Assessing the genetic diversity of Myrsine umbellata (Primulaceae) in Brazilian Atlantic Forest remnants - an important step towards reforestation efforts

Micheli Sossai Spadeto Thais Lazarino Maciel Tatiana Tavares Carrijo Marcia Flores da Silva Ferreira Milene Miranda Praça Fontes About the authors

Abstract

The investigation of genetic diversity in natural populations of species that show potential for use in reforestation programs is a key step in making management decisions. However, reforestation programs with native species in Brazil are still rarely based on a genetic understanding of the seed matrices used for seedling production. This is also the case for Myrsine umbellata, a dioecious shrub within the family Primulaceae that has been used in reforestation programs in Brazil, mainly due to its high production capacity of fruits attractive to the avifauna. The goal of this study was to measure intra- and interpopulational genetic diversity in natural populations of M. umbellata in six forest remnants of the Atlantic Forest using ISSR markers. The results revealed that the intrapopulational genetic diversity was greater than the genetic diversity among the studied populations. For this reason, the cultivation of seedlings from seeds obtained in more than one population seems the most appropriate strategy for reforestation purposes. Even though the most isolated populations are also the ones with highest genetic structure, all populations of M. umbellata included in this study revealed to be an important germplasm bank conserved in situ.

Key words:
Capororoca; ISSR markers; Myrsinoideae; tropical forests

Resumo

Investigar a diversidade genética em populações naturais de espécies com potencial para uso em programas de reflorestamento é um passo importante para a tomada de decisões relacionadas ao planejamento e manejo dos plantios. No entanto, programas de reflorestamento com espécies nativas no Brasil raramente são realizados com base no conhecimento do genótipo das matrizes doadoras das sementes utilizadas para produção de mudas. Este é o caso de Myrsine umbellata, um arbusto dioico pertencente à família Primulaceae, que vem sendo utilizado em programas de reflorestamento no Brasil principalmente por apresentar elevada capacidade de produção de frutos atrativos à avifauna. O objetivo deste estudo foi mensurar a diversidade genética intra e interpopulacional em populações naturais de M. umbellata em seis remanescentes florestais da Floresta Atlântica, utilizando marcadores ISSR. Os resultados revelaram que a diversidade genética intrapopulacional foi maior que a diversidade genética entre as populações estudadas. Por essa razão, o cultivo de mudas a partir de sementes obtidas em mais de uma população parece ser a estratégia mais apropriada para reflorestamento. Mesmo considerando que as populações mais isoladas são aquelas mais estruturadas, todas as populações de M. umbellata incluídas neste estudo revelaram ser importantes bancos de germoplasma conservados in situ.

Palavras-chave:
Capororoca; marcadores ISSR; Myrsinoideae; tropical forests

Introduction

Anthropogenic actions have been modifying the structure of tropical forests, particularly the Atlantic Forest domain and its associated ecosystems (Magnago et al. 2014Magnago LF, Edwards DP, Edwards FS, Magrach A, Martins SV & Laurance WF (2014) Functional attributes change but functional richness is unchanged after fragmentation of Brazilian Atlantic forests. Journal of Ecology 102: 475-485.). These changes lead to a reduction in population density among species affecting their reproduction, pollination and the gene flow between and within populations (De Lacerda et al. 2013De Lacerda AEB, Roberta NE & Sebbenn AM (2013) Modeling the long-term impacts of logging on genetic diversity and demography of Hymenaea courbaril. Forest Science 59: 15-26.). The consequent forest fragmentation process may lead to the reduction of genetic variability between populations due to the isolation of populations and their individuals, and the increase of self-fertilizations and correlated mattings (Young & Boyle 2000Young AG & Boyle TJ (2000) Forest fragmentation. In: Young A, Boshier D & Boyle T (eds.) Forest conservation genetics. CABI Publishing, Wallingford. Pp. 123-135.; Duminil et al. 2016Duminil J, Dainou K, Kaviriri DK, Gillet P, Loo J, Doucet JL & Hardy OJ (2016) Relationships between population density, fine-scale genetic structure, mating system and pollen dispersal in a timber tree from African rainforests. Heredity 116: 295-303.). In this context, the use of genetic markers in population diversity studies of tree species is a powerful tool to help with decision-making on management and conservation.

