Vasconcelos Rayssa Gomes, Hernández-Rangel Sandra Marcela, Farias Izeni Pires, Hrbek Tomas, Sampaio Paulo de Tarso Barbosa, Medeiros Raquel da Silva, Cunha-Machado Antonio Saulo
Programa de Pós-Graduação em Ciências de Florestas Tropicais (PPG-CFT), Instituto Nacional de Pesquisas da Amazônia (INPA), Manaus, Amazonas, Brazil.
Laboratório de Evolução e Genética Animal (LEGAL), Universidade Federal do Amazonas (UFAM), Manaus, Amazonas, Brazil.
Mol Biol Rep. 2024 Dec 30;52(1):92. doi: 10.1007/s11033-024-10192-6.
Native to the Amazon region, Copaifera multijuga Hayne is a large tree (≈ 36 m in height) that is heavily exploited for extraction of its oleoresin. Many studies have addressed the phytochemical properties and applications of this raw material; however, there are few initiatives that have focused on the genetic characterization of native populations of this species. To this end, our objective was to develop microsatellite markers for C. multijuga, which were previously unavailable, and apply them to the characterization of a native population located in the Adolpho Ducke Forest Reserve (Manaus, Amazonas, Brazil).
Using next-generation sequencing technology on the Ion Torrent PGM™ platform, 19 pairs of microsatellite primers were designed and developed. For the characterization and validation, we used a group of 47 C. multijuga trees. After preliminary tests, amplification conditions were standardized for 14 loci. The CmH05 locus was excluded from the analyses for being monomorphic, and the remaining loci were used to estimate key genetic parameters for the species, such as observed (Ho) and expected heterozygosity (He), total number of alleles (A), fixation index (f), and polymorphic information content (PIC), among others. The population showed levels of genetic diversity that were higher than 0.63 (Ho: 0.67; He: 0.64) and an average number of 5.4 alleles. According to the PIC estimates, all loci were considered highly (9) or moderately informative (3), except for CmH11, which had a value that was below 0.2.
The 13 developed microsatellites were efficient in characterizing the genetic diversity of C. multijuga and may be advantageous in future investigations aimed at defining effective conservation strategies for the species.
多花苦配巴原产于亚马逊地区，是一种大型树木（高度约36米），因其树脂的提取而被大量开发利用。许多研究探讨了这种原材料的植物化学特性和应用；然而，很少有研究关注该物种原生种群的遗传特征。为此，我们的目标是开发多花苦配巴的微卫星标记（此前尚无此类标记），并将其应用于对位于阿道夫·杜克森林保护区（巴西亚马孙州玛瑙斯）的一个原生种群的特征分析。
利用Ion Torrent PGM™平台上的下一代测序技术，设计并开发了19对微卫星引物。为了进行特征分析和验证，我们使用了一组47株多花苦配巴树。经过初步测试，对14个位点的扩增条件进行了标准化。CmH05位点因呈单态性而被排除在分析之外，其余位点用于估计该物种的关键遗传参数，如观察杂合度（Ho）、期望杂合度（He）、等位基因总数（A）、固定指数（f）和多态信息含量（PIC）等。该种群的遗传多样性水平高于0.63（Ho：0.67；He：0.64），平均等位基因数为5.4个。根据PIC估计，除CmH11位点的值低于0.2外（该位点信息性低），所有位点均被认为具有高信息性（9个）或中等信息性（3个）。
所开发的13个微卫星在表征多花苦配巴的遗传多样性方面是有效的，并且可能有利于未来旨在为该物种确定有效保护策略的研究。
