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草业学报 ›› 2022, Vol. 31 ›› Issue (1): 131-144.DOI: 10.11686/cyxb2020481

• 研究论文 • 上一篇    

全基因组水平蒺藜苜蓿反转录转座子IRAP分子标记开发及应用

尹晓凡(), 魏娜, 郑淑文, 刘文献()   

  1. 兰州大学草地农业生态系统国家重点实验室,兰州大学农业农村部草牧业创新重点实验室,兰州大学草地农业教育部工程研究中心,兰州大学草地农业科技学院,甘肃 兰州 730020
  • 收稿日期:2020-10-27 修回日期:2020-12-28 出版日期:2021-12-01 发布日期:2021-12-01
  • 通讯作者: 刘文献
  • 作者简介:Corresponding author. E-mail: liuwx@lzu.edu.cn
    尹晓凡(1995-),女,新疆乌鲁木齐人,硕士。E-mail: yinxf18@lzu.edu.cn
  • 基金资助:
    甘肃省自然科学基金(20JR10RA622);兰州大学中央高校基本科研业务费专项资金(lzujbky-2020-18)

Genome-wide development and utilization of LTR retrotransposon-based IRAP markers in Medicago truncatula

Xiao-fan YIN(), Na WEI, Shu-wen ZHENG, Wen-xian LIU()   

  1. State Key Laboratory of Grassland Argo-ecosystems,Key Laboratory of Grassland Livestock Industry Innovation,Ministry of Agriculture and Rural Affairs,Engineering Research Center of Grassland Industry,Ministry of Education,College of Pastoral Agriculture Science and Technology,Lanzhou University,Lanzhou 730020,China
  • Received:2020-10-27 Revised:2020-12-28 Online:2021-12-01 Published:2021-12-01
  • Contact: Wen-xian LIU

摘要:

由于苜蓿品种间遗传差异日益缩小,通过传统形态学鉴定表型愈加困难。在苜蓿育种过程中,利用分子标记可大大提高育种效率和品种鉴定。反转录转座子长末端重复序列(LTR)广泛分布在植物基因组中,基于LTR的分子标记具有丰富的多态性和高信息量等优势,被广泛用于物种品种鉴定、评价种质资源多样性等方面。本研究在全基因组水平鉴定和设计了大量蒺藜苜蓿LTR反转录转座子扩增多态性(IRAP)标记,并利用其对国内外40个紫花苜蓿种质资源进行遗传多样性分析。结果表明,根据设计开发出的431个IRAP引物,并按照其染色体位置信息组合获得69对IRAP引物。利用筛选出的37对多态性引物组合共扩增出325个等位位点,平均每个标记可产生8.8个等位位点;多态性条带比率(PPB)为50%~100%,平均值为79.9%;多态性信息含量(PIC)的范围为0.34~0.88,平均值为0.69。基于遗传相似系数(GS)对供试品种采用算术平均值非加权组平均法(UPGMA)进行聚类分析,以0.82为阈值可将40份供试材料分为4类,其分组结果与不同材料地理分布信息以及STRUCTURE分析相对一致。本研究首次在蒺藜苜蓿全基因组水平上开发了IRAP分子标记并在紫花苜蓿种质资源中进行评价与应用,获得的大量IRAP分子标记可对后续苜蓿品种的鉴定保护以及遗传背景分析提供技术支撑。

关键词: 苜蓿, 反转录转座子, IRAP标记, 遗传多样性

Abstract:

Due to the narrowing of genetic differences between alfalfa (Medicago sativa) varieties, it is more difficult to identify phenotypes by traditional morphological methods. In the process of alfalfa breeding, molecular markers can greatly improve the breeding efficiency and variety identification. Reverse transcriptional transposon long terminal repeat sequence (LTR) are widely distributed in plant genomes. Molecular markers based on LTR are characterized by rich polymorphism and high information content, and are widely used in species identification and germplasm resource diversity evaluation. In this study, a large number of inter-retrotransposon amplified polymorphism (IRAP) markers were identified and designed at the genome-wide level in Medicago truncatula and used to analyze the genetic diversity of 40 alfalfa germplasm lines from within China and abroad. This approach produced 431 IRAP primers according to the design standards and the chromosomal location, from which 69 pairs of IRAP primers were obtained. A total of 325 alleles were amplified by 37 pairs of polymorphic primers; The average number of alleles was 8.8 per marker. Percentage of polymorphic bands (PPB) ranged from 50% to 100% with a mean of 79.9%. The polymorphic information content (PIC) ranged from 0.34 to 0.88, with an average of 0.69. Based on the genetic similarity coefficient (GS), unweighted pair-group method with arithmetic means (UPGMA) was used for cluster analysis of the tested varieties. With 0.82 as the threshold, 40 samples of the tested germplasm lines could be divided into 4 categories. The grouping results were relatively consistent with geographic distribution information and STRUCTURE analysis of the 40 lines. In this study, IRAP molecular markers were developed at the whole genome level in M. truncatula for the first time and then evaluated and applied to investigate germplasm lines of alfalfa. A large number of the IRAP molecular markers obtained could provide technical support for the identification, protection and genetic background analysis of alfalfa varieties.

Key words: alfalfa, retrotransposon, IRAP marker, genetic diversity