<figure><img src="image_353.png" /></figure>

### S.151.3 Exploring the role of local bee diversity on the evolution and spatial distribution of floral uniformity in tropical trees

Thais Vasconcelos¹, Lena Heinrich¹, Aline Martins¹

1 Department of Ecology and Evolutionary Biology, University of Michigan, Ann Arbor, MI, USA

Floral phenotypes, as any other plant trait, may present geographical structuring based on biotic and abiotic conditions that favor certain traits to occur more often in sympatry due to convergence. Recently, it has been suggested that angiosperms and their main pollinators, the bees, may present a spatial mismatch in their patterns of diversity, with areas where bee species richness peak being areas where angiosperm species richness is relatively low and vice versa. We here explore how this spatial mismatch may impact the geographical distribution of floral phenotypes of bee pollinated plant lineages. First, we describe the spatial mismatch and explore whether this pattern leads to a decrease in the proportion of bee pollinated angiosperms in areas where bees are less diverse. Second, we investigate how local floral phenotype diversity in bee pollinated plants may be impacted by a less diverse, but not necessarily less abundant, pollinator community, specifically testing whether floral phenotypes would tend to be more morphologically uniform through time in these lineages. We discuss the results in the context of coevolution between bees and angiosperms and how this relationship has modulated the convergent and divergent evolution of floral traits across time and space.

### S.151.5 Plant quantitative genetics to understand floral trait diversity

Maria Clara Castellanos¹, Andrés Romero-Bravo¹

1 University of Sussex, Brighton, UK

Plants can evolve rapidly after changes in their pollinators and this is believed to be an important force behind the evolution of floral diversity. Both floral morphological and nectar traits can be expected to play important adaptive roles, with nectar being offered as a reward that varies depending on pollinator preference,

and morphology optimizing the mechanical aspects of pollen transfer and access to rewards. Yet the response of different floral traits to novel natural selection can vary. Floral morphology is often expected to show high phenotypic integration to maintain pollination accuracy, while nectar traits can be environmentally sensitive. Genetic correlations and phenotypic plasticity can therefore play important roles that are not fully understood, particularly for nectar traits. We explored this in the context of recent pollinator change, using *Digitalis purpurea* to study the potential to respond to selection of traits that often show convergent patterns of evolution in response to pollination-mediated selection. *D. purpurea* shows rapid recent convergent evolution of corolla morphology but not nectar traits after a range expansion with hummingbirds added as pollinators. We studied phenotypic plasticity, heritability, evolvability, and integration of morphology and nectar in wild populations and in a multi-population common garden. Morphological traits showed higher heritable variation than nectar traits, and the proximal section of the corolla, which regulates access to nectar and shows rapid change in introduced populations, presented higher evolvability and lower integration than all other traits. Nectar was more plastic than morphology, driven by highly plastic sugar concentration. Nectar production rate showed high evolvability and potential to respond to selection. Our results explain patterns of rapid evolution in this species and provide novel insights on the causes and constraints in the evolution behind the extraordinary diversity of flowers.

### S.151.6 Genetic mechanisms of convergent floral color evolution in Pedicularis

Yue Yang¹, Jianjun Jin¹, Deren Eaton¹

Ecology, Evolution and Environmental Biology Department, Columbia University, New York, USA

Floral color, one of the conspicuous phenotypic traits in plants, exhibits a wide spectrum of variations. Color variations are mainly determined by the types, combinations and concentrations of different pigments. The underlying genetic pathways of pigment production have been well-studied and are relatively conserved across species. However, the evolutionary and genetic mechanisms driving the interspecific floral color variations, particularly in lineages such as *Pedicualris*, remain elusive. *Pedicularis* has over 500 species worldwide and over two-thirds are distributed,
ed in Hengduan Mountain, a recognized biodiversity hotspot. This plant genus has an exceptional diversity in floral color with a range encompassing white, yellow, red, purple and their combinations. A particular intriguing aspect of this genus is its floral color convergence, where similar color patterns recur in multiple independent lineages. It is unclear how independent mutations (genetic convergence), inheritance of ancestral polymorphisms and introgressions contribute to the observed convergence. Moreover, if genetic convergence is a major factor, its scale and scope is uncertain - occurring at a specific site, within a particular gene, along a pathway or in a genetic net-

work. In our study, we analyzed transcriptome data of 83 samples across 57 *Pedicularis* species to build a phylogenetic tree using both coalescent-based and concatenated-based methods. We reconstructed ancestral colors in internal nodes using different MK-based models. We investigated gene tree-species tree discordances and the prevalence of hemiplasy in floral color evolution. By applying diverse phylogenetic comparative methods, we aim to detect potential evolutionary and genetic mechanisms underlying floral color evolution. This study will contribute to our understanding of the evolution and function of convergent floral phenotypes.

# S.152 MOUNTAIN BIODIVERSITY AND EVOLUTION. SESSION 1

## S.152.1 Applications of spatial phylogenetic to unravel biodiversity patterns and evolutionary history of the Himalayan flora

Saddam Saqib¹,²,³*, Wajid Zaman⁴, Fazal Ullah¹, Zhi-Duan Chen², You-Cai Xiong¹

1 State Key Laboratory of Herbage Improvement and Grassland Agro-ecosystems, College of Ecology, Lanzhou University, Lanzhou, China. 2 State Key Laboratory of Plant Diversity and Specialty Crops, Chinese Academy of Sciences, Beijing, China. sadamsaqib. qau@gmail.com, zhiduan@ibcas.ac.cn. 3 University of Chinese Academy of Sciences, Beijing, China. 4 Department of Life Sciences, Yeungnam University, Gyeongsan, Gyeongbuk, Republic of Korea

Mountainous ecosystems are prime biodiversity reservoirs and also serve as hub of raw materials and minerals. Various factors such as climate, landscape, ecological gradients and topographic factors play role in maintaining biodiversity in mountainous regions such as Himalaya. Himalaya is one the world highest mountains range and known as biodiversity hot spot. The region is affected by strong elevational gradients, drastic altitudinal climate variations over very short distances which generate various environmental niches that leads to simple geometric constraints on species distribution boundaries resulting in divergence and speciation. This study focused on general trends and recent advances in highlighting biodiversity hotspots and its evolutionary history. Our study observed that

use of species richness, phylogenetic diversity, phylogenetic endemism, and weighted endemism can help to understand biodiversity hotspots in the study area. On the other hand, phylogenetic patterns such as use of net related index and nearest taxon index revealed phylogenetic clustering and over dispersion within study region. Furthermore, our study highlighted that spatial distribution and phylogenetic tree by using β diversity metrics, help to understand phytogeogions, indicator species and evolutionary distinctions within the Himalayan flora. Based on these factors, our study recommends that integrative, multifacteted and policy backed approaches are required for understanding of the Himalayan flora. Our study will be helpful in examining ecosystem drivers and provide novel insights into the maintenance and conservation of other mountain regions.

## S.152.2 Floristic and evolution of the “sky islands” of East Africa

Shengwei Wang¹,², Neng Wei¹,², Tao Wang¹,², Qingfeng Wang¹,²

1 Wuhan Botanical Garden, Chinese Academy of Sciences. 2 Sino-Africa Joint Research Center, Chinese Academy of Sciences

The East African mountains are one of the world's crucial biodiversity hotspots, and their distinctive "sky island" structure makes them a Garden of Eden for the evolution of plant diversity. Long-term studies have fo-
<figure><img src="image_354.png" /></figure>

cused on the diversity and evolution of the Afroalpine
flora, and there is a lack of research on the floristic his-
tory of the entire montane system. Recent plant cata-
logs from East African mountains reveal the extremely
high diversity of plants here, with about 7,000 species
in 193 families and 1,544 genera, of which more than
15% are endemic to East Africa. The spatial and tempo-
ral phylogenetic patterns of the mountain flora reveal
that the East African mountains are a convergence of
ancient and young plants, serving as a museum and
cradle of plant diversity in East Africa. Habitat distur-
bances caused by Pleistocene climate oscillations
have led to colonization, dispersion, extinction, and
evolution of species. Especially the dispersal of habi-
tats of montane forest species to intervening lowlands
during the Pleistocene glacial period facilitated spe-
cies exchange between mountains. Meanwhile, the
temporal differences in mountainous geological dy-
namics, such as volcanism and uplift, coupled with the
island effect, intensify the differentiation of populations
among mountains. Our research shows that the East
African mountains are experiencing a gradual upward
trend in the number of exotic plants, which poses a
serious problem of homogenization of native plants.
Plant diversity in the East African mountains faces vul-
nerability and urgently needs more conservation and
research attention.

S.152.3 Mountain radiation
across the Andes and the
Himalaya-Hengduan Mts.
driven by evolutionary
confluence in a shrub clade,
Berberis

Chih-Chieh Yu¹, Qiu-Yue Zhang¹, Wen-Na Ding¹,², Richard H. Ree³, Yao-Wu Xing¹

1 CAS Key Laboratory of Tropical Forest Ecology, Xi-
shuangbanna Tropical Botanical Garden, Chinese
Academy of Sciences, Mengla, China. 2 WSL, Swiss
Federal Research Institute for Forest, Snow and
Landscape, Birmensdorf, Switzerland. 3 Life Scienc-
es Section, Negaunee Integrative Research Center,
Field Museum, Chicago, IL, USA

The high-altitude regions of the Andes and the Heng-
duan-Himalaya Mts. are home to some of the world's
most remarkable plant radiations. However, little is
known whether these radiations are experienced sim-
ilar or different diversification history. The high-altitude
regions of the Andes and the Hengduan-Himalaya
Mts. are home to some of the world's most remarkable

plant radiations. However, little is known whether these
radiations are experienced similar or different diversi-
fication histories. In this study, we investigated the or-
igin of the radiation of *Berberis*, an Andes and Heng-
duan-Himalaya Mts. (HHM) centered shrub clade. Our
results revealed that the net diversification rate of the
HHM steadily rised since the Miocene, whilethe rate
of net diversification in the Andes remained relatively
flat during the same period. We recognized multiple
trait-environment matchings evolved from the HHM
unlike the Andes, and found such relationships were
stepwise evolved along the diversification history of
an alpine deciduous clade. Here, we unraveled a dis-
tinct but little-tested scenario for mountain radiation,
in which sequential assembly of multiple enablers (i.e.,
key trait evolution, mountain uplift, and paleo-climate
change) synergistically function as a trigger to main-
tain and promote the radiation.

S.152.4 Different levels
of competition explain
species richness differences
between two biodiversity
hotspots

Martha Kandziora¹, Diana L. A. Vasquez²,
Christian Brochmann³, Abel Gizaw³, Petr
Sklenář¹, Filip Kolář¹,², Nicolai M. Nürk⁴, and
Roswitha Schmickl¹,²

1 Department of Botany, Charles University, Prague, Czech Republic. 2 Institute of Botany, The Czech Academy of Sciences, Průhonice, Czech Republic. 3 Natural History Museum, University of Oslo, Oslo, Norway. 4 Department of Plant Systematics, Bayreuth Centre of Ecology and Environmental Research, University of Bayreuth, Bayreuth, Germany

Niche packing and niche specialisation are two cen-
tral factors modulating biodiversity patterns across
latitudes and altitudes. Niche packing is when species
niches are finely packed in a multi-dimensional niche
space while occurring in the same geographic space.
Specialised species on the contrary should have nar-
row, separate niches and are highly adapted to certain
environmental conditions. Both phenomena enable
species to coexist by avoiding competition via resource
partitioning. Here, we combine phylogenomics of six
plant lineages, with occurrence records to compare
niche and range metrics - size and overlap - of species
between the highly diverse tropical alpine habitats of
the South American Andes and the less species-rich al-
pine habitats on the eastern African mountains. Com-
<figure><img src="image_355.png" /></figure>

paring species niche sizes between both continents using a multidimensional climate space, we found that overall niche size is larger in the Andes than in the mountains of eastern Africa. Additionally, range-overlapping species in the Andes showed generally less overlap in niche space than in Africa. Taken together, we hypothesise that biodiversity differences between the two tropical alpine ecosystems might be caused by different degrees of competition.

## S.152.5 From the Andes to the Cascades—biogeography and evolution across western South and North American mountain ranges in Cistantheae

Anri Chomentowska¹, Sophie Dauerman¹, Patrick Sweeney², Iris Peralta³,⁴, C. Matt Guilliams⁵, Monica Arakaki⁶, Joseph Holtum⁷, Matthew Ogburn⁸, Lillian Hancock⁹, Erika Edwards¹,²

1 Department of Ecology and Evolutionary Biology, Yale University, Connecticut, USA. 2 Peabody Museum, Yale University, Connecticut, USA. 3 Facultad de Agronomía, Universidad Nacional del Cuyo, Mendoza, Argentina. 4 Instituto Argentino de Investigaciones de las Zonas Áridas, Mendoza, Argentina. 5 Santa Barbara Botanic Garden, California, USA. 6 Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, Lima, Perú. 7 College of Science and Engineering, James Cook University, Queensland, Australia. 8 Department of Biology, Southern Utah University, Utah, USA. 9 Department of Ecology and Evolutionary Biology, Brown University, Rhode Island, USA

The plant family Montiaceae (Caryophyllales) has an affinity for desert or alpine habitats and exhibits significant evolutionary lability in life history and climate niche. Lability in life history traits has been shown to cluster especially in one South and North American sublineage, the Cistantheae; this clade is distributed in desert and mountain regions from the Andes to the Cascade mountains, but is absent from equatorial regions (amphitropic distribution). Exploring evolutionary patterns in this lineage has been stymied thus far by unresolved phylogenies, exacerbated by decades of taxonomic confusion due to the presence of intraspecific variation in morphology. We aimed to clarify phylogenetic relationships within Cistantheae and delimiting species using various next-generation sequencing techniques, with the ultimate goal of understanding its

biogeographical history and biome transitions. We sequenced 460 plant specimens involving 103 putative species (including outgroup taxa) collected largely from Chile, Argentina, Peru, and USA; the sequencing reads were assembled using a whole-genome reference of the species *Cistanthe longiscapa*. The consequent phylogeny supported genus-level relationships in the Cistantheae, and clarified species relationships in its subsections. Phylogenomic and population genetic approaches showed signatures of gene flow between select populations in smaller geographical ranges; we also inferred multiple long-distance dispersals between continents, with South America acting as the source pool for movements into North America. These analyses support several phylogenetic hypotheses from previous studies and reveal instances where species have been over-split and need taxonomic revision. The phylogeny also raises interesting implications about life history strategies and climate niche evolution, and their role in desert and mountain radiations. Finally, Cistantheae includes multiple rare and highly distinctive species that are vulnerable to extinction—highlighting the necessity for the conservation of their montane habitats and the importance of describing the clade's biodiversity.

## S.152.6 Phylogenomic analysis of “quinas” and “pseudoquinas” (Cinchoneae - Rubiaceae) from Peru

Eder Chilquillo¹*, Joaquina Albán¹, Monica Arakaki¹, Carla Maldonado², Pedro Romero³, Jorge L. Ramirez³, Isabel Cruz¹, Julio C. Torres¹, Edgardo M. Ortiz⁴

1 Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, Jesús María, Lima, Perú. 2 Herbario Nacional de Bolivia, Instituto de Ecología, Universidad Mayor de San Andrés, La Paz, Bolivia. 3 Facultad de Ciencias Biológicas, Universidad Nacional Mayor de San Marcos, Lima, Perú. 4 Techni-cal University of Munich, Freising, Germany

In its current circumscription, the Cinchoneae tribe includes 9 genera of neotropical distribution. Its species occur mostly in the Andes in South America, with few reaching Central America. Preliminary phylogenetic studies have suggested the monophyly of Cinchoneae but intrageneric relationships are still debatable. In this study, we sampled 8 genera of Cinchoneae (with ca. 50% of species for *Cinchona* and *Laden-*)
bergia) and obtained multiple single-copy nuclear loci (ca. 207 genes) by using the "Angiosperm353 universal probe set", which was complemented with a taxonomic review of Cinchoneae. Phylogenetic inferences were realized with multispecies pseudo-coalescent (ASTRAL III) and gene concatenation analysis (ML). Our results strongly support the monophyly of the tribe and most of the genera, except for *Ladenbergia*. Furthermore, *Ciliosemina*, *Ladenbergia* and *Remijia* formed a clade, although the position of

*Ciliosemina* (=*Remijia pedunculata*) and *Ladenbergia muzonensis* is still elusive. The position of *Ladenbergia muzonensis* is intriguing due to its intermediate floral morphology, which resembles both *Remijia* and *Ladenbergia* species. Additionally, our phylogeny also supports the recognition of a new species in *Cinchona*. Finally, our results show that sequencing data using the probe set designed for multiple gene capture is a useful tool for phylogenetic reconstructions in taxonomically complex groups.

# S.153 USING HIGH-THROUGHPUT SEQUENCING TO UNDERSTAND PLANT EVOLUTION ON ISLANDS IN A DATA-RICH ERA: NEW INSIGHTS FOR NEW CHALLENGES. SESSION 2

## S.153.1 Do we have enough data yet? Phylogenomics of a rapid island species radiation: an example from New Zealand *Myosotis* (*Boraginaceae*)

Heidi M. Meudt¹, Sofie Pearson²,³, Weixuan Ning²,⁴, Jessica M. Prebble⁵, Jennifer A. Tate²

1 Museum of New Zealand Te Papa Tongarewa, PO Box 467, Cable St, Wellington, New Zealand. 2 School of Natural Sciences, Massey University, Private Bag 11222, Palmerston North, New Zealand. 3 Present address: The University of Queensland, Warwick, QLD, Australia. 4 Present address: Iowa State University, Ames, IA, USA. 5 Manaaki Whenua - Landcare Research, PO Box 69040, Lincoln, New Zealand

Island systems represent natural laboratories for studies of species radiations, but they often present several challenges for species delimitation. New Zealand forget-me-nots (*Myosotis*, *Boraginaceae*) are a typical example where the c. 50 species are morphologically divergent but often lack sufficient genetic diversity to distinguish among the identifiable lineages. Many of these *Myosotis* species are geographically restricted in alpine areas, uncommon, threatened, and have high taxonomic and conservation priority. Several previous studies have attempted to identify species boundaries and groups using numerous data sources,

including single-gene sequences (ITS, ETS, and plastid markers), AFLPs, and microsatellites. Here we present phylogenomic results based on Angiosperms353 baits, whole plastomes, and nrDNA to address long-standing questions in the southern hemisphere lineage of *Myosotis*. We sampled 300 individuals distributed across New Zealand representing all species and subspecies, and included representatives from the two native Australian species. Specifically, we address whether these genetic-level datasets identify taxonomically useful groups (subgeneric, species, subspecies) that are recognized by morphological attributes, including vegetative and reproductive characteristics and pollen. Additional questions we aim to address using these datasets include: Do species form monophyletic groups? Do geographic patterns exist? Can the data be used to delimit species as well as place individuals whose identity was uncertain based on their morphological characteristics? Overall, the phylogenomic data reflect earlier studies that show a recent, rapid species radiation of *Myosotis* in New Zealand. Although the backbone of the phylogenies generally have short branches with low support, collectively the phylogenomic data presented here are more useful than previous studies in identifying species groups. Finally, although this phylogenomic study does not fully overcome all of the challenges regarding species delimitation of rapid island species radiations, it nevertheless makes an important contribution to an integrative taxonomic revision of the southern hemisphere species of *Myosotis*.
<figure><img src="image_356.png" /></figure>

## S.153.2 Evolution in isolation: sympatric and hybrid speciation on a remote oceanic island

Theodore S. Brook¹,², Kyle Macleod¹, Martin Bidartondo¹,², Ian Hutton³, Sue Bower⁴, Alex Papadopulos⁵, Trevor Wilson⁶, Vincent Savolainen¹

¹Department of Life Sciences, Imperial College London, London, UK. ²Ecosystem Stewardship, Royal Botanic Gardens, Kew, London, UK. ³LHI Museum, Lord Howe Island, Australia. ⁴Bower Bush Works, Sydney, Australia. ⁵Environment Centre of Wales, Bangor University, Bangor, UK. ⁶Botanic Gardens of Sydney, Sydney, Australia

Speciation with gene flow – sympatric speciation – has often been considered improbable. Recent evidence, which is becoming more readily available with next generation sequencing data, has revealed that it can occur and, in some scenarios, may be relatively common. The flora of Lord Howe Island (LHI) provides a unique system to study this process due to its isolation and small size, enabling us to rule out allopatry in many cases. A putative example of sympatric speciation on LHI is a small radiation of Coprosma (Rubiaceae), which has produced up to seven species across two independent colonisations. Alongside producing an integrated taxonomic revision of this genus, we are using a transcriptomic data to test the hypotheses that these species evolved in sympatry and that hybrid speciation has produced at least two of the (putative) species. The latter could provide some of the first substantial evidence for the syngameon hypothesis, the proposed role of elevated levels of hybridisation upon colonisation of a new environment in enabling rapid adaptive diversification under disruptive/divergent selection. This phylotranscriptomic approach involves constructing reference transcriptomes for each species, mapping RNA-seq reads from samples across each species' range, and producing a phylogenetic tree and network of the genus on LHI. This is complimented by a range of population genetic analyses to determine the stage of speciation and transcriptomic regions associated with differentiation in this radiation. Given the strong evidence for sympatric speciation among other genera on LHI (Howea and Metrosideros), further study into Coprosma allows us to determine how general sympatric speciation is across the island and thus how relevant this process is generating biodiversity on LHI and beyond.

## S.153.3 Repeated dispersals and in-situ radiations shaped cactus diversity of the Antilles

Lucas C. Majure¹, Duniel Barrios², Yuley Encarnación¹, Alan Franck¹, Peter Breslin³

¹Florida Museum, University of Florida, Gainesville, Florida, U.S.A. ²Universidad de La Habana, Cuba ³University of Arizona, Tucson, U.S.A.

