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Phenology, Seed Germination, and Genetics Explains the Reproductive Strategies of Diospyros lasiocalyx (Mart.) B. Wall

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Diospyros lasiocalyx (Mart.) B. Wall. is a dioecious tree species found in the threatened and fragmented Brazilian Cerrado. Its fleshy fruits, which are consumed extensively by wildlife, make the species ideal for use in restoration. This study includes a ten-year phenological observation, germination tests, and analysis of molecular markers to understand the reproductive strategy of D. lasiocalyx based on samples from three populations in two Cerrado ecosystems in São Paulo State, Brazil. The within population reproductive phenological pattern presented low synchrony, indicating a lack of sexual reproduction. The vegetative phenological pattern was well pronounced, with the peak of senescence occurring at the end of the dry season, just before leaf flushing. A small number of unique genets were found, indicating asexual reproduction and low genotypic diversity in the studied populations. The absence of inbreeding and the presence of ramets (stems) suggest that reproduction mainly occurs by apomixis and vegetative propagation, with some mating among unrelated stems and a low germination rate. Our study indicates that D. lasiocalyx can colonize harsh environments such as the Cerrado using alternative asexual reproductive strategies, while simultaneously attracting many seed dispersers.

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  1. Albernaz LC, Paula JE, de Romero GAS, Silva MRR, Grellier P, Mambu L, Espindola LS (2010) Investigation of plant extracts in traditional medicine of the Brazilian cerrado against protozoans and yeasts. J Ethnopharmacol 131:116–121

  2. Balloux F, Lehmann T (2003) The population genetics of clonal and partially clonal diploids. Genetics 164:1635–1644

  3. Batalha MA, Martins FR (2004) Reproductive phenology of the cerrado plant community in Emas National Park (Central Brazil). Aust J Bot 52:149–161

  4. Billotte N, Lagoda PJL, Risterucci AM, Baurens C (1999) Microsatellites-enriched libraries: applied methodology for the development of SSR markers in tropical crops. Fruits 54:277–288

  5. Borchert R (1999) Climatic periodicity, phenology, and cambium activity in tropical dry forest trees. IAWA J 20:239–247

  6. Borchert R, Rivera G, Hagnauer W (2002) Modification of vegetative phenology in a tropical semi-deciduous Forest by abnormal drought and rain. Biotropica 34:27–39

  7. Brando PM, Durigan G (2005) Changes in cerrado vegetation after disturbance by frost (São Paulo state, Brazil). Plant Ecol 175:205–215

  8. Brzosko E, Wróblewska A, Ratkiewicz M (2002) Spatial genetic structure and clonal diversity of island populations of lady's slipper (Cypripedium calceolus) from the Biebrza National Park (Northeast Poland). Mol Ecol 11:2499–2509

  9. Caetano APS, Simão DG, Carmo-Oliveira R, Oliveira PE (2013) Diplospory and obligate apomixis in Miconia albicans (Miconieae, Melastomataceae) and an embryological comparison with its sexual congener M. chamissois. Plant Syst Evol 299:1253–1262

  10. Caetano APS, Cortez PA, Teixeira SP, Oliveira PE, Carmello-Guerreiro SM (2018) Unusual diversity of apomictic mechanisms in a species of Miconia, Melastomataceae. Plant Syst Evol 304:343–355

  11. Camargo MGG, de Carvalho GH, Alberton BDC, Reys P, Morellato LPC (2018) Leafing patterns and leaf exchange strategies of a cerrado woody community. Biotropica 50:442–454

  12. Carrillo-Angeles I, Mandujano MC, Golubov J (2011) Influences of the genetic neighborhood on ramet reproductive success in a clonal desert cactus. Popul Ecol 53:449–458

  13. Cavers S, Degen B, Caron H, Lemes MR, Margis R, Salgueiro F, Lowe AJ (2005) Optimal sampling strategy for estimation of spatial genetic structure in tree populations. Heredity 95:281–289

  14. Chaves CL, Sebbenn AM, Baranoski A, Goez BD, Gaino APSC, Ruas CF, Ruas E, Ruas PE (2017) Gene dispersal via seeds and pollen and their effects on genetic structure in the facultative-apomictic Neotropical tree Aspidosperma polyneuron. Silvae Genetica 65(2):46–57

