杧果PARMS反应体系的建立与优化  

Establishment and Optimization of PARMS Reaction System of Mango

作　　者：刘静萍 肖熙鸥[2] 周开兵[1] 柳凤[2] 张国辉 普金安 张翠英 沐云松 Liu Jin gping;Xiao Xiou;Zhou Kaibing;Liu Feng;Zhang Guohui;Pu Jinan;Zhang Cuiying;Mu Yunsong(College of Horticulture,Hainan University,Haikou,570000;Key Laboratory of Biology of Tropical Fruit trees of the Ministry of Agriculture,Key Laboratory of Postharvest Physiology and Fresh-keeping of Tropical Horticultural products of Hainan Province,Institute of South Subtropical Crops,Chinese Academy of Tropical Agricultural Sciences,Zhanjiang,524091;Huaping County Organic Late Ripening Mango Research Center,Huaping,674800;Economic Crop Workstation,Xinping Yi and Dai Autonomous County,Xinping,653404)

机构地区：[1]海南大学园艺学院,海口570000 [2]中国热带农业科学院南亚热带作物研究所,海南省热带园艺产品采后生理与保鲜重点实验室,农业部热带果树生物学重点实验室,湛江524091 [3]华坪县有机晚熟芒果研究中心,华坪674800 [4]新平彝族傣族自治县经济作物工作站,新平653404

出　　处：《分子植物育种》2023年第21期7071-7077,共7页Molecular Plant Breeding

基　　金：国家重点研发计划项目(2019YFD1001103);海南省自然科学基金项目(320QN319);中国热带农业科学院基本科研业务费专项(1630062022014)共同资助。

摘　　要：五引物扩增受阻突变体系(penta-primer amplification refractory mutation system,PARMS)是基于PCR反应的最新基因分型技术,构建杧果PARMS分型体系对杧果种质资源分类和品种选育均有重要意义。本研究以杧果基因组DNA为模板,对PARMS体系的反应体积、引物浓度、DNA浓度进行调整优化,以期建立最佳的杧果PARMS分型反应体系,同时利用48份杧果种质对优化体系进行了验证。结果表明:杧果PARMS分型反应体系的最佳体积为4μL:包含2μL 2×PARMS Master Mix,1μL DNA模板(稀释100倍),0.28μL特异性扩增引物混合物(100μmo L/L),0.72μL dd H_2O。从可靠性和稳定性可以看出,优化的反应体系可将48份杧果种质资源显著分型。该反应体系的建立可在遗传多样性分析、指纹图谱构建、分子标记辅助育种等多领域对杧果展开研究。The five primer amplification hindered mutation system(Penta-primer amplification refractory mutation system,PARMS)is the latest genotyping technique based on PCR reaction.The construction of mango PARMS genotyping system is of great significance for mango germplasm classification and variety selection.In this study,using mango genomic DNA as a template,the reaction volume,primer concentration and DNA concentration of PARMS system were adjusted and optimized in order to establish the best mango PARMS typing reaction system.48 mango germplasm were used to verify the optimized system.The results showed that the optimum volume of mango PARMS typing reaction system was 4μL:2μL 2×PARMS Master Mix,1μL DNA template(diluted 100 times),0.28μL specific primer mixture(100μmoL/L),0.72μL ddH2O.From the reliability and stability,it can be seen that the optimized reaction system can significantly classify 48 mango germplasm resources.The establishment of this reaction system can be used to study mango in many fields,such as genetic diversity analysis,finger-print construction,molecular marker-assisted breeding and so on.

关 键 词：杧果(Mangifera indica L.) SNP PARMS 体系建立 

分 类 号：S667.7[农业科学—果树学]