Cactaceae are a diverse subset (ca. 1800 spp.) of the arid flora of the Americas. Major diversity centers exist in Mexico, Andean South America and eastern Brazil. The Caribbean biodiversity hotspot, including the Bahamas, and the Greater and Lesser Antilles also houses substantial diversity of the family, especially species of seasonally dry tropical forest, with around 110 species of cacti found there. Most of those species (ca. 91) are endemic to the islands, thus providing a model system for studying island biogeography and the origins of Caribbean biodiversity. Previous work has provided insight into the possible ways in which this diversity was generated, however, no broadly sampled phylogeny is yet available to test these patterns. We generated a near comprehensive dataset of Antillean species of Cactaceae, as well as close relatives based on chloroplast genomes, to test the age and origin of Caribbean cacti. Numerous putative dispersals from the continent occurred during the late Pliocene and throughout the Pleistocene, which led to the tremendous diversity of cacti we now see in the islands. In-situ radiations were most prominent on the larger islands of Cuba and Hispaniola, and those two islands also were the most influential for the further spread of cacti across the rest of the Antilles. Cactaceae provide an intriguing piece to the puzzle for understanding Caribbean biogeography, and Pleistocene climatic events appear to have played a major role in their diversification, a pattern comparable to parts of the mainland distribution of the family.

## S.153.4 Historical biogeography of two Caribbean clades of bromeliads

Natalia Ruiz-Vargas¹, Julián Aguirre-Santoro², Roberta Mason-Gamer¹
<figure><img src="image_357.png" /></figure>

1 Department of Biological Sciences, University of Illinois at Chicago, Chicago, USA. 2 Universidad Nacional de Colombia, Bogotá, Colombia

*Pitcairnia* (*Pitcairnioideae*) and *Wittmackia* (*Bromelioidae*) are two genera in Bromeliaceae with high Caribbean endemism. *Pitcairnia* is the second largest genus in the family, distributed from Mexico to northern Argentina and the only one with presence in Africa. There are approximately 15 species reported in the Caribbean islands. *Wittmackia* is distributed in Brazil and the Caribbean with two separate radiations into the islands, one to the Grater Antilles that diversified into 17 different species, and another to the Lesser Antilles that resulted in one species with widespread distribution. Using a target Sequence Capture approach with the Bromeliad 1776 baits set we analyze the relationships within the Caribbean group and discuss biogeographic implication. Our analysis indicate that *Pitcairnia* colonized the Caribbean from northern South America in one single successful event and later diversified and that species diversity has been overestimated likely because of high phenotypic plasticity in the group. The colonization of the Grater Antilles by *Wittmackia* is also the result of a single event, in this case from Brazil, that resulted in a Puerto Rican and a (mostly) Jamaican clade.

## S.153.5 Correlates of inter- and intra-island speciation in Aeonium (Crassulaceae), a species-rich Macaronesian radiation

Thibaud F.E. Messerschmid¹, Stefan Abrahamczyk², Ángel Bañares-Baudet³, Miguel A. Brilhante⁴, Urs Eggli⁵, Philipp Hühn⁶, Joachim W. Kadereit⁶,⁷, Patrícia dos Santos⁴,⁸, Jurriaan M. de Vos⁸, Gudrun Kadereit¹,⁷

1 Botanischer Garten München-Nymphenburg, München, Germany. 2 Staatliches Museum für Naturkunde Stuttgart, Stuttgart, Germany. 3 Universidad de La Laguna, San Cristóbal de La Laguna, Spain. 4 Universidade de Lisboa, Lisboa, Portugal. 5 Sukkulenten-Sammlung Zürich/Grün Stadt Zürich, Zürich, Switzerland. 6 Johannes Gutenberg-Universität Mainz, Mainz, Germany. 7 Ludwig-Maximilans-Universität München, München, Germany. 8 Universität Basel, Basel, Switzerland

The *Aeonium* alliance (*Crassulaceae*) is the most species-rich and ecologically diverse plant radiation on the Canary Islands. In such radiations, speciation can

take place either within islands or following dispersal between islands. By quantifying intra- and inter-island speciation events in the evolution of *Aeonium* and exploring their correlates, we hypothesized that (1) intra-island diversification resulted in stronger ecological divergence of sister lineages, and that (2) taxa on islands with a longer history of habitation by *Aeonium* show stronger ecological differentiation and produce fewer natural hybrids. We studied the biogeographical and ecological setting of diversification processes in *Aeonium* with a fully sampled and dated phylogeny inferred using a ddRADseq approach. Ancestral areas and biogeographical events were reconstructed in BioGeoBEARS. Eleven morphological characters and three habitat characteristics were taken into account to quantify the morphological and ecological divergence between sister lineages. In lineages that diversified within single islands, morphological and ecological divergence was stronger than in lineages derived from inter-island diversification, but only the difference in morphological divergence was significant. Those islands with the longest history of habitation by *Aeonium* had the lowest percentages of co-occurring and hybridizing taxon pairs compared to islands colonized by *Aeonium* more recently. Our findings illustrate the importance of both inter- and intra-island speciation in island radiations, of which only the latter is potentially sympatric speciation.

## S.153.6 The draft genome of the cucumber tree (Dendrosicyos socotranus, Cucurbitaceae) as a genomic basis of adaptive evolution on islands

Gentaro Shigita¹,², Edgardo M. Ortiz¹, Alina Höwener³, Hiroshi Ezura², Hanno Schaefer¹

1 Technical University of Munich, Freising, Germany. 2 University of Tsukuba, Tsukuba, Japan. 3 Ludwig Maximilian University of Munich, Munich, Germany

The island syndrome is a recurring pattern in ecological and morphological differences between insular species and their continental relatives, typically involving loss of defensive structures, increased woodiness, and modifications in floral and reproductive traits. Although this phenomenon has long been treated as a textbook example of adaptive evolution, the underlying genetic mechanisms remain largely unknown. The cu-
cumber tree (Dendrosicyos socotranus Balf.f.) is endemic to Socotra island, Yemen, and the only arborescent species among ~1,000 Cucurbitaceae species. Its anatomical features demonstrate adaptation to Socotra's seasonally dry habitat by storing water in the parenchyma of its swollen stems, allowing extended photosynthesis during the dry season. Moreover, the species attracts additional attention due to its vulnerable status on the IUCN Red List and its cultural significance for local communities as a source of fodder and medicine. Here, we report the first genomic exploration of *D. socotranus*. Based on k-mer spectrum analyses, the species is estimated to be diploid, with a ~400 Mb genome. Using a combination of Nanopore long-reads and Illumina short-reads, the genome was assembled into 181 contigs, totaling 405.9 Mb with an N50 of 4.9 Mb, and 97.7%

BUSCO completeness. Genome annotation revealed that 65.4% of the genome consists of repeats and contains 19,241 protein-coding genes. Comparative genomics with 22 other cucurbits identified 7,592 gene families conserved across all examined cucurbits, as well as 19 gene families unique to *D. socotranus*. Functional annotation of these 19 gene families found one gene family presumably associated with self-incompatibility, suggesting a potential shift in the reproductive manner as observed in other insular species. Further analyses of this genome and comparison with its closest continental relatives will provide valuable insights into the genetic mechanisms that make *D. socotranus* outstanding in the family and facilitate our understanding of adaptive evolution on islands.

# S.154 CYCADS AS EMERGING MODELS IN EVOLUTIONARY BIOLOGY AND SYMBIOSIS

## S.154.1 The Cycas genome and the early evolution of seed plants

Yang Liu¹,²,³⁴, Sibo Wang¹,³⁴, Linzhou Li¹,³⁴, Ting Yang¹,³⁴, Shanshan Dong²,³⁴, Tong Wei¹,³⁴, Shengdan Wu³,³⁴, Yongbo Liu⁴,³⁴, Yiqing Gong², Xiuyan Feng⁵, Jianchao Ma⁶, Guanxiao Chang⁶, Jinling Huang⁵,⁶,⁷, Yong Yang⁸, Hongli Wang¹,⁹, Min Liu¹, Yan Xu¹,⁹, Hongping Liang¹,⁹, Jin Yu¹,⁹, Yuqing Cai¹,⁹, Zhaowu Zhang¹,⁹, Yannan Fan¹, Weixue Mu¹, Sunil Kumar Sahu¹, Shuchun Liu², Xiaoan Lang²,¹⁰, Leilei Yang², Na Li², Sadaf Habib²,¹¹, Yongqiong Yang¹², Anders J. Lindstrom¹³, Pei Liang¹⁴, Bernard Goffinet¹⁵, Sumaira Zaman¹⁵, Jill L. Wegrzyn¹⁵, Dexiang Li¹⁰, Jian Liu⁵, Jie Cui¹⁶, Eva C. Sonnenschein¹⁷, Xiaobo Wang¹⁸, Jue Ruan¹⁸, Jia-Yu Xue¹⁹, Zhu-Qing Shao²⁰, Chi Song²¹, Guangyi Fan¹, Zhen Li²², Liangsheng Zhang²³,²⁴, Jianquan Liu²⁵,

Zhong-Jian Liu²⁶, Yuannian Jiao²⁷, Xiao-Quan Wang²⁷, Hong Wu²⁸, Ertao Wang²⁹, Michael Lisby³⁰, Huanming Yang¹, Jian Wang⁹, Xin Liu¹, Xun Xu¹, Nan Li², Pamela S. Soltis³¹, Yves Van de Peer¹⁹,²²,³², Douglas E. Soltis³¹,³³, Xun Gong⁵, Huan Liu¹, Shouzhou Zhang²

1 State Key Laboratory of Agricultural Genomics, BGI-Shenzhen, Shenzhen, China. 2 Key Laboratory of Southern Subtropical Plant Diversity, Fairy Lake Botanical Garden, Shenzhen & Chinese Academy of Sciences, Shenzhen, China. 3 State Key Laboratory of Grassland Agro-Ecosystems, College of Ecology, Lanzhou University, Lanzhou, China. 4 State Environmental Protection Key Laboratory of Regional Eco-process and Function Assessment, Chinese Research Academy of Environmental Sciences, Beijing, China. 5 Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China. 6 Key Laboratory of Plant Stress Biology, State Key Laboratory of Crop Stress Adaptation and Improvement, Henan University, Kaifeng, China. 7 Department of Biology, East Carolina University, Greenville, NC, USA. 8 College of Biology and Environment, Nanjing Forestry University, Nanjing, China. 9 College of Life Sciences, University of Chinese Academy of Sciences, Beijing, China. 10 Nanning Botanical Garden, Nanning, China. 11 School of Life Sciences
<figure><img src="image_358.png" /></figure>

es, Sun Yat-sen University, Guangzhou, China. 12 Sichuan Cycas panzhihuaensis National Nature Reserve, Panzhihua, China. 13 Global Biodiversity Conservancy, Chonburi, Thailand. 14Department of Entomology, China Agricultural University, Beijing, China. 15 Department of Ecology and Evolutionary Biology, University of Connecticut, Storrs, CT, USA. 16 Guangdong Provincial Key Laboratory for Plant Epigenetics, Longhua Institute of Innovative Biotechnology, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, China. 17 Department of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, Denmark. 18 Shenzhen Agricultural Genome Research Institute, Chinese Academy of Agricultural Sciences, Shenzhen, China. 19 College of Horticulture, Academy for Advanced Interdisciplinary Studies, Nanjing Agricultural University, Nanjing, China. 20 State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, China. 21 Chengdu University of Traditional Chinese Medicine, Chengdu, China. 22 Department of Plant Biotechnology and Bioinformatics, Ghent University, VIB UGent Center for Plant Systems Biology, Gent, Belgium. 23 College of Agriculture and Bio-technology, Zhejiang University, Hangzhou, China. 24 Hainan Institute of Zhejiang University, Sanya, China. 25 The College of Life Sciences, Sichuan University, Chengdu, China. 26 Key Laboratory of Orchid Conservation and Utilization of National Forestry and Grassland Administration at College of Landscape Architecture, Fujian Agriculture and Forestry University, Fuzhou, China. 27 State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Beijing, China. 28 College of Life Sciences, South China Agricultural University, Guangzhou, China. 29 National Key Laboratory of Plant Molecular Genetics, Chinese Academy of Sciences Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, China. 30 Department of Biology, University of Copenhagen, Copenhagen, Denmark. 31 Florida Museum of Natural History, University of Florida, Gainesville, FL, USA. 32 Department of Biochemistry, Genetics and Microbiology, University of Pretoria, Pretoria, South Africa. 33 Department of Biology, University of Florida, Gainesville, FL, USA. 34 e-mail: yang.liu0508@gmail.com; yvpee@psb.vib-ugent.be; dsoltis@ufl.edu; gongxun@mail.kib.ac.cn; liuhuan@genomics.cn; shouzhouz@szbg.ac.cn

Cycads represent one of the most ancient lineages of living seed plants. Identifying genomic features uniquely shared by cycads and other extant seed plants, but not non-seed-producing plants, may shed light on the origin of key innovations, as well as the early diversification of seed plants. Here, we report the 10.5-Gb reference genome of Cycas panzhihuaensis, complemented by the transcriptomes of 339 cycad species. Nuclear and plastid phylogenome-

ic analyses strongly suggest that cycads and Ginkgo form a clade sister to all other living gymnosperms, in contrast to mitochondrial data, which place cycads alone in this position. We found evidence for an ancient whole-genome duplication in the common ancestor of extant gymnosperms. The Cycas genome contains four homologues of the fitD gene family that were likely acquired via horizontal gene transfer from fungi, and these genes confer herbivore resistance in cycads. The male-specific region of the Y chromosome of C. panzhihuaensis contains a MADS-box transcription factor expressed exclusively in male cones that is similar to a system reported in Ginkgo, suggesting that a sex determination mechanism controlled by MADS-box genes may have originated in the common ancestor of cycads and Ginkgo. The C. panzhihuaensis genome provides an important new resource of broad utility for biologists.

## S.154.2 Paleogenomic data on karyotype evolution in the gymnosperms support Ginkgo and cycads as sisters and both as sister to Pinaceae

Pengchuan Sun¹, Susanne S. Renner², Jianquan Liu¹

1 College of Life Sciences, Sichuan University, Chengdu, China. 2 Department of Biology, Washington University, Saint Louis, USA

Chromosome-scale genomes provide a basis for inferring ancestral linkage groups and processes of karyotype evolution. We have reconstructed the karyotypes of nine representatives of the major groups of gymnosperms, using an approach that starts from retained intact chromosomes and syntenic blocks. The results show that the most recent common ancestor of the gymnosperms had 24 unique protochromosomes and that their fusion can be traced from the Pinales ancestor to extant species of (((Pinus, Cycas, Ginkgo), ((Cupressus, Metasequoia), (Taxus, and Torreya))). The karyotypes of *Gnetum* and *Welwitschia*, by contrast, are extremely modified. Recent assessments of cycad and Ginkgo male and female reproductive organ development, pollen ontogeny and germination as well as ovule development, female gametophyte formation, and ovule integument differentiation (Offer et al. Critical Rev. in Plant Science 2023) support the (*cycad*, Ginkgo) sister relationship.
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## S.154.3 The Cuba Conundrum: Insights into Caribbean Zamia (Zamiaceae, Cycadales) evolution

Michael Calonje¹, Ramona Oviedo Prieto², Gabriel Brull Puebla³, Lisbet Gonzalez-Oliva², Ramiro Chaves², M. Patrick Griffith¹, Javier Francisco-Ortega⁴,¹, Dayana Salas-Leyva⁵, Mario Coiro⁶,⁷, Alan W. Meerow¹,⁸

1 Montgomery Botanical Center, Coral Gables, Florida, USA. 2 Instituto de Ecología y Sistemática, Habana, Cuba. 3 Empresa Nacional para la Protección de la Flora y la Fauna, Habana, Cuba. 4 Florida International University, Florida, USA. 5 University of Cambridge, Department of Biochemistry, Cambridge, UK. 6 Department of Palaeontology, University of Vienna. 7 Ronin Institute for Independent Scholarship, Montclair, USA. 8 Arizona State University, College of Life Sciences, Tempe, AZ, USA

We used SSR and Single Copy Nuclear Gene molecular markers to explore patterns of genetic variation across 130 wild populations of Caribbean zamias sampled throughout the entirety of their geographic range. We report the results of several genetic analyses with an emphasis on populations occurring in Cuba, the country with the highest species richness (6 currently accepted spp.) and morphological diversity in this group. We present the following findings: 1) The strongest genetic separation is between zamias in Florida and those occurring in the rest of Caribbean, 2) Cuban populations appear closest to the ancestral Caribbean zamia population and other Caribbean Islands where colonized from Cuba, 3) Bahamian populations are closest genetically to Cuban populations and there is a close relationship between populations in Dominican Republic and Puerto Rico and between populations in Jamaica and the Cayman Islands, 4) Hispaniola was colonized from Puerto Rico rather than from Cuba, explaining the unusual distribution of the genus on the island, where it is restricted to Eastern Dominican Republic, 5) 80% of Florida populations have Heterozygote excess indicating a recent bottleneck event, likely due to the massive starch industry in Florida. 6) Zamia lucayana, a species endemic to Long Island in the Bahamas, is more closely related to Cuban Zamia populations than to other Bahamian populations, 7) Cuban populations are the most morphologically distinct in the Caribbean, but are incredibly genetically admixed, suggesting a high degree of genetic connectivity between them in the recent

past. 5) there is a high morphological convergence between genetically distinct populations in the Caribbean, and this extreme conflict between morphological resemblance and genetic similarity makes them a challenging group for morphology-based traditional species delimitation.

## S.154.4 Symbiotic microbial and metabolic diversity and age estimates of coralloid roots: deciphering cycad's resilience secrets

Angélica Cibrián-Jaramillo¹, Francisco Barona-Gómez²

1 Naturalis Biodiversity Center, Leiden, the Netherlands. 2 Institute of Biology, Leiden University, the Netherlands.

Cycads are known to host symbiotic cyanobacteria, including Nostocales species, as well as other symbiotic bacterial taxa within their specialized coralloid roots. Even if it is reasonable to expect that Nitrogen-fixing microbiomes contribute towards the remarkable adaptability of cycads, it remains to be elucidated if (i) these bacteria share a phylogenetic origin and/or common genomic functions that allow them to engage in facultative symbiosis; (ii) how the metabolic products of these microbiomes mediate and sustain 'symbiotic' interactions amongst conserved bacteria and with the cycad host; and (iii) what is the role of these communities, and of the whole coralloid root, on cycad's longevity, which remains to be determined. In this presentation, we provide an integrated view on the cycad's coralloid root, including its microbiome composition and related metabolic products, as well as the anatomy of the host itself, obtained after a broad range of analytical approaches. These include phylogenometagenomics and functional analysis of synthetic sub-communities, to evaluate evolutionary and functional relationships; metabolomics and imagining DESI mass spectrometry, to provide mechanistic insights underlying the symbiosis functions; and q-FISH to measure telomers lengths and laser ablation tomography to analyze internal anatomy of colonized and non-colonized coralloid roots. Results derived from our integrated approach support a role of coralloid roots on cycad's adaptive resilience, central to the notion of developing cycads as an evolutionary plant model and with a bearing on conservation strategies of this highly endangered plant group.
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**S.154.5 Enemy-free space in cycad-herbivore interactions: a case study of Zamia cycads and Eumaeus butterflies**

Melissa RL Whitaker¹

1 East Tennessee State University, Johnson City TN USA

Cycads support fascinating insect herbivore faunas composed of mutualists like brood site pollinators, as well as parasitic herbivores such as folivorous caterpillars. While most research on cycad-insect interactions has focused on insect pollination, parasitic herbivores also significantly affect the fitness of many cycads, with sometimes dire consequences for cycad conservation and management. These herbivore associations are strongly influenced by cycads' defensive traits, particularly their unique chemical defenses, requiring significant adaptation on the part of the insects to be able tolerate, excrete, and/or detoxify cycads'phyto-toxins. For this reason, the majority of parasitic cycad herbivores are obligate cycad specialists. For example, the neotropical butterfly genus, *Eumaeus*, is a wholly cycadivorous lineage that feeds primarily on *Zamia* cycads and provides an emerging model system for investigating cycad-herbivore interactions. Recent research in into *Zamia-Eumaeus* interactions includes genomic investigations of both the insects and plants, toxicity and survival assays, and chemical analyses. In this presentation, I synthesize recent research on *Zamia-Eumaeus* interactions (both published and in progress), including results from preference-performance assays conducted with cycad-specific toxins, and descriptive and experimental investigations into the importance of higher trophic levels driving cycad-insect interactions. I evaluate these results in the framework of enemy-free space, using a multi-trophic approach to understand the evolution and ecology of cycads' fascinating symbioses with insects.

**S.154.6 Multi-modal pollination signaling in cycads, an ancient plant lineage**

Shayla Salzman¹

1 University of Georgia, Athens, USA

Cycads are an ancient lineage of dioecious gymnosperm that overwhelmingly rely on insect mutualists for pollination services, providing an excellent opportunity for studies into the ecology and evolution of insect pollination, plant-insect signaling, and the maintenance of mutualisms. Specialized cycad-beetle pollination has existed since at least the early Jurassic, placing it before the ecological dominance of angiosperms and the accompanying diversification of major lineages of pollinators such as butterflies and bees (Cai et al 2018). Entire lineages of beetles exclusively live on and pollinate cycads in what has been suggested to be a classic case of co-evolution. Chemical communication (scent) has been shown to be important for the maintenance of these mutualisms (Salzman et al. 2021) and for plant reproduction through 'push-pull pollination' (Terry et al. 2007), the most ancient pollination mechanism yet documented (Salzman et al 2020), yet scent is not the only signal influencing pollinator behavior (Salzman et al. 2023). These entomophilous plants produce dense woody reproductive cones without the overt visual signals commonly employed by angiosperms and must rely on other signals and cues for communication with their insect partners. As such, they are thermogenic plants, producing internal heat within their reproductive structures that in turn alters the carbon dioxide, humidity, and scent of the cones. All of the plant produced primary and secondary metabolites that are perceived by pollinators are used in decision making. I discuss plant signaling and morphology in an ecological and evolutionary framework to understand extant and ancient pollination.

