade Gochnatieae + Wunderlichieae + Cyclolepis and Tarchonanthoideae as sister group of Carduoideae (not Dicomeae). A few tribes are polyphyletic under their current circumscription (Wunderlichieae, Neurolaeneae, Bahieae, Madieae). The evolution of Compositae is marked by at least ten whole genome duplication and numerous hybridisation events, which may explain the incongruences found between different datasets and methods of phylogenomic reconstruction (e.g., coalescent-based Astral vs. concatenated supermatrix). This phylogeny will allow new insights about morphological evolution, diversification, and biography of the Compositae.

## S.181.6 The Global Compositae Database: challenges and opportunities

Mauricio Bonifacino<fnref n="1" />, John C. Semple<fnref n="2" />, Morgan R. Gostel³, Nicola G. Bergh⁴, Benoit Loeuille⁵, Paola de Lima Ferreira⁶, Alexander N. Sennikov⁷, Nádia Roque⁸, Gustavo Heiden⁹, Guy Nesom¹⁰, Liliana Katinas¹¹

Compton Herbarium, South African National Biodiversity Institute, Newlands, Cape Town, South Africa. 5 Royal Botanic Gardens, Kew. 6 Aarhus University, Aarhus, Denmark. 7 Botanical Museum, Finnish Museum of Natural History, University of Helsinki, Finland. 8 Federal University of Bahia, Brazil 9 Embrapa Clima Temperado, Pelotas, Brazil. 10 Academy of Natural Sciences, Philadelphia, Pennsylvania, USA. 11 División Plantas Vasculares, Museo de La Plata, Pcia. Buenos Aires, Argentina

Online taxonomic databases are fundamental resources for a post-2020 framework for the Global Strategy for Plant Conservation, particularly Target 1 of that effort – “an online flora for all known plants.” Launched in 2019 as a revitalization of the Global Compositae Checklist, the Global Compositae Database is a freely accessible, expert-curated, online taxonomic database of all names for the largest angiosperm family Compositae (Asteraceae) that is generously supported by the World Register of Marine Species (WoRMS) and maintained by taxonomic specialists from The International Compositae Alliance (TICA). The team behind the data curation includes researchers from different backgrounds, with taxonomic expertise across the many tribes currently recognized in the family. This is a formidable task, involving almost 170,000 names for ca. 33,000 accepted species, and so TICA is actively recruiting additional taxonomic specialists i. The development of this database highlights the importance of collaboration and is being coordinated, in part, through a larger collaboration with the World Flora Online, for which TICA serves as the Taxonomic Expert Network (TEN). The database is hosted on the WoRMS platform, accessible via the TICA website (www.compositae.org), has a friendly interface, and provides tremendous flexibility in terms of editing and adding taxonomic data. Current challenges include a lack of specific resources to curate the database, and a general shortage of expert taxonomists. Among the groups in Compositae with most up-to-date information are the basal grade of the family, and various elements of the subfamily Asteroideae. A broader geographic participation of TICA members and taxonomic specialists, together with a series of workshops aimed at facilitating the access and usage of the database, will help increase momentum in advancing this important resource.

<footnote n="1">1 Facultad de Agronomia & Facultad de Ciencias, Universidad de La República, Montevideo, Uruguay.</footnote>

<footnote n="2">2 Department of Biology, University of Waterloo, Waterloo, Ontario Canada 3 Botanical Research Institute of Texas, Fort Worth, Texas, USA. 4 The</footnote>
# S.182 PRIORITISE, PLAN, ACT AND MONITOR – PROMOTING AN INTEGRATED APPROACH TO THREATENED TREE CONSERVATION

## S.182.1 Strengthening the population of threatened tree species in the Colombian Caribbean region

Maria Paula Contreras¹

1 Horticulture and Science Department, Cartagena Botanical Garden, Colombia

The Colombian Caribbean region houses three of the most threatened ecosystems and areas in the country: the tropical dry forest, the mangrove forests, and the Sierra Nevada de Santa Marta. Among the main threats are non-sustainable cattle farming, unplanned urban development that causes deforestation, and climate change. Aiming to strengthen the population of Caribbean threatened species, *Pachira quinata* (VU), *Libidibia punctata* (VU), *Hdroanthus coralibe* (VU), *Clavija sanctae-martae* (VU), *Aspidosperma polyneuron* (EN), *Pelliciera benthamii* ex. *Pelliciera rhizophorae* (VU), and *Pterocarpus acapulcensis* (VU), studies in their distribution, propagation and phenology were made. No actions to conserve and strengthen their populations have been reported in the last decade, and the non-scientific public usually does not know their conservation status and importance. Few places that host the native population are under conservation status (public or private). However, there are currently no actions focused on their conservation and propagation in nurseries at a large scale. The Cartagena Botanical Garden's science and horticulture teams have worked together to strengthen the population of the previous mention native threatened tree species. The Garden strives to conserve, restore, and enrich the ecosystems through long-lasting actions involving local communities. Several field trips to coastal and middle-elevation localities have allowed seed collection, phenological annotations and the recognition of potential planting areas. The Garden has successfully established a partnership with local nurseries, one in the northern Caribbean municipality of Colombia, La Guajira, and the other

in Magdalena, where the vulnerable endemic to Colombia specie, *Clavija sanctae-martae*, is the priority. Suitable landowners and changes in land use for planting actions are presented as one of the most challenging aspects. The result includes more than 10,000 planted plants, established propagation protocols, distribution niche models and regional capacity building.

