Reproductive strategies in Cerrado plants: outcrossing vs. apomixis and the organization of plant-pollinator interaction webs in Central Brazil

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Reproductive strategies in Cerrado plants: outcrossing vs. apomixis and the organization of plant-pollinator interaction webs in Central Brazil Paulo Eugênio Oliveira Universidade Federal de Uberlândia - MG

Cerrado: Neotropical savanna areas in Central Brazil Felipe Ribeiro

Cerrado Physiognomies Gallery forest Campos Cerrado Cerradão

Deforestation and land use in Cerrado Klink & Moreira 2002

Cerrado as a conservation hot spot Forestry 0,07% Agriculture 11,35% Cultivated pastureland 41,56% Urban areas 1,90% others 0,59% Native areas 44,53%

Floristic basis 376 sites Ratter et al, 2003 Bridgewater et al. 2004 Woody species = 953 spp Herbs/woody ratio= 3-4/1 No species occurred in all sites Gallery forest and riverine woody species = 1 900 spp Total = > 12000 spp High β diversity and species turn-over Mendonça e at (1998), Ratter et al (2003), Felfili et al (2001), Sano et al. (2008)

Cerrado Determinants Seasonal climate Fire Nutrient poor soils

Water use and seasonality Herbaceous plants with shallow root system Woody species with deep roots

Fire Accumulated biomass and fire

Traditional ideas suggested fire and environmental restrictions would render sexual reproduction of woody species very difficult and would favor asexual reproduction

Malvaceae (Gottsberger 1986) 1986) Woody hummingbirds and bat pollinated species with specialized flowers Herbaceous bee pollinated promiscuous groups Gallery forest Campos Cerrado Cerradão

Diversity of pollination systems Oliveira & Gibbs 2000

Breeding systems of woody species in Netropical areas Cerrado Dry Secondary Cloudy Dry Lowland forest forest forest forest rainforest BSB Costa Rica Venezuela Venezuela Mexico Costa Rica Dioecy 15 22 24 31 20 23 Monoecy 5 10 0 3 10 11 Self-incompat. 66 54 64 26 53 53 Autogamous 14 14 12 43 17 13 Outcrossing 81 76 88 57 73 75 Oliveira & Gibbs 2000

Distribution of Vochysia (Vochysiaceae) in Cerrado plant formations. V. elliptica V. pumila V. pyramidalis V. rufa V. thyrsoidea V. tucanorum Polinizadores, vicariancia, fenologia e processo de especiação Oliveira & Gibbs 1994 - self-incompatible - dependent on large bees for fruit set. - guild of plants (including other groups) and large bees

Sphingids Guilds and interaction web Plants Amorim 2008

Average proboscis lenght (cm) Blind connections: association between hawkmoths and plants in Cerrado. Average flower lenght (cm) Amorim 2008

Cláudia Inês da Silva Pollen load on Xylocopa bees Xylocopa (Neoxylocopa) grisescens Lepeletier 1,75cm 1,25 cm Xylocopa (Neoxylocopa) frontalis (Olivier) 1,75cm 1,25 cm Xylocopa (Neoxylocopa) suspecta (Moure & Camargo) Cláudia Inês Silva

Xylocopa frontalis Xylocopa frontalis Xylocopa suspecta Xylocopa suspecta Xylocopa grisecens Xylocopa grisescens Xylocopa hirsutissima Xylocopa hirsutissima 30 plant spp. intensely used Cláudia Inês Silva

Wasps 50 Floral visitors on Matayba guianensis (Sapindaceae) in Cerrados of Uberlândia Bees 110! Beetles 37 Ana Coelho Carvalho, Unpublished data

Guilds, redundancy and resilience Obligatory outcrossing: dependency on pollinators. Specialized relationships are rare. Generalist guilds? Redundancy and stability? Guilds and resilience after disturbance. Adjust to high β-diversity?