References: Cai et al. 2018. Current Biology 28: 2806-2813. Salzman et al. 2020. Science Advances 6: eaay6169. Salzman et al. 2021. Frontiers in Plant Science 12: 639368. Salzman et al. 2023. Current Biology 33: 1654-1664. Terry et al. 2007. Science 318: 70.
# S.155 KEY MORPHOLOGICAL INNOVATIONS IN FLOWER EVOLUTION

## S.155.1 Development of sexually deceptive petal spots

Farahnoz N. Khojayori¹, Udhaya Ponraj¹, Xiao Wang¹, Róisín Fattorini², Roman Kellenberger¹, and Beverley J. Glover¹

¹University of Cambridge, Cambridge, UK. ²University of Liverpool, Liverpool, UK

Gorteria diffusa, a South African daisy, has at least 15 distinct populations that differ in their floral morphology, specifically in the number, complexity, and position of petal spots. However, all morphs are polinated by the same bee-fly, Megapalpus capensis. Floral morphs with simple spots lead to foraging and resting by the bee-fly, while those with complex petal spots mimic the female bee-fly, tricking the males into pseudo-copulation. Our research suggests multiple genetic pathways may have been co-opted and integrated to produce an effective bee-fly mimic. We are comparing these pathways across the populations of *G. diffusa* to identify processes underlying the evolution of this novel character.

## S.155.2 Macroevolutionary patterns of floral diversification

Jürg Schönenberger¹, Julian Herting², Andrea López-Martínez³, Marion Chartier¹, Susana Magallón³, Maria von Balthazar¹, Hervé Sauquet²

¹University of Vienna, Vienna, Austria. ²National Autonomous University of Mexico, Mexico City, Mexico. ³Botanic Gardens of Sydney, Sydney, Australia

Since crown group angiosperms originated more than 140 Mya, flowers have diversified into a multitude of floral forms. Their diversification was shaped by the combination of extrinsic physical (e.g., climate) and biological (e.g., pollinators) factors as well as by their intrinsic structural and genetic constraints. The continuing progress in phylogenetic and

phylogenomic analyses in angiosperms, together with results from comparative floral development and morphology, provide a unique opportunity to understand macroevolutionary patterns of floral diversification. In this study, we trace the evolution and diversification of various floral traits related to the three main organ categories (perianth, androecium, and gynoecium) and key aspects of floral structure (e.g., merism, phyllotaxis, and union of organs). We base our analyses on an extensive, referenced and continuously updated and curated dataset of floral morphology, which we compiled over the past decade in the collaborative database PROTEUS, as part of the eFLOWER initiative. The current dataset comprises 30 floral traits scored for 1201 species, representing all currently recognized angiosperm families. We apply ancestral state reconstructions using parsimony, maximum likelihood, and Bayesian inference, exploring different evolutionary models, to trace the evolution and diversification of pivotal floral traits. In addition, we reconstruct and re-evaluate ancestral flowers at the crown node and additional deep nodes in the angiosperm phylogeny. We find that floral diversification was shaped both by modification and innovation and that similar evolutionary trajectories were often explored repeatedly and independently in different parts of the angiosperm phylogenetic tree. Our study complements earlier and ongoing studies from the fields of paleobotany, comparative morphology, and evo-devo in their attempts to answer key questions on the early evolution and diversification of flowers in angiosperms.

## S.155.3 Developmental patterning of head-like inflorescences in Asteraceae

Paula Elomaa¹, Teng Zhang¹, Feng Wang¹, Andrew Owens², Mikolaj Cieslak², Przemysław Prusinkiewicz²

¹Department of Agricultural Sciences, Viikki Plant Science Centre, University of Helsinki, Finland. ²Department of Computer Science, University of Calgary, Canada
<figure><img src="image_361.png" /></figure>

Asteraceae (sunflower family) represents one of the largest and most diverse families of plants. The inflorescences of Asteraceae mimic solitary flowers but are in fact compressed, head-like structures typically comprised of multiple florets with specialized functions. This unique architecture has been considered as a key innovation behind the evolutionary success of the family. Flower heads are also iconic examples of geometric regularity found in nature; the involucral bracts and florets are arranged in regular left- and right-curving spirals whose numbers follow the mathematical Fibonacci series. We have explored the growth dynamics and patterning of the enlarged inflorescence meristem in the model plant *Gerbera hybrida* and shown with DR5 auxin reporter lines how the high spiral numbers emerge. The molecular data was integrated into a computational model to explain phyllotactic patterning of Asteraceae heads. We have extended our studies to explore how head vasculature is associated with organ phyllotaxis. By applying synchrotron-based micro-CT imaging, we have explored the vascular networks in heads of diverse Asteraceae species. The observed diversity and irregularities of vasculature stand in contrast with the regularity of phyllotactic patterns, confirming that phyllotaxis in heads in not driven by the vasculature. We discuss how our recent discoveries on meristem patterning relate to distinct scenarios of the evolutionary origin of Asteraceae heads.

## S.155.4 Gene Duplication Facilitates but does not Trigger Floral Color Pattern Diversification

Mark D. Rausher¹

¹ Duke University, Durham, USA

The genus *Clarkia* exhibits a tremendous diversity of floral color patterns. Patterns consist of a small number of pattern elements, including background petal color, spots, sprays, white patches. The presence/absence, positions and sizes of these elements frequently differ between very closely related species. In general, different pattern elements are controlled by different paralogs of R2R3Myb proteins that have arisen via duplication, and similar pattern elements (e.g. spots) are controlled by different paralogs in different species. The spatial locations of elements are determined by the expression domains of the genes coding for these proteins. While gene duplication clearly has facilitat-

ed the diversification of color patterns, this does not necessarily imply that duplication events actually triggered his diversification. By exploring the presence of different paralogs in *Clarkia* and related genera, we have found that the R2R3Myb duplications occurred long before diversification of color patterns, indicating a substantial lag time between duplication and the association of these genes with color pattern elements. Duplication events thus did not trigger pattern diversification. Instead, this lag suggests that pattern diversification did not occur until environmental conditions made it advantageous to have novel patterns.

## S.155.5 A generalized polarity-based modeling system for plant lateral organ morphogenesis and evolution

Jie Cheng¹,², Xu Yao¹,², Xukun Li¹,²,³, Tan Qin¹,²,³, Dian Yu¹,²,³, Xiaofeng Yin¹,², Hongyan Shan¹,², Christopher Whitewoods⁴, Enrico Coen⁵, Hongzhi Kong¹,²,³

¹ State Key Laboratory of Plant Diversity and Specialty Crops, CAS Center for Excellence in Molecular Plant Sciences, 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 Sainsbury Laboratory, University of Cambridge, Cambridge UK. ⁵ Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park, Norwich, UK

Shape and form are basic attributes of objects. Exploring how the various shapes of Organisms were made through development and evolution is the key to understanding biodiversity. Peltate organs, such as the prey-capturing traps of carnivorous plants and necessary-bearing petals of ranunculaceous species, are widespread in nature and have intrigued and perplexed scientists for centuries. Shifts in the expression domains of adaxial/abaxial genes have been shown to control leaf peltation in some carnivorous plants, yet the mechanisms underlying the generation of other peltate organs remain unclear. Here, we show that formation of various peltate ranunculaceous petals was also caused by shifts in the expression domains of adaxial/abaxial genes, followed by differentiated regional growth sculpting the margins and/or other parts of the organs. By inducing parameters to specify the time, position, and degree of the shifts and growth, we fur-
<figure><img src="image_362.png" /></figure>

ther propose a generalized modeling system, through
which various unifacial, bifacial, and peltate organs can
be simulated. These results demonstrate the existence
of a hierarchical morphospace system and pave the
way to understand the mechanisms underlying plant
organ diversification. Based on this generalized organ
modelling system, we are extending it into a more pow-
erful framework available for multiple organs as well as
complex biological structures.

S.155.6 Developmental genetics of corolla tube formation: a key morphological innovation during angiosperm evolution

Yao-Wu Yuan²

1 Department of Ecology & Evolutionary Biology, University of Connecticut, USA

About one third of all angiosperm species produce
flowers with petals fused into a corolla tube. Differ-
ences in the length, width, and curvature of the co-
rolla tube have led to an endless variety of different-

ly shaped flowers that attract specialized groups of
pollinators (e.g., beefflies, hawkmoths, hummingbirds,
nectar bats). As such, the corolla tube is considered
a key morphological innovation that contributes to
plant diversification and speciation; yet it remains
one of the least understood plant organs from a de-
velopmental genetics perspective. In this talk, I pres-
ent our work towards understanding this problem
using monkeyflowers (Mimulus) as a model system.
Through genetic analyses of several Mimulus mutants
with split corolla tubes, characterization of corolla tube
ontogenesis in the wild type and mutants, and a se-
ries of transgenic experiments, we have established
a regulatory pathway and a new conceptual model
for the developmental genetic control of corolla tube
formation. At the heart of this model is auxin-induced
synchronized growth between the bases of the petal
primordia and the inter-primordial regions during ear-
ly stages of flower development. Upstream of this core
module is the tasiRNA-ARF3/4 pathway that regulates
auxin homeostasis in the synchronized growth zone;
downstream is an organ boundary gene (MINAC1) that
suppresses localized tissue growth if not repressed by
auxin signaling. We expect that this conceptual model
and the regulatory pathway (i.e., tasiRNA-ARF3/4-aux-
in-MINAC1) will serve the foundation for many interest-
ing studies on corolla tube development and evolu-
tion in the coming years.

S.156 RELICT PLANT VULNERABILITY TO CLIMATE CHANGE

S.156.1 Algerian relict trees under global and climat change: statuts, threats and conservation

Sarmoum Mohamed¹, Abdoun Fatiha²

1 Laboratory of Plant Physiology and Soil Culture, Faculty of Nature and Life Sciences, Ibn Khaldoun University, PO Box 78 Zaaroura, Tiaret, Algeria. mohamed.sarmoum@univ-tiaret.dz. 2 Laboratory of Plant Ecology and Environment, Faculty of Biological Sciences, University of Science and Technology Houari Boumediene, BP 32 El Alia, Bab Ezzouar, Alger, Algeria.

In the context of global and climatic changes which
prevail today, many trees are threatened with dis-
appearance. Algeria is a vast country which is lo-
cated between two large geographical areas: Med-

iterranean (Northern Algeria) and desert (Southern
Algeria). This situation has generated a remarkable
diversity of habitats and ecosystems. Unfortunately,
global and climatic changes have negatively af-
fected this diversity (deforestation, grazing, desert-
ification, forest dieback, etc.). Among the important
elements of this biodiversity, relict trees occupy an
important place in the landscapes. Having lived in
different geological eras, these trees are witness-
es to the environmental changes that have taken
place over millions of years. They also constitute a
natural heritage of great ecological, economic and
cultural value which must be preserved for future
generations. In this work, we present the state of
knowledge on the relict trees of Algeria, their eco-
logical and biogeographical status, the threats and
the problems linked to their conservation under
global and climatic changes.
<figure><img src="image_363.png" /></figure>

## S.156.2 Diversity of Taxaceae in Northeastern China during Middle-Late Jurassic

Chong Dong¹, Gongle Shi¹, Fabiany Herrera², Yongdong Wang¹, Patrick S. Herendeen³, Peter R. Crane⁴,⁵

1 Nanjing Institute of Geology and Palaeontology, Chinese Academy of Sciences, Nanjing, China.

2 Field Museum of Natural History, Chicago, USA.

3 Chicago Botanic Garden, Glencoe, USA. 4 Oak

Spring Garden Foundation, Upperville, USA. 5 Yale

University, New Haven, USA

Taxaceae (the yew family) are a small family of conifers comprising about 25 extant species in five genera, including *Taxus*, *Amentotaxus*, *Torreya*, *Pseudotaxus*, and *Austrotaxus*. Their axillary ovulate structures are characterized by bearing a single terminal seed with a fleshy aril, which makes the family very different from other extant conifers. Taxaceae have a long fossil history with the earliest occurrences from the earliest Jurassic (Hettangian, 201–197 Ma) of southern Sweden. Recently, we discovered well-preserved, diverse Taxaceae fossils from the Middle- Late Jurassic Daohugou Biota (~165–158 Ma) in eastern Inner Mongolia, northeastern China. These include a least four species that could be assigned to *Amentotaxus*, *Taxus*, *Torreya* and probably an extinct genus different from all extant genera of Taxaceae. Among the new fossils, *Amentotaxus ningchengensis* sp. nov. and *Taxus daohugouensis* sp. nov. are based on leafy shoots with attached seed-bearing structures, and they resemble those of extant *Amentotaxus* and *Taxus* respectively in vegetative as well as ovulate features. *Torreya daohugouensis* sp. nov. is based on leafy shoots with well-preserved cuticles. It is also comparable to extant *Torreya* in morphology of leaves as well as detailed cuticle structures. The new fossils from the Dahugou Biota show that the yew family had been diverse in Northeastern China since the Middle-Late Jurassic and this pattern of diversification is also seen in early Cretaceous floras of Mongolia. *Amentotaxus*, *Taxus* and probably *Torreya* appeared early in the evolutionary history of the family. The Asian fossils also indicate that *Amentotaxus* and *Taxus* may have undergone little morphological change since the Middle-Late Jurassic.

## S.156.3 Meta-analysis of genomic vulnerability of relict plants under rapid climate change in East Asia

Wen-Hao Li¹, Zheng-Zhang Zhu¹, Yun-Peng Zhao¹

¹Systematic & Evolutionary Botany and Biodiversity Group, MOE Key Laboratory of Biosystem Homeostasis and Protection, College of Life Sciences, Zhejiang University, Hangzhou, China

Relict species form an essential component of contemporary biodiversity as representatives of a patrimony nested in the Tree of Life. Rarity and restricted distributions obviously place relict species in special demographic jeopardy of extinction, especially in the face of ongoing climatic changes and other ecological perturbations (Habel et al. 2010). East Asia harbors hundreds of relict species, as several long-term climatically stable refugia provided chances of species-level persistence (Tang et al. 2018). However, climatic-derived genetic diversity loss of relict species in the coming decades has few reported. In this study, we evaluated the population-level climatic vulnerability of 10 relict taxa by genetic-environment associated loci data, considering local adaptation by heterogenous habitats. We aimed to (1) identify the most fragile areas with highest genomic vulnerability, highlighting prior local populations for conservation; (2) assess effects of rescue via migration with or without distance limitation; (3) reveal the traits/genetic basis that enable populations overcome the adaptive challenges. Our work quantified the maladaptation and migration to understanding the extinct risks of relict plants, further help target future conservation actions.

**References:** Habel, J. C., Assmann, T., Schmitt, T., & Avise, J. C. (2010). Relict species: from past to future. In Relict species: phylogeography and conservation biology (pp. 1–5). Springer Berlin Heidelberg. Tang, C. Q., Matsui, T., Ohashi, H., Dong, Y. F., Momohara, A., Herrando-Moraira, S., ... & López-Pujol, J. (2018). Identifying long-term stable refugia for relict plant species in East Asia. *Nature Communications*, *9*(1), 4488.
<figure><img src="image_364.png" /></figure>

## S.156.4 The origin, speciation, and adaptive divergence of the relict genus Pterocarya under the climate change

Yi-Gang Song¹, Tian-Rui Wang¹, Zi-Jia Lu¹, Hong-Hu Meng², Si-Si Zheng¹, Gregor Kozlowski¹,³,⁴

1 Eastern China Conservation Centre for Wild En-
danger ed Plant Resources, Shanghai Chenshan
Botanical Garden, Shanghai, China. 2 Plant Phylo-
genetics and Conservation Group, Center for In-
tegrative Conservation, Xishuangbanna Tropical
Botanical Garden, Chinese Academy of Sciences,
Kunming, China. 3 Department of Biology and Bo-
tanic Garden, University of Fribourg, Fribourg, Swit-
zerland. 4 Natural History Museum Fribourg, Fri-
bourg, Switzerland

The integration of past, present and future spa-
tial-temporal patterns is crucial in all research areas
of biogeography and conservation biology. Com-
bined with the phylogenomics, biogeography, and
adaptive genomic variation, we explored the origin,
speciation, hybridization, and adaptive evolution of
the small Cenozoic relict genus Pterocarya that live
in riparian areas with six to eight species. Our results
clearly confirm the division of the genus into two
monophyletic sections: the Platyptera section and
the Pterocarya section. The climate changes during
the Early Miocene era coped with the divergence
between the two sections. The climate cooling and
formation of Gobi Desert in the Middle Miocene ep-
och caused a split in the continuous distribution of
the genus and formation of the disjunction. Focus on
East Asia, the phylogeographic break of Pterocarya
hupehensis conformed to the boundary of the Si-
no-Himalayan and Sino-Japanese forests sub-king-
doms. However, in contrast to the nuclear gene, two
chloroplast clades within the eastern lineage further
diverged along the Yangtze River, and the extensive
pollen flow may be the primary factors. Three ge-
netic lineages within P. macroptera: the Qinling-Da-
ba-Tianmu Mountains, Western Sichuan, and North-
west Yunnan lineages, which showed significant
signals of isolation by distance (IBD) and isolation
by environment (IBE). The identified GEA SNP-related
genes were involved in chemical defence and gene
regulation and may exhibit higher genetic variation
to adapt to the environment. Gradient forest anal-
ysis revealed that the genetic variation was mainly
shaped by temperature-related variables. A limited

adaptive potential was suggested by the high lev-
els of genetic vulnerability in marginal populations.
Environmental gradient mainly shaped the popula-
tion differentiation of P. macroptera. Marginal pop-
ulations may be at high risk of extinction, and thus
proactive management measures, such as assist-
ed gene flow, are required to ensure the survival of
these populations.

## S.156.5 Local belief as a conservation driver of golden larch (Pseudolarix amabilis) in China

Cheng Jin, Yuan Chen¹, Siwei Hu, Lihua Zhou¹, Yongchuan Yang¹

1 Key Laboratory of the Three Gorges Reservoir Region's Eco-Environment, Ministry of Education, Chongqing University, Chongqing, China

Pseudolarix amabilis is a specific mono-species
genus plant in China, and one of the most interna-
tionally influential ancient relict plants in China. P.
amabilis is facing various threats such as habitat
fragmentation, population isolation, and number
dwindle. Fortunately, it has been pointed out that
the village fengshui forest provides shelter for the
natural population of P. amabilis. However, how
can the village fengshui forest protect the natural
population of P. amabilis remains to be explored
in depth. Therefore, based on the methods of field
investigation and community interview, this study
conducted a comprehensive investigation of the
recruitment status, conservation behavior, and
disturbance degree for the natural population of
P. amabilis. At the same time, we took the natural
population of P. amabilis in Linjiatang as an exam-
ple to reveal the protection and management par-
adigm of P. amabilis fengshui forest and explore its
role in the effective protection of P. amabilis. The
results showed that DBH structure of P. amabilis
in fengshui forest was more continuous, and the
seedling density was higher, compared with other
habitat types. Fengshui and taboo culture carried
by shuikou forest, cemetery forest, and yangji tree
are the internal reasons why the fengshui forest
can effectively protect the natural population of
P. amabilis. The development of rural tourism has
further promoted the protection of the P. amabilis
fengshui forest. Our results can provide a basis for
making an effective protection paradigm for the
natural population of P. amabilis.
<figure><img src="image_365.png" /></figure>

S.156.6 Deep phylogeographic divergence of beaked hickory calls for transboundary conservation in a biodiversity hotspot

Jie Liu¹

1 CAS Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences

The conservation of endangered plants is a cru-
cial step for the maintenance of biodiversity in
a changing world. However, the lack of compre-
hensive phylogeographic information for most
endangered species greatly impedes effective
action, especially for species distributed across
national borders. Beaked hickory (Carya sinensis),
which is endemic to China and Vietnam, part of
the Indo-Burma biodiversity hotspot, exemplifies
these issues. Here, based on comprehensive field
sampling, we used three genomic datasets (i.e.,

plastid, mitochondrial, and nrDNA) and ecologi-
cal niche modeling to reveal its genetic diversity,
phylogeographic structure, and population histo-
ry, shedding insights into conservation. Three dis-
tinct clades were resolved, i.e., Clade A in Vietnam,
Clade B in China, and Clade C covering both coun-
tries. Genetic diversity is low at species and with-
in-clade levels, but multiple private haplotypes
were observed in each clade. The three clades
diverged between 6 and 8 Mya, making this a far
older species than others within *Carya*, and each
likely had a separate glacial refugium. Bottlenecks
were detected in all three clades after the LGM,
probably driven by the late Pleistocene glacial os-
cillations, but Clade C experienced a recent (~800
years ago) expansion which might have been an-
thropogenic. From this, we recommend treating
the three clades as distinct conservation units for
*C. sinensis* and propose a precise evidence-based
*in* and *ex situ* conservation management strategy,
as part of a practical transboundary conservation
framework. This sets a model for transboundary
conservation through joint efforts between na-
tions, for other threatened species using a similar
integrative approach.

