麻竹EST-SSR标记开发及其对慈竹变异类型的分析研究

EST-SSR Molecular Marker Development from Dendrocalamus latiflorus and Its Application on Genetic Diversity Analysis of Variation Types from Bambusa emeiensis

摘要:

利用麻竹(Dendrocalamus latiflorus)已有的EST 序列,开发了一批EST-SSR 分子标记,并用于慈竹(Bambusa emeiensis)栽培变异类型的多态性研究。结果表明,在麻竹9574 个EST 中找到了331 个EST,含有381 个SSR 位点,SSR 出现的频率为3.98%。EST-SSR 的重复类型共有59 个,其中2 个核苷酸的重复最多,占63.8%, 其次是3 个核苷酸的重复序列。根据含有SSR 的EST 序列设计出了44 对引物,对慈竹及其变异类型进行了扩增效率、多态性及通用性检测,其中37 对引物能够扩增出稳定且清晰的条带,且扩增具有多态性,扩增片段长度主要集中在100~1000 bp,引物有效率达84.1%。遗传相似性分析结果显示,所检测样品间遗传距离为0.263~0.840,平均为0.552,其中‘黑笋慈竹’与‘琴丝慈竹’、‘蛇头慈竹’与‘龙头慈竹’之间的遗传距离较近,而‘罗汉慈竹’与其他样品的遗传距离最远。

关键词:麻竹;EST-SSR;分子标记;慈竹;栽培变异类型;遗传距离

Abstract:

EST-SSR molecular markers were developed from ESTs of Dendrocalamus latiflorus, the genetic diversity of variation types from Bambusa emeiensis was studied by using EST-SSR markers. There were 381 SSR sites locating in 331 ESTs collected from 9574 ESTs of D. latiflorus, with SSR frequency of 3.98%. There were 59 EST-SSR types, in which dinucleotide type was the most with 63.8%, followed by trinucleotide type (35.7%). Based on the EST sequences containing SSR site, forty pairs of EST-SSR primer were designed for detecting amplification efficiency, polymorphism and transferability in cultivated variation types of B. emeiensis. The results showed that 37 primer pairs, accounting for 80.43%, could amplify stable and clear bands with polymorphisms, and the length of amplified fragments mainly ranged from 100 bp to 1000 bp. Genetic distance among cultivated variation types of B. emeiensis ranged from 0.263 to 0.840 with mean of 0.552. There were close genetic distance between B. emeiensis ‘Heisunci’ and B. emeiensis ‘Qinsici’, B. emeiensis ‘Shetouci’ and B. emeiensis ‘Longtouci’, respectively, and B. emeiensis ‘Luohanci’ was far away from others.

Key words:Dendrocalamus latiflorus;EST-SSR;Molecular marker;Bambusa emeiensis;Cultivated variation type;Genetic distance