## S.182.2 Natural drivers leading Conifers to extinction

Cañadas, E.M.¹, Calderisi, G.², Lorite, J.¹, Cogoni, D.², Fenu, G.²

1 Department of Botany, University of Granada, Granada, Spain. 2 Department of Life and Environmental Sciences

Currently, over 28% of species are at risk of extinction, which could rise to 50% by 2100 since the rate of extinction has been particularly dramatic for the last 50 years. Conifers stand out as a highly endangered group, with a percentage of threatened species of ca. 34%. Reversing the situation is crucial since coniferous species, which are mostly trees, constitute key pieces for more than 39% of the world's forests and for the provision of ecosystem services globally. Deforestation and other man-made hazards, including climate change, threaten conifers which, like other gymnosperms, are also doomed to extinction due to natural causes. In this context, we explore intrinsic or natural traits (both biological and ecological) leading conifers to extinction in order to support decision-making on biodiversity conservation. Furthermore, we delved into the largest conifer family, *Pinaceae*. For this, we downloaded from the IUCN Red List database all the 610 species from the *Pinopsida* Class, assigning the global Red List category. For each species we also compiled traits related to altitude, adult plant size, and seed size. Moreover, for the *Pinaceae* family (225 species), we collected traits about leaves, seed cones, pollen cones, and seed wing, among others. Then we assessed the re-
<figure><img src="image_422.png" /></figure>

relationship between extinction risk (with values ranging between 1 (LC) and 5 (CR)) and traits, finding that the most threatened conifers are smaller in size, have smaller seeds, and inhabit a narrow altitudinal range at mid-mountain. Pinaceae showed similar trends, but the most endangered species had heavier seeds and smaller leaves in this case. Our results could guide conservation efforts for these plants custodians of an ancient and long evolutionary history since species with certain identified traits could be more vulnerable to moving to a higher threat category if no action is taken.

## S.182.3 Tools and approaches for identifying and filling gaps in knowledge for the conservation of exceptional tree species

Valerie C. Pence¹, Emily B. Bruns¹,², Megan Philpott¹, Melissa Duda¹,³

1 Center for Conservation and Research of Endangered Wildlife, Cincinnati Zoo & Botanical Garden, Cincinnati, USA. 2 Botanic Gardens Conservation International--US, San Marino, USA. 3 Northwestern University, Evanston, USA

Exceptional species cannot be conserved in conventional seed banks and will require other approaches for long-term, ex situ conservation. Cryobiotechnologies can be harnessed to preserve exceptional species, but currently these methods are not being implemented on the needed scale. Our recent analysis of the literature has shown that there are significant gaps in knowledge of cryopreservation and the supporting in vitro technologies in their application to exceptional species, particularly with exceptional tree species. While a high proportion of exceptional species are trees, a much lower proportion of trees are represented in both the cryopreservation and in vitro literature, with a particular gap in tropical trees. Filling these gaps and scaling up application will require developments in both science and technology. Currently, developing cryopreservation and in vitro protocols is primarily an experimental process, and information and tools are needed to develop greater predictability and efficiency. Deeper analyses and understanding of reported research are needed, and the creation and analysis of large datasets of information could facilitate this. The Comparative In Vitro

Database and the Comparative Cryopreservation Database will be introduced as platforms for these data that will align them with taxonomic, morphological, and climatic data to evaluate patterns and move toward the goal of predictability in developing new protocols. The companion Media Comparison Tool has also been developed, which provides rapid visualization and comparison of the components of tissue culture media as a way of analyzing past research and selecting or developing new formulations for species new to in vitro culture. These tools should both help identify more specific information gaps and help prioritize research for more efficiently applying cryopreservation and in vitro technologies for the ex situ conservation of exceptional tree species.

Acknowledgments: Research supported in part by a grant from the Institute of Museum and Library Services.

## S.182.4 Unveiling the extinction risk of Brazilian trees

Eduardo Fernandez¹,²,³, Eduardo Amorim²,³, Fernanda Wimmer²,³, Gláucia Crispim²,³, Lucas Arguello²,³, Lucas Jordão²,³, Marcio Verdi¹,²,³, Mário Gomes²,³, Monira Bicalho²,³, Fernanda Fraga²,³, Camila Nardy Delgado²,³, Carlos Ferreira²,³, Vicente Calfo²,³, Gustavo Martinelli¹,²,³

1 Rio de Janeiro Botanic Garden Research Institute, Rio de Janeiro, Brazil. 2 Brazilian National Centre for Plant Conservation, Rio de Janeiro Botanic Garden Research Institute, Rio de Janeiro, Brazil. 3 IUCN Species Survival Commission Brazil Plant Stand-Alone Red List Authority, Rio de Janeiro, Brazil

