中国樱桃(Prunus pseudocerasus Lindl.)IRAP分子标记技术体系的建立  

Establishment of IRAP Molecular Marker System of Prunus pseudocerasus Lindl

作　　者：刘厚宇[1,2,3] 李顺雨 吴敏芳 乔光 吴亚维[5] 宋莎[5] LIU Houyu;LI Shunyu;WU Minfang;QIAO Guang;WU Yawei;SONG Sha(The Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),Institute of Agro-bioengineering and College of Life Sciences,Guizhou University,Guiyang 550025,China;College of Forestry,Guizhou University,Guiyang 550025,China;Agricultural and rural area Bureau of Qingzhen Guiyang,Guizhou Province 550004,China;Fruit industry Development Center of Weining,Bijie,Guizhou Province 553100,China;Institute of Fruit Tree of Guizhou Academy of Agricultural Sciences,Guiyang 550006,China)

机构地区：[1]贵州大学农业生物工程研究院/生命科学院,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]贵州大学林学院,贵阳550025 [3]贵州省清镇市农业农村局,贵阳550004 [4]贵州省威宁自治县果业发展中心,贵州威宁553100 [5]贵州省农业科学院果树科学研究所,贵阳550006

出　　处：《种子》2020年第9期31-35,40,共6页Seed

基　　金：贵州省精品水果现代农业产业技术体系建设项目(GZCYTX2020-10);贵州农业攻关项目(黔科合支撑[2016]2520)。

摘　　要：基于中国樱桃LTR类反转录转座子的反转录酶(RT)序列信息设计引物,并根据扩增谱带的数量、多态性和可重复性等指标筛选IRAP-PCR引物;采用5因素4水平L 16(45)的完全随机正交试验,对影响PCR体系的主要因子进行优化,建立了中国樱桃IRAP分子标记反应体系。结果表明,筛选出9条引物,可以扩增222个清晰位点,位点的平均多态性比率为90.09%。PCR的最佳反应体系为25μL,含2.0 mmol·L^-1 MgCl 2、1.0 U Taq酶、0.2 mmol·L^-1引物、20 ng模板DNA、0.3 mmol·L^-1 dNTP,采用8%聚丙烯酰胺凝胶电泳检测。In this study,the primers were designed based on the reverse transcriptase(RT)coding sequence of LTR(long terminal repeat)-type reverse transcriptional transposons of Prunus pseudocerasus,and IRAP-PCR primers were screened according to the number,polymorphism and repeatability of the amplified bands.A completely random orthogonal test with 5 factors and 4 level L 16(45)was performed to optimize the main factors affecting the PCR system and established the IRAP molecular marker reaction system of P.Pseudocerasus.The results showed that 222 obvious loci could be amplified by 9 primers,and the average polymorphism rate of the loci was 90.09%.The optimal PCR reaction system was 25μL,containing 2.0 mmol·L^-1 MgCl 2,1.0 U Taq enzyme,0.2 mmol·L^-1 primers,20 ng DNA template,0.3 mmol·L^-1 dNTP,and detected with 8%polyacrylamide gel electrophoresis.

关 键 词：中国樱桃 IRAP 技术体系 优化 

分 类 号：S662.5[农业科学—果树学]