马铃薯SSR引物的开发、特征分析及在彩色马铃薯材料中的扩增研究  

Development and characterization analysis of potato SSR primers and the amplification research in colored potato materials

作　　者：张霞 于卓[1] 金兴红 于肖夏[1] 李景伟 李佳奇 ZHANG Xia;YU Zhuo;JIN Xing-Hong;YU Xiao-Xia;LI Jing-Wei;LI Jia-Qi(Agronomy College,Inner Mongolia Agricultural University,Hohhot 010019,Inner Mongolia,China)

机构地区：[1]内蒙古农业大学农学院,内蒙古呼和浩特010019

出　　处：《作物学报》2022年第4期920-929,共10页Acta Agronomica Sinica

基　　金：内蒙古自治区科技重大专项(ZDZX2018019);内蒙古农业大学科技成果转化专项资金动植物品种培育项目(YZGC2017006);内蒙古自治区马铃薯种业技术创新中心项目资助。

摘　　要：彩色马铃薯较普通栽培马铃薯具有更丰富的营养物质,特别是富含抗氧化物质花青素,是近年来育种家研究的焦点。迄今为止开发的马铃薯SSR引物数量有限,特别是彩薯相关的SSR引物。本研究基于马铃薯全基因组序列利用MISA软件对SSR位点进行了分析。研究发现,在马铃薯全基因组序列中共获得218,997个SSR位点,平均3.39 kb出现一个SSR位点。单核苷酸是主要重复类型,占总SSR的62.05%,其次是二核苷酸和三核苷酸,所占比例为22.39%和13.11%。6种核苷酸类型的重复次数分布在5~746次,以5~10次为主,占总SSR的60.9%。在检测到的全部SSR中,共获得215种基元类型,除六核苷酸重复类型外,其他5种核苷酸重复类型的优势基元均以含有A/T-的基序为主。SSR基序长度主要分布在12~20 bp之间,占全部SSR的43.19%。通过Primer 5共设计出100对SSR引物,利用彩薯双亲的DNA初步筛选出48对可以扩增出条带的引物,有效扩增率达48%。进一步验证引物的有效性,随机选择F2群体的6个单株进行PCR扩增,筛选出条带清晰稳定,多态性较高的引物26对,平均多态性比率为72.52%。本研究结果表明马铃薯全基因组SSR位点数量丰富、类型多样,具有中等的多态性。开发的引物多态性较高,可用于彩色马铃薯SSR标记的开发,遗传多样性分析和指纹图谱构建等方面的研究,为进一步挖掘花青素含量相关基因提供科学依据。Compared with commonly cultivated potato,colored potato is rich in nutrition,especially in anthocyanins,which is the focus of breeding research in recent years.To date,the number of potato SSR primers developed is limited,especially those related to colored potato.In this study,SSR loci were analyzed using MISA software based on the whole genome.The results were as follows:a total of 218,997 SSR loci were obtained from potato genome with an average of 3.39 kb.Mononucleotide was the main repeat type,accounting for 62.05% of the total SSR,followed by dinucleotide and trinucleotide,accounting for 22.39% and 13.11%,respectively.The repeats of the six nucleotide types ranged from 5 to 746,mainly from 5 to 10,accounting for 60.9% of the total SSRs.A total of 215 motif types were obtained in all detected SSRs.The dominant motifs of the other five nucleotide repeat types were mainly A/T-containing motifs,except for hexanucleotide repeat type.The length of SSR motifs ranged from 12 bp to 20 bp,accounting for 43.19% of all SSRs.A total of 100 SSR primers were designed by Primer 5 software,among which 48 primers were preliminarily screened using parents’genomic DNA of the colored potato,with an effective amplification rate of 48%.Then,six F2 individuals were randomly selected for PCR amplification,and 26 primers with clear and stable bands and high polymorphism were finely screened,with the average polymorphism rate of 72.52%.In summary,the number and types of SSR loci in potato genome was abundant and diverse,and the polymorphism was moderate.The developed primers were highly polymorphic,which can be used in the development of SSR markers,genetic diversity analysis,and SSR fingerprint of colored potato,providing a scientific basis for further mining genes related to anthocyanin content.

关 键 词：马铃薯基因组 彩色马铃薯 SSR 引物开发 

分 类 号：S532[农业科学—作物学]