S.157 PLANT RESPONSES TO STRESSES AND CLIMATE CHANGE. SESSION 7

S.157.1 Predicting the effects of climate change on seed germination: implications for woodland regeneration and tree seed sourcing

Roberta L.C. Dayrell¹, Charlotte Seal¹, Efisio
Mattana¹, Kaitalin White¹, Pablo Gómez-
Barreiro¹, Ted Chapman¹

1 RBG Kew, Millennium Seed Bank, Ardingly, UK

Expanding and connecting woodlands to capture
atmospheric carbon and enhance resilience to en-
vironmental change is garnering support as a cli-
mate change mitigation action. This requires an
unprecedented increase in tree planting, which de-
mands the efficient use of limited native seed re-
sources. Thus, there is a need to develop improved

practices to overcome hurdles posed by the vari-
able quality, dormancy states, and germination re-
quirements of native seeds. Moreover, as the climate
changes, species germination requirements may no
longer be fulfilled or be met at an unfavourable time
for seedling establishment, impacting woodland re-
generation and ecosystem diversity and services.
We aimed to investigate intra-specific variation in
seed germination traits of two native broad-leafed
species of forestry importance in the UK: *Alnus gluti-
nosa* and *Betula pubescens*. We tested seeds from
six populations of each species collected in the
wild and stored at the RBG Kew's Millennium Seed
Bank, to represent the latitudinal gradient across
the country. Germination tests were conducted on
collections of each population at a range of con-
stant temperatures, both with and without a cold
stratification pre-treatment to alleviate dormancy.
The collected data were then used in a modelling
approach to estimate thermal thresholds for seed
germination (base, optimum and ceiling tempera-
<figure><img src="image_366.png" /></figure>

tures; sub- and supra-optimal thermal time) and determine the requirement for cold stratification for all collections. In addition, we modelled germination parameters under different climate change scenarios to identify plant populations at the greatest risk. Finally, we explored the potential of morphological traits serving as predictors of germinative behaviour. This information helps evaluate the impact of climate change on woodland regeneration and allows for informed adaptive 'climate-smart' seed sourcing to enhance reforestation practices. The results also provide insights on how climate and provenance shape the functional diversity of tree seeds.

### S.157.2 Revealing within tree sensitivity to drought in temperate forest species differing in shade tolerance

Francisco J. Cano¹,², Rosana López³,
Jesús Rodríguez-Calcerrada³, Charles R.
Warren⁴, Luis Gil³, Ismael Aranda¹

1 Instituto de Ciencias Forestales (INIA), CSIC, Madrid, Spain. 2 ARC Centre of Excellence for Translational Photosynthesis, Hawkesbury Institute for the Environment, Western Sydney University, NSW, Australia. 3 Department of Natural Systems and Resources, Universidad Politécnica de Madrid, Madrid, Spain. 4 School of Life and Environmental Sciences, University of Sydney, Sydney, Australia

Droughts are becoming a recurrent event worldwide, which reduce carbon uptake and increase tree mortality risk threatening several ecosystem functions and services. Global change is increasing the intensity, duration, and frequency of droughts due to a combined effect of scatter precipitations, atmosphere warming and soil erosion. However, droughts do not affect equally to all forest tree species due to species-specific differences in drought resilience and niche partition. Furthermore, leaf physiology, anatomy and morphology acclimate within the tree canopy to changes in light intensity and quality, evaporative demand and soil water availability. However, not many studies analyse within tree canopy inter-specific drought tolerance and plasticity, which in turn may constrain the adaptive strategies of the species shade tolerance. Here, we measured leaf gas exchange and hydraulics within the canopies of four forest tree species differing in shade and drought tolerance. Trees were in transects of increasing soil depth at the Natural World Heritage

Site of "Hayedo de Montejo", a relict European forest growing under Mediterranean conditions, and measurements were done during very dry and wet years. We show that shaded leaves are more susceptible to water stress than sun-developed leaves, and that decreased photosynthesis is due to the combined effect of reduced stomatal and mesophyll conductances and to a lesser extent by biochemical constraints, while top canopy leaves are mainly limited by the stomatal closure. This results in earlier lower carbon balance of the more shaded leaves within the canopy, what explains the earlier leaf shedding of the more shaded leaves. More shade tolerant species were more affected by drought along the canopy, while more drought tolerant ones were able to maintain modest gas exchange rates but only in sun-exposed leaves. Climate change may promote open forests which may benefit the more drought albeit less shade tolerant species.

### S.157.3 Strategies for enhancing urban air quality through vegetation

Barbara Baesso Moura¹,², Yasutomo
Hoshika¹,²,³, Jacopo Manzini⁴, Elena
Marra¹,³, Elena Paoletti¹,²,³

1 Institute of Research on Terrestrial Ecosystems, National Research Council, esto Fiorentino, Italy. 2 National Biodiversity Future Center, Palermo, Italy. 3 Italian Integrated Environmental Research Infrastructures System, Tito Scalo (Potenza), Italy. 4 Department of Agricultural, Food, Environmental and Forestry Science and Technology, University of Florence, Firenze, Italy

Cities face diverse environmental challenges, making it crucial to identify the most significant ones for each city to design effective urban strategies. Implementing urban green infrastructure is a promising approach that can significantly improve public health by reducing air pollution. Vegetation controls pollutant flow and distribution, with some tree species being more effective at trapping pollutants than others. Key traits for improving air quality include canopy density, foliage longevity, water-use strategy, and high stomata conductance, which is essential for gaseous pollutants capture. However, trees can also emit volatile organic compounds (VOCs), which react to form ground-level ozone (O3). Therefore, avoiding tree species that emit high VOC levels is important, particularly in residential areas. Advanced models and tools can help estimate
<figure><img src="image_367.png" /></figure>

ecosystem services and support urban greenery management. These tools assist public institutions and citizens in selecting species that maximize air pollution reduction. The effectiveness of urban greenery varies with fragmentation, green cover, and plant species. In conclusion, while beneficial vegetation traits enhance pollutant capture, they do

not guarantee suitability for all contexts. Selecting tree species with high pollution mitigation potential does not always yield desirable results. Plant species must be adapted to local conditions, and stress tolerance should be prioritized in urban environments, especially roadside areas facing multiple stressors.

## S.158 TELLING PLANT SPECIES APART WITH DNA. SESSION 2

### S.158.1 Comparative analysis of complete plastome sequences of Tulipa L. (Liliaceae Juss.) species from Kazakhstan

Almerekova S.¹, Yermagambetova M.¹, Turuspekov Y.¹, Abugalieva S.¹

1 Laboratory of Molecular Genetics, Institute of Plant Biology and Biotechnology, Almaty, Kazakhstan

The *Tulipa* L. genus belongs to the Liliaceae Juss. family, consisted around 100 species all over the world. *Tulipa* species are native to southern Europe, North Africa, the Middle East, and Central Asia. The mountainous regions of Central Asia (including Kazakhstan) are considered the primary center of *Tulipa* origin. According to Ivaschenko (2019) 42 wild *Tulipa* species are found in Kazakhstan, 18 of them listed in the Red Book of Kazakhstan. These plants have an important economic, floricultural, ecological significances, cultural value in many regions of the World. Wild species are important genetic resources for breeding and play a significant role in ecosystems, especially for pollinators and insects. Ten *Tulipa* species from Kazakhstan were studied based on complete plastome genome sequences using NovaSeq 6000 (Illumina) platform. All the sequenced *Tulipa* species from Kazakhstan were deposited to the National Center for Biotechnology Information GenBank and publicly available. *Tulipa* species plastomes contained 136 genes, including 90 protein-coding genes, 38 transfer RNA, and 8 ribosomal RNA genes (including duplicated genes). The forward, reverse, and palindromic repeat elements and mononucleotide, dinucleotide, trinucleotide, tetranucleotide, pentanucleotide and

hexanucleotide repeats were detected in four *Tulipa* species from Kazakhstan. Furthermore, tandem repeats in each of the *Tulipa* plastome were identified. Comparative analysis of plastome genome sequences revealed the genes with high polymorphism, which can be used as candidate DNA barcode markers for further studies.

Acknowledgements: This research is funded by the Science Committee of the Ministry of Education and Science of the Republic of Kazakhstan (Grant No. AP14870612).

### S.158.2 Large scale DNA sequencing of plant biodiversity in global biodiversity hotspots – the China plant barcode of life

De-Zhu Li¹, Meng-Yuan Zhou¹, Chun-Xia Zeng¹, Jun-Bo Yang¹, Hong-Tao Li¹, Lian-Ming Gao², Peng-Fei Ma¹, Jin-Mei Lu¹, Cen Guo³, Ting-Shuang Yi¹, Peter M. Hollingsworth⁴

1 Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming Yunnan, China. 2 CAS Key Laboratory for Plant Diversity and Biogeography in East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming Yunnan, China. 3 Center for Integrative Conservation & Yunnan Key Laboratory for the Conservation of Tropical Rainforests and Asian Elephants, Xishuangbanna Tropical Botanical Garden, Mengla, Yunnan, China. 4 Royal Botanic Garden Edinburgh, Edinburgh, UK.

Vascular plants dominate terrestrial ecosystems and provide food, drugs, timber, fibers, fuels and ornamen-
<figure><img src="image_368.png" /></figure>

tals for human beings. Our understanding of plant diversity remains a substantial challenge and identification becomes particularly difficult where material is juvenile, fragmented, or processed. The combination of plastid rbcL+matK sequences was proposed to be the standard barcode for land plants in 2009, followed by our proposal to incorporate the nuclear ribosomal ITS into barcoding practice. Because of the development of next-generation sequencing, it is possible to enhance the plant barcode, focusing on increasing discriminatory power via either gene capture of nuclear markers or genome skimming. Genome skimming has the advantage of using existing protocols, being backward compatible with first-generation barcode, and the depth of sequence coverage can be increased as sequencing costs fall. We made a practical trial of routine recovery of rDNA and plastome sequences from herbarium collections. We sequenced specimens up to 80 years old by using as little as 500 pg of degraded starting DNA, and it can be performed with limited sample destruction. We gathered genome-skimming data of 25,583 individuals representing 20,260 species of 431 genera in 473 families of vascular plants, mainly from biodiversity hotspots in China, to build up a large-scale reference database for the China plant barcode of life initiative. Analyses of recalcitrant genera (or subgenera, or species complex) showed that organelle-scale barcode, together with rDNA sequences increased the discriminatory power by 10-15%. However, using Skmer, a method for estimating genomic distances between the query and reference genome skimming sequences, after removing the rDNA and plastome sequences, the discriminatory power was further increased. Furthermore, using deep genome skimming, particularly where a reference genome is available for the genus, a large number of single-copy orthologs can be acquired, opening up a new avenue for nuclear DNA barcodes.

## S.158.3 Plant Species Identification by Multispecies Coalescent (MSC) Method Using Single-Copy Orthologs (SCOs)

Zheng-Shan He<fnref n="1" />, De-Zhu Li<fnref n="1" />, Jun-Bo Yang<fnref n="1" />

Standard barcodes and ultra-barcodes face challenges in delimitation and discrimination of closely related species with deep coalescence, hybrid speciation, gene flow or low sequence-variation. Single copy orthologs (SCOs) have been recommended as standardized nuclear markers in metazoan DNA taxonomy. Here, we assessed the performance of SCOs in identifying recently diverged species in *Cymbidium* subgenus *Jensoa* which has been poorly settled by ultra-barcode. More than 90% of target 9094 reference SCOs inferred from three genomes of *Cymbidium* were successfully retrieved for all 11 representative species in subg. *Jensoa* by ALiBaSeq from as low as 5x depth whole genome shotgun sequences. Species tree reconstructed from multiple refined SCO matrices under multi-species coalescent model successfully discriminated all species and discerned wrongly identified or labeled species. Plentiful and refined SCOs matrices obtained by implementing our pipeline facilitate not only phylogenetic study, but also high-resolution species diagnosing. Biparentally inherited SCOs as multi-locus marker not only advances the force of DNA barcoding, but also facilitates an eventual transition to species-tree-based barcoding strategies. We also test the performance of SCOs in other taxa among embryophytes to screen sevel universal and capable SCOs as the new barcodes to advance the plant species identification under the multispecies coalescent.

## S.158.4 DNA barcoding of Chinese and Japanese Morus alba (Moraceae) documented by genome resequencing

Chun-Xia Zeng<fnref n="1" />, Jun-Bo Yang<fnref n="1" />

1 Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming Yunnan, China;

In Pharmacopoeia of the People's Republic of China 2020th edition, Mori Folium, Mori Cortex, Mori Ramulus, Mori Fructus, however, botanical origin of all of these four medicines are limited to only one species, *M. alba*. The Japanese Pharmacopoeia Eighteenth Edition also limit the botanical origin of Mori Cortex to *M. alba*. Because of polyploidization and hybridization occured frequently in *Morus*, we found that not a few individuals and specimens of *Morus* plants are difficult to identify because of their intermedi-

<footnote n="1">1 Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan, China. Heilongtan, Kunming, Yunnan, China. Email: dzl@mail.kib.ac.cn; jbyang@mail.kib.ac.cn</footnote>
<figure><img src="image_369.png" /></figure>

ate morphological characteristics between species especially *M. alba* and other species. For the safety and standardization of medication, it is particularly necessary to answer that the medicinal plant *Morus alba* is really a "good" species or not? The previous phylogentic analyses of ITS and cpDNA have been reported most of the *Morus* species collected in China were grouped into a clade lacking for phylogenetic resolution. Genome skimming has the potential for generating large data sets for DNA barcoding and wider biodiversity genomic studies, particularly via the assembly and annotation of full chloroplast (cpDNA), nuclear ribosomal DNA (nrDNA) sequences and single copy orthologs (SCOs). Based on phylogenetic analyses of plastome sequences and ITS sequences, most *Morus* species grouped into a large clade lacking for phylogenetic resolution. Except for *M. notabilis*, other 8 species all should be considered as the botanical origin of *Mori Cortex*. However, based on 272 SCOs, *Morus alba* could be a monophyletic species. Plant species identification is difficult because of the main reasons being incomplete lineage sorting, artificial cultivation, natural hybridization and chloroplast capture. It appears that nuclear genome data have a vital role in species identification and are expected to be used as next-generation nuclear barcodes.

**S.158.5 Whether cryptics are the reason for adulteration in plant herbal products? – case studies from Indian subcontinent**

Umapathy Senthilkumar¹, Chatterjee Abhinav², Kaliappan Ilango³, Parani Madasamy⁴

1 DNA Barcoding Laboratory, Department of Botany, Madras Christian College (Autonomous), Chennai, Tamil Nadu, India. 2 Department of Biotechnology, School of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India. 3 Department of Pharmaceutical Sciences, College of Pharmacy, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India. 4 Department of Genetic Engineering, School of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India.

Authentic species of Parpataka (*Hypertelis & Oldenlandia*) and Gokshura (*Tribulus terrestris*) are well known due to their synergistic effect with other herbs for anti-pyretic activity and urinary dis-

orders respectively. The vernacular conflicts and its co-existence with other morphologically similar species lead to unintentional adulteration in the trade. To validate both i.e., Parpadaka and Gokshura, the samples were purchased from herbal raw drug stores across India for Gokshura and within southern India for Parpataka. These samples were further validated using *rbcL* gene of chloroplast for Parpataka and ITS2 region of nuclear genome for Gokshura respectively. DNA from the market samples was extracted and amplified with the selected barcodes and validated using Bayesian phylogenetic framework in MrBayes 3.2. Trade analysis of "parpadaka" samples revealed five different species of Molluginaceae and a species of *Oldenlandia* in the trade. The tree showed nearly 72.5% of *Hypertelis umbellata*, found in Indian trade for the first time and thus were achieved as a new report to India from this study since there was no report on its natural occurrence in any regional flora. Out of the remaining 27.5%, four different species of *Mollugo s.l* resulted into 17.5%. In Gokshura, *Tribulus lanuginosus* had replaced the *T. terrestris* (an authentic species) with 40.7% and *T. subramanyamii*, an endemic species had replaced it with 18.5%. Our results show that, market samples of Parpataka sold in southern India contains few substitutes, though among them, *H. umbellata* showed the cryptic nature with its counterpart i.e., *H. cerviana*, was predominant and completely replaced the authentic substitute in the indigenous trade. Similarly, in case of Gokshura, *Tribulus* had showed high degree of cryptic nature along with its counterpart, i.e., *T. subramanyamii* that evident by means its prevalence in the trade.
**S.159** ETHNOBOTANY AND CONSERVATION OF MEDICINAL PLANTS. SESSION 5

**S.159.1 From chlorine to biodiversity: a project for the naturalization of urban ponds.**

Bárbara Martínez Escrich¹, Mónica López Martínez¹

1 IMGEMA- Royal Botanic Garden of Córdoba, Spain

"From chlorine to Biodiversity", a short phrase that titles the project that is being developed in the Royal Botanic Garden of Córdoba (Spain). The main objective of this initiative is the naturalization of urban ponds, removing chlorinated water and introducing autochthonous aquatic species and charophytes meadows in order to create a hot spot of biodiversity. In this way is not only possible to eliminate chemical compounds harmful to the environment and reduce water consumption but also ecological restoration points are created in cities and villages. One of the greatest strengths of this project is that the species used are autochthonous aquatic macrophytes and algae. After the five years that the project has been developed and having naturalized 22 spaces, it has been possible to establish a protocol that imitates the natural processes that occur in the ponds of the countryside, it is possible to establish a trophic chain that allows the self-regulation of the new ecosystems. More than 1500 people have participated in the programme and 44 students have completed their internships within the project. Furthermore, naturalized sources have been revealed as a novel and attractive instrument to explain the complex concept of Biodiversity. The educative programme offered by the Royal Botanic Garden of Córdoba conceives the ponds as ecosystems that allow the observation of associated flora and fauna, as well as the interactions that occur between them. The growing pollution of cities, the scarcity of water and climate change can be tackled through urban policies and management strategies that considers green structures as backbones of the design of cities and villages. Experience has shown us that citizen participation is essential. The creation of

synergies with associations and collectives helps to drive the transformation of these urban green infrastructures.

**S.159.2 Ethnobotanical significance of Zingiberaceae in southern Western Ghats of Kerala, India**

Valukattil P. Thomas¹, Thundiathu T. Binoy¹

1 Catholicate College, Pathanamthitta, Kerala, India

India has a rich diversity of Zingiberaceae and constitutes one of the ten largest families in the country. The family is considered one of the important families in monocotyledons because of their economic, medicinal and ornamental values. Ethnobotanical studies on Zingiberaceae of Kerala identified 26 taxa with ethnobotanical significance and were used by various tribal communities in Kerala, India. The study covered 34 tribal communities distributed over 14 districts of Kerala and informants were interviewed during house-to-house surveys. The information from tribal practitioners was compared with results obtained during earlier visits. Taxonomic studies were carried out and voucher specimens were deposited at CATH. The specimens are conserved in the Ginger House of Catholicate College Botanical Garden (CCBG). The collected data were analyzed using the informant consensus factor (Fic), fidelity level and use value (UV). The plants were utilized in traditional medicine, food, fodder, religious purposes, etc. The Fic values of various species were analyzed and calculated for 22 categories. The highest values of Fic for Zingiberaceae were calculated in the case of the ENT category (1.0) and the lowest value was estimated for gynaecological ailments (0.28). The highest use value was determined for *Curcuma longa* (0.056). The highest fidelity level was reported for *Curcuma longa* (92.4%) followed by *C. amada* (87.1%) and *Zingiber officinale* (86.3%). The documentation of traditional knowledge and conservation of the valuable plants are very important, and the information collected will serve as baseline data for future phytochemical and pharmacological research to develop new drugs and services.
<figure><img src="image_370.png" /></figure>

**S.159.3 Phytochemical studies and anti-diabetic activities of Plumbago zeylanica, Neolitsea cassia and Wrightia tinctoria at Palamalaihill**

E.G. Wesely¹, L. Thamaraiselvi¹

1 Department of Botany, Arignar Anna Government Arts College, Namakkal, India

Medicinal plants have been identified and used in traditional medicine all over the world since the dawn of civilization. Medicinal plants are a rich source of novel drugs, which are used in traditional medical systems (Bauri et al., 2015). More than 50,000 plant species are reported to be used for medicinal purposes around the world. Diabetes mellitus (DM) is a metabolic disorder caused by an insufficient amount of insulin in the body, either during its production or action, or both (Mukhtar et al., 2019). If DM is not treated properly, serious complications such as muscular tissue damage, retinopathy, neuropathy, nephropathy, heart complications, and ulceration problems can occur. Insulin and glucagon are pancreatic hormones that regulate blood glucose levels in a normal range based on the body's needs (Arden and Ramsey, 2015). Although synthetic drugs are primarily used to treat diabetes, they have significant and irreversible side effects, and their accessibility to the general public is also difficult due to their high cost (Amin et al., 2013). Little research has been done on the anti-diabetic properties of ethanolic extract of leaves and stems *Plumbago zeylanica*, *Neolitsea cassia* and *Wrightia tinctoria*. Due to the rising prevalence of diabetes, antihyperglycemic drugs are in high demand. Because of the minimal side effects and medicinal importance of ethanolic extract of *Plumbago zeylanica*, *Neolitsea cassia* and *Wrightia tinctoria*, the current study is being conducted to conduct anti-diabetic studies. The results of the study suggest that the ethanol extract of the leaves and stems has antidiabetic properties. Oral hypoglycaemics such as acarbose, which belongs to the α-amylase inhibitor class, have been shown to inhibit alpha-amylase. As a result, in the assays, acarbose was used as a positive control.

