[1]马杰,屈雯,陈春艳,等.基于转录组序列的羊肚菌EST-SSR标记开发与遗传多样性分析[J].江苏农业学报,2020,(05):1282-1290.[doi:doi:10.3969/j.issn.1000-4440.2020.05.027]
　MA Jie,QU Wen,CHEN Chun-yan,et al.Development of EST-SSR markers based on transcriptome sequencing of Morchella spp. and its genetic diversity analysis[J].,2020,(05):1282-1290.[doi:doi:10.3969/j.issn.1000-4440.2020.05.027]
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基于转录组序列的羊肚菌EST-SSR标记开发与遗传多样性分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2020年05期
页码:
1282-1290
栏目:
园艺
出版日期:
2020-10-31

文章信息/Info

Title:
Development of EST-SSR markers based on transcriptome sequencing of Morchella spp. and its genetic diversity analysis
作者:
马杰1; 2; 屈雯2; 陈春艳1; 王磊2; 马维1; 刘针杉2; 马俊1; 杨珊1; 丁丽3; 高强4; 孙勃2
（1.毕节市农业科学研究所，贵州毕节551700；2.四川农业大学园艺学院，四川成都611130；3.毕节市职业技术学院，贵州毕节551700；4.毕节市植保站，贵州毕节551700）
Author(s):
MA Jie1; 2; QU Wen2; CHEN Chun-yan1; WANG Lei2; MA Wei1; LIU Zhen-shan2; MA Jun1; YANG Shan1; DING Li3; GAO Qiang4; SUN Bo2
(1.Bijie Institute of Agricultural Sciences, Bijie 551700, China；2.College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China；3.Bijie Vocational and Technical College, Bijie 551700, China；4.Plant Protection Station of Bijie City, Bijie 551700, China)
关键词:
羊肚菌; 分子标记; EST-SSR; 转录组; 遗传多样性
Keywords:
Morchella spp.; molecular marker; expressed sequence tag-simple sequence repeats (EST-SSR); transcriptome; genetic diversity
分类号:
S646.701
DOI:
doi:10.3969/j.issn.1000-4440.2020.05.027
文献标志码:
A
摘要:
利用羊肚菌（Morchella spp.）转录组数据开发EST-SSR(表达序列标签-简单重复序列)标记，并对33份羊肚菌材料进行遗传多样性分析。结果表明，转录组测序共获得73 781条Unigene序列，其中25 461条Unigene序列中含有41 814个简单重复序列（Simple sequence repeats, SSR）位点，SSR位点发生频率为34.51%，平均分布距离为2.51 kb。优势重复序列类型为单核苷酸，占总SSR位点数量的51.39%，其次为三核苷酸和二核苷酸，分别占总SSR位点数量的28.04%和13.35%。A/T、AG/TC、AGC/TCG分别是单核苷酸、二核苷酸、三核苷酸的优势重复基元。以33份羊肚菌和1份酵母菌为材料，从61对SSR引物中筛选出15对多态性引物，经扩增得到130条多态性条带，多态性比例为100%。遗传多样性分析结果显示，平均每个位点的等位基因数（Na）为4.866 7个，平均有效等位基因数（Ne）为2.124 2个，平均多样性指数（I）为0.958 3，平均观察杂合度（Ho）为0.191 1，平均期望杂合度（He）为0.495 1，平均Nei’s基因多样性指数为0.483 5，表明筛选出的15对SSR引物的扩增产物具有较好的遗传多样性。通过非加权组平均法(Unweighted pair-group method with arithmetic means, UPGMA)聚类分析，可将33份羊肚菌材料分为4类。试验结果可为羊肚菌的种质资源鉴定、品种鉴别和分子标记辅助育种等研究提供依据。
Abstract:
Expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed using transcriptome data of Morchella spp., and the genetic diversities of 33 varieties of Morchella spp. were analyzed. The results showed that a total of 73 781 Unigene sequences were obtained using transcriptome sequencing. There were 41 814 simple sequence repeats (SSR) loci in 25 461 Unigene sequences, the occurrence frequency of SSR locus was 34.51%, the average distribution distance was 2.51 kb. Single nucleotide was the advantage repetitive sequence type, which accounted for 51.39% of the total SSR loci, followed by tri-nucleotides and di-nucleotides, which accounted for 28.04% and 13.35%, respectively. A/T, AG/TC, AGC/TCG were the predominant repetitive motifs in single nucleotides, di-nucleotides, tri-nucleotides, respectively. 15 pairs of polymorphic primers were selected from 61 pairs of SSR primers, using 33 varieties of Morchella spp. and one variety of yeast as material. 130 polymorphic bands were obtained through amplification, with a polymorphic rate of 100%. Results of genetic diversity analysis showed that the average number of alleles (Na) was 4.866 7, the average number of effective alleles (Ne) was 2.124 2, the average diversity index (I) was 0.958 3, the average observed heterozygosity (Ho) was 0.191 1, the average expected heterozygosity (He) was 0.495 1, and the average Nei’s gene diversity index was 0.483 5, indicating that the amplified products of the selected 15 pairs of SSR primers had a good genetic diversity. 33 varieties of Morchella spp. could be divided into four groups by unweighted pair-group method with arithmetic means (UPGMA) cluster analysis. The results can provide basis for germplasm resources identification, variety identification and molecular marker assisted breeding of Morchella spp.

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备注/Memo

备注/Memo:
收稿日期：2020-03-31基金项目：贵州省科技厅项目（黔科合支撑
[2016]2598-1号）；贵州省人才办项目（RCJD2020-7）；中央补助地方科技基础条件专项（黔科合条中补地
[2015]4003）；毕节市科技局项目（毕科联合字zy
[2019]1号、毕科合字
[2017] 46号）作者简介：马杰（1981-），男，山西大同人，硕士，高级农艺师，研究方向为食用菌种质资源与栽培技术研究。（Tel）0857-8333150；（E-mail）115705717@qq.com通讯作者：孙勃，（E-mail）bsun@sicau.edu.cn
更新日期/Last Update: 2020-11-16