The assessment of genetic diversity of a species at the intra- and interpopulational level allows inferences about the genetic structure of the population, gene flow and potential genetic bottlenecks, due to habitat fragmentation or other spatial or temporal barriers (Mondini et al. 2009Mondini L, Noorani A & Pagnotta MA (2009) Assessing plant genetic diversity by molecular tools. Diversity 1: 19-35.). In addition, genetic studies at population level can be applied to identify individuals with satisfactory genetic characteristics to be used as seed matrices to be collected for recovery and restoration of degraded areas (Rodrigues et al. 2009Rodrigues RR, Lima RAF, Gandolfi S & Nave AG (2009) On the restoration of high diversity forests: 30 years of experience in the Brazilian Atlantic Forest. Biological Conservation 142: 1242-1251.). For these purposes, it is essential to increase the knowledge about genetic diversity of interesting species for reforestation.

In the context of reforestation, Myrsine umbellata Mart. (Primulaceae) stands out as a facilitating species in areas of natural regeneration, which supplies fruit and shelter for the avifauna (Backes & Irgang 2002Backes P & Irgang B (2002) Árvores do sul: guia de identificação e interesse ecológico. Editora Palloti, Santa Maria. 325p.; Pascotto 2007Pascotto MC (2007) Rapanea ferruginea (Ruiz & Pav.) Mez. (Myrsinaceae) como importante fonte alimentar para as aves em uma mata de galeria no interior do estado de São Paulo. Revista Brasileira de Zoologia 24: 735-741.). Also, M. umbellata is the second most widely distributed species of Myrsine in South America (Sánchez-Tapia et al. 2018Sánchez-Tapia A, Garbin ML, Siqueira MF, Guidoni-Martins KG, Scarano FR & Carrijo TT (2018) Environmental and geographical space partitioning between core and peripheral Myrsine species (Primulaceae) of the Brazilian Atlantic Forest. Botanical Journal of the Linnean Society 187: 633-652.). The members are dioecious shrubs, with ornithochoric seed dispersal (Jung-Mendançolli et al. 2005Jung-Mendaçolli SL, Bernacci LC & Freitas MF (2005) Myrsinaceae. In: Wanderley MGL, Shepherd GJ, Melhem TS & Giulietti AM (eds.) Flora fanerogâmica do estado de São Paulo. Instituto de Botânica, São Paulo. Vol. 4, pp. 279-300.). These attributes made this species, and its close relatives in the genus Myrsine, important components in reforestation programs in Brazil. However, until now seed obtainment has been carried out without considering the genetic variability of natural populations.

Inter Simple Sequence Repeat (ISSR) is a DNA marker widely used to estimate the genetic diversity of natural plant populations (e.g., Sheng et al. 2017Sheng F, Chen S, Tian J, Li P, Qin X, Wang L, Luo S & Li J (2017) Morphological and ISSR molecular markers reveal genetic diversity of wild hawthorns (Crataegus songorica K. Koch.) in Xinjiang, China. Journal of Integrative Agriculture 16: 2482-2495.; White et al. 2018White LAS, Silva AVC, Alvares-Carvalho SV, Silva-Mann R, Arrigoni-Blank MF, Souza EMS, Almeida CS, Nizio DAC, Sampaio TS & Blank AF (2018) Genetic diversity of a native population of Myrcia ovata (Myrtaceae) using ISSR molecular markers. Genetics and Molecular Research 17: gmr18022.). This marker was previously applied to study the genetic diversity of M. coriacea (Sw.) R. Br. ex Roem. & Schult. (Paschoa et al. 2018Paschoa RP, Christ JA, Valente CS, Ferreira MFS, Miranda FD, Garbin ML & Carrijo TT (2018) Genetic diversity of populations of the dioecious Myrsine coriacea (Primulaceae) in the Atlantic Forest. Acta Botanica Brasilica 32: 376-385.). Here we used the same molecular marker to measure the genetic diversity in natural populations of M. umbellata found in forest remnants of Atlantic Forest. Our main goal was to improve the knowledge of genetic diversity in the selected species in order to guide practices that allow the use of M. umbellata in reforestation programs.

Material and Methods

Populations studied and leaf samples collection

Healthy young leaves of 63 individuals of Myrsine umbellata were collected at six different locations in the Atlantic Forest (Fig. 1) in the state of Espírito Santo following a previous study on another species in the same genus (Paschoa et al. 2018Paschoa RP, Christ JA, Valente CS, Ferreira MFS, Miranda FD, Garbin ML & Carrijo TT (2018) Genetic diversity of populations of the dioecious Myrsine coriacea (Primulaceae) in the Atlantic Forest. Acta Botanica Brasilica 32: 376-385.). Samples were taken only from individuals which had a height of 4 to 5 m, a diameter at breast height (DBH) of at least 30 cm, and had clearly reached reproductive age. The leaf samples were stored individually in paper bags, sealed, labelled, and kept on silica gel until transported to the laboratory.