**S.159.4 Anti-inflammatory activity of Curcuma putii rhizome extracts in HaCaT cell via NF-κB nuclear translocation inhibition**

Pannita Karnjanasrimak¹, Suparerk Borwornpinyo²,⁴, Saroj Ruchisansakun¹, Warawuth Wannalo⁴, Tanawadee Khumpanied⁴, Chawisa leekuakul⁴, Kedchin Kearawuttanakul⁴, Mutita Kongphet⁴, Sitthivut Charoensutthivarakul³

1 Department of Plant Science, Faculty of Science, Mahidol University, Thailand. 2 Department of Bio-technology, Faculty of Science, Mahidol University, Thailand. 3 Department of Bioinnovation, Faculty of Science, Mahidol University, Thailand. 4 Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok, Thailand

Curcuma is a genus in the Zingiberaceae family. Previous studies have demonstrated their diverse medicinal benefits, including antioxidant, anti-cancer, antimicrobial and anti-inflammatory activities. The objective of this research is to investigate the anti-inflammatory activity of the rhizomes of *Curcuma putii*, a species discovered in Thailand in 2019. To determine the anti-inflammatory activity, the inhibition of nuclear translocation of a transcription factor is employed by disrupting the nuclear translocation of NF-κB from cytoplasm into nucleus. Dried rhizomes of *Curcuma putii* were extracted using six different solvents including ethanol, methanol, acetone, hexane, ethyl acetate and dichloromethane. Initially, cytotoxicity experiments through the MTT assay were performed and found that concentration of 50 µg/mL is the highest concentration in all solvents determined to be non-toxic to human immortalized keratinocyte HaCaT cells. In addition, the NF-κB nuclear translocation activity was performed and found that the extracts from all six solvents at 50 µg/mL non-significantly inhibited against HaCaT cells. This serves as a direction for future studies on the anti-inflammatory potency and other bioactivity of *Curcuma putii*.
<figure><img src="image_371.png" /></figure>

S.159.5 Biogeography of
Medicinal Plants registered
in the State of Hidalgo,
Mexico

Valdés-Vásquez I.F.¹, Monroy-Ata A.¹,
Cuervo-Robayo A.P.², Villaseñor J.L.³,
Defossez E.⁴, Morrone J.J.⁵, Bye-Boettler
R.A.⁶, Calmé S.⁷, Reyes-Chilpa R.⁸, Osorio-
Olvera L.O.⁹, Ramírez-Barahona S.A.³,
Beltrán-Rodríguez L.A.¹⁰, Couturier-
Stéphane R.A.¹¹, Mas J.F.¹², Aguilar-
Contreras A.¹³, Cristians-Niizawa S.⁶, De
Nova-Vázquez J.A.¹⁴, Yáñez-Arenas C.A.¹⁵,
Morales-Manilla L.M.¹⁶, Bollo-Manent M.¹⁶,
Ortiz-Bermúdez E.³, Estrada-Márquez
A.S.³, Heike-Vibrans L.¹⁷, Torrescano-Valle
N.¹⁸, Espinosa-Organista D.N.¹⁹, Santiago-
Alvarado M.¹⁹ and Guevara-Fefer P.²⁰

1 Unidad de Investigación en Ecología Vegetal, FES Zaragoza Universidad Nacional Autónoma de México. México. 2 Laboratorio Nacional CO-NAHCYT de Biología del Cambio Climático, Instituto de Biología UNAM, Mexico. 3 Departamento de Botánica, Instituto de Biología UNAM, México. 4 Laboratory of Functional Ecology, Institute of Biology, University of Neuchâtel, Neuchâtel, Switzerland. 5 Departamento de Biología Evolutiva, Facultad de Ciencias UNAM, México. 6 Laboratorio de Etnobotánica, Jardín Botánico IB UNAM, Mexico. 7 Department of Biology, Université de Sherbrooke, Québec, Canada. 8 Departamento de Productos Naturales, Instituto de Química UNAM, México. 9 Departamento de Ecología de la Biodiversidad, Instituto de Ecología UNAM, Mexico. 10 Laboratorio de Etnobotánica Ecológica, IB UNAM, Mexico. 11 Laboratorio de Análisis Geoespacial, Instituto de Geografía UNAM, Mexico. 12 Laboratorio de Análisis Espacial del CIGA, ENES Morelia UNAM, Michoacán, Mexico. 13 Unidad de Investigación de Plantas Medicinales, Herbario Medicinal IMSS, México. 14 Instituto de Investigación de Zonas Desérticas, Universidad Autónoma de San Luis Potosí, Mexico. 15 Laboratorio de Ecología Geográfica, Unidad Académica Sisal, Facultad de Ciencias, ENES Mérida UNAM, Yucatán, Mexico. 16 Laboratorio Universitario de Drones, (CIGA) ENES Morelia UNAM, Michoacán, Mexico. 17 Departamento de Botánica, Colegio de Postgraduados, Universidad Chapinango, Estado de México, Mexico. 18 Departamento de Conservación de la Biodiversidad, El Colegio de la Frontera Sur Unidad Chetumal, Quintana Roo, Mexico. 19 Unidad de Investigación en Biogeografía, Sistemática y Conservación, FES Zaragoza UNAM, Mexico.

20 Laboratorio de Fitoquímica,
Facultad de Ciencias UNAM, Mexico

Mexico is the second country with the greatest diversity of medicinal plant species in the world, after China. In Central Mexico, the state of Hidalgo is in an area with high floristic diversity, that is in the Mexican Transition Zone, between the Neartic and Neotropical Regions, composed of six types of vegetation: cloud forest, semi-evergreen tropical forest, xeric scrub, coniferous and fir forest, pine-oak forest and induced grassland. The objective of this work was to develop a specialized database of medicinal plants registered in the state of Hidalgo, through taxonomic, phytochemical, biogeographic, and ecological parameters, to make thematic maps on geographic distribution patterns, analyze ecological relationships of secondary metabolites. with climatic variables from biogeographic and physiographic provinces, as well as evaluating floristic richness by documenting the composition between nine ecoregions. Likewise, a mini drone was used to carry out multistage *in situ* sampling of aerial photography, with which 21 areas were characterized in strata: 10 geocultural regions of Nahua, Otomi, and Tepehua ethnic groups, 9 ecoregions, 6 types of vegetation, 4 biogeographic provinces and 3 physiographic provinces. Also, based on the registration of 400 species of medicinal plants, belonging to 80 botanical families, and consulting international databases, species distribution models were designed. The results show thematic maps with geographical distribution patterns of species characteristic of each type of vegetation, organized by ecoregions, for plants for medicinal use to fine-scale. Also, with spatial analysis, it was found that altitudinal gradients influence the diversity of second phytochemicals at different spatial scales. Finally, the state of Hidalgo constitutes 1.1% of the Mexican territory, with a record of 400 species for therapeutic use and 4,000 similar plants that are distributed throughout the country, so 1% of the country is home to 10% of the medicinal plants' diversity.
<figure><img src="image_372.png" /></figure>

# S.160 POLYPLOIDY AND HOMOPLOID HYBRIDIZATION AS EVOLUTIONARY DRIVERS

## S.160.1 Reticulate genetic set-up of endangered species and its consequences for conservation strategies.

Marcus A. Koch¹

1 Centre for Organismal Studies, Heidelberg, Heidelberg University, Heidelberg, Germany

Gladiolus palustris (marsh Gladiolus) of wet grasslands is an extremely rare and highly endangered species in Central Europe. Ongoing loss of habitat, population fragmentation, drought, and higher mean annual temperatures caused by global warming have all contributed to a severe decline in its population over the past few decades. Additionally, hybridization with other species, such as *G. imbricatus* and *G. illyricus*, and genetic depletion may pose a significant threat to the species' survival. The focus here is set to characterize major gene pools of the species in southern and southwestern Germany and illustrate that past hybridization and introgression in Central Europe are more extensive than previously thought, posing a challenge to conservation strategies targeting taxonomically defined species. A parallel exploration into the water plant *Nuphar pumila*, a post-glacial relic in Central Europe, sheds light on the diverse signs of introgression from the widespread *Nuphar lutea*. These genetic imprints serve as historical markers, reflecting centuries-old processes and changes in the landscape. Contrary to expectations, hybrid gene pools did not invariably result in intermediate morphologies. Instead, introgression appears to have played a pivotal role in enabling these species to adapt to evolving environmental conditions, such as heightened nitrogen levels and water temperatures. Given the longstanding presence of these populations in the region and their vital contribution to the active gene pool, the evidence suggests a need to champion and safeguard these populations. In light of these findings, it is proposed that conservation efforts should also prioritize naturally occurring hybrids and introgressed populations. Recognizing

their resilience and adaptive capabilities, these populations emerge as crucial custodians of biodiversity, embodying the intricate dance between genetic diversity and environmental change.

## S.160.2 Investigating conserved floral scent profiles acting as a driver of hybridization in Heuchera

Nicholas J. Engle-Wrye¹, Ryan A. Folk¹

1 Department of Biological Sciences, Mississippi State University, Mississippi State, USA

Little is known about environmental drivers of hybridization, but its phylogenetic distribution across plants is heterogeneous, suggesting that plant traits play an important role. Because animal vectors are responsible for gene flow in most plants, conservatism of biotic niche could explain why some plants are more prone to hybridization, a hypothesis yet to be tested. The taxonomic tribe Heuchereae (Saxifragaceae) is a well-characterized system for pollinator interactions, with studies in *Asimitellaria* and *Lithophragma* suggesting the VOCs comprising floral scent are hypervariable at the population level and responsible for pollination selectivity, leading to divergent pollinator assemblages. High levels of pollinator attractant divergence may be responsible for the relatively low levels of hybridization observed in these groups. *Heuchera* flowers, by contrast, are thought to be pollination generalists and *Heuchera* species hybridize frequently. Measurement of floral volatiles, yet to be characterized for *Heuchera*, would offer insight into whether divergence of pollinator attractants can more broadly explain hybridization rate differences across taxa. Given that pollinator-mediated gene flow primarily hinges on VOCs in the relatives of *Heuchera*, we investigated whether high rates of hybridization observed in *Heuchera* are associated with low interspecific divergence of VOCs, using as a system the hybrid zone between *H. americana* and *H. richardsonii*. We optimized a GC-MS (gas chromatography-mass spectrometry) protocol to character-
<figure><img src="image_373.png" /></figure>

ize VOCs. To identify floral visitation and effective pollinators, we conducted pollination observations at 47 different populations over two years. GC-MS data from ~110 *Heuchera* representing 21 populations, analyzed with a suite of statistical methods, suggests that VOC compound classes and individual compounds are largely shared within the hybrid complex, while other *Heuchera* that do not hybridize with these species have distinct species-specific compounds. Pollination observations confirmed shared effective pollinators in the hybrid zone and the overall importance of VOCs in shaping gene flow.

## S.160.3 Genomics of hybridisation in British native flowering plants

Meng Lu¹,², Peter M. Hollingsworth², Alex D. Twyford¹,²

1 Ashworth Laboratories, Institute of Ecology and Evolution, The University of Edinburgh, Edinburgh, UK.
2 Royal Botanic Garden Edinburgh, Edinburgh, UK

Hybridization can have various evolutionary outcomes, from homogenizing the differences between populations to generating phenotypic and genotypic novelty. However, the majority of these important phenomena are evaluated in isolated case studies of evolutionary model species chosen for their notable hybridization outcomes, and how well the current findings represent generalities of natural hybridization in a wider range of taxa remains unknown. As such, this project aims to explore the general patterns of natural hybridization in the British flora using genomic approaches. The British flora is very well-studied with extensive records of hybrids, rendering it the ideal system for understanding natural hybridization at a broad scale. Specifically, we evaluate the extent of introgression in native flowering plants, using model-based demographic inference on whole genome resequencing data. Six conspicuous hybridizing species pairs were selected, each pair from a distinct taxonomic family, including genera that are well known for their common natural hybrids, such as *Primula* and *Geum*. The results support models with asymmetrical gene flow in focal pairs. Additionally, using a demographically explicit genome scan approach, we were able to estimate heterogeneous gene flow across the studied genomes and identify candidate barri-

er regions to gene flow. By studying hybridization outcomes of species pairs from diverse taxonomic lineages, we hope to provide a comparative perspective to understand how introgression is affected by intrinsic factors, such as parental species divergence.

## S.160.4 Genetic integration of species diagnostic characters and their evolutionary responses to selection within hybridizing oaks

Ilga M. Porth¹

1 Department of Forest and Wood Sciences, Laval University, Quebec, Canada

The European oak species complex *Quercus petraea/robur* represents the best studied system for temperate tree speciation and population dynamics. Due to their extensive historical introgression, genetic clustering follows a continuous transition between the species, with only few species-specific clusters rather than strong species differentiation. Investigation of GxE and species' integrity found that GxE decreases with increasing species integrity, and therefore GxE interactions across the studied species discriminant leaf traits were strongly driven by the level of introgression between species across tested forest stands. Using a marker-based relationship matrix, we estimated coefficients of genetic integration for each trait, with those leaf traits being the least integrated that showed the highest species discriminatory power. Historical introgression seemed highest within cooler and wetter environments, while organismal modularity allowed for the independent evolution in oak species' morphology relating to ecological adaptation. This was corroborated by networks of genetic correlations. Hence, low within-species genetic integration of traits allows for the highest difference in evolutionary response to selection between species. Thus, less integrated traits can effectively drive *Quercus* species divergence, and more specifically, the evolutionary response to selection for leaf veins' function could explain the difference in water use efficiency and gas exchange between the different oak species. Our study highlights the importance of knowledge
<figure><img src="image_374.png" /></figure>

about the genetic, epistatic, and plastic effects associated with species diagnostic traits when considering conservation units in oak.

## S.160.5 Deciphering complex reticulate evolution of Asian Buddleja (Scrophulariaceae)

Fengmao Yang¹,², Jia Ge¹,²

1 Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan, China. 2 Yunnan Key Laboratory for Integrative Conservation of Plant Species with Extremely Small Populations, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan, China

Species of the genus *Buddleja* in Asia are mainly distributed in the Sino-Himalayan region and form a challenging taxonomic group, with extensive hybridization and polyploidization. A phylogenetic approach to unravelling the history of reticulation in this lineage will deepen our understanding of the speciation in biodiversity hotspots. For this study, we obtained 80 accessions representing all the species in the Asian *Buddleja* clade, and the ploidy level of each taxon was determined by flow cytometry analyses. Whole plastid genomes, nuclear ribosomal DNA, single nucleotide polymorphisms and a large number of low-copy nuclear genes assembled from genome skimming data were used to investigate the reticulate evolutionary history of Asian *Buddleja*. Complex cytonuclear conflicts were detected through a comparison of plastid and species trees. Gene tree incongruence was also analysed to detect any reticulate events in the history of this lineage. Six hybridization events were detected, which are able to explain the cytonuclear conflict in Asian *Buddleja*. Furthermore, PhyloNet analysis combining species ploidy data indicated several allopolyploid speciation events. A strongly supported species tree inferred from a large number of low-copy nuclear genes not only corrected some earlier misinterpretations, but also indicated that there are many Asian *Buddleja* species that have been lumped mistakenly. Divergent time estimation shows two periods of rapid diversification (8–10 and 0–3 Mya) in the Asian *Buddleja* clade, which might coincide with the final uplift of the Hengduan Mountains and Quaternary climate fluctuations, respectively. This study presents a

well-supported phylogenetic backbone for the Asian *Buddleja* species, elucidates their complex and reticulate evolutionary history and suggests that tectonic activity, climate fluctuations, polyploidization and hybridization together promoted the diversification of this lineage.

## S.160.6 Sub-Genome Influence on Tree Topology in the Tribe Brassiceae of the Mustard Family (Brassicaceae)

R. Shawn Abrahams¹, Shawn Thomas², Tatiana Arias, Jacob Washburn², J. Chris Pires

1 Department of Ecology & Evolutionary Biology, Yale University, New Haven, CT, USA. Department of Biochemistry, Purdue University, West Lafayette. r.shawnabrahams@gmail.com. 2 Division of Biological Sciences and Bond Life Sciences Center, University of Missouri, Columbia, Missouri, USA

The Brassiceae tribe of the Mustard Family has a complex evolutionary history of whole genome duplication and hybridization. These mechanisms contribute to phenotypic plasticity and adaptability of the tribe, while also confounding attempts to understand the relationships among the species of the tribe. To account for these complications, we utilize a genome-guided phylotranscriptomic method that leverages genomic synteny to inform phylogenomic inference. With this method, we infer a nuclear tree with a novel topology that places the *Crambe* clade as sister to both the *Nigra* and *Rapa/Oleracea* group species. Further investigation of single copy genes from the ancestral sub-genome of the tribal hexaploidy identified differences in topology among genes derived from different parental genomes. These findings help explain why we see various topologies based on the data type method of inference when attempting to reconstruct clade relationships.
# S.161 PALEOBOTANY AND PALEOPALYNOLOGY: DIVERSITY, TAXONOMY, AND PALEOECOLOGY

## S.161.1 Balkan Neanderthals: the Late Pleistocene palaeoecological sequence of Pešturina Cave (Niš, Serbia)

Juan Ochando¹,², José S. Carrión¹,³, Donatella Magri², Ana B. Marín-Arroyo⁴, Federico Di Rita², Manuel Munuera⁵, Fabrizio Michelangeli⁶, Gabriela Amorós¹, Stefan Milošević⁷, Katarina Bogićević⁸, Vesna Dimitrijević⁷,⁹, Draženko Nenadić⁸, Mirjana Roksandic¹⁰,¹¹, Dušan Mihailović⁷

¹Department of Plant Biology (Botany Area), Faculty of Biology, University of Murcia, Murcia, Spain. ²Sapienza University of Rome, Department of Environmental Biology, Rome, Italy. ³Evolutionary Studies Institute, University of Witwatersrand, South Africa. ⁴Grupo I+D+i EvoAdapta, Departamento Ciencias Históricas, University of Cantabria, Santander, Spain. ⁵Department of Agricultural Engineering, Polytechnic University of Cartagena, Cartagena, Spain. ⁶Department of Biological, Geological and Environmental Sciences, University of Bologna, Bologna, Italy. ⁷Department of Archaeology, Faculty of Philosophy, University of Belgrade, Belgrade, Serbia. ⁸Department of Palaeontology, Faculty of Mining and Geology, University of Belgrade, P.O. Box 227, Belgrade, Serbia. ⁹Laboratory for Bioarchaeology, Faculty of Philosophy, University of Belgrade, Belgrade, Serbia. ¹⁰Department of Anthropology, University of Manitoba, Winnipeg, Manitoba, Canada. ¹¹Department of Anthropology, University of Winnipeg, Winnipeg, Manitoba, Canada

The Central Balkans are a key biogeographical region in Southern Europe, influenced by a central European-Mediterranean climate, which acted as a refugium for flora and fauna, and favoured the dispersion of Neanderthals and migration of modern human populations during Late Glacial Period. This study presents pollen analyses of sediment and hyaena coprolites from Pešturina Cave in Serbia to reconstruct the vegetation landscapes faced by Balkan Neanderthals and early Anatomically Modern Humans between MIS 5e-3. Between MIS 5e-5c (archaeological layers 4c and 4b) and MIS 5b-5a (layer 4a), semi-forested environments prevailed, characterized by *Pinus*, deciduous *Quercus*,

*Tilia* and other angiosperm woody taxa, accompanied by heliophytes such as *Artemisia* and Poaceae. During MIS 4-3 (layers 3-2), the vegetation was dominated by *Artemisia*-Poaceae steppes with *Quercus* patches, conifers and legumes. Overall across the sequence, pollen assemblages are highly diverse and include a number of deciduous trees and sclerophylls. In addition, the occurrence of several herbaceous taxa reinforces the view that the Balkans were outstanding for endemicity. Neanderthals and early Upper Palaeolithic hominins lived in a highly diverse refugium, offering multiple opportunities for survival during the warm interstadials and, more critically, the cold stadials of the Pleistocene.

## S.161.2 Paleohistories for Conservation Practice from Quaternary Paleoecology of the Iberian Peninsula

José S. Carrión¹, Manuel Munuera², Juan Ochando³,¹, Gabriela Amorós¹

¹University of Murcia, Spain. ²University Polytechnic of Cartagena, Spain. ³University La Sapienza, Roma, Italy

The Quaternary flora and vegetation records of the Iberian Peninsula provide insights for biological conservation strategies in the context of large projects like Rewilding Europe and other initiatives aimed at restoring ecosystem dynamics and functionalities from pre-anthropogenic periods. It is crucial to integrate information derived from the identification and characterization of glacial plant refugia, considering their significance as biodiversity sinks and sources of geographic expansion during critical moments in the glacial history of the European continent. It is important to note that many of these refuges are more hydrorefuges than therm-refuges, as aridity conditions became the limiting factor in many stadial episodes across extensive territories of the southern Mediterranean peninsulas. Likewise, it is worth considering that refuge areas were located on emerged coastal platforms and intra-
<figure><img src="image_375.png" /></figure>

mountain valleys at medium altitudes. From a structural perspective, many pre-anthropogenic Quaternary forests were influenced by herbivory, a factor that has been overlooked for decades and is now emerging as a consideration in the renaturation of protected or pilot areas. During the Holocene, it is important to analyze cultural collapses resulting from state changes in the paleoecosystem caused by anthropogenic disturbances. An example of ecocide could be the Argaric collapse. Finally, the dynamics of Quaternary vegetation aid in understanding the explanatory patterns of the floristic and ecological configuration of current plant communities, while also allowing the establishment of plausible scenarios in the near future under the influence of climate change and environmental crisis.

modern forest types occurring in the ecoregion as a second approach to understand changes in the forest distribution. Our results highlight the complexity of Iberian ecosystems during the LGM, including mixed distribution of temperate and mediterranean climates as detected by modern analogues. We also found a similar bioclimatic zonation of the Iberian Peninsula in both LGM and modern climates, with the northwestern regions experiencing a relative stronger Atlantic influence. Additionally, available pollinic records were used to validate the potential vegetation of the Iberian Peninsula during LGM. Our approach allowed us to establish new hypotheses about the LGM distribution of ecosystem types in southern Europe, to be used in historical and ecological biogeography.

