基于e-PCR的小麦、山羊草属SSR电子指纹图谱的开发  

Development of the Wheat SSR Electronic Fingerprint Based on e-PCR

作　　者：杨晟 尹誉蓉 王长彪 任永康 牛瑜琦 韩斌 唐朝晖 YANG Sheng;YIN Yurong;WANG Changbiao;REN Yongkang;NIU Yuqi;HAN Bin;TANG Zhaohui(College of Agricultural,Shanxi Agricultural University,Jinzhong,Shanxi 030801,China;College of Life Sciences,Shanxi Agricultural University,Taiyuan,Shanxi 030031,China)

机构地区：[1]山西农业大学农学院,山西晋中030801 [2]山西农业大学生命科学学院,山西太原030031

出　　处：《麦类作物学报》2023年第5期551-561,共11页Journal of Triticeae Crops

基　　金：山西省重点研发计划项目(201903D211004-2-2);山西农业大学生物育种工程项目(YZGC092);山西农业大学学术恢复科研专项(2020xshf06)。

摘　　要：电子指纹图谱的绘制可对电泳结果进行规范化、可视化操作,进而提高小麦分子标记的流通性,加速小麦育种进程。本研究用MicroSAtellite(MISA)软件对小麦属、山羊草属共61个叶绿体和13个线粒体的全基因组SSR标记进行搜索,发现SSR位点的出现频率与基因组类型有关,线粒体、叶绿体基因组均以2次重复的五核苷酸和六核苷酸重复基序为主,同一种类型的基序重复次数与SSR数目成反比;基于电子PCR技术(e-PCR)并结合MapChart软件共筛选出30对引物,其中来源于二倍体和四倍体山羊草属叶绿体全基因组的引物分别有16和2对,来源于四倍体和六倍体小麦属叶绿体全基因组的引物分别有5和2对,来源于六倍体小麦属线粒体全基因组的引物有5对,最后用MapChart软件绘制电子指纹图谱。本研究采用了一种新的SSR引物设计和筛选方法,完成了小麦属、山羊草属叶绿体和线粒体共74个全基因组序列SSR分子标记的初步开发和电子指纹图谱的绘制,为引物开发提供了新思路,有利于提高分子标记的流通性以及小麦种质资源亲缘关系的鉴定。The electronic fingerprint can be used to standardize and visualize the electrophoresis results,to improve the circulation of wheat molecular markers and accelerate wheat breeding process.In this study,we used MicroSAtellite(MISA)software to search SSR markers in 61 chloroplast genomes and 13 mitochondrial genomes in Triticum L.and Aegilops L.species,and found that the frequency of SSR loci was related to chromosome type.Mitochondrial and chloroplast genomes were dominated by two repetitions of pentanucleotide and hexanucleotide repetitions,and the motif repeat number of the same type was inversely proportional to the number of SSR locus.A total of 30 pairs of primers were screened based on electronic PCR(e-PCR)technology,among which 16 and 2 pairs of primers were derived from the whole chloroplast genomes of diploid and tetraploid Aegilops L.,and 5 and 2 pairs of primers were derived from the whole chloroplast genomes of tetraploid and hexaploid Triticum L.,respectively;Five pairs of primers were derived from the whole mitochondrial genomes of hexaploid Triticum L..Finally,electronic fingerprints were mapped with MapChart software.In this study,a new SSR primer designing and screening method was used to develop SSR markers and electronic fingerprints of 74 whole genome sequences of chloroplasts and mitochondria of Triticum L.and Aegilops L.were developed,which provided a new idea for the development of primers,and was beneficial to improve the circulation of molecular markers and the identification of genetic relationships of wheat germplasm resources.

关 键 词：小麦属 山羊草属 e-PCR SSR分子标记 电子指纹图谱 

分 类 号：S512.1[农业科学—作物学] S330