Anthurium Reproductive Morphology and Evolution

Reconstructing the Evolutionary History of Reproductive Characters in Anthurium (Araceae) Benjamin Durrington Sofia Wolfson Mentor: Dr. Mónica Carlsen

® REU Coordinator: Dr. David Bogler Overview I. Introduction II. Research Goals III.Methods IV. Results & Discussion V. Conclusion VI. Future Research I. Introduction Anthurium (Araceae) • 912 species (and counting) • Large Neotropical distribution • Present in a variety of ecosystems • Variable vegetative and reproductive morphology • Usually bee, beetle or fly-pollinated • Epiphytic, lithophytic, or terrestrial • Some members economically important

https://earth.google.com/ I. Introduction Classification

• Croat & Sheffer (1983) proposed 18 sections and 2 series • Carlsen & Croat (2013) generated molecular phylogeny with 17 highly supported clades • Series Multinervia, series Pachyneurium, and sections Calomystrium, Leptanthurium, Tetraspermium, Dactylophyllium, and Polyphyllium supported by Carlsen & Croat Carlsen & Croat (2013)

I. Introduction Characters and Classification Grayum (1990) • For classification, reproductive morphology is generally more useful than vegetative morphology • Reproductive morphological changes  reproductive isolation  speciation Carlsen & Croat (2013) • “Reproductive characters…seem to be more reliable to characterize clades within Anthurium”

II. Research Goals Research Goals

1. Use reproductive morphology to characterize clades proposed by Carlsen & Croat (2013) 2. Evaluate the importance of reproductive character shifts in the evolution of Anthurium

A. huixtlense A. formosum A. flexile A. acaule A. willdenowii aroidpictures.fr III. Methods Materials and Methods: Data Collection

A. armeniense Reproductive morphology: • 32 characters • 98 species A. oxyphyllum Pollen: • 15 characters • 60 species III. Methods Inflorescence Data Collection

A. antioquiense

Observation of specimen under A. andicola A. olfersianum dissecting scope

Berries Spadix Epiphytic bryophyte III. Methods Reproductive characters (32) Inflorescence (18) Fruit (6) Flower (8) Quantitative (6) Qualitative (12) Quantitative (2) Qualitative (4) Quantitative (6) Qualitative (2) • Peduncle length • Inflorescence length/leaf length • Berry • Berry color • Flowers/ • Stamens • Spathe length • Peduncle thickness /petiole length • Shape of right spiral habit • Spathe width thickness • Berry width berry tip • Flowers/ • Presence • Stipe length • Peduncle length/petiole length • Berry seed left spiral of tepal • Spadix length • Spathe shape abundance • Flowers/ protuber- • Spadix diameter • Spathe 3D-shape • Spathe principle ances • Spathe disposition at anthesis persistence spiral • Spathe color when plant is • Flowers/se • Presence of punctations on in fruit condary spathe spiral • Presence of stipe • Flower • Spadix shape length • Spadix position • Flower • Spadix color width

A. moodeanum A. muyunense A. buganum A. wagenurianum A. crenatum III. Methods Collecting Pollen Data

Pollen from herbarium specimens

Kimwipes™ packets Removal of anthers Eppendorf tubes

Pollen from pickled collections

Dehydration series

Sputter-coater (Denton Vacuum Desk V)

Scanning electron microscope (JCM 5000 Critical-point dryer (Emitech K850) Neoscope) III. Methods Pollen Characters (15) Quantitative (5) Qualitative (10) • Lumen size • Pollen unit • Pollen range of sizes • Pollen shape • Pollen size • Pollen symmetry • Pollen range (variability) • Pollen polarity • Aperture number • Pollen aperture type • Pollen aperture shape • Exine ornamentation • Muri type • Presence of crystals • Other characters

A. digitatum A. besseae A. friedrichsthalii III. Methods Materials and Methods: Data Analysis • Mesquite software (Maddison & Maddison 2011) used to map character states onto molecular phylogeny (Carlsen & Croat 2013), for 47 different characters • Trace Character History function used under Maximum Parsimony • Synapomorphies identified for each clade IV. Results Results: Clade synapomorphies

