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46416个Unigene中检出含有二核苷酸~六核苷酸重复类型的SSR位点60238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。
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 60238 SSR loci including dinucleotide to hexanucleotide repeats were detected from 46416 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
作者
钱丽华
严建立
吴晓疆
阮松林
尹舒雅
崔海瑞
QIAN Li-hua;YAN Jian-li;WU Xiao-jiang;RUAN Song-lin;YIN Shu-ya;CUI Hai-rui(Hangzhou Academy of Agricultural Sciences,Hangzhou 310024,China;Institute of Nuclear-Agricultural Sciences,Zhejiang University,Hangzhou 310058,China)
出处
《江苏农业学报》
CSCD
北大核心
2023年第5期1120-1131,共12页
Jiangsu Journal of Agricultural Sciences
基金
杭州市重点研发项目(202002A03)。
关键词
黄精
转录组
SSR标记开发
资源分析
Polygonatum
transcriptome
simple sequence repeat(SSR)marker development
resource analysis