  15. Clark PL, Evans FC (1954) Distance to nearest neighbor as measure of spatial relationship in population. Ecology 35:445–456

  16. Condit R, Ashton PS, Baker P, Bunyavejchewin S, Gunatilleke S, Gunatilleke N, Hubbell SP, Foster RB, Itoh A, La Frankie JV, Lee H, Losos SE, Manokaran N, Sukumar RE, Yamakura T (2000) Spatial patterns in the distribution of tropical tree species. Science 288:1414–1418

  17. Creste S, Tulmann N, Figueira A (2001) Detection of single sequence repeat polymorphism in denaturing polyacrylamide sequencing gels by silver staining. Plant Mol Biol Report 19:299–306

  18. De Paula PC, da Silva SM, Faria KF, Frézard F, Moreira PS, Foubert K, Lopes JCD, Campana VPR, Rocha MP, Silva AF, Silva CG, Pieters L, Almeida VL (2019) Journal of Ethnopharmacology 232:155–164

  19. Degen B, Petit R, Kremer A (2001) SGS – spatial genetic software: a computer program for analysis of spatial genetic and phenotypic structures for individuals and populations. J Hered 92:447–448

  20. Dering M, Chybicki IJ, Raczka G (2015) Clonality as a driver of spatial genetic structure in populations of clonal tree species. J Plant Res 128:731–745

  21. Deus FF, Vale VS, Schiavini I, Oliveira PE (2014) Biodiversity of reproductive ecological groups in semideciduous season forests. Bioscience Journal 30:1855–1902

  22. Dias ACC, Serra AC, Sampaio DS, Borba EL, Bonetti AM, Oliveira PE (2018) Unexpectedly high genetic diversity and divergence among populations of the apomictic Neotropical tree Miconia albicans. Plant Biol 20:244–251

  23. Dick CW, Hardy OJ, Jones FA, Petit RJ (2008) Spatial scales of pollen and seed-mediated gene flow in tropical rain forest trees. Trop Plant Biol 1:20–33

  24. Dorken ME, Eckert CG (2001) Severely reduced sexual reproduction in northern populations of a clonal plant, Decodon verticillatus (Lythraceae). J Ecol 89:339–350

  25. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

  26. Durigan G, Baitello JB, Franco GADC, Siqueira MF (2004) Plantas do cerrado Paulista: imagens de uma paisagem ameaçada. Páginas & Letras, São Paulo

  27. Durigan G, Siqueira MF, Franco GADC (2007) Threats to the Cerrado remnants of the state of the São Paulo. Brazil, Scientia Agricola 64:355–363

  28. Escobar DE, Silveira FAO, Morellato LPC (2018) Timing of seed dispersal and seed dormancy in Brazilian savanna: two solutions to seasonality. Ann Bot 121:1197–1209

  29. Finger A, Kettle CJ, Kaiser-Bunbury CN (2012) Forest fragmentation genetics in a formerly wildespread island endemic tree: Vateriopsis seychellarum (Dipterocarpaceae). Mol Ecol 21:2369–2382

  30. Gaino APSC, Silva AM, Moraes MA, Alves PF, Moraes MLT, Freitas MLM, Sebbenn AM (2010) Understanding the effects of isolation on seed and pollen flow, spatial genetic structure and effective population size of the dioecious tropical tree species Myracrodruon urundeuva. Conserv Genet 11:1631–1643

  31. Garcia AS, Bressan EA, Ballester VR, Figueira A, Sebbenn AM (2019) High rates of pollen and seed flow in Hymenaea stigonocarpa on a highly fragmented savanna landscape in Brazil. New For 1:1–16

  32. Gottsberger G, Silberbauer-Gottsberger I (2006) Life in the cerrado: a south American tropical seasonal vegetation. Reta Verlag, Ulm

  33. Goudet J (2002) Fstat (version a computer program to calculate F-statistics. J Hered 86:485–486

  34. Hardy OJ, Vekemans X (2002) SPAGeDI: a versatile computer program to analyses spatial genetic structure at the individual or population levels. Mol Ecol Notes 2:618–620

  35. Hardy OJ, Maggia L, Bandou E, Breyne P, Caron H, Chevallier MH, Degen B (2005) Fine-scale genetic structure and gene dispersal inferences in 10 Neotropical tree species. Mol Ecol 15:559–571

