Identification and development of microsatellite (SSRs) makers of Exbucklandia (HAMAMELIDACEAE) by high-throughput sequencing
Identification and development of microsatellite (SSRs) makers of Exbucklandia (HAMAMELIDACEAE) by high-throughput sequencing
复制标题
通过高通量测序鉴定和开发 Exbucklandia (HAMAMELIDACEAE) 的微卫星 (SSR) 标记
DOI：
10.1007/s11033-019-04800-z
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发表时间：
2019-06-01
影响因子：
2.8
通讯作者：
Liao, Wenbo
中科院分区：
文献类型：
--
作者：
Huang, Cuiying;Yin, Qianyi;Liao, Wenbo
金缕梅科（Hamamelidaceae）为虎耳草目（Saxifragales），约有6亚科30属140种，其中大部分为第三纪残遗植物。鹿草属（Exbucklandia）是金缕梅科鹿草亚科（Exbucklandioideae）中唯一的一个属，仅有2-4种。其中E.长瓣花T. Chang是中国特有植物，已被《中国高等植物生物多样性红色名录》列为濒危物种，但有分类学家提出E. longipetala应并入E. tonkinensis（Lecomte）H. T.昌目前，可能由于缺乏有效的分子标记，对该属植物的地理分布研究几乎空白。本研究对大肠杆菌E.基于高通量测序技术，对越南鳖进行了测序，获得了约6 G的原始数据，进一步从头组装成303，481个重叠群。在此基础上，从13,596个重叠群中鉴定出15,326个SSR，并成功设计了10,660个SSR的引物。共合成了139对引物，其中106对引物在6个毛兰属个体中扩增出预期的扩增产物，其中24对引物在毛兰属3个居群中表现出多态性。因此，预期和观察到的杂合性在0.097-0.717和0.098-0.583之间。在此基础上，进一步开展白木香属植物的遗传多样性和种群结构的研究，有助于了解白木香属植物的系统起源和地理分布格局。
Hamamelidaceae (Saxifragales, previously Rosales) comprises approximately six subfamily, 30 genera and 140 species, most of which are Tertiary relicts. Exbucklandia is the only genus of the subfamily Exbucklandioideae, Hamelidaceae, containing only 2-4 species. Of them, the species E. longipetala H. T. Chang is endemic to China and listed as endangered in The Biodiversity Red List of China: Higher Plant, yet some taxonomists put forward that E. longipetala should be merged into E. tonkinensis (Lecomte) H. T. Chang. Currently, there was nearly no phylogeographic studies on this genus possibly due to the deficiency of efficient molecular markers. In this study, we sequenced the genome of E. tonkinensis based on high throughput sequencing technology, and obtained approximately 6 G raw data, which was further de novo assembled into 303,481 contigs. Based on them, 15,326 SSRs were identified from 13,596 contigs, and primers were successfully designed for 10,660 SSRs. A total of 139 paired primers were synthesized, 106 of them were successfully amplified in six Exbucklandia individuals with expected PCR product size, and 24 demonstrated to be polymorphic among three Exbucklandia populations. Accordingly, the expected and observed heterozygosity were between 0.097-0.717 and 0.098-0.583. Based on these efforts, future researches on genetic diversity and population structure of Exbucklandia can be performed to understand its phylogenetic origins and phylogeographic pattern.