### S.161.3 Ecosystems of North-Western Iberian Peninsula in the Last Glacial Maximum based on paleoecological modelling

Víctor González-García¹, Eduardo

Fernández Pascual¹, Borja Jiménez-Alfaro¹

1 Biodiversity Research Institute-IMIB (CSIC-Univ. Oviedo-Princ. Asturias), Mieres, Spain

Southern European peninsulas provided refugia for temperate and mediterranean ecosystems during the Last Glacial Maximum (LGM). Despite pollen data, fossils and distribution modelling have been all used to understand vegetation during the LGM, we still lack a spatially explicit approach to map ecosystem types from modern climatic analogues. Here, we develop a framework for mapping ecosystem diversity of North-Western Iberian Peninsula during the LGM. First, we classified the Iberian Peninsula in climatic regions based on LGM climatic models and k-means unsupervised classification. The selected regions were sampled to conduct distribution models that were extrapolated to modern climates of Europe to detect modern climatic analogues. The areas with the highest climatic similarity to each one of the Iberian LGM regions were characterized with recent vegetation data to estimate the relative dominance of functional ecosystem types under modern climate conditions. In addition, we performed distribution models for

### S.161.4 Palynology of the Canaan Peak Formation in Utah (United States of America)

Uxue Villanueva-Amadoz¹, Brayan Bustamante², Jeffrey G. Eaton³

1 Estación Regional del Noroeste, Instituto de Geología, UNAM, Hermosillo, Mexico. E-mail: uxue@geologia.unam.mx. 2 DICTUS, Universidad de Sonora, Hermosillo, Mexico. 3 Natural History Museum of Utah, Salt Lake City, Utah, USA

The Canaan Peak Formation, overlying the Campanian Kaiparowits Formation, in southern Utah (United States of America) is mostly conglomeratic unit with mudstone intervals containing plant debris and vertebrate remains (Bowers, 1972). This author indicated a lower or middle Campanian age for this formation on the basis of palynological assemblage studied by R.H. Tschudy due to its similarity to other palynological zonation in Montana, however, taxa are not determinative for this age. New palynological sampling from this dark grey mudstone interval yielded well-preserved palynomorphs, mainly composed by pollen grains (genera *Alisporites*, *Carolinapollis*, *Fraxinoipollenites*, *Holkopollenites*, *Inaperturopollenites* and *Taxodiaceae*pollenites, *Discoidites parvistriatus*, *Rhoipites globosus*, *Tschudypollis retusus*, among more common taxa) and scarce spores (*Cyathidites* minor, *Laevigatosporites hardtii*). The presence of *Discoidites parvistriatus* whose first appearance is the late Maastrichtian and K-taxa *Tschudypollis retusus* suggests a late Maastrichtian age.
<figure><img src="image_376.png" /></figure>

## S.161.5 Early angiosperms: New insights from a pre-Aptian pollen flora of Portugal

Julia Gravendyck¹,², François-Nicolas Krencker¹, Rute Coimbra³, Ulrich Heimhofer¹

1 Institute of Geology, Leibniz University Hannover, Hannover, Germany. 2 Bonn Institute for Organis- mic Biology, University of Bonn, Bonn, Germany. 3 Department of Geoscience, University of Aveiro, Aveiro, Portugal

The timing of the origin of angiosperms is a long-standing topic of debate in plant evolution. Different disciplines and approaches provide high- ly variable age estimates. It is generally accepted, that early macro- and mesofossils showing un- equivocal angiosperm features occur in lower Ap- tian deposits. Workers employing palynology, i.e., the extremely durable, abundant, and widespread pollen record, have suggested much earlier, but highly disputed ages. To date, it remains question-

able how far back pollen with unequivocally an- giospermous features can push back the age for the origin of (early) angiosperms. Here we present new 'early records' from pre-Aptian coastal marine strata from the Lusitanian Basin of Portugal that are known for their rich and unique fossil assemblages and excellent pollen preservation. Based on stron- tium isotope stratigraphy, we provide a refined high-resolution stratigraphic framework for existing and newly studied sections to better constrain the age assignment of our new palynological findings of subsequent palynological findings. Employing a new fluorescence-based screening method paired with the standard light-microscope approach, we try to find the needle in the haystack. We document challenges and solutions to finding the extremely rare angiosperm pollen and illustrate new finds of early angiosperms with super-resolution micros- copy (Airy Scan). The rare but exciting finds of four triaperturate pollen grains from the Early Barremi- an (124.63–126.24 Ma, GTS2020) of Portugal indicate the presence of eudicot angiosperms in pre-Aptian strata and provide a new, well-dated calibration point for phylogenetic approaches.

## S.162 IAPT EARLY CAREER INVESTIGATOR SYMPOSIUM: THE EDGE OF THE UNKNOWN IN PLANT, ALGAE, AND FUNGAL SYSTEMATICS. SESSION 2

### S.162.1 Molecular and morphological diversification of the family Inocybaceae (Agaricales, Fungi) in tropical Africa

Hyppolite L. Aïgnon¹, Yu-Guang Fan², Martin Ryberg³, Nourou S. Yorou¹

1 Research Unit Tropical Mycology and Plants-Soil Fungi Interactions, Faculty of Agronomy, Universi- ty of Parakou, Parakou, Benin. 2 Key Laboratory of Tropical Translational Medicine of Ministry of Edu- cation Tropical Environment and Health Laborato- ry, School of Pharmacy, Hainan Medical University, Haikou, China. 3 Systematic Biology program, De-

partment of Organismal Biology, Uppsala Universi- ty, Norbyvägen, Uppsala, Sweden.

Over the past decade, Africa has recorded the high- est annual rate of forest loss with 3.9 million hectares destroyed. This forest destruction causes the extinc- tion of many species and at this rate of destruction, many species can go extinct and perhaps without being known. Whereas, despite the scientific com- munity's efforts to estimate and catalog fungal di- versity, worldwide only a small fraction (ca. 3%) of fungal diversity has been named and described. In recent years, with the progress of science, mycolo- gy has undergone a rapid development. Thus, since 2017, we have been interested to the diversity of Ino- cybaceae in Tropical Africa through the description and documentation of new species with an update on known species. The results show that in Africa, the
diversity of Inocybaceae is about 80 species against 1,100 species in the world and the members of Inocybaceae are one of the most dominant in the soil of the woodlands and gallery forests after Russulaceae Lotsy. Nine new species are described and published in three genera like *Inocybe* (*I. beninensis*, *I. flavipes*, and *I. fuscobrunnea* and *I. pallidiangulata*), *Inosperma* (*I. africanum*, *I. bulbomarginatum*, *I. flavobrunneum*, and *I. macrocarpa*) and *Malloyce* (*Malloyce africana*). We are convinced that there are still enough unknown Inocybaceae species and we must redouble our efforts in sampling and also the exploration of new habitats in Africa.

## S.162.2 Reconstructing an allo-decaploid: identifying the ancestors of Fragaria cascadensis

Sebastian Mortimer¹, Aaron Liston¹

1 Oregon State University, Oregon, USA

Allopolyploidy is widespread among plant lineages and is an important driver of speciation. Understanding how allopolyploid species have formed through reticulate evolution is a challenging task. How can we infer the ancestry of extant allopolyploids when many ancestral diploids and lower-level polyploid ancestors are extinct? Moreover, genomic signal is often eroded in the allopolyploid genome through homeologous exchange. Using a suite of genomic tools, we developed a pipeline for identifying the progenitor lineages of high-level allopolyploids (6x and above). We test our pipeline on *Fragaria cascadensis*, a recently described allo-decaploid which is narrowly endemic to a single mountain range in western North America. The species has acquired five subgenomes through sequential interspecific hybridization events likely taking place in Beringia in the last 1 million years. Using whole genome alignments between extant diploid and polyploid strawberry species, a phylogenetic hypothesis testing framework, and k-mer based inferences of transposon activity, we reconstruct the allopolyploid ancestors and identify putative diploid progenitors of the decaploid strawberry *F. cascadensis*. Our new genomic evidence suggests a revision of the current ancestry hypothesis. We characterize the extent of asymmetric subgenome evolution and compare the decaploid strawberry to closely related octoploid species. Finally, we consider the implications of a new allo-decaploid hypothesis on the biogeography, systematics, and evolution of mating systems in polyploid *Fragaria*. We anticipate that our pipeline for reconstructing the origin of polyploids will be informative in other lineages with complex allopolyploids.

## S.162.3 New approaches in the study of the sexual reproduction of the green microalga *Trebouxia: when free-living means more sex*

Enrico Boccato¹, Fabio Candotto Carniel¹, Mauro Tretiach¹

Department of Life Sciences, University of Trieste, Trieste, Italy

The aero-terrestrial green microalgae of the genus *Trebouxia* Puymaly are the most frequent photobionts of lichens, but our comprehension of their sexual life is very limited, because in the lichen symbiosis it is typically suppressed by the fungal partner. In this study, innovative techniques were applied to characterise the formation and destiny of zoospores in four axenically cultured *Trebouxia* species (*T. decolorans*, *T. vagua*, *T. gelatinosa*, and *T. angustilobata*), selected to represent three of the four primary phylogenetic clades(A, A, I, and S, respectively)recognized within the genus. Frequent zoosporogenesis was achieved by increasing the concentration of glucose and peptone in the growth solid medium. By light transmitted and scanning electron microscopy(SEM)on samples treated with theCO₂ critical point dryer, we identified two distinct zoospore morphotypes, conserved among the major clades of the genus: spherical to sub-spherical vs. broadly ellipsoidal to ovoid. By observing zoospores stained with OsO₄ at the SEM in backscattered electrons mode, we could easily identify lipid bodies, as confirmed by the detection of Os by energy dispersive X-ray spectrometry, whose presence varied among species. These lipid bodies, which serve as energy sources in motile gametes of other green algae, led us to hypothesise that the heteromorphic zoospores of *Trebouxia* could represent true gametes. Sexual reproduction was corroborated in *T. decolorans* by confocal observations, through the assessment of the DNA content at cell population level, thus providing the first insights into the ploidy of this organism. Finally, cells with four flagella, which might be interpreted as the first direct evidence of a kariogamic
<figure><img src="image_377.png" /></figure>

event, were documented by light and SEM micros-
copy in populations of zoospores isolated by cyto-
fluorimetry. The role of heteromorphic zoospores in
the life cycle of *Trebouxia* and the implications of
sexual reproduction in the photobionts of lichens will
be thoroughly discussed.

S.162.4 To the mountains
and beyond: diversity and
systematics of Agarista
(Ericaceae) in the Americas

Claudenice H. Dalastra¹, Peter Fritsch²,
Morgan Gostel², Gustavo Heiden³

1 PPG Botany, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil. 2 Botanical Research Institute of Texas, Fort Worth, EUA. 3 Embrapa Clima Temperado, Pelotas, Brazil dalastra.ch@gmail.com

Agarista is a little-known and understudied genus of subshrubs to trees in the blueberry family, comprising about 35 species. One species is distributed along the mountains of tropical Africa and the islands of the western Indian ocean, and the other 34 species are endemics from tropical mountains and temperate lowlands of the Americas. Despite previous monographic and cladistics studies providing an excellent framework of Agarista circumscription through a phyletic approach, sampling for phylogenetic assessment has been sparse. Only 6% of the Agarista species are represented in phylogenetic, biogeographic and evolutionary studies based on molecular data. Thus, this research provides a systematic study of Agarista focusing on the American species, especially in the Brazilian diversity center. The study comprises morphological data, taxonomic revision, molecular phylogenetic analysis, evolutionary character reconstruction, and biogeographic analysis. We are testing the monophyly of Agarista and reconstructing a phylogenetic hypothesis for its evolution. The American species are distributed among five main geographic areas previously defined and each of these areas comprises an endemic set of species. On this hand, we are also in search to understand how morphological characters evolved within the group and, based on biogeographic analysis, test hypotheses of past connections among these currently isolated set of species. Herbaria reviews included in person consults to 31 American herbaria, plus the herbaria revised online through their specific websites. Typifications were made for 17 names in Agarista. Four

fieldwork expeditions allowed collecting 15 samples
of distinct taxonomic entities. We confirm the oc-
currence of 34 species in the Neotropics, 24 of them
Brazil, being 22 endemics to this country. Addition-
ally, three new species were described. Molecular
analysis is in progress with the Botanical Research
Institute of Texas in view to clarify the evolutive and
biogeographical history of the genus based on the
Angiosperms-353 kit.

S.162.5 Phylogenomics of
the tropical African genus
Hymenocoleus Robbr.
(Rubiaceae)

Lovisa Thilén¹, Olivier Lachenaud²,³, Sylvain Razafimandimbison⁴, Catarina Rydin¹,⁵

1 Stockholm University, Stockholm, Sweden. 2 Meise Botanic Garden, Meise, Belgium. 3 Herbarium et Bibliothèque de Botanique Africaine, Université Liège de Bruxelles, Brussels, Belgium. 4 Swedish Museum of Natural History, Stockholm, Sweden. 5 Bergius Foundation, Stockholm, Sweden

Hymenocoleus is a small genus in the coffee fam-
ily (Rubiaceae). It belongs to the pantropical tribe
Palicoureeae of the subfamily Rubioideae, and the
species of the genus are herbs or undershrubs re-
stricted to rainforests in tropical Africa. Infrageneric
relationships in Hymenocoleus have received limit-
ed attention since its description and phylogenet-
ic relationships have never been investigated, but
the genus has undergone a recent taxonomic re-
vision, recognising 19 species. Several species are
newly described, and some are rare and as far as
known endemic to narrow geographic ranges. Here
we present the first phylogeny of Hymenocoleus, in-
ferred using a phylogenomics approach. We used
the Angiosperms353 probe set to capture hundreds
of low-copy nuclear genes and inferred a species
tree using coalescent-based methods. The result
is a fully resolved, highly supported species-level
phylogeny of Hymenocoleus. Data produced using
the Angiosperms353 probe set here showed capa-
ble of resolving relationships in the genus, includ-
ing some infraspecific relationships. Moreover, the
topology proved robust to different analytical ap-
proaches, and use of different data partitions and
filtering schemes. Hymenocoleus libericus is sister to
the remaining species, however, incomplete lineage
sorting is affecting the deepest splits in the genus
and it is possible that cytonuclear discordance will
<figure><img src="image_378.png" /></figure>

be detected in future studies, for this node as well as a few others. It is further interesting to note that there is morphological support for several clades in the species tree.

**S.162.6 Evolution and megaspore morphology in Isoetes (Lycophyta)**

Eva Larsén¹, Catarina Rydin¹

1 Department of Ecology, Environment and Plant Sciences, Stockholm University, Sweden

*Isoetes* L. (*Isoetaceae*, *Isoetales*) is a cosmopolitan genus of heterosporous lycopods with an ancient and diverse evolutionary history that presumably reached its zenith in the Paleozoic. The genus belongs to the order *Isoetales*, which appeared in the Late Devonian and had its greatest diversity and ecological dominance in the late Carboniferous when arborescent lepidodendrids were widespread in the coal swamps of Euramerica. But living alongside and outliving their tree-shaped relatives were also several genera of smaller and mostly unbranched isoetaleans that were common in the Triassic and continued to diversify throughout the Mesozoic and Cenozoic. The age of the living clade has however never been satisfactorily clarified and no fossils have been phylogenetically placed within the extant clade. With little morphological (and molecular) divergence among species, megaspore morphology may be one of the best hopes for resolution of relationships in the group as well as relationships among extant and extinct forms. We have studied the megaspore morphology, ornamentation and surface texture/structure, in 74 phylogenetically placed samples representing 59 species of *Isoetes* and discuss evolutionary implications of the results. The samples were studied with scanning electron microscopy, and ornamentation and surface structure/texture were mapped onto a phylogeny constructed based on molecular data from the same samples whenever possible. While there are general features common to all megaspores of the family (trilete with an outermost siliceous coating) there is in addition ample morphological variation, of which some appears clade-specific.
## S.163 GENOMICS OF LOCAL ADAPTATION.
SESSION 2

### S.163.1 Genome-wide differentiation along elevation corresponds to the Mediterranean and cold temperate climate niches in lichens

Edgar L.Y. Wong¹,², Henrique Valim¹,², Imke Schmitt¹,²,³

1 Senckenberg Biodiversity and Climate Research Centre, Frankfurt am Main, Germany. 2 LOEWE Centre for Translational Biodiversity Genomics, Frankfurt am Main, Germany. 3 Goethe University Frankfurt, Institute of Ecology, Evolution and Diversity, Frankfurt am Main, Germany

Lichens are known to withstand extreme environmental heterogeneity such as seasonal and diurnal temperature differences, and regular hydration-desiccation cycles. Many species also have a wide distribution range across multiple climate zones. However, little is known about the mechanisms and genomic basis of environmental adaptation in lichen-forming fungi. In this talk, we present genomic work on *Umbilicaria* species, which are obligate lichen-forming fungi that grow on rocks. Using samples collected (1) from different mountainous gradients in Europe and USA; (2) from low-elevation Mediterranean zone to high-elevation cold temperate zones; and (3) with different reproductive modes (asexual or sexual), we compared patterns of genomic differentiation and signatures of adaptation to climate niches. Specifically, we ask: (1) How do population genomic features of different species vary along replicated elevational gradients? (2) Which genomic regions are associated with the climate niche transition from the Mediterranean to the cold temperate zone? (3) Which bioclimatic variables are important predictors of population genomic change at the boundary between the two climate zones? (4) Do species with

contrasting reproductive modes show different genomic features? Some of our results include clear genomic differentiation between samples collected from the Mediterranean versus the cold temperate climate niche. Cline analyses showed one bioclimatic variable (winter temperature) as a strong selective pressure that led to allele fixation. Coupled with genomic scans for selective sweeps and subsequent gene ontology analyses, we present strong evidence that this bioclimatic variable is a major driver of genomic differentiation and hence adaptation. Overall, our results enrich our knowledge of fungal genomic functions related to climatic niche, population genomics, and the response of long-lived, sessile species to environmental heterogeneity.

### S.163.2 Local adaptation-induced evolutionary trap in alpine plants under climate change

Li Li¹, Jie Zhang¹, Victoria L. Sork²,³, Matthew C. Fitzpatrick⁴, Hong Liao¹, Gang Wang⁵, Yan-Bo Sun¹,⁶, Qing-Jun Li¹ and Jian-Li Zhao¹*

1 Ministry of Education Key Laboratory for Trans-boundary Ecosecurity of Southwest China, Yunnan Key Laboratory of Plant Reproductive Adaptation and Evolutionary Ecology and Centre for Invasion Biology, Institute of Biodiversity, School of Ecology and Environmental Science, Yunnan University, Kunming, Yunnan, China. 2 Department of Ecology and Evolutionary Biology, University of California, Los Angeles, CA, USA. 3 Institute of the Environment and Sustainability, University of California, Los Angeles, CA, USA. 4 Appalachian Laboratory, University of Maryland Center for Environmental Science, Frostburg, Maryland, USA. 5 CAS Key Laboratory of Tropical Forest Ecology, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla, Yunnan, China. 6 Laboratory for Conservation and Utilization of Bio-resources, Yunnan University, Kunming, Yunnan, China. *e-mail: jianli.zhao@ynu.edu.cn.
<figure><img src="image_379.png" /></figure>

Evolutionary traps occur when populations deviate from local adaptation and become maladapted under rapid climate change. Alpine plants, with genetically distinct ecotypes in sky islands, are susceptible to being trapped by rapid climate change. We employed landscape genomics to test a local adaptation-induced evolutionary trap in the alpine perennial plant *Roscoeia tibetica* (Zingiberaceae) with three distinct ecotypes along altitudinal gradients in the Hengduan Mountains. Strong local adaptation to alpine environments was observed through isolation-by-environment and rapid decreases in effective population size following interglacial niche contraction. Genomic offset increased with climate warming and was positively correlated with the intensification of local adaptation. Instead of high-elevation ecotypes, an evolutionary trap was indicated in the low-elevation forest ecotype, which showed the strongest local adaptation and niche loss, requiring the greatest shift in allele frequencies to keep pace with future climate change, and will likely experience the largest population decline. Therefore, stronger local adaptation induces evolutionary traps more easily and aggravates the harm to alpine plants facing rapid climate warming. Our study provides the first empirical case of local adaptation-induced evolutionary traps for alpine plants and highlights that it is crucial for alpine biota conservation to consider and disarm evolutionary traps under climate change.

**S.163.3 GWAS analysis**
reveals genetic basis of
some phenological and
morphological traits in
Chouardia litardierei
(Hyacinthaceae)

Sara Laura Šarančić¹, Nikolina Pleić²,³, Krešimir Križanović⁴, Ivan Radosavljević¹

1 Department of Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia. 2 Department of Medical Biology, School of Medicine, University of Split, Split, Croatia. 3 University of Applied Sciences ASPIRA, Split, Croatia. 4 Department of Electronic Systems and Information Processing, Faculty of Electrical Engineering and Computing, University of Zagreb, Zagreb, Croatia

Chouardia litardierei (Hyacinthaceae) is a bulbous, perennial species of the western parts of the Balkan Peninsula. It is characterized by a pronounced ecological plasticity since its populations are distributed across contrasting habitat types: karst poljes, wet meadows

near the seashore, and dry mountain slopes. Thus, *C. litardierei* presents a valuable model for addressing one of the central questions of evolutionary biology: what genomic mechanisms underlie local adaptation and ecological divergence? Here we present the results of a genome-wide association study (GWAS) that aimed at elucidating the genetic architecture of some complex traits in studied species. For this purpose, we set the common garden experiment with 214 individuals from nine selected populations, three representing each assumed ecotype. We performed comprehensive morphometric and phenological analyses and genotyped all individuals using a genotyping-by-sequencing approach. Finally, we combined all the obtained results in a comprehensive study to characterize the genotype-phenotype relations of selected traits. For the analysis, we used 24,660 SNPs, five reproduction-related morphological, and five phenological traits. All traits were considered polygenic, and GWAS analyses were performed assuming an additive genetic model. For each association analysis, we considered two different statistical approaches: the frequentist single-locus models and Bayesian multi-locus models. Within the single-locus models, for each trait, we fitted a standard linear mixed model (LMM) in GEMMA, as well as a Poisson linear mixed model in GMMAT for traits that represented count data, to correctly model the trait's distribution. Within the multi-locus models, we fitted a Bayesian sparse linear mixed model (BSLMM) and a latent Dirichlet Process Regression (DPR) model for each trait. Results were visualized using Manhattan and QQ plots. Multiple candidate loci were detected for different traits, and the results are being discussed.