  36. Hedrick F (2005) A standardized genetic differentiation measured. Evolution 59:1633–1638

  37. Hoffmann WA (1998) Post-burn reproduction of woody plants in a neotropical savanna: the relative importance of sexual and vegetative reproduction. J Appl Ecol 35:422–433

  38. Hoffmann WA, Solbrig OT (2003) The role of topkill in the differential response of savanna woody species to fire. For Ecol Manag 180:273–286

  39. Jacquemyn H, Brys R, Honnay O, Hermy M, Roldán-Ruiz I (2006) Sexual reproduction, clonal diversity and genetic differentiation in patchily distributed populations of the temperate forest herb Paris quadrifolia (Trilliaceae). Oecologia 147:434–444

  40. Judson OP, Normark BB (1996) Ancient asexual scandals. Trends in Ecology and Evolution 11:41–46

  41. Kalinowski ST, Taper ML, Marshall TC (2007) Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Mol Ecol 16:1099–1106

  42. Keeley JE, Pausas JG, Rundel PW, Bond WJ, Bradstock RA (2011) Fire as an evolutionary pressure shaping plant traits. Trends Plant Sci 16:406–411

  43. Keuroghlian A, Eaton DP, Desbiez ALJ (2009) The response of landscape species, white-lipped peccaries, to seasonal resource fluctuations in tropical wetland, the Brazilian Pantanal. International Journal of Biodiversity and Conservation 14:87–97

  44. Köppen W (1948) Climatologia; con un estudio de los climas de la tierra. Fondo de Cultura Economica, México

  45. Loiselle BA, Sork VL, Nason J, Graham C (1995) Spatial genetic structure of a tropical understory shrub, Psychotria officinalis (Rubiaceae). Am J Bot 82:1420–1425

  46. Lowman MD (1992) Leaf growth dynamics and herbivory in five species of Australian rain-forest canopy trees. J Ecol 80:433–447

  47. Maguire JD (1962) Speed of germination aid in selection and evaluation for seedling emergence and vigor. Crop Sci 2:176–177

  48. Marçal-Júnior O (2009) Impact of humanity on tropical ecosystems: an overview. In: Del-Claro K, Oliveira PS, Rico-Gray V (eds) Tropical biology and conservation management, 2nd edn, pp 1–22

  49. Max JCM, Melo ACG, Honda EA, Durigan G, Malícia LC, Souza MBM, Cardoso MM, Bôas OV, Ramos VS, Contiéri WA (2007) Plano de manejo da Floresta Estadual de Assis. Instituto Florestal 30:1–80

  50. Mendes-Rodrigues C, Oliveira PE (2012) Polyembryony in Melastomataceae form Brazilian Cerrado: multiple embryos in a small word. Plant Biol 14:845–853

  51. Miranda-Melo AA, Martins FR, Santos FAM (2007) Estrutura populacional de Xylopia aromatica (Lam.) Mart. e de Roupala montana Aubl. em fragmentos de cerrado no Estado de São Paulo. Revista Brasileira de Botânica 30:501–507

  52. MMA (2015) Mapeamento do Uso e Cobertura do Cerrado: Projeto TerraClass Cerrado 2013. MMA, Brasília, p 67

  53. Moraes MA, Kubota TYK, Rossini BC, Marino CL, Freita MLM, Moraes MLT, Silva AM, Cambuim J, Sebbenn AM (2018) Long-distance pollen and seed dispersal and inbreeding depression in Hymenaea stignocarpa (Fabaceae: Caesalpinioidae) in the Brazilian savannah Ecol Evol 8:7800–7816

  54. Morellato LPC, Alberti LF, Hudson IL (2010) Applications of circular statistics in plant phenology: a case studies approach. Phenological Research 1:339–359

  55. Morellato LPC, Alberton B, Alvarado ST, Borges B, Buisson E, Camargo MGG, Cancian LF, Carstensen DW, Escobar DFE, Leite PTP, Mendoza I, Rocha NMWB, Soares NC, Silva TSF, Staggemeier VG, Streher AS, Vargas BC, And Peres CA (2016) Linking plant phenology to conservation biology. Biol Conserv 195:60–72

  56. Morellato LPC, Abernethy K, Mendoza I (2018) Rethinking tropical phenology: insights from long-term monitoring and novel analytical methods. Biotropica 50:371–373