Figure 1
a-c. Geographic locations of the six studied natural populations of Myrsine umbellata - a. map of Brazil and natural distribution of the Atlantic Forest (Fundação SOS Mata Atlântica 2019Fundação SOS Mata Atlântica (2019) Atlas dos Remanescentes Florestais da Mata Atlântica. Relatório técnico 2017-2018. Fundação SOS Mata Atlântica, São Paulo. 35p.); b. state map of Espirito Santo; c. localities of the studied populations[ blue: population 1 (municipality of Dores do Rio Preto, locality Macieira in the Caparaó National Park - 20º41’20”S, 41º50’43”W); pink: population 2 (municipality of Ibitirama, locality on rural property - 20º32’29”S, 41º40’02”W); dark green: population 3 (municipality of Iúna, locality Serra do Valentim - 20º20’45”S, 41º32’09”W ); light green: population 4 (municipality of Castelo, locality Forno Grande State Park - 20º36’13”S, 41º11’05”W); yellow: population 5 (municipality of Domingos Martins, locality Pedra Azul State Park - 20º21’48”S, 40º39’33”W); orange: population 6 (municipality of Santa Teresa, locality on rural property 19º56’08”S, 40º36’01”W) ].

DNA extraction and ISSR marker analysis

The total genomic DNA was isolated and purified using the extraction methodology by Doyle & Doyle (1990Doyle JJ & Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13-15.), with modifications following Ferreira & Grattapaglia (1998Ferreira ME & Grattapaglia D (1998) Introdução ao uso de marcadores moleculares em análise genética. 3ª ed. Embrapa Cenargen, Brasília. 220p.). DNA quality and concentration were verified using a nanoDrop TM 2000 spectrophotometer (Thermo Scientific). Integrity was confirmed with gel electrophoresis in 0.8% agarose gel.

A screening was performed with 32 ISSR primers obtained from British Columbia University, Vancouver, Canada and, based on the highest number of polymorphic fragments presented and the quality of amplification, the ten best were selected (Tab. 1). We selected ISSR markers due to the absence of SSR (codominant) markers for this species. It is has been shown that polymorphism analysis based on ISSR generates robust and highly polymorphic data for both intraspecific and interspecific analyses within the genus. In addition, ISSRs are considered neutral genome markers, an important feature for studying patterns of genetic dispersion, diversity and genetic structure in natural populations (Paschoa et al. 2018Paschoa RP, Christ JA, Valente CS, Ferreira MFS, Miranda FD, Garbin ML & Carrijo TT (2018) Genetic diversity of populations of the dioecious Myrsine coriacea (Primulaceae) in the Atlantic Forest. Acta Botanica Brasilica 32: 376-385.).

Table 1
The ten ISSR primers selected for amplification of DNA fragments from the six studied Myrsine umbellata populations. Descriptive analysis of the primers with the respective sequences, total number of bands, total number of polymorphic bands and percentage of polymorphism.

Volumes of 20 µL containing 8.3 mM enzyme mix for PCR (Promega®) 1X, 0.5 µM primer, 1 unit of Taq polymerase and 60 ng of DNA were used for amplification. Amplification conditions were: denaturation 5 minutes at 94 ºC, followed by 40 cycles of 1 minute at 94 ºC; 1 minute at 55 ºC and 1 minute at 72 ºC, with a final extension phase of 2 minutes at 72 ºC. The fragments obtained from the PCR reactions were separated by horizontal gel electrophoresis in 1.5% agarose gel containing 0.02 µL/mL ethidium bromide and TBE running buffer (Tris-base, boric acid and EDTA), at 90 volts for about 1 hour and 15 minutes. After electrophoresis, gels were visualized with an imaging system (BioRad Gel DocTM EZ Imager) in acordance to Carvalho et al. 2020Carvalho MS, Ferreira MFS, Oliveira WBS, Marçal TS, Guilhen JHS, Mengarda LHG & Ferreira A (2020) Genetic diversity and population structure of Euterpe edulis by REML/BLUP analysis of fruit morphology and microsatellite markers. Crop Breeding and Applied Biotechnology 20: e31662048., with modification.

