[1]孙叶,刘红,马辉,等.兰属植物EST-SSR标记开发及应用[J].江苏农业学报,2020,(03):681-688.[doi:doi:10.3969/j.issn.1000-4440.2020.03.021]
　SUN Ye,LIU Hong,MA Hui,et al.Development and application of EST-SSR marker in Cymbidium[J].,2020,(03):681-688.[doi:doi:10.3969/j.issn.1000-4440.2020.03.021]
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兰属植物EST-SSR标记开发及应用()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2020年03期
页码:
681-688
栏目:
园艺
出版日期:
2020-06-30

文章信息/Info

Title:
Development and application of EST-SSR marker in Cymbidium
作者:
孙叶1; 2; 刘红2; 马辉2; 单东方2; 曹宏2; 包建忠2; 陈秀兰2; 赵国琦1
（1.扬州大学动物科学与技术学院，江苏扬州225009；2.江苏里下河地区农业科学研究所，江苏扬州225007）
Author(s):
SUN Ye1; 2; LIU Hong2; MA Hui2; SHAN Dong-fang2; CAO Hong2; BAO Jian-zhong2; CHEN Xiu-lan2; ZHAO Guo-qi1
(1.College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China;2.Institute of Agricultural Sciences of the Lixiahe District in Jiangsu Province, Yangzhou 225007, China）
关键词:
兰属植物; EST-SSR标记; 遗传多样性; 亲缘关系
Keywords:
Cymbidium; expressed sequence tag-simple sequence repeat (EST-SSR) marker; genetic diversity; genetic relationship
分类号:
S682
DOI:
doi:10.3969/j.issn.1000-4440.2020.03.021
文献标志码:
A
摘要:
本研究利用兰属植物杂交种转录组测序数据开发EST-SSR标记，分析标记的多态性及兰属种质的遗传多样性，为兰属植物种质资源的创新和分类研究提供参考。结果表明，转录组测序分析获得113 780条Unigene，从中共识别到23 709个SSR位点，SSR位点出现频率为20.84％。从设计的200对引物中筛选出20对具有多态性，并且扩增条带大小与预期相符的EST-SSR标记引物，对48份兰属种质材料进行PCR扩增，平均多态位点数为3.0，共检测到81.00个等位基因，平均每对引物检测到4.05个等位基因。观测杂合度平均值为0.320 8，期望杂合度平均值为0.507 0；Shannon’s信息指数的变化范围为0.233 8～1.472 4，平均值为0.932 1，多态信息含量为0.110 3～0.662 2。对4个参试兰属植物种群进行遗传多样性分析，春兰和大花蕙兰的F1杂交种群与大花蕙兰种群亲缘关系较近，与春兰种群的亲缘关系较远。聚类分析结果表明，遗传相似系数为0.72时，48份种质聚成7类，春兰和大花蕙兰的F1代杂交种群大多聚入第I类，春兰种群聚入第V类，第II类、第III类、第IV类、第VII类均为大花蕙兰种群。
Abstract:
Expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed based on transcriptome sequencing, the polymorphism of markers and the genetic diversity of Cymbidium germplasms were analyzed in order to provide reference for the innovation and classification of resources. The results showed that 113 780 unigenes were obtained from transcriptome sequencing, a total of 23 709 SSR loci were detected in the unigene with the frequency of 20.84%. In addition, 20 pairs of EST-SSR primers with polymorphism were selected from 200 primers, and the size of amplified bands was in accordance with the expectation. Moreover, 48 Cymbidium germplasms were amplified by PCR, and the average number of polymorphic loci was 3.0. A total of 81.00 alleles were observed, with an average of 4.05 alleles per locus. The mean values of observed heterozygosity and expected heterozygosity were 0.320 8 and 0.507 0, respectively. Shannon′s information index ranged from 0.233 8 to 1.472 4, and the average value was 0.932 1. The polymorphic information content(PIC) ranged from 0.110 3 to 0.662 2. The F1 population of Cymbidium georingii and Cymbidium hybridum was close to the population of C. hybridum, and far away the population of C.georingii. Cluster analysis results indicated that 48 germplasms were grouped into seven groups at the genetic similarity coefficient of 0.72, the F1 population of C.georingii and C. hybridum was grouped into class I, the population of C.georingii was grouped into class V, and class II, class III, class IV and class VII were the population of C. hybridum.

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备注/Memo

备注/Memo:
收稿日期：2019-12-05基金项目：江苏省农业科技自主创新基金项目［CX（17）2012］；江苏省农业种质资源保护与利用平台资助项目（JSGB2018-01）作者简介：孙叶（1978-），女，江苏武进人，硕士，副研究员，主要从事花卉遗传育种研究。(E-mail)sunye9999@126.com通讯作者：赵国琦，(E-mail)gqzhao@yzu.edu.cn
更新日期/Last Update: 2020-07-14