摘要
本研究基于茄子自交系WT和L6-5重测序数据设计SNP特异引物,通过对反应体积、DNA用量和引物浓度等参数进行优化,建立了茄子PARMS(Penta-primer amplification re-fractory mutation system)SNP分型体系。研究结果表明最佳反应体系为6μL,其中1μL DNA模板(10~100 ng/μL),3μL PARMS mix(2×),0.45μL引物混合物(100μmol/L),1.55μL ddH2O。利用WT和L6-5的F2群体验证该体系的稳定性,获得了较好的SNP分型结果。本研究建立的PARMS-SNP分型体系,为开展茄子种质资源遗传多样性分析、基因定位、指纹图谱构建和分子标记辅助选择等提供了技术支持。
In the present study,the PARMS(penta-primer amplification re-fractory mutation system)reaction system for eggplant was established through optimization of the reaction volume,primer concentration and template DNA amount.The genomic DNA of the inbreed line of WT and L6-5 as template.And the primers of the SNPs was designed according to the genome re-sequenced date of WT and L6-5.The result showed that the optimal PARMS reaction system for 96-well plate format was as follows:total volume 6μL,containing 1μL template DNA(10~100 ng/μL),3μL PARMS mix(2×),0.45μL primer assay mix(100μmol/L)and 1.55μL ddH2 O.With the optimal PARMS reaction system,high quality results of PARMS-SNP genotyping were produced on the F2 generation plant of WT and L6-5 which indicated the stability of the system.The optimized PARMS reaction system can be used in genetic study for eggplant,such as genetic diversity analysis,gene mapping,fingerprint map construction and marker-assisted selection.
作者
肖熙鸥
赵秋芳
林文秋
欧雄常
冯恩友
莫定鸣
Xiao Xi’ou;Zhao Qiufang;Lin Wenqiu;Ou Xiongchang;Feng Enyou;Mo Dingming(South Subtropical Crop Research Institute Chinese Academy of Tropical Agricultural Sciences,Zhanjiang,524091;Key Laboratory for Post-harvest Physiology and Technology of Tropical Horticultural Products of Hainan Province,South Subtropical Crop Research Institute Chinese Academy of Tropical Agricultural Sciences,Zhanjiang,524091;Zhanjiang Academy of Agricultural Sciences,Zhanjiang,524091)
出处
《分子植物育种》
CAS
北大核心
2021年第18期6074-6079,共6页
Molecular Plant Breeding
基金
国家自然科学基金面上项目(31872117)
中国热带农业科学院基本科研业务费专项资金(1630062017014)
茄子物种品种资源保护费(125163006000160001)共同资助。
