基于IRAP标记的沙子空心李遗传多样性评价及指纹图谱构建  

Evaluation of genetic diversity and construction of DNA fingerprint of Prunus salicina Lindl. ’Shazikongxinli’ based on IRAP markers

作　　者：王贵 李蕊蕊 吴茂宏 任菲宏 王丽丽[4] 乔光[1] WANG Gui;LI Ruirui;WU Maohong;REN Feihong;WANG Lili;QIAO Guang(The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountain Region(Ministry of Education)/College of Life Sciences,Guizhou University,Guiyang 550025,China;Tongren Academy of Forestry,Tongren 554300,China;Tongren Academy of Agricultural Sciences,Tongren 554300,China;Guizhou Academy of Agricultural Sciences,Guiyang 550006,China)

机构地区：[1]贵州大学生命科学学院/山地植物资源保护与种质创新教育部重点实验室,贵阳550025 [2]贵州省铜仁市林业科学院,铜仁554300 [3]贵州省铜仁市农业科学院,铜仁554300 [4]贵州省农业科学院,贵阳550006

出　　处：《华中农业大学学报》2023年第1期1-11,共11页Journal of Huazhong Agricultural University

基　　金：贵州省科技计划项目(黔科合支撑[2020]1Y023);贵州省科技计划项目(黔科合服企[2019]4013);中央引导地方科技发展资金项目(黔科中引地[2021]4010)。

摘　　要：通过走访调查,选取92株具有优良性状的沙子空心李种质,根据李反转录转座子的RT序列开发IRAP标记引物,设计L16(43)的正交试验优化10μL IRAP-PCR主要影响因素的含量;利用18条IRAP引物对92份沙子空心李进行遗传多样性分析,构建供试种质的指纹图谱。结果显示:最优的10μL IRAP-PCR反应体系为10^(-5)mol/L IRAP引物l.3μL,PCR Mix 5.0μL、模板DNA(30 ng/μL) l.0μL,PCR循环40次。18条IRAP引物共扩增出189个位点,多态性位点180个,等位基因数、有效等位基因数、Nei’s基因多样性指数和Shannon指数均值分别为1.930、1.426、0.261及0.405。采用邻接法将92份沙子空心李聚类为4个类群。确定IRAP引物Ty3-2及Ty3-6为核心引物,可高效构建92份供试种质的指纹图谱。Ninty-two accessions of‘Shazikongxinli’germplasm with elite traits were selected by visiting and surveying to evaluate the genetic diversity of Prunus salicina Lindl.‘Shazikongxinli’IRAP primers were developed based on the conserved regions of RT sequences of plum reverse transcription transposons Ty1-copia and Ty3-gypsy.L_(16)(4^(3)) orthogonal experiments were designed to optimize the main factors affecting content of IRAP-PCR with 10μL volume.18 IRAP primers were used to analyze the genetic diversity and construct the fingerprint of the germplasm tested.The results showed that the optimized 10μL IRAP-PCR reaction system was 1.3μL of 5-10 mol/L IRAP primers,5.0μL of PCR mixture,1.0μL of template DNA(30 ng/μL),and the number of PCR cycles was 40 times.A total of 189 loci were amplified from 18 IRAP primers with good repeatability and high polymorphism,including 180 polymorphic loci.The average value of observed number of alleles(N_(a)),effective number of alleles(N_(e)),Nei’s gene diversity(H_(e)) and Shannon’s information index(I) for all primers was 1.930,1.426,0.261 and 0.405,respectively.The 92 accessions tested were clustered into 4 groups using the neighbor-joining method.The IRAP primers Ty3-2 and Ty3-6 were identified as core primers,which can be used to efficiently construct the fingerprints of 92 accessions tested.

关 键 词：沙子空心李 种质资源 分子标记 IRAP 指纹图谱 遗传多样性 种质创新 

分 类 号：S662.3[农业科学—果树学]