Cattleya walkeriana, one of the most improved Brazilian Cattleyas, is a popular tropical orchid endemic from Brazil and currently endangered. In the present study, for the first time microsatellite markers were developed for C. walkeriana and their transferability was tested for the species C. loddigesii and C. nobilior. The markers were used for genotyping 26 C. walkeriana specimens from different growers and from different levels of improvement. The transferability was successful, with five polymorphic loci transferred to C. loddigesii and six polymorphic loci to C. nobilior. Eight loci were polymorphic, revealing a maximum of two to ten alleles per locus in C. walkeriana and two to four and two to five in C. loddigesii and C. nobilior, respectively. There was no significant linkage disequilibrium in the studied loci. For C. walkeriana, the observed and expected heterozygosities ranged from 0 to 0.963 and from 0.138 to 0.841, respectively. These markers identified polymorphisms and may be used to study the genetic diversity, gene flow or hybridization of these species.

Genetic studies in tropical tree species have found signs of decreased genetic diversity and increased levels of inbreeding and spatial genetic structure (SGS) in fragmented and exploited populations. The aim of this paper was to investigate genetic diversity, structure, and intrapopulation SGS using eight microsatellite loci for three Orbignya phalerata populations that have undergone different intensities of seed harvesting. From each population, we georeferenced and sampled 30 seedlings, 30 juveniles, and 30 adult trees. The total number of alleles over all loci (k), and observed (H o ) and expected heterozygosity (H e ) presented lower values for the population experiencing more intense fruit harvesting than less heavily exploited populations, suggesting that fruit harvesting may decrease genetic diversity. Null alleles were detected in practically all loci among seedlings, juveniles, and adults in all populations, indicating that the estimates of H o , H e , and fixation index (F) are biased. When corrected for null alleles (F Null ), the fixation index decreased for all samples, resulting in significantly higher than zero results for seedlings of all populations, but not for juveniles and adults of all populations. The comparison of F Null values between cohorts in the most heavily exploited population (ESP) suggests that inbred individuals are eliminated between seedling and adult stages. Significant SGS was detected up to 60 m in all populations, which indicates short distance seed dispersal. Genetic differentiation (G' ST ) between pairwise populations was related to spatial distance between populations, with the greatest difference between more distant populations.

Abstract. Studies on population genetics are the key to designing effective in situ management plans for tree species, in particular, those subjected to pressure from anthropogenic processes, such as forest fragmentation and logging. To investigate genetic diversity, inbreeding and intrapopulation spatial genetic structure (SGS) in a fragmented population of the insect-pollinated tropical tree, Esenbeckia leiocarpa, we developed specific microsatellite markers for this species and mapped and sampled 100 individuals in a forest plot. Two issues were addressed in particular: (i) the level of genetic diversity, inbreeding and effective population size, (ii) whether intrapopulation spatial genetic structure exists. Among the 14 loci developed, we only used the three that presented polymorphism to estimate the genetic parameters. Genetic diversity was low, whereby the average number of alleles per locus (A) was 3.3 and observed (H 0 ) and expected heterozygosities (H e ) were 0.336 and 0.298, respectively. The average fixation index was significantly higher than zero (F = 0.112), suggesting inbreeding. Significant SGS was found up to 7 m and between 31 to 38 m, indicating that trees growing within these distances may be related. Estimates of the effective population size indicated that the 100 sampled trees correspond to 14 individuals that are neither related nor inbred. Our results suggest that the microsatellite markers developed in this study are suitable for studies on genetic diversity and structure, mating systems, gene flow and SGS in this species.

ABSTRACT. The aim of this study was to evaluate the genetic diversity among native plants and some individuals obtained from crosses with unknown genealogy of C. walkeriana as well as C. loddigesii and C. nobilior and to advance towards solving the question of the genetic purity of the "Orchidglade" clone. Eight microsatellite loci were used to evaluate the genetic diversity between individuals of C. walkeriana. Microsatellites were not efficient in determining the genetic diversity between C. walkeriana groups (native and improved). The difficulty in determining the genetic distance between the different genotypes can be attributed to the complex mating system of the species and to a weak genetic barrier that facilitates the development of hybrids. Our analysis revealed smaller genetic distances between the "Orchidglade", "Equilab", "Kenny" and "Pedentive" clones and the species C. loddigesii and C. nobilior. Native C. walkeriana plants were genetically more distant from the C. loddigesii and C. nobilior species.Keywords: orchid improvement, genetic variation, genetic differentiation, Cattleya loddigesii, Cattleya nobilior.Diversidade genética entre clones famosos nativos e melhorados de Cattleya walkeriana Gardner RESUMO. O objetivo desta pesquisa foi o de avaliar a diversidade genética entre plantas nativas e indivíduos de genealogia desconhecida de C. walkeriana, bem como C. loddigesii e C. nobilior, e também avançar na solução do dilema da origem do clone "Orchidglade" de C. walkeriana. Oito locos microssatélites foram utilizados para avaliar a diversidade genética entre indivíduos de C. walkeriana. Os marcadores microssatélites não foram eficientes na determinação da diversidade genética entre os grupos C. walkeriana (nativas e melhoradas). A dificuldade em determinar a distância genética entre os genótipos diferentes pode ser devida a um sistema complexo de reprodução das espécies e devido a uma fraca barreira reprodutiva facilitando o desenvolvimento de híbridos. Nossa análise revelou menores distâncias genéticas entre os clones Orchidglade, "Equilab", "Kenny" e "Pedentive" e as espécies C. loddigesii e C. nobilior. As C. walkeriana nativas se mostraram geneticamente mais distantes das espécies de C. loddigesii e C. nobilior. Palavras-chave: melhoramento de orquídeas, variação genética, diferenciação genética, Cattleya loddigesii, Cattleya nobilior.