Apomixis in Cerrado Polyembryony and apomixis Specific Grupos groups Específicos Salomão & Allem 2001 Allem 2003 Melastomataceae 39% Miconieae Eriotheca pubescens Importance of apomixis 63% Goldenberg & Shepherd 1998 Oliveira et al. 1992 Martins & Oliveira 2003 Mendes-Rodrigues et al. 2005

Questions: How common is polyembryony among cerrado plants? Is polyembryony a diagnostic feature to estimate importance of apomixis in Cerrado plants? What is the origin of apomictic taxa? How widespread are polyembryony and apomixis in Cerrado environment? Consequences and perspectives? Apomixis in Cerrado plants CNPq research grant (2005-2007)

Polyembryony analysis Family Species Method Mean SD N SE Min. Max Mono Poly %M %P Anacardiaceae Astronium fraxinifolium Embryos 1,00 0,00 253,00 0,00 1,00 1,00 253,00 0,00 100,00 0,00 Anacardiaceae Astronium fraxinifolium Seedlings 1,01 0,08 697,00 0,00 1,00 2,00 692,00 5,00 99,28 0,72 Anacardiaceae Lithraeae molleoides Seedlings 1,00 0,00 1.335,00 0,00 1,00 1,00 1.335,00 0,00 100,00 0,00 Anacardiaceae Myracrodruon urundeuva Embryos 1,01 0,11 250,00 0,01 1,00 2,00 247,00 3,00 98,80 1,20 Anacardiaceae Myracrodruon urundeuva Seedlings 1,01 0,10 486,00 0,00 1,00 2,00 481,00 5,00 98,97 1,03 Apocynaceae Aspidosperma cylindrocarpum Seedlings 1,00 0,00 221,00 0,00 1,00 1,00 221,00 0,00 100,00 0,00 Bignoniaceae Jacaranda cuspidifolia Seedlings 1,01 0,11 404,00 0,01 1,00 3,00 402,00 2,00 99,50 0,50 Bignoniaceae Tabebuia avellanedae Seedlings 1,00 0,00 346,00 0,00 1,00 1,00 346,00 0,00 100,00 0,00 Bignoniaceae Tabebuia caraiba Seedlings 1,00 0,00 132,00 0,00 1,00 1,00 132,00 0,00 100,00 0,00 Bignoniaceae Tabebuia chysotricha Embryos 2,18 1,12 606,00 0,05 1,00 7,00 192,00 414,00 31,68 68,32 Bignoniaceae Tabebuia chysotricha Seedlings 1,66 0,74 563,00 0,03 1,00 5,00 270,00 293,00 47,96 52,04 Bignoniaceae Tabebuia ochracea Seedlings 1,36 0,48 45,00 0,07 1,00 2,00 29,00 16,00 64,44 35,56 Bignoniaceae Tabebuia roseo-alba Seedlings 1,01 0,08 676,00 0,00 1,00 2,00 672,00 4,00 99,41 0,59 Bignoniaceae Tabebuia serratifolia Embryos 1,26 0,57 46,00 0,08 1,00 4,00 36,00 10,00 78,26 21,74 Bignoniaceae Tabebuia serratifolia Seedlings 1,18 0,38 200,00 0,03 1,00 2,00 165,00 35,00 82,50 17,50 Caesalpiniaceae Copaifera langsdorfii Seedlings 1,00 0,04 577,00 0,00 1,00 2,00 576,00 1,00 99,83 0,17 Caesalpiniaceae Hymenaea courbaril Seedlings 1,00 0,03 1.173,00 0,00 1,00 2,00 1.172,00 1,00 99,91 0,09 Caesalpiniaceae Senna sylvestris Seedlings 1,00 0,05 346,00 0,00 1,00 2,00 345,00 1,00 99,71 0,29 Clusiaceae Kielmeyera coriaceae Embryos 1,00 0,00 62,00 0,00 1,00 1,00 62,00 0,00 100,00 0,00 Clusiaceae Kielmeyera grandiflora Embryos Seeds 1,00 0,00 from 200,00 10 different 0,00 1,00 1,00 200,00 0,00 100,00 0,00 Clusiaceae Kielmeyera variabilis Embryos plants 1,00 0,00 for each 50,00 species 0,00 1,00 in 1,00 50,00 0,00 100,00 0,00 Cocholospermaceae Cocholospermum regium Embryos 1,00 0,00 62,00 0,00 1,00 1,00 62,00 0,00 100,00 0,00 Connaraceae Rourea induta Embryos the 1,03triângulo 0,16 40,00 mineiro 0,03 1,00 region 2,00 39,00 1,00 97,50 2,50 Erythroxylaceae Erythroxylum daphnites Embryos 1,00 0,00 139,00 0,00 1,00 1,00 139,00 0,00 100,00 0,00 Euphorbiaceae Manihot anomala Embryos 1,00 0,00 60,00 0,00 1,00 1,00 60,00 0,00 100,00 0,00