Brazil is the epitome of plant diversity and champion the global ranking of tree species' richness. However, this astonishing diversity confronts a looming crisis as human-induced activities intensify. Rampant habitat conversion for agriculture and cattle ranching, coupled with extensive overexploitation, imperils Brazil's tree flora. With great responsibility to protect its irreplaceable natural heritage, Brazil has recognized that assessing species' extinction risk is a vital step in setting up effective conservation strategies. The Brazilian National Centre for Plant Conservation (CNCFlora), from Rio de Janeiro Botanic Garden Research Institute (JBRJ), is designated nationally and internationally as the Brazil's plants Red List Authority and adopts the standards and procedures recommended by IUCN for
<figure><img src="image_423.png" /></figure>

at-risk species detection. Driven by initiatives tailored to deliver conservation assessments of trees, such as the Global Tree Assessment (GTA) from Botanic Gardens Conservation International (BGCI), CNCFlora/ JBRJ has conducted full IUCN conservation assessments of over 3,851 Brazilian trees between 2015-2023. Our results show that 37% (1,276) of the assessed tree species are Threatened (187 as CR, 794 as EN and 295 as VU). NT and LC categories were assigned to 1,943 species, while 633 species were deemed as DD. Brazil has thousands of threatened endemic trees, and while advances in detect at-risk plants increased in recent years, only two native tree species have species-specific national Conservation Action Plans in-place. Most threatened trees in Brazil are found within two global biodiversity hotspots (Mata Atlântica and Cerrado), and in the Amazon, where selective logging is pressing timber species towards extinction. A comprehensive evaluation of Brazil's entire tree diversity is imperative to strategically guide interventions and prevent irreversible losses, emphasizing the immediate need to extend conservation measures to safeguard endemic trees for the enduring preservation of Brazil's tree biodiversity.

## S.182.5 A collaborative programme to protect, conserve and enhance the natural capital of the Mexican forests

Tiziana Ulian¹,² Michael Way¹, Maraeva Gianella¹, Silvia Bacci¹, Elizabeth Bell¹, Elena Castillo-Lorenzo¹, Efisio Mattana¹, Cesar M. Flores Ortiz³, Daniel Cabrera Santos³, Isela Rodríguez Arevalo³, Oswaldo Tellez, Rafael Lira³, Gabina Sol Quintas⁴, Jazmin Cobos Silva⁴, Estrella Chevez⁴, Elisa Peresbarbosa Rojas⁴, Maria Toledo⁵, Tarin Toledo⁵, Mauricio Diazgranados¹,⁶, Patricia Davila Aranda³

¹Royal Botanical Garden, Kew, Wellcome Trust Millennium Building, Wakehurst, Ardingly, West Sussex, UK. ²University of Turin, Turin, Italy. ³Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Tlalnepantla, Estado de Mexico, Mexico. ⁴Pronatura Veracruz A.C., Coatepec, Veracruz, Mexico. ⁵Instituto de Ecología A.C., Veracruz, Mexico. ⁶New York Botanical Garden, Bronx, NY, USA

Mexico is a megadiverse country, hosting 23,314 species of vascular plants, 40% of which are endemic. Its ecosystems range from arid to wet and tropical to temperate. It is rich in cultural diversity and is home to one of the most important biocultural legacies of the world, resulting in more than 9,000 plants with known uses. The Royal Botanic Gardens, Kew (RBG Kew) has been collaborating in Mexico with the Facultad de Estudios Superiores, Iztacala of the National Autonomous University of Mexico (Fes-I UNAM) for over 20 years to conserve the rich flora of Mexico through seed banking, research, and training. Seeds are preserved at Fes-I UNAM, with duplicates stored at Kew's Millennium Seed Bank at Wakehurst, in the UK, under an Access and Benefit Sharing Agreement. Since 2015, RBG Kew and Fes-I UNAM have developed a programme to protect, conserve and enhance the natural capital of the Mexican forests, built around a detailed understanding of native tree seeds and their regeneration under future climatic change scenarios. We have published the first checklist of native trees of Mexico, with information on the distribution, uses and conservation status of 2,885 species. This information is used to prioritise taxa for seed collecting and conservation: seeds are either conserved ex-situ through conventional seed banking or propagated for in situ conservation. When propagated, trees are donated to local communities to support their livelihoods or used for reforestation in urban and peri-urban areas and degraded forests. This programme has been delivered through several projects funded by different sources and in collaboration with the NGO Pronatura Veracruz and the Instituto de Ecología A.C. Local communities are involved in the programme activities through a participatory approach and by applying gender equality and social inclusion principles.

## S.182.6 Nutritional status of Pinus sylvestris and Quercus pyrenaica is improved in heterospecific neighbourhoods

Tallón Elena, Marina, Carmen Urea, M. Esther Pérez, Enrique Andivia.

¹Departamento de Biodiversidad, Ecología y Evolución, Facultad de Ciencias Biológicas, Universidad Complutense de Madrid, Madrid, España

Tree species admixture is considered as a key management strategy for improving forest eco-