  57. Motta-Junior JC, Martins K (2002) Frugivorous diet of the maned wolf, Chrysocyon brachyurus in Brazil: ecology and conservation. In: Levey DJ, Silva WR, Galleti M (eds) Seed dispersal and frugivory: ecology, evolution and conservation, pp 291–303

  58. Myers N, Mittermeier RA, Mittermeier CG, Fonseca GAB, Kent J (2000) Biodiversity hotspots for conservation priorities. Nature 403:853–858

  59. Oliveira PE (1996) Dioecy in the cerrado vegetation of Central Brazil. Flora 191:235–243

  60. Olson MS, Hamrick JL, Moore R (2016) Breeding systems, mating systems, and genomics of gender determination in angiosperm trees. In: Groover A, Cronk Q (eds) Comparative and evolutionary genomics of angiosperm trees. Plant genetics and genomics: crops and models, vol 21. Springer, Cham, pp 139–158

  61. Pausas JG, Pratt RB, Keeley JE, Jacobsen AL, Ramirez AR, Vilagrosa A, Paula S, Kaneakua-Pia IN, Davis SD (2015) Towards understanding resprouting at the global scale. New Phytol 209:945–954

  62. Pires JPDA, Silva AGD, Freitas L (2014) Plant size, flowering synchrony and edge effects: what, how and where they affect the reproductive success of a Neotropical tree species. Austral Ecology 39:328–336

  63. Potascheff CM, Oddou-Muratorio S, Klein EK, Figueira A, Bressan EA, Oliveira PE, Lander T, Sebbenn AM (2019) Stepping stones or stone dead? Fecundity, pollen dispersal and mating patterns of roadside Qualea grandiflora Mart. Trees. Conserv Genet 20:1–13

  64. R Development Core Team (2019) A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Accessed 7 April 2019

  65. Ramos DM, Valls JFM, Borghetti F, Ooi MKJ (2019) Fire cues trigger germination and stimulate seedling growth of grass species from Brazilian savannas. Ann Bot 106:1–12

  66. Ranal MA, Santan DG, Schiavini I (2010) Are there germination patterns for cerrado species? In: Del-Claro K, Oliveira PS, Rico-Gray V, AAA B, Bonet A, Scarno FR, FJM G, Sampaio MV, Morris MR, Ramirez N, Marçal Júnior O, Macedo RHF, Marquis RJ, Coelho L, Martins RP, Rodrigues SC, Luttge U (eds) Encyclopedia of life support systems. Tropical biology and conservation management, savannah ecosystems. UNESCO/EOLSS, Oxford

  67. Ratter JA, Bridgewater S, Ribeiro JF (2003) Analysis of the floristic composition of the Brazilian cerrado vegetation. III: comparison of the woody vegetation of 376 areas. Edinburgh Journal of Botany 60:57–109

  68. Reys P, Camargo MGG, Teixeira AP, Assis MA, Grombone-Guaratini MT, Morellato LPC (2013) Estrutura e composição florística entre borda e interior de um cerrado sensu stricto e sua importância para propostas de recuperação. Hoennea 40:437–452

  69. Santana VM, Baeza MJ, Blanes MC (2013) Clarifying the role of fire heat and daily temperature fluctuations as germination cues for Mediterranean Basin obligate seeders. Ann Bot 111:127–134

  70. Sebbenn AM (2006) Sistema de reprodução em espécies arbóreas tropicais e suas implicações para a seleção de árvores matrizes para reflorestamentos ambientais. In: Higa AR, Silva L (eds) Pomares de sementes de espécies nativas. Fundação de Pesquisas Florestais do Paraná– FUPEF, Curitiba

  71. Silva CRS, Albuquerque PSB, Ervedosa FR, Mota JWS, Figueira A, Sebbenn AM (2011) Understanding the genetic diversity, spatial genetic structure and mating system at the hierarchical levels of fruits and individuals of a continuous Theobroma cacao population from the Brazilian Amazon. Heredity 106:973–985

  72. Silvertown J (2008) The evolutionary maintenance of sexual reproduction: evidence from the ecological distribution of asexual reproduction in clonal plants. Int J Plant Sci 169:157–168