**S.163.4 Perennial Grass**
**Ecotype Ecological and**
**Genetic Responses to**
**"Home" and "Away"**
**Microbial Inocula is**
**Ecotype-Specific**

Eli Hartung¹, Soumyadev Sarkar², Anna Kazarina¹, Kian Fogarty¹, Brad Olson¹, Matt Galliart³, Jack Sytsma¹, Ari Jumponen¹, Sonny Lee¹, Loretta Johnson¹

1 Division of Biology, Kansas State University. 2 University of Arizona. 3 Fort Hays State University

Andropogon gerardii (big bluestem) is a dominant grass of the US Great Plains and its broad distribution
<figure><img src="image_380.png" /></figure>

across a steep rainfall gradient has given rise to locally adapted wet and dry ecotypes. While abiotic factors that drive local adaptation have been well-studied, little is known about the role of soil microbiomes in local adaptation. We used reciprocal inoculation to investigate how home and away microbiomes affect local adaptation. We collected seed and A. gerardii rhizosphere soils from prairies in Kansas (dry ecotype, 580 mm rainfall year-1) and Illinois (wet ecotype, 1167 mm rainfall year-1). We cultured microbes from roots and native soils, and reciprocally inoculated wet and dry microbes weekly into wet and dry ecotypes growing in common garden soil for 12 weeks in greenhouse. We measured plant form and function weekly for a variety of ecological responses. We extracted mRNA from leaves to analyze transcriptome responses. Genetic background between ecotypes determined most of our results. In addition to ecotype, inoculation increased biomass, especially that of the dry ecotype by ~30%. Furthermore, ecotypes had higher chlorophyll absorbance when grown with local soil microbiomes suggesting biotic local adaptation. Gene expression differed between ecotypes with ~7000 genes differentially expressed between ecotypes. However, when ecotypes were inoculated with their local soil microbes, with whom they share evolutionary history, inoculum strongly affected diverse aspects of plant growth and development. We conclude that plant microbial interactions for ecological and genetic responses strongly depend on ecotype context.

**S.163.5 Patterns and Consequences of Subgenomic Variation in Invasive Johnsongrass (Sorghum halepense)**

Michael R. McKai, Fae D. Bramblepelt, Bishnu Adhikari, Nathaniel Hofford, M'Kalya Motley, Bryan N. MacNeill, Sontosh K. Deb, Andrew Busby, Alaina Dawkin, Allison Hayes, William A. Laycock, Caitlin McCann, Susie McLaughlin, Kayln Pate¹

*Department of Biological Sciences, The University of Alabama, Tuscaloosa, Alabama, USA*

Johnsongrass (Sorghum halepense (L.) Pers.) is an invasive, polyploid grass found throughout disturbed and agriculture areas of temperature, subtropical, and tropical regions. This species is thought to be

an allotetraploid formed through the hybridization of sorghum (Sorghum bicolor (L.) Moench) and Sorghum propinquum (Kunth) Hitchc. Since its introduction into the United States in the early 1800s and secondary introduction into Arizona, Johnsongrass has spread throughout much of the Southeast, lower Great Plains, portions of the Midwest, Southwest, and California. The movement of this invasive has been facilitated by the transport of agricultural products and the development of major railway and highway systems where Johnsongrass thrives in the disturbed areas surrounding fields, railroads, and highways. In this study, we assembled four genomes, resequenced 38 accessions at either 20X or 40X coverage, and low-coverage whole genome shotgun sequenced approximately 50 additional populations of Johnsongrass from diverse regions of the United States. Using these data, we assess patterns of overall genetic diversity between subgenomes within individuals, among accessions, and across populations to determine how subgenomic variation might drive adaptation to varying habitats. We also explore homoeolog retention and loss patterns, leveraging phylogenetic relationships to identify potential loci contributing to Johnsongrass's success as an invasive species in the United States. Furthermore, we examine the abundance and dispersion of homoeologous exchange across the Johnsongrass genome and identify the extent to which different loci have either moved to homoeologous chromosomes or have been lost. Ultimately, this research sheds light on the impact of diploidization on the diversity of U.S. Johnsongrass germplasm, offering insights into how polyploidy influences patterns of invasion.

**S.163.6 Repeated adaptation of Arabidopsis to challenging environments**

Filip Kolář¹, Sonia Celestini¹, Veronika Konečná¹, Magdalena Bohutínská¹

1 Department of Botany, Faculty of Science, Charles University, Prague, Czech Republic

Repeated adaptation provides valuable insight into the predictability of evolutionary change through replicated natural experiments. We investigated the strength and genomic underpinnings of repeated local adaptation of two outcrossing Arabidopsis species (*A. arenosa*, *A. lyrata*) towards challenging environments. Based on structured sampling and genome sequencing of 50 ecologically divergent
populations, transplant experiments and popula-
tion genomics, we discovered rapid multi-parallel
adaptation towards different environmental gra-
dients (high elevations, toxic soils) and detected
significant over-representation of shared adaptive
candidate genes for each environment. Although
vast majority of the parallel candidates sourced on
shared allelic variation, we also discovered a small
fraction of genes exhibiting convergent evolution
from independent *de novo* mutations. In addition,
a significant number of genes was identified re-
peatedly as candidates to two or more different
environments (e.g. challenging soils and alpine),
suggesting spatio-environmentally fluctuating se-

lection may further contribute to the maintenance
of diverse pools of adaptive standing variation.
Then, we complemented our investigations with
additional published gene lists and our genome
scans from other Brassicaceae genera and found
that the extent of genomic parallelism significant-
ly decreased with increasing divergence between
the compared lineages. This relationship was ex-
plained by decreasing probability of repeated
selection on the same standing or introgressed
alleles. We therefore conclude that genetic diver-
gence between populations, species and genera,
affecting the pool of shared variants, is an import-
ant factor in the predictability of genome evolution.

**S.164** GRASSROOTS MULTIDISCIPLINARY AND INTEGRATIVE RESEARCH ON THE GLOBAL DIVERSITY OF THE GRASSES (POACEAE). SESSION 1

S.164.1 Grasses through
space and time:
An overview of the
biogeographical and
macroevolutionary history
of Poaceae

Robert J. Soreng¹, Timothy J. Gallaher²,
Paul M. Peterson¹, Konstantin
Romaschenko¹, Fernando O. Zuloaga³,
De-Zhu Li⁴,⁵, Lynn G. Clark⁶, Christopher D.
Tyrrell⁷, Cassiano A. D. Welker⁸, Elizabeth A.
Kellogg⁹, Jordan K. Teisher¹⁰

¹Department of Botany, National Museum of Natural History, Smithsonian Institution, Washington, DC 20013-7012, USA. ²Department of Natural Science, Herbarium Pacificum, Bishop Museum, Honolulu, HI, USA. ³Instituto de Botánica Darwinion (CONICET-ANCEFN), Casilla de Correo 22, San Isidro, Buenos Aires, Argentina. ⁴Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China. ⁵University of Chinese Academy of Sciences, Beijing, China. ⁶Department of Ecology, Evolution, and Organismal Biology, Iowa State University, Ames, IA, USA. ⁷Botany Department, Milwaukee Public Museum, Milwaukee, WI, USA.

8 Instituto de Biologia, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil. 9 Donald Danforth Plant Science Center, St Louis, MO, USA. 10 Missouri Botanical Garden, St. Louis, MO, USA.

Grasses are widespread on every continent and occur
in all terrestrial biomes. The dominance and spread of
grasses and grassland ecosystems led to significant
changes in Earth's climate, geochemistry, and biodi-
versity. Abundant chloroplast DNA sequence data, and
advances in placing grass fossils within the family al-
low for phylogenetic reappraisal of the family's origins,
timing, geographic spread, and factors promoting di-
versification. We constructed a time-calibrated grass
phylogeny and inferred ancestral areas using chloro-
plast DNA sequences from nearly 90% of extant grass
genera. With a few notable exceptions, the phylogeny
is well resolved to the subtribal level and in agreement
with our current classification. Using this phylogenetic,
temporal, and spatial framework, we review the evolu-
tion and biogeography of the family and present bio-
geographical hypotheses concerning its origins, evolu-
tionary tempo, and diversification. Extant lineages of the
family began to diversify around the Late/Early-Creta-
ceous boundary (crown 98.54 Ma) in West Gondwana.
All subfamily crowns (except Pharoideae) evolved be-
tween the Late-Cretaceous to Late-Eocene, with most
tribe crowns between Mid-Eocene to Early-Oligocene,
and stems of most genera between the Mid-Oligocene
and Late-Miocene. The splitting of Gondwana appears
<figure><img src="image_381.png" /></figure>

responsible for the initial vicariance of Anomochlooideae from the rest of the family: It's genera lack true spikelets and are endemic to and the only subfamily (or tribe) with a crown (80 Ma) in South America. Africa was the center of origin for much of the early diversification of the family (Spikelet and Bistigmatic clades, Puelioideae, Core-Grass-clade, BOP and Oryzoideae, PACMAD and ACMAD clades, Panicoideae, Aristidoideae, Arundinoideae, and Chloridoideae). Bambusoideae and Micrairoideae exhibit Indomalayan origins. Pooideae and tribes exhibit Palearctic origins. Ehrhartoideae and Cyperochloeae are tribes of Australian origins. No tribes have North American origins. Key questions to answer are the roles of Antarctica, extinction, unequal-rates of sequence evolution (woody-bamboos, and annuals), and whether Aristidoideae or Panicoideae evolved first.

## S.164.2 Nuclear phylogenomics of grasses (Poaceae) supports classification and reveals repeated reticulation

Grass Phylogeny Working Group III (Arthan W., Baker W.J., Barrett M.D., Barrett R.L., Bennetzen J., Besnard G., Bianconi M.E., Birch J.L., Catalán P., Christin P.-A., Clark L.G., Columbus J.T., Crayn D.M., Davidse G., Dunning L., Duvall M.R., Fisher A.E., Fjellheim S., Forest F., Gillespie L.J., Hackel J., Haevermans T., Hodkinson T.R., Huang C.-H., Huang W., Humphreys A., Jobson R.W., Kellogg E.A.E., Larridon I., Li D.-Z., Liu J.-X., Ma H., Macfarlane T.D., Maurin O., McKain M., McLay T.G.B., Moreno-Aguilar M.F., Murphy D.J., Peterson P., Saarela J.M., Simpson L., Snow N.W., Soreng R.J., Thompson E.J., Traiperm P., Verboom G.T., Vorontsova M.S., Walsh N.G., Washburn J.D., Watcharamongkol T., Waycott M., Welker C.A.D., Xanthos M., Zhang L., Zizka A., Zuntini A.R.)

Grasses (Poaceae) have over 11,800 species and are central for human livelihoods and terrestrial ecosystems. Knowing their relationships and evolutionary history is key to comparative research and crop

breeding. We inferred a comprehensive nuclear species tree of grasses by combining new and published sequences, including paralogs, for 331 genes from genome, transcriptome, target enrichment and shotgun data. Our 1,153-tip tree covers 78% of grass genera and all but two small tribes. We compared it to a 910-tip plastome tree. The nuclear phylogeny matches that of the plastome at most deep nodes, with only a few instances of incongruence. Gene tree-species tree reconciliation suggests that reticulation events occurred repeatedly in the history of grasses. We provide a robust framework for completing the grass tree of life to support research on grass evolution, including modes of reticulation, and genetic diversity for sustainable agriculture.

## S.164.3 How can grass taxonomy secure the future of Madagascar?

Maria S. Vorontsova¹

1 Accelerated Taxonomy, Royal Botanic Gardens, Kew, Richmond, Surrey, UK

Twenty million years before human arrival on Madagascar, the ancient ancestors of Fatakamanga or the Blue Bamboo, also known as *Lecomtella madagascariensis*, made their journey to the island. The only remaining population of this isolated lineage is restricted to the mountains of the Andringitra National Park. The list of Andringitra micro-endemics whose presence justifies protected area funding also includes the world's only grass that imitates Erica shrubs: *Panicum cupressifolium*. But savannas at lower elevations are not included in biodiversity protection campaigns and not believed to be worthy of protection. Landscape genomics of *Loudetia simplex* which dominates vast expanses of savanna are helping us reconstruct ecosystem history to help plan conservation. Traditional Malagasy livelihoods are based around herds of zebu dependent on the guilds of forage grasses forming grazing lawn assemblages, including the recently revised endemics *Panicum luridum* (*Panicum* sensu stricto), and *Panicum umbellatum* with *Brachiaria subrostrata* (which need to be given new generic names within the Melinidinae in the Panicoideae). Agriculture professionals are unaware of native forages, and we are working to promote their use. The most beautiful baskets are woven in the Fandriana region from harsh bunchgrasses called Haravola and Haravola namboa, which are threatened by expanding rice
<figure><img src="image_382.png" /></figure>

cultivation. One is a sorghum relative *Lasiorhachis viguieri*, and the other is a mystery likely affiliated with the South African *Phacelurus franksiae*. None of these natural history facts were known or connected to national policy before the start of the Madagascar grass project in 2011. Placing grasses at the centre, we are working to piece together useful stories combining herbarium taxonomy, ethnobotany, phylogenetics, and functional ecology interfacing with social science and biome modelling, livestock development, and conservation. This talk will present an overview of how Madagascar's grasses are being braided together to support the country's future.

**S.164.4 Advances on a new classification for koelerioid grasses based on molecular studies and morphology (Poaceae: Pooideae: Aveninae)**

Alejandro Quintanar¹, Robert J.

Soreng², Paul M. Peterson², Konstantin

Romaschenko², Carlos Aedo¹, Patricia

Barberá³

¹ Real Jardín Botánico de Madrid CSIC, Madrid, Spain. ² National Museum of Natural History, Smithsonian Institution, Washington DC, USA. ³ Department of Africa and Madagascar, Missouri Botanical Garden, St. Louis, USA

Analyses of molecular data accumulated over nearly two decades have revealed extensive phylyy affecting many of the genera that make up the oats subtribe, Aveninae, a group of subfamily Pooideae grasses of global economic importance. Within Aveninae, Koeleriinae clades A and B (koelerioid grasses) contain most of the biodiversity in the subtribe, including mostly perennial lineages, such as *Trisetum* s.l. or *Koeleria* (both worldwide) and other American genera like *Sphenopholis* and *Graphephorum*, and a series of annuals like *Rostraria* and *Trisetaria* from the Mediterranean (except *Limnodea*). We present our latest results on the systematic relationships of the koelerioids resulting from the largest sampling of these grasses carried out to date. Our new phylogenetic trees have improved our understanding of the evolution of this diverse group and led to drastic revisions of their classification. Amendments have involved the publication of new genera and the expansion,

reduction or resurrection of others. A notable case of reduction is *Trisetum*, before considered to host ca. 70 perennial species, and now restricted to only three species, one of them annual. In contrast, *Koeleria*, another large perennial genus from cold or temperate regions of all hemispheres, has grown from 25 to nearly 100 species after absorbing part of the diversity of *Trisetum*. *Aegialina*, *Acrospelion*, and *Cinnagrostis* were resurrected. The detection of past reticulation events forming *Graphephorum* and parts of *Rostraria* suggest the importance of the hybridization as driving force of evolution in these groups, a common feature with other lineages of supertribe Poodae.

**S.164.5 Decoding hybridization events in temperate Loliinae grasses: a nuclear single-copy gene perspective**

María Fernanda Moreno-Aguilar¹, Juan Viruel², Aminael Sánchez-Rodríguez³, Itziar Arnelas³, Juan Camilo Ospina⁴, Gloria Martínez-Segarra⁵, Diana Calderón³, Juan Antonio Devesa⁵, Pilar Catalán¹,⁶

¹ Departamento de Ciencias Agrarias y del Medio Natural. Escuela Politécnica Superior de Huesca. Universidad de Zaragoza, Huesca, Spain. mmoreno@uniza.es. ² Royal Botanic Gardens, Kew, Richmond. UK. ³ Departamento de Ciencias Biológicas y Agropecuarias, Universidad Técnica Particular de Loja, Loja. Ecuador. ⁴ Facultad de Ciencias Agrarias. Universidad Nacional de Jujuy. San Salvador de Jujuy, Jujuy, Argentina. ⁵ Departamento de Botánica, Ecología y Fisiología Vegetal, Universidad de Córdoba, Spain. ⁶ Grupo de Bioquímica, Biofísica y Biología Computacional (UNIZAR), Unidad Asociada al CSIC, Zaragoza. Spain.

Resolving the phylogeny of recently evolved groups that have experienced recurrent introgressions and polyploidizations throughout their history is challenging. Evolutionary reconstruction based on a large set of biparentally inherited single-copy nuclear genes is considered a suitable approach to recover relationships that might otherwise be obscured by nuclear ribosomal genes in concerted evolution or biased by maternally inherited plastomes. Here we present the first single-copy gene phylogeny of the ecologically and economically important grass subtribe Loliinae (*Festuca* and related
<figure><img src="image_383.png" /></figure>

genera) using 241 nuclear coding loci captured from the Angisoperm353 probe set and a large sample of 137 representative taxa of Loliinae, covering all of its 22 evolutionary lineages. We have further completed genome-skimming sampling of other nuclear (35S rDNA gene) and organellar (plastome) sequences to elucidate the reticulate history of the Loliinae from multiple and complementary genome sources. Concatenated maximum likelihood and multispecies coalescent trees of single-copy genes showed well-supported relationships that were generally consistent across analyses and with previous taxonomic and phylogenetic findings but also revealed high levels of gene discordance. Hybridization and topological incongruence tests between the nuclear and plastome-based trees confirmed the rampant introgression experienced by Loliinae at deep and shallow nodes, detecting hybridization in four broad-leaved (Subulatae-Hawaiian, Schendorus, Tropical-South African, Mexico-Central-South American) and five highly-diversified fine-leaved (American II, Exaratae-Loretia, Aulaxyper, Afroalpine, American-Neozeylandic) Loliinae lineages. The levels of intragenomic discordance could have been magnified by the prevalence of allopolyploids in the Loliinae and methodological bias in the selection of orthologs; however, our nuclear and plastome trees have revealed key hybrid origins in these grasses.

## S.164.6 Taxonomy of Digitaria (Panicoideae, Poaceae) grasses to support weed and forage management in Madagascar

Nantenaina H. Rakotomalala<fnref n="1" />,
Lorna MacKinnon², Fenitra N.
Randrianarimanana<fnref n="1" />, Sami Touafchia³,
Thomas Haevermans³, George
Burton², Rafael F. de Almeida², Jean
A. Randriamampianina⁴, Mijoro
Rakotoarinivo⁵, Maria S. Vorontsova²

The Poaceae family is species-rich and functionally significant for Madagascar's ecosystems and agri-systems. As part of an interdisciplinary project on grass diversity and agricultural interactions, we revise the classification of the genus *Digitaria* in Madagascar. This complex group poses identification challenges due to strong similarities between species. Over 50% of the 14,000 Madagascar's Poaceae herbarium collections are housed in Paris, presenting a constraint for Malagasy botanists aiming to study grass microscopic features that are not discernible in digital images. Consequently, I undertook trips to both Paris and London to access the essential resources, supplementing fieldwork efforts. Through examination of herbarium materials and field collections, we identified 30 species in Madagascar, including seven endemics, and excluding five species previously believed to occur in the region. Noteworthy are a likely new species from central and northern Madagascar and a new record of *D. ternata*. Three names were placed in synonymy: endemics *D. psammophila* and *D. planiculmis* are synonyms of the non-endemic *D. didactyla*, while *D. madagascariensis* is a synonym of the endemic *D. fulva*. A key to the species is provided along with illustrations, descriptions, and distribution information. This revision updates the *Digitaria* data in the Weed Identification and Knowledge in the Tropical and Mediterranean Areas portal. Using the updated species concepts, ethnobotanical analyses revealed 11 *Digitaria* species considered to be weeds affecting agriculture and used as forages in the highlands. Weedy traits of *D. humbertii* include its entangling growth habit and the ability to infiltrate cropping systems. Phylogenetic analysis indicates independent evolution of weedy species in the paraphyletic *Digitaria* clade. In order to disseminate these findings, we created an illustrated field guide in English and Malagasy, developed collaboratively with national agricultural institutions and the Ministry of Agriculture to help farmers recognize native forages and identify weed control methods.