Data analysis

To analyze the molecular data, the monomorphic and polymorphic bands were coded as absent (0) or present (1), respectively. The polymorphic loci of the 10 selected primers were used to generate a matrix of binary data of individuals (rows) by bands (columns). The Sorensen-Dice coefficient (Dice 1945Dice LR (1945) Measures of the amount of ecological association between species. Ecology 26: 297-307.) was used to calculate genetic dissimilarity among individuals. Dissimilarity averages were used to run a grouping analysis of the individuals following the UPGMA method (Unweighted Pair-Group Method Average).

From this matrix, the values of ΦST between the pairs of populations were estimated in the program GENES (Cruz 2013Cruz CD (2013) Programa Genes: Biometria. Editora UFV, Viçosa. 382p.). The values of Φ are analogous to the traditional F statistic, so that increasing positive values of ΦST (between 0 and 1) indicate increasing genetic differentiation between populations.

The consistency and stability of the formed clusters were calculated using the bootstrap reliability index, generating a dendrogram from 1,000 permutations. The analysis of molecular variance (AMOVA, Excoffier et al. 1992) was performed to estimate variance within and between populations. All analyses were carried out in the program GENES (Cruz 2013Cruz CD (2013) Programa Genes: Biometria. Editora UFV, Viçosa. 382p.).

The inference of genetic groups of individuals from the M. umbellata populations was performed using a Bayesian Monte Carlo Markov Chain (MCMC) approach (Excoffier & Heckel 2006) using the multilocus genotypes of individuals to detect probable genetic (K) groups, assuming a mixed populations model. The analyses were later carried out in Structure version 2.3.4 (Pritchard et al. 2000Pritchard JK, Stephens M & Donnelly P (2000). Erence of population structure using multilocus genotype data. Genetics 155: 945-959.; Falush et al. 2007Falush D, Stephens M & Pritchard JK (2007) Inference of population structure using multilocus genotype data: dominant markers and null alleles. Molecular Ecology Notes 7: 574-578.). To find the best K, we used the ΔK method of Evanno et al. (2005Evanno G, Regnaut S & Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Molecular Ecology 14: 2611-2620.), as implemented in Structure Harvester (Earl & VonHoldt 2011Earl DA & Von Holdt BM (2011) Structure Harvester: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conservation Genetics Resources 4: 359-361.).

Results

The combination of the 10 used primers revealed a total of 127 polymorphic loci, reaching 98.7% of the amplified loops. Five to 18 bands were detected (Tab. 1), and high percentages of polymorphism were observed within each sample (Tab. 2).

Table 2
Intrapopulation polymorphism for ISSR. N = Number of individuals; N.T.L. = Total number of loci; N.l.M. = Number of monomorphic loci.

The genetic differentiation value of the populations (ΦST) was 0.2801. The AMOVA showed that intrapopulational genetic diversity is larger (71.92%) than interpopulational diversity (28.07%). Estimates of ΦSTbetween pairs of populations (Tab. 3) showed that populations 5 and 6 were the most divergent from the others. Populations 1, 2, 3, 4 and 5 presented the lowest averages of ΦST, even though population 1 was the most structured one of them in relation to the others; populations 2, 3 and 4 were the closest.

Table 3
Genetic differentiation values (ΦST) calculated for pairs of populations (upper diagonal) and geographic distances between populations of Myrsine umbellata (lower diagonal) estimated in km.

Despite the high structuring the results verified, it was possible to detect two groups of populations through the dendogram analysis based on the UPGMA method. Populations 1 and 2 formed a large group with populations 3 and 4, being denominated Group 1, while populations 5 and 6, which are geographically farther from the remaining ones, fell into a second group, Group 2 (Fig. 2).

Figure 2
Dendogram obtained by the UPGMA method based on the genetic distances expressed by the Sorensen- Dice coefficient for six populations of Myrsine umbelatta. P1.1 to P1.8 = individuals 1 to 8 of the population 1; P2.1 to P2.9 = individuals 1 to 9 of the population 2; P3.1 to P3.9 = individuals 1 to 9 of the population 3; P4.1 to P4.12 = individuals 1 to 12 of the population 4; P5.1 to P5.10 = individuals 1 to 10 of the population 5; P6.1 to P6.14 = individuals 1 to 14 of the population 6.