  73. Souza-Silva JC, Ribeiro JF, Fonseca CEL, Antunes NB (2001) Germinação de sementes e emergência de plântulas de espécies arbóreas e arbustivas que ocorrem em Matas de Galeria. In: Ribeiro JF, Fonseca CEL, Sousa-Silva JC (eds) Cerrado: caracterização e recuperação de Matas de Galeria. Embrapa Cerrados, Brasília

  74. Spoladore J, Mansano VF, Lemes MR, De-Freitas LCD, Sebbenn AM (2017) Genetic conservation of small populations of the endemic tree Swartzia glazoviana (Taub.) Glaz. (Leguminosae) in the Atlantic Forest. Conserv Genet 118:1105–1117

  75. Stoeckel S, Grange J, Fernandez-Manjarrez JF, Bilger I, Frascaria-Lacoste N, Mariette S (2006) Heterozygote excess in a self-incompatible and partially clonal Forest tree species – Prunus avium L. Mol Ecol 15:2109–2118

  76. Tambarussi EV, Boshier D, Vencovsky R, Freitas MLM, Sebbenn A (2015) Paternity analysis reveals significant isolation and near neighbour pollen dispersal in small Cariniana legalis Mart. Kuntze populations in the Brazilian Atlantic Forest. Ecology and Evolution 5(23):5580–5600

  77. Tambarussi EV, Menezes LC, Ibañes B, Antiqueira LMOR, Dequigiovanni G, Moreno MA, Ferraz EM, Zucchi MI, Veasey EA, Vencovsky R (2016) Microsatellite markers for Cattleya walkeriana Gardner, an endangered tropical orchid species. Plant Genetic Resources 3:1–4

  78. Untergasser A, Cutcutache I, Koressaar TYJ, Faircloth BC, Remm M, Rozen SG (2012) Primer3—new capabilities and interfaces. Nucleic Acids Res 40:e115

  79. Vandepitte K, Ródan-Ruiz I, Honnay O (2009) Reproductive consequences of mate quantity versus mate diversity in wind-pollinated plant. Acta Oecology 35:548–553

  80. Vekemans X, Hardy OJ (2004) New insights from fine-scale spatial genetic structure analyses in plant populations. Mol Ecol 13:921–935

  81. Ventura A, Berengut G, Victor MAM (1966) Características edafoclimáticas das dependências do Serviço Florestal do Estado de São Paulo. Silvicultura em São Paulo 4:57–139

  82. Vogado NO, de Camargo MGG, Locosselli GM, Morellato LPC (2016) Edge effects on the phenology of the guamirim, Myrcia guianensis (Myrtaceae), a cerrado tree, Brazil. Tropical Conservation Science 9:291–312

  83. Wallnöfer B (2018) A revision of Neotropical Diospyros (Ebenaceae): part 11. Annalen des Naturhistorischen Museums in Wien 120:145–226

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We would like to thank Francisco Carlos Antoniolli and Edivaldo Furlan for assistance in sample collection. Bruna Ibanes Aguiar was supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes: Finance Code 001); Nara O. Vogado received scholarships from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Alexandre M. Sebbenn, Paulo Y. Kageyma and L.P.C. Morellato were supported by research productivity fellowships granted by CNPq. We would also like to thank Fazenda São José da Conquista and the Instituto Arruda Botelho (IAB), the Ecological Station of Itirapina and State Forest of Assis for allowing us to conduct fieldwork. The phenology observations were supported by São Paulo Research Foundation (FAPESP) grants: #2007/59779-6 and #2009/54208-6; FAPESP-Microsoft Research #2013/50155-0 and #2010/52113-5 and FAPESP-VALE-FAPEMIG #2010/51307-0 and is a contribution from the Laboratory of Phenology at UNESP. Finally, we thank Dr. Evelyn R. Nimmo for editing the English of the manuscript.

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  1. Paulo Yoshio Kageyama is deceased. This paper is dedicated to his memory.

    • Paulo Yoshio Kageyama

Correspondence to Bruna Ibanes Aguiar.

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Aguiar, B.I., Sebbenn, A.M., Tarazi, R. et al. Phenology, Seed Germination, and Genetics Explains the Reproductive Strategies of Diospyros lasiocalyx (Mart.) B. Wall. Tropical Plant Biol. 13, 23–35 (2020). https://doi.org/10.1007/s12042-019-09243-1

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  • Genotypic diversity
  • Microsatellite markers
  • Population genetics
  • Tropical tree species
  • Vegetative and reproductive phenology