[1]钱丽华,严建立,吴晓疆,等.基于滇黄精转录组序列的SSR标记开发及其在黄精属资源分析中的应用[J].江苏农业学报,2023,(05):1120-1131.[doi:doi:10.3969/j.issn.1000-4440.2023.05.004]
　QIAN Li-hua,YAN Jian-li,WU Xiao-jiang,et al.Development of simple sequence repeat (SSR) markers based on transcriptome sequences of Polygonatum kingianum and their application in analysis of Polygonatum germplasm resources[J].,2023,(05):1120-1131.[doi:doi:10.3969/j.issn.1000-4440.2023.05.004]
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基于滇黄精转录组序列的SSR标记开发及其在黄精属资源分析中的应用()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2023年05期
页码:
1120-1131
栏目:
遗传育种·生理生化
出版日期:
2023-08-31

文章信息/Info

Title:
Development of simple sequence repeat (SSR) markers based on transcriptome sequences of Polygonatum kingianum and their application in analysis of Polygonatum germplasm resources
作者:
钱丽华1; 严建立1; 吴晓疆2; 阮松林1; 尹舒雅1 ; 崔海瑞2
（1.杭州市农业科学研究院，浙江杭州310024；2.浙江大学原子核农业科学研究所，浙江杭州310058）
Author(s):
QIAN Li-hua1; YAN Jian-li1; WU Xiao-jiang2; RUAN Song-lin1; YIN Shu-ya1; CUI Hai-rui2
（1.Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China;2.Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310058, China）
关键词:
黄精; 转录组; SSR标记开发; 资源分析
Keywords:
Polygonatum; transcriptome; simple sequence repeat (SSR) marker development; resource analysis
分类号:
S567.2
DOI:
doi:10.3969/j.issn.1000-4440.2023.05.004
文献标志码:
A
摘要:
本研究基于滇黄精转录组序列开发简单重复序列（SSR）标记并将其应用于黄精属资源分析。设计合成了45对SSR引物，经PCR扩增验证，选择其中20对SSR引物对75份黄精属资源进行分析。结果表明，共在46 416个Unigene中检出含有二核苷酸～六核苷酸重复类型的SSR位点60 238个，序列SSR发生频率为2278%，平均分布距离约707 kb；SSR位点中的主导类型是二核苷酸和三核苷酸重复，分别占5006%和3489%。测试的45对SSR引物中有34 对（7556%）可扩增SSR条带。筛选的20对引物共扩增出153个条带，多态率为9869%，每对引物扩增条带400～1400个，平均765个，不同SSR标记的多态性信息含量为0626～0973，平均为0870。75份材料的等位基因数和遗传相似系数分别为7.00～5200个和0531～0941，平均值分别为2465个和0689，显示出丰富的遗传多样性。基于SSR标记分析的聚类图显示，在遗传相似系数0666处可将供试材料分为4类，较好地反映了供试材料的分类归属。此外，还发现5份多花黄精材料具有特异性的SSR条带扩增或缺失，可作为不同多花黄精材料鉴定的重要分子依据。本研究开发的SSR标记多态性较高，能够有效揭示黄精属种质资源的遗传多样性，对于丰富黄精分子标记种类、构建遗传图谱、促进种质资源的评价与育种应用、开展特定性状的辅助选择等研究都具有重要的意义。
Abstract:
In this study, simple sequence repeat (SSR) markers were developed based on transcriptome sequence data of Polygonatum kingianum and were utilized in germplasm resource analysis of Polygonatum Forty-five pairs of SSR primers were designed and synthetized, then 20 of them were selected by PCR amplification verification test to analyse 75 Polygonatum resources The results showed that, a total of 60 238 SSR loci including dinucleotide to hexanucleotide repeats were detected from 46 416 Unigenes in P kingianum transcriptome, with an occurring frequency of 2278% and an average distribution distance of 707 kb Dinucleotide repeat and trinucleotide repeat were the dominant types, with frequencies of 5006% and 3489% of all SSRs, respectively Among 45 pairs of SSR primers, 34 of them showed amplification ability, which accounted for 7556% A total of 153 bands were amplified by the selected 20 pairs of primers, with a polymorphism rate of 9869% Each pair of primer could amplify 400-1400 bands, with an average of 765 bands The polymorphic information content varied from 0626 to 0973 for different SSR markers, with an average value of 0870 Number of allele genes and genetic similarity coefficients of 75 materials were 700-5200 and 0531-0941 respectively, and the average values were respectively 2465 and 0689, which displayed rich genetic diversity A dendrogram constructed based on SSR markers showed that, the test materials could be classified into four cluster groups with the genetic similarity coefficient of 0666, which presented a good reflection of affiliation in taxonomy of tested Polygonatum materials In addition, five P cyrtonema materials showed specific present or absent SSR bands, which could be used as important molecular basis for identification of different materials The SSR markers developed in this study were highly polymorphic and could be used to effectively reveal the genetic diversity of the Polygonatum germplasm resources The markers are of great significance for enriching molecular marker types, constructing genetic linkage map, promoting the evaluation and breeding application of germplasm resources, and carrying out assisted selection of specific traits in Polygonatum

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备注/Memo

备注/Memo:
收稿日期：2022-10-27 基金项目：杭州市重点研发项目（202002A03）作者简介：钱丽华（1975-），女，浙江杭州人，高级农艺师，主要从事中药材生物技术和遗传育种研究。（Tel）0571-87093826；（E-mail）358843884@qq.com 通讯作者：崔海瑞，（E-mail） hrcui@zju.edu.cn
更新日期/Last Update: 2023-09-13