Polyembryony and apomixis Our survey: 70 species 27 families Total (+ Salomão & Allem): 125 species 39 families 57% Polyembryonic 43% 71% Monoembryonic 29% Polyembryonic Monoembryonic Apomictics?

Polyembryonic and apomictic? Polyembryonic > 5 % seeds 6% Monoembryonic 71% 23% 90,00 80,00 70,00 60,00 50,00 40,00 30,00 20,00 Species ranked by frequency of apomictic seeds Polyembryonic < 5 % seeds 10,00 0,00 0 20 40 60 80 100 120 140 5%

Polyembryonic species and breeding systems Family Species Polyembryony % Source BS Anacardiaceae Astronium fraxinifolium Poli 0,72 Original data Dioecious Anacardiaceae Myracrodruon urundeuva Poli 2,00 Original data Dioecious Anacardiaceae Tapirira guianensis Poli 2,00 Salomão e Allem 2001 Dioecious Apocynaceae Hancornia speciosa Poli 5,00 Salomão e Allem 2001 SI Bignoniaceae Jacaranda cuspidifolia Poli 0,50 Original data ni Bignoniaceae Tabebuia chysotricha Poli 68,32 Original data Apomictic Bignoniaceae Tabebuia ochracea Poli 35,56 Original data SI/Apomictic Bignoniaceae Tabebuia roseo-alba Poli 0,59 Original data SI Bignoniaceae Tabebuia serratifolia Poli 21,74 Original data SI/Apomictic Burseraceae Commiphora lepthophloeos Poli 2,00 Salomão e Allem 2001 ni Caesalpiniaceae Copaifera langsdorfii Poli 0,17 Original data SI Caesalpiniaceae Hymenaea courbaril Poli 0,09 Original data SI Caesalpiniaceae Senna sylvestris Poli 0,29 Original data SI Connaraceae Rourea induta Poli 2,50 Original data SI Fabaceae Machaerium acutifolium Poli 2,22 Original data ni Fabaceae Ormosia arborea Poli 0,25 Original data ni Flaucortiaceae Casearia sylvestris Poli 0,89 Original data SC Hippocrateaceae Salacia campestris Poli 2,08 Original data ni Hippocrateaceae Salacia crassifolia Poli 1,94 Original data ni Malpighiaceae Byrsonima basiloba Poli 3,00 Salomão e Allem 2001 ni Malvaceae Eriotheca pubescens Poli 81,82 Original data Apomictic Malvaceae Guazuma ulmifolia Poli 1,21 Original data SI Melastomataceae Clidemia hirta Poli 12,53 Original data Apomictic Melastomataceae Miconia albicans Poli 1,51 Original data Apomictic Melastomataceae Miconia ferruginata Poli 24,74 Original data Apomictic Mimosaceae Acacia polyphylla Poli 2,00 Original data Mimosaceae Inga laurina Poli 41,33 Original data Apomictic? Myrtaceae Campomanesia pubescens Poli 0,12 Original data SI/SC Myrtaceae Eugenia calycina Poli 1,31 Original data ni Myrtaceae Eugenia hegeliana Poli 3,40 Original data ni Myrtaceae Eugenia punicifolia Poli 0,45 Original data SC Rhamnaceae Zizyphus joazeiro Poli 2,00 Salomão e Allem 2001 SC Rubiaceae Genipa americana Poli 3,00 Salomão e Allem 2001 Dioecious Sapindaceae Magonia pubescens Poli 3,00 Salomão e Allem 2001 ni Vochysiaceae Callisthene fasciculata Poli 0,22 Original data SI Vochysiaceae Callisthene major Poli 0,57 Original data SI 36 species 17 families At least 11 SI Contrasting breeding system results in some putative apomictics