The Bayesian structure analysis confirmed Group 1 and Group 2 based on the change rate in Ln (K), statistic ΔK, indicating a convergence for 2 Bayesian groups, K = 2 (Fig. 3a,c). The two-dimensional graphic representation of genetic distances among individuals of all populations (Fig. 3b) corroborated the results of population clustering (Fig. 2). Population 1 presented the largest relative distance from the other populations within its group, while population 5, belonging to Group 2, was the closest to the populations of Group 1. On the other hand, individuals from population 6 were the most isolated.

Figure 3
a-c. Grouping analyses of the six populations of Myrsine umbellata in Structure - a. graph showing K(2), following the criteria proposed by Evanno et al. (2005)Evanno G, Regnaut S & Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Molecular Ecology 14: 2611-2620. to define the correct number of groups, based on the rate of change in Ln(k). The vertical bar along the X-axis represents the sample and along the Y-axis the association coefficient of a sample for a subgroup; b; two-dimensional graphical representation of the genetic distances between the individuals of the sampled populations; c. graphical representation of Bayesian analysis in the software Structure. K = 2 shows two groups: group 1 - green (population 1, population 2, population 3 and population 4); group 2 - red (population 5 and population 6).

Discussion

To adopt strategies of conservation or management of any species in situ, it is necessary to know the structure and genetic diversity contained in target populations. The use of the ISSR markers to access the genetic diversity ofMyrsine umbellatahas proven to be satisfactory in the present study, since it was possible to measure the level of polymorphism in the studied populations (98.7%) with the use of the 10 selected ISSR markers. Studies on different tree species have demonstrated the efficiency of ISSR in the initial assessment of genetic diversity for natural populations, such as those developed for Larix melinii Rupr. (Pinaceae; 98.83%, Zhanget al. 2013Zhang L, Zhang HG & Li XF (2013) Analysis of genetic diversity in Larix gmelinii (Pinaceae) with RAPD and ISSR markers. Genetics and Molecular Research 12: 196-207.), Erythrina velutina Willd. (Fabaceae; 98.0%, Gonçalveset al. 2014Gonçalves LO, Pinheiro JB, Zucchi MI & Silva-Mann R (2014) Caracterização genética de mulungu (Erythrina velutina Willd.) em áreas de baixa ocorrência. Revista Ciência Agronômica 45: 290-298.) or Myrsine coriacea (93%, Paschoaet al. 2018Paschoa RP, Christ JA, Valente CS, Ferreira MFS, Miranda FD, Garbin ML & Carrijo TT (2018) Genetic diversity of populations of the dioecious Myrsine coriacea (Primulaceae) in the Atlantic Forest. Acta Botanica Brasilica 32: 376-385.).

The greatest diversity was found within the populations (71.92%) in relation to the diversity among them (28.07%), indicating historical gene flow between them. This high intra-population genetic variability ofM. umbellatawas already expected, since dioecious species have a high rate of recombination through obligatory cross-fertilization (Hamricket al. 1992Hamrick JL, Godt MJW & Scherman-Broyles SL (1992) Factors influencing levels of genetic diversity in woody plant species. New Forests 6: 95-124.). For species conservation purposes, the existence of genetic variability in the population allows the evolution of new genetic combinations, conferring greater capacity for evolution and adaptation to environmental changes (Shihariet al. 2013Shihari JM, Verma B, Kumar N, Chahota RK, Singh V, Rathour R, Singh SK & Sharma TR (2013) Analysis of molecular genetic diversity and population structure in sea buckthorn (Hippophae spp L.) from north-western Himalayan region of India. Journal of Medicinal Plants Research 7: 3183-3196.). Moreover, this genetic variability is of fundamental importance not only for the species conservation, but also to guarantee vigor and resistance in the progeny. This factor becomes important when selecting matrices for reforestation purposes (Paschoaet al.2018Paschoa RP, Christ JA, Valente CS, Ferreira MFS, Miranda FD, Garbin ML & Carrijo TT (2018) Genetic diversity of populations of the dioecious Myrsine coriacea (Primulaceae) in the Atlantic Forest. Acta Botanica Brasilica 32: 376-385.).

The statistic ΔK indicated two groups that represent contrasting regions in relation to the altitude: we found a higher polymorphism in the populations of Group 1, which occur in higher altitudes (Pop 3 and Pop 1), while the populations of Group 2, located in lower regions (Pop 5 and Pop 6), presented somewhat lower polymorphic percentages. We also observed that population 6 was the most isolated; besides being located below 1,000 meters of altitude, this population is also geographically located farthest from the others. Another relevant observation is that, although most sampled individuals were grouped according to their origin, the individuals P2.1 and P4.1 were grouped with population 3 (Fig. 2).