<footnote n="1">1 Royal Botanic Gardens Kew, Antananarivo, Madagascar. 2 Royal Botanic Gardens Kew, London, UK. 3 Museum National d'Histoire Naturelle Paris 4 National Centre for Applied Research on Rural Development (FOFIFA), Madagascar. 5 University of Antananarivo, Madagascar</footnote>
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# S.165 MORPHOLOGICAL, ANATOMICAL, AND EVOLUTIONARY BASIS OF HABIT TRANSITIONS IN PLANTS

## S.165.1 Apocynaceae wood evolution matches key morphological innovations

Vicky Beckers¹,², Mary Endress³, Pieter Baas¹, Erik Smets¹,², Frederic Lens¹,²

1 Naturalis Biodiversity Center, Leiden, The Netherlands. 2 Leiden University, Leiden, The Netherlands. 3 University of Zurich, Zurich, Switzerland

Apocynaceae are a wonderfully diverse group, widely studied for their variable evolutionary processes. The stem is one of the most functionally important plant parts, yet a wood anatomical overview of this family is missing. Here, we present a wood anatomical overview of the Apocynaceae, including reconstructions of wood evolution. To do so, we revised over 200 published wood anatomical descriptions and made original sections and light microscopic descriptions for another ~50 species. Changes in wood anatomical characters through time were visualized with ancestral state reconstructions on a selection of 26 informative traits for 147 woody species included in the family's most recent molecular phylogeny. We performed tests for correlated evolution on a combined dataset of anatomical and published morphological traits to identify potential synnovations and key traits associated with growth form adaptations. The most important finding was a shift towards a syndrome of derived wood anatomical traits within the paraphyletic apocynoid grade that include an increased presence of vessel multiples, vessel dimorphism, laticifers, cambial variants, and paratracheal axial parenchyma. These traits separate the grade into early- and later diverging tribes, and continue in the monophyletic Periplocoideae, Secamonoideae, and Asclepiadoideae. When we compared these trait reconstructions with morphology traits evolution, we found that key morphological innovations co-occur with wood characters on consecutive nodes in the family's phylogeny. In contrast to these abrupt trait changes, vessel elements and fibers gradually shorten along the phylogenetic backbone of the family. This correlates to a general plant size reduction, regardless of the many transi-

tions towards (phylogenetically) derived woodiness and a scandent or erect growth form. In conclusion, there are clear evolutionary transitions in the wood anatomy of Apocynaceae -many associated with climbers - representing structural adaptations that are part of a cascade of morphological evolutionary changes across the family.

## S.165.2 Ontogeny of inter- and intra-xylary phloem in eudicotyledons

Kishore S. Rajput¹

1 Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, India

Migration of aquatic autophytes to terrestrial conditions compelled them to evolve a new strategy of specialised tissues for mechanical support and conduction, i.e., secondary xylem and phloem respectively. From them, the phloem is a relatively delicate tissue that plays a vital role in the translocation of photosynthates. In most of the eudicots, the phloem develops external to the vascular cambium and the xylem internal to it. Besides their position external to the cambium, in a small fraction of eudicots, it also occurs within (interxylary phloem that develops from the single ring of vascular cambium) and inside (intraxylary) the secondary xylem. Based on their unique ontogeny, interxylary phloem is classified variously (Strychnos, Combretum, Azima and Calycopteris types) by earlier researchers. The present study documents several additional examples that do not fit into any of the types mentioned above (i.e., it develops in a later stage by dedifferentiation of parenchyma and not from the cambium). Similarly, the intraxylary phloem located on the pith margin either initiates from procambial derivatives, ground meristem, from the adjacent pith cells or by developing intraxylary cambium. Little information is available on the formation of intraxylary cambium and its unifacial (deposition of the secondary xylem and phloem in the same direction) or bifacial (xylem externally and phloem internally) nature but
<figure><img src="image_385.png" /></figure>

detailed information on its behaviour remains unexplored. Several such examples of intraxylary CAMBium with intermediate variants are also discussed herewith and their possible functions are attributed to defence against insect injury and herbivory based on pieces of experimental evidence available in the literature.

**S.165.3 Just keep growing! Woodiness as a simple extension of herbaceous habit.**

Kamil E. Frankiewicz¹,²,³,⁴, Carmen R. Marcati³, Anthony R. Magee¹, Alexei A. Oskolski⁵,⁶

1 Compton Herbarium, South African National Bio-diversity Institute, Private Bag X7, Claremont, Cape Town, South Africa. 2 Bolus Herbarium, Department of Biological Sciences, Faculty of Science, University of Cape Town, Private Bag X3, Rondebosch, South Africa. 3 Laboratório de Anatomia da Madeira, Departamento de Ciência Florestal, Solos e Ambiente, Faculdade de Ciências Agronômicas, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil. 4 Plant Cytogenetics and Molecular Biology Group, Institute of Biology, Biotechnology and Environmental Protection, Faculty of Natural Sciences, University of Silesia in Katowice, Katowice, Poland. 5 Department of Botany and Plant Biotechnology, University of Johannesburg, P.O. Box 524, Auckland Park, Johannesburg, South Africa. 6 Komarov Botanical Institute of the Russian Academy of Science, St. Petersburg, Russia

Transitions between herbaceous and woody habits is a key to angiosperm ecological and evolutionary success. Some correlations between habits and climatic niches have been established, such as that the increase in herbaceous life forms since the Eocene was a strategy to evade seasonal frost, or that reversals to woodiness during the Miocene climate aridification enhanced resistance to drought-induced embolism. While these relationships are well-understood, intrinsic (organismal) traits governing angiosperm habit adaptability remain elusive. We take advantage of our extensive sampling to delve into the anatomical distinctions between woody and herbaceous plants. Do these differences extend beyond quantity of secondary xylem? Are there any qualitative traits universally distinguishing herbs from woody plants? What makes angiosperms so adaptable when their closest extant relatives (gymnosperms) are constrained to a narrow range of life forms? First,

we observe that across diverse angiosperms, spanning magnoliids, asterids, and rosids, the rays of secondary woody may simply prolongate patterns from primary xylem, without qualitative changes. Then, we show that quantitative traits such as vessel area and vessel wall thickness develop at comparable rates in closely related herbaceous and secondarily woody species (i.e., shrubs derived from herbaceous ancestors). These underscore that woodiness may be a mere prolongation of patterns present in herbaceous ancestors. Finally, we propose a novel perspective: habit shifts are linked to changes in wood lignin content, influencing stem mechanical properties and, consequently, maximal height. We argue that this could foster morphological disparification contributing to angiosperms unparalleled diversity. In summary, our observations emphasize that angiosperm habit adaptability results from their ability to alter cambial activity and, possibly also, the amount of lignin produced and deposited in xylem. Simultaneously, while wood anatomical traits are crucial for adaptations to specific niches, they do not seem to be primarily related to life forms.

**S.165.4 Developmental processes behind habit transitions from lianas to trees and shrubs in vascular plants**

Marcelo R. Pace¹, Angélica Quintanar Castillo¹, Camila Alcántara¹

1 Universidad Nacional Autónoma de México, Instituto de Biología, Departamento de Botánica

The ancestral habit of woody plants is inferred as self-supporting i.e., either erect shrubs or trees. The vascular system in self-supporting woody plants fulfills the roles of water and photosynthate transport, mechanical support, storage, and longevity. Lianas, however, do not support themselves, therefore their stem anatomy, released from this role, undergo modifications towards other critical purposes. First, lianas have some of the most efficient hydraulic systems, with less fibers, wider and more abundant vessels per unit area. Second, liana stems undergo selection for flexibility, thus their stems have wider and taller rays, sometimes more abundant axial parenchyma, and the wholesale restructuring of the vascular architecture (vascular variants), which intermixes parenchymatous and lignified tissues. Interestingly, within sev-
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eral ancestrally lianescent lineages, self-supporting plants have re-evolved, when occupying semi-arid regions. In these unique cases, the vascular system, therefore, must provide support to keep the stem upright, which is reflected in the reversal to a secondary xylem with narrower vessels, smaller rays and more fibers, and sometimes with changes in the axial parenchyma abundance. Critically, when the self-supporting habit is regained, vascular variants in their stems may even be lost or greatly reduced. Conversely, if the emergent self-supporting habit is succulent, these plants co-opt the ancestral vascular variant for increased water storage, representing a case of exaptation. In this talk, we discuss the re-emergence of self-supporting plants from ancestrally lianescent lineages using case studies in Bignoniaceae, Convolulaceae, and Malpighiaceae.

**S.165.5 Evolution of Development of Vascular Variants in a Large Lineage of Neotropical Woody Vines: Paullinieae (Sapindaceae)**

Joyce G. Onyenedum¹, Israel L. Cunha Neto¹, Chun Su¹, Pedro Acevedo-Rodríguez¹

1 Department of Environmental Studies, New York University, New York, NY, USA. 2 Department of Botany, Smithsonian National Museum of Natural History, Washington, DC, USA

Climbing woody vines-- "lianas" -- are the snakes of the plant kingdom, capable of wrapping around host trees in search of light at the top of the forest canopy. Intriguingly, this lifeform has evolved independently multiple times, and in accordance, lianas have independently evolved significant modifications to their stem anatomy to meet the unique demand to twist without breaking. Unlike the stems of most trees and shrubs, comprised of a core of wood (xylem) sheathed by inner bark (phloem), lianas often have "vascular variants", expressed as aberrations in the typical distribution and abundance of xylem and phloem. In this study, we sought to elucidate the evolution of development of vascular variants in a large neotropical liana tribe, Paullinieae (Sapindaceae), comprised of 11 vascular variants across 495 species. We here leverage the newly developed molecular phylogeny of Paullinieae, comprised of 75% species of the tribe, together with robust developmental anatomy dataset to address

the following questions: (1) How many times did each type of vascular variant evolve? (2) Do vascular variants have systematic value in defining clades? (3) How did changes in the mode and tempo (heterochrony), spatial reorganization (heterotopy), tissue identity transformations (homeosis), and the re-purposing of old tools (exaptation) contribute to the diversity of vascular architecture in lianas? This study represents the power of an integrative approach, bridging developmental anatomy, within a robust phylogenetic context, to elucidate the evolution of development of complex traits.

**S.165.6 Cambial activity contrasts between trees and lianas in temperate and tropical regions**

Veronica Angyalossy¹, André C. Lima², Lui Agostinho Teixeira¹, Marcelo R. Pace³, Carmen R. Marcati², Ray F. Evert⁴

1 Departamento de Botânica, Instituto de Biociências, Universidade de São Paulo, São Paulo, Brazil. 2 Departamento de Ciência Florestal, Solos e Ambiente, Faculdade de Ciências Agronômicas, Universidade Estadual Paulista, Botucatu, Brazil. 3 Departamento de Botânica, Instituto de Biología, Universidad Nacional Autónoma de México, Ciudad de México, Mexico. 4 Department of Botany, University of Wisconsin, Madison, USA

The vascular cambium typically has a seasonal activity, both in trees and lianas, either in the tropics or in temperate regions. This seasonal activity is generally annual for both habits. Photoperiod is one of the main triggers of the resume of cambial activity after dormancy, but this is in concert to a rise in temperatures in the temperate regions and the end of the dry season in the tropics. Both the onset and offset of cambial activity occur within the favorable growing season, i.e., summer in the temperate region and rainy season in the tropics, indicating that cambial rhythm anticipates the seasons that come ahead. However, lianas typically have a much larger inter-specific variation than trees. Within a study under the same conditions, some lianas had only 2 months activity, others 5 month and others yet had the cambial activity throughout the year. We explore here the best methods to detect cambial activity, the most reliable initial and terminal growth rings markers, specificities in growth ring formation in lianas and how vascular variants of lianas may also greatly influence cambial activity.
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# S.166 PHYLOGENOMICS AND SYSTEMATICS OF LILIACEAE

## S.166.1 Plastome structure, phylogenetics, and historical biogeography of Liliaceae subfamily Streptopoideae

Kenneth M. Cameron¹, Shenyi Wang¹, Pan Li²

1 Department of Botany, University of Wisconsin-Madison, USA. 2 Zhejiang University, Hangzhou, China

Liliaceae always have been a challenge to classify and the taxonomy of one subfamily, Streptopoideae, also has been plagued with controversy. As currently circumscribed Streptopoideae contain three genera: Prosartes, Scoliopus, and *Streptopus* with 17 species distributed across the Northern Hemisphere. A few of the species even exhibit inter-continental disjunctions. Although several higher-level molecular phylogenetic studies have included a handful of species from this subfamily as placeholders, the whole picture of finer scale phylogenetics among these plants has never been studied adequately. We resolved the phylogenetic relationships, estimated divergence times among clades, and reconstructed historical biogeography within Streptopoideae and several outgoup taxa using newly sequenced de novo assembled and annotated whole chloroplast genomes. We achieved a high level of sampling for the ingroup (15 of 17 species), and also considered a moderate level of intra-specific sampling for the two species, *S. amplexifolius* and *S. lanceolatus*, with the widest geographic distribution. We compared and contrasted the structure of Streptopoideae plastomes, demonstrating that they have relatively large genomes, but with same gene arrangement that characterizes other Liliaceae. A relatively well resolved and supported phylogenetic reconstruction supports the North American genera *Prosartes* and *Scoliopus* sharing a recent common ancestor, with *Streptopus* as sister to them. The sub-family diverged from its shared ancestor with *Tricyrtis* at least 57 Mya in the late Paleocene of either Western North America or East Asia. The species of *Streptopus* in North America appear to have migrated to the east from Asia more recently. Relationships within each genus are fully resolved, but mostly supported only by

short branch lengths. One individual of *S. lanceolatus* from the Pacific Northwest showed evidence of chloroplast capture from sympatric *S. amplexifolius*. These results indicate that alternative molecular methods targeting the nuclear genome will be needed to further tease apart the evolutionary history of this group.

## S.166.2 Phylogenomics, historical geography, adaptive radiation, and quantitative floral convergence in lilies (Lilium)

Thomas J. Givnish¹, Mark Skinner², Ivana Rešetnik³, Nursel Ikinci⁴, Ricardo Kriebel⁵, Alan R. Lemmon⁶, Emily Moriarty Lemmon⁷, Yundong Gao⁸

1 Department of Botany, University of Wisconsin-Madison, Madison, USA. 2 Skinner & Associates, Portland, USA. 3 Department of Biology, University of Zagreb, Zagreb, Croatia. 4 Department of Botany, Abant Izzet Baysal University, Bolu, Turkey. 5 Department of Botany, California Academy of Sciences, San Francisco, USA. 6 Department of Scientific Computing, Florida State University, Tallahassee, USA. 7 Department of Biological Science, Florida State University, Tallahassee, USA. 8 Department of Botany, Chinese Academy of Sciences, Chengdu, China

Lilies – with ca. 125 species in *Lilium* and the former genus *Nomocharis* – are restricted to the Northern Hemisphere, with centers of diversity in East Asia, southern Europe, the Caucasus, and eastern and Western North America. Species vary widely in habitat and in floral form, orientation, color, and fragrance. To identify relationships within *Lilium* and reconstruct its adaptive radiation and geographic spread, we conducted the first phylogenomic analysis of relationships in the genus by sequencing, assembling, and analyzing more than 400 single-copy nuclear loci and whole plastomes of almost all extant species. The maximum-likelihood plastome phylogeny fails to resolve any of the traditional, morphology-based sections as monophyletic, with two origins for North American § *Pseudolirium*, two for Himalayan/Tibetan § *Nomocharis*, four for East Asian § *Sinomartagon*, two for Asian § *Leucolirion*, two for Asian
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§ *Archelirion*, three for European and Central Asian § *Martagon*, two for East Asian § *Daurolirion*, and two for European § *Liriotypus*. The nuclear phylogeny, however, resolves most sections as monophyletic, except for polyphyletic § *Sinomartagon* and § *Leucolirion*. Reconstructions of historical biogeography point to an origin in East Asia or Himalayas/Tibetan Plateau, with subsequent dispersals into North America, the Himalayas/Tibetan Plateau, Europe, Formosa, Japan, and the Philippines. Within the North American clade § *Pseudolirium*, we found strong quantitative convergence in flower shape, orientation, and color associated with pollination by hummingbirds, sphingid moths, and bees, and apparent retention of ancestral characters in species pollinated by butterflies.

### S.166.3 A monograph of Chinese lilies

Yundong Gao¹, Melvyn Herbert²

1 CAS Key Laboratory of Mountain Ecological Restoration and Bioresource Utilization & Ecological Restoration and Biodiversity Conservation Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, China. 2 Llwynteg, Bwlchllan, Nr Lampeter, Ceredigion, UK.

Lilies are amongst the most fabulous of cultivated flowers, being important subjects for both the amateur gardener and professional growers supplying the fresh cut flower trade across the globe. Compared to the great multitude of hybrid cultivars that have been developed the naturally occurring wild species are relatively few, just over a hundred of these being mainly concentrated in the northern temperate zone. Of the major distributional areas, Eastern Asia (mainly China) exhibits the greatest diversity of wild lily species, boasting over three-quarters of the taxa currently known to science. In addition to possessing the largest number of species, the Chinese lilies also display great diversity at the sub-generic level. At least 5 out of 7 of the sections that are commonly accepted are represented in the Chinese flora (though differences of opinion regarding classification within the genus do remain). Recent technological and scientific innovations, along with easier access to remote areas, mean that it is now possible to resolve many of the outstanding taxonomic puzzles relating to the lilies of China that remain from earlier times and also to incorporate the newly recognized species into a fully updated catalog of these wonderful plants. In this work, the make a substantial revision of all the *Lilium* species that are currently known to occur in China. By tracing the history of their discovery, verifying the relevant literature (i.e.,

the protologue and other related materials) and locating the type specimens; and combining these with the molecular evidence accumulated by the present, as well as the findings of over a decade of field investigation, this monograph aims to provide a fully updated catalog and enumeration of Chinese native lilies including each species' history, details of the type specimen, morphological description, distribution information, all accompanied by field images.

### S.166.4 Phylogenomics of Liliaceae revisited

Mingyue Ye¹, Meizhen Wang¹, Kenneth Cameron², Pan Li¹,*

1 Laboratory of Systematic & Evolutionary Botany and Biodiversity, College of Life Sciences, Zhejiang University, Hangzhou, China. 2 Department of Botany, University of Wisconsin, Madison, USA. *panli@zju.edu.cn

The lily family (Liliaceae), which is distributed mainly across the temperate Northern Hemisphere, is of great horticultural, culinary, and medical importance. It is also a family with a long history of taxonomic uncertainty. Challenges in accurate species identification persist and phylogenetic relationships among genera within the family continue to be unresolved and/or weakly supported due to the use of limited molecular markers with insufficient variability. Here, plastome and transcriptome data were generated and combined with previously published data for this family, providing a total of 86 plastomes and 88 transcriptomes (plus one WGS data) covering all 15 currently recognized genera for analyses. Both plastid and nuclear phylogenomic analyses strongly confirm the recognition of four subfamilies: Tricyrtidoideae with four genera; Medeoloideae with two genera (*Clintonia* and *Medeola*); Lilioideae with eight genera; and a distinct subfamily, Calochortoideae, for *Calochorpus*, which is sister to Medeoloideae and Lilioideae. Within Lilioideae, both plastid and nuclear phylogenomic analyses show that the Lilieae clade of (*Notholirion*, (*Cardiocrinum*, (*Fritillatia*, *Lilium}*)) is sister to the Tulipeae clade of (*Gagea*; (*Tulipa*, (*Erythronium*, *Amana}*)). However, plastid and nuclear trees show different topologies within Tricyrtidoideae, which is (*Tricyrtis*, (*Streptopus*, (*Prosartes*, *Scoliopus}*)) in the plastid tree but (*Tricyrtis*, (*Prosartes*, (*Streptopus*, *Scoliopus}*)) in the nuclear tree. This cytonuclear conflict is likely due to ancient hybridization. Our results provide a solid framework for future studies on the evolution of Liliaceae.
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**S.166.5 Updated molecular phylogenetic analysis, dating and biogeographical history of the lily family (Liliaceae: Liliales)**

Joonhyung Jung¹, Joo-Hwan Kim¹

1 Department of Life Science, Gachon University

Liliaceae sensu APG IV include ca. 600 species, and have been circumscribed into three sub-families, Lilioideae, Calochortoideae and Streptopoideae. Molecular phylogenetic approaches to this family have produced dynamic changes in the generic circumscriptions. We conducted molecular phylogenetic analyses, time estimations and biogeographical analyses to confirm generic relationships, discuss their circumscription for classification and clarify the evolutionary history of the family. A phylogenetic analysis is presented as a combined data set of ca. 6.1 kb from four plastid coding gene sequences for 142 taxa representing all the genera of Liliaceae. Medeoloideae were newly defined as an independent subfamily from Lilioideae. *Tricyrtis* of Calochortoideae is embedded in Streptopoideae, despite its unique characteristics. The crown age of Liliaceae was calculated at ca. 85 Mya, and Liliaceae are considered to have originated in temperate Asia in the late Cretaceous and to have expanded their distribution via dispersal with the occurrence of repetitive vicariance events during their evolution.

**S.166.6 Taxonomic revision of Tricyrtis (Liliaceae) and a new species Tricyrtis lingyunyanensis in Taiwan**

Yu-Huan Chen¹, Ting-Hsuan Huang¹, Chang-Tse Lu², Rong-Show Shen¹

1 Department of Horticultural Science, National Chiayi University, Chiayi City, Taiwan. 2 Department of Biological Resources, National Chiayi University, Chiayi City, Taiwan

*Tricyrtis* Wall. (Liliaceae) is a perennial herb, including approximately 20 species, endemic to East Asia. *Tricyrtis* exhibit similar morphologies; thus, some species display infraspecific morphological variation and ambiguous boundaries within species. In this study, we examined the morphological characteristics, elevation, and physiology to revise this genus in Taiwan. As a result, we confirmed five species and one variety: i.e., *T. formosana* Baker, *T. lasiocarpa* Matsumura, *T. lasiocarpa* var. ovatifolia (S.S. Ying) Y.H. Chen & T.H. Huang, stat. nov., *T. lingyunyanensis* Y.H. Chen & T.H. Huang, sp. nov., *T. ravenii* C.I Peng & Tiang, and *T. suzukii* Masamune. *Tricyrtis lingyunyanensis* Y.H. Chen & T.H. Huang is a new species found only in the north of Wunfong Mountain, Chiayi Country, Taiwan, at an elevation of approximately 1100 m. The unique feature is that dried capsules fall off from the base of the capsule, and bulbils develop at the nodes of the aerial stem aboveground. Furthermore, we discovered bulb formation in this genus in Taiwan and described the growth patterns of its underground parts. This study seeks to provide a detailed assessment of the morphology of each species by offering descriptions and precise comparative photographs.