Polyembryony without apomixis Callisthene major Callisthene species (Vochysiaceae) R. Carmo-Oliveira & L.N. Custódio unpublished data Callisthene fasciculata

Callisthene major

Callisthene fasciculata Qualea multiflora

Polyembryony and sporophitic apomixis Eriotheca pubescens Mendes-Rodrigues et al. 2005 Plant biology 7:533-540

Eriotheca gracilipes Eriotheca pubescens

Frequency (%) Handroanthus (syn. Tabebuia) Bignoniaceae 45 40 35 30 25 20 15 10 5 0 Embryos Seedlings 1 2 3 4 5 6 7 Number Embryos and seedlings per seed of Tabebuia chrysotricha (embryos n = 606 seeds; seedlings n = 640 seeds). Costa et al. 2004 Sampaio et al. 2007

Apomixis and Neopolyploidy Eriotheca pubescens 2n = 40 2n = 80 2n= 276 Oliveira et al. 1992 Sampaio et al. 2008

Apomixis without polyembryony Miconia albicans Goldenberg & Shepherd 1998 Caetano 2007 Cortez et al. 2008

How widespread are apomictic taxa? Handroanthus ochraceus Eriotheca pubescens Miconia albicans Between the 125 most frequent woody species in Cerrado (ca. 1000 spp.) Bridgewaters et al. 2004

Breeding systems mosaics? Eriotheca pubescens 2n=80 A B 2n=40 Monoembryonic individuals Polyembryonic individuals 100 Km Handroanthus ochraceus (syn. Tabebuia ochracea)

Species Breeding system and distribution in the Melastomataceae 40 84% 35 30 wide restrict 60% 25 20 75% 40% 15 10 5 16% 25% 0 Apomictis Self-incompatible Self compatible Breeding system Apomictics widespread Mila et al. in preparation

Questions: How common is polyembryony among cerrado plants? Is polyembryony a diagnostic feature to estimate importance of apomixis in Cerrado plants? What is the origin of apomictic taxa? Hybridization? Polyploidy? ± How widespread are polyembryony and apomixis in Cerrado environment? Widely distributed apomictics Consequences: apomixis as a reproductive alternative?

Disturbance and reduced gene flow Caryocar brasiliense Foto: Rogério Gribel Collevatti, R.G., Grattapaglia, D., & Hay, J.D. 2001. Population genetic structure of the endangered tropical tree species Caryocar brasiliense, based on variability at microsatellite loci. Molecular ecology 10: 349-356 Collevatti, R.G., Grattapaglia, D., & Hay, J.D. 2001. High resolution microsatellite based analysis of the mating system allows the detection of significant biparental inbreeding in Caryocar brasiliense, an endangered tropical tree species. Heredity 86 (1): 60-67

Miconia albicans The future of Cerrado? - Independent of pollinators - Bird dispersed fruits and viable seeds - But no pollen for bees

Renata Carmo-Oliveira Felipe W. Amorim Diana Sampaio Ana Paula Caetano Ana Paula Mila Rosana Romero Luciana N. Custódio Pietro K. Maruyama Clesnan Mendes-Rodrigues Daniela Simão Peter E. Gibbs