As long distance wind pollination is unlikely, we can infer that dispersion is the agent responsible for the observed level of diversity among populations. In addition, the present study suggests that fruit dispersion is impaired not only by geographic distance, but also by the variance in altitude between populations since birds have a metabolic cost to maintain body temperature as the ambient temperature increases or decreases (Chatelainet al. 2013Chatelain M, Halpin CG & Rowe C (2013) Ambient temperature influences birds’ decisions to eat toxic prey. Animal Behaviour 86: 733-740.). A similar result was found in a study on Cedrela fissilis Vell. (Meliaceae), where long-distance dispersion was considered the most important process that influences the current distribution of the species (Mangaraviteet al. 2016Mangaravite E, Vinson CC, Rody HVS, Garcia MG, Carniello MA, Silva RS & Oliveira LO (2016) Contemporary patterns of genetic diversity of Cedrela fissilis offer insight into the shaping of seasonal forests in eastern South America. American Journal of Botany 103: 1-10.).

Despite the high observed diversity within populations, our analyses indicated the existence of genetic diversity among populations, presenting high values ​​of ΦST. The lowest value of genetic differentiation among pairs of populations was found between populations 2 and 3 (0.11), which are also the closest geographically, located only 28.7 km apart from each other. The greatest differentiation was in turn between geographically more distant populations (2 and 5, genetic differentiation of 0.40, geographic distance of 119 km). The degree of genetic differentiation may provide an estimate of the gene flow among the studied populations, which is a determinant of their genetic structure. The low differentiation between the populations ofM. umbellata indicates high gene flow between them, which suggests that genetic drift is neutralized. These values ​​also show a relationship between genetic and geographic distances in populations. This is expected, since the more distant the populations, the more different they are in allelic frequencies and phenotypic traits of genetic origin, although there is often no strict correlation.

Although the similarity of the genetic diversity values ​​(ΦST) between the population pairs is remarkable (Tab. 2), populations 5, 6 and 1 are the most genetically diverse when compared to the others. This is confirmed by the graphical representation of the genetic distances between the trees of the sampled populations (Fig. 3b), where individuals 21, 49 and 62 are the most genetically distant. These estimates indicate that seed collection should be performed in these matrices, since the more divergent the selected parents are, the greater the variability of their progenies (Manfioet al. 2012Manfio CE, Motoike SY, Resende MDV, Santos CEM & Sato AY (2012) Avaliação de progênies de macaúba na fase juvenil e estimativas de parâmetros genéticos e diversidade genética. Revista Pesquisa Florestal Brasileira 32: 63-68.). This is of great importance when the seeds are destined for forest restoration (Sebbenn 2002Sebbenn AM (2002) Número de árvores matrizes e conceitos genéticos na coleta de sementes para reflorestamentos com espécies nativas. Revista do Instituto Florestal 14: 115-132.). Taking into account that the presence of genetic structuring is a considerable prerequisite for the efficient management of plant species, our results suggest that the studied populations could be considerable seed banks for reforestation projects.

Conclusion

Our study revealed that intrapopulational genetic diversity is higher than interpopulational genetic diversity in the sampled populations of Myrsine umbellata. The high intrapopulational genetic variability recorded for the species can cause high morphological heterogeneity among seedlings obtained from the same population. In this scenario, it seems appropriate to obtain seeds in more than one studied population thinking of reforestation purposes. Considering the interpopulational genetic diversity, the geographically more isolated populations are more genetically structured, but all populations analyzed are important germplasm banks of M. umbellata conserved in situ.

Acknowledgements

The authors are grateful to Fundação de Amparo à Pesquisa do Espírito Santo - FAPES (61860808/2013), for research financial support; to Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq, for the scholarship to M.F.S. Ferreira (311950/2016-7) and productivity grant T.T. Carrijo (05821/2016-4). This study was partially funded by the Coordernação de Aperfeiçoamento de Pessoal de Nível Superior -Brasil (CAPES) - Finance Code 001.

References

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Publication Dates

  • Publication in this collection
    18 Jan 2021
  • Date of issue
    2021

History

  • Received
    20 Feb 2019
  • Accepted
    24 Sept 2019
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