Anthurium_andicola Character: Anthurium_parvispathum Anthurium_berriozabalense Tepals_protuberances_( Anthurium_lancetillense presence) Anthurium_clarinervium Anthurium_leuconeurum Parsimony Anthurium_umbrosum reconstruction ( Anthurium_pedatoradiatum Anthurium_sagittatum Unordered) [Steps: 6] Anthurium_trilobum 0 Anthurium_balaoanum 1 Anthurium_dolichostachyum Anthurium_sp_1 Anthurium_cuspidatum Anthurium_urbanii Anthurium_longicaudatum Anthurium_mindense Anthurium_microspadix Anthurium_furcatum Anthurium_macleanii Anthurium_pedatum Anthurium_tremulum Anthurium_antioquiense Anthurium_formosum Anthurium_hoffmannii Anthurium_obtusilobum Anthurium_veitchii Anthurium_curvispadix Anthurium_huixtlense Anthurium_dressleri Anthurium_subsignatum Anthurium_crystallinum Anthurium_sp_2 Anthurium_brownii Anthurium_besseae Anthurium_willifordii Anthurium_oxyphyllum Anthurium_napaeum Anthurium_carchiense Anthurium_palenquense Anthurium_lancea Anthurium_cf_michelii Anthurium_maculosum Anthurium_michelii Anthurium_fornicifolium Anthurium_spectabile Anthurium_bradeanum Anthurium_halmoorei Clade 9: pointed berries Anthurium_schottianum Anthurium_spathiphyllum Anthurium_protensum Anthurium_crenatum Anthurium_venosum Anthurium_gracile Anthurium_vittariifolium Anthurium_sp_4 Anthurium_bakeri Anthurium_lancifolium Anthurium_sp_3 Anthurium_margaricarpum Anthurium_ovatifolium Anthurium_punctatum Anthurium_wendlingeri Anthurium_morae Anthurium_bicollectivum Anthurium_peltigerum Anthurium_friedrichsthalii Anthurium_citrifolium Anthurium_laciniosum Anthurium_cf_obtusum Anthurium_obtusum Anthurium_scandens Anthurium_chiapasense Anthurium_lucens Anthurium_verapazense Anthurium_brevipedunculatum Anthurium_clavigerum Anthurium_sinuatum Anthurium_digitatum Anthurium_longissimum Anthurium_thrinax Anthurium_eminens Anthurium_croatii Anthurium_polydactylum Anthurium_kunthii Anthurium_acaule Anthurium_cordifolium Anthurium_willdenowii Anthurium_cordatum Anthurium_grandifolium Anthurium_dominicense Anthurium_bellum Anthurium_longipes Anthurium_radicans Anthurium_bromelicola Anthurium_harrisii Anthurium_lucidum Anthurium_olfersianum Anthurium_coriaceum Anthurium_parvum Anthurium_flexile Clade 10: purple berries Anthurium_clidemioides

Modified from GYCHACHS MrB Anthurium_only Tepal protuberances: present or absent IV. Results Results: Best characters

• Fifty-eight synapomorphic character states identified • Twenty-nine (62%) useful characters • 100% fruit characters, 67% general inflorescence characters, 63% flower characters, and 40% pollen characters yielded synapomorphies • Berry color, spadix color, peduncle/petiole length ratio, and berry width were most important IV. Results

Each clade was supported by 1-9 synapomorphies

Clades A, 3, 4 and 7 were best supported IV. Results Clade A Clade 3

A. digitatum A. flexile

25 µm

http://sura.ots.ac.cr/ Synapomorphy: orange, pointed berries Synapomorphy: large pollen grains Clade 7 Clade 13

A. huixtlense A. bakeri

Synapomorphy: spinulose muri Synapomorphy: white/lavender spadix Conclusion Since all 17 clades could be identified with reproductive character shifts, we conclude that reproductive changes have played a major role in the diversification of Anthurium, as proposed by Grayum (1990).

Future Studies

• Increase sample size, given that current datasets include only ca. 7-11% of Anthurium species (some characters may be more or less important when sampling size is increased) • Include more characters • Redefine some character states (e.g. colors) • Pollination and fruit dispersal could be compared to morphological data Acknowledgements • Angie Macias • Dr. Tom Croat • Washington University – Microscopy & Digital Imaging Core • REU Students • Missouri Botanical Garden and Staff • National Science Foundation – REU Program