由基因序列开发番茄枯萎病抗性基因I-2的共显性分子标记.pdfVIP

HEREDITAS (Beijing) 2008 7 , 30(7): 926―932 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2008.00926 I-2 1 1, 2 于拴仓 , 邹艳敏 1. , 100097 2. , 100037 摘要: 根据I-2 的基因序列设计特异扩增引物对I-2/5F 和I-2/5R, 扩增I-2 基因3 132 ～3 765 bp 之间片段, 基因 型为I-2 / I-2 的材料03F-7 可扩增出633 bp 的条带, 而基因型为i-2/ i-2 的材料Moneymaker 可扩增出693 bp 的 条带, 杂合型材料可扩增出以上2 个条带。通过这两个特异扩增片段的克隆和测序证明, 抗病材料扩增的633 bp 片段为I-2 基因的3 132 ～3 765 bp 之间的序列, 而感病等位基因中出现大量的碱基突变和60 bp 片段插入。利 用引物对I-2/5F 和I-2/5R, 可区分纯合抗病材料、杂合抗病材料和纯合感病材料, 从而建立了I-2 基因的共显性 分子标记。在此基础上, 利用该标记对 16 个主要番茄品种进行基因型鉴定, 8 个品种含有I-2 基因, 其中 1 个品 种基因型为I-2 / I-2, 其他品种为I-2 / i-2 。通过一次PCR 和一次Hind Ⅲ酶切建立了I-2 和Tm-22 双基因检测体 系, 为多基因鉴定及标记辅助选择提供了有力工具。 关键词: 番茄; 枯萎病; I-2 基因; Tm-22 基因; 分子标记 A co-dominant molecular marker of fusarium wilt resistance gene I-2 derived from gene sequence in tomato 1 1, 2 YU Shuan-Cang , ZOU Yan-Min 1. National Engineering Research Center for Vegetable, Beijing 100097, China; 2. College of Life Science, Capital Normal University, Beijng 100037, China Abstract: Sequence-specific PCR primers, I-2/5F and I-2/5R were designed according to the sequence from 3 132 bp to 3 765 bp in I-2 gene. With them a 633 bp fragment was amplified from 03F-7 with a genotype of I-2 / I-2, 693 bp fragment from Moneymaker with a genotype of i-2/ i-2, and both fragments from Tebao with heterozygous genotype I-2 / i -2. These two specific fragments were cloned and sequenced. The results from multiple sequence alignment showed that the 633bp fragment from 03F-7 was identical with the sequence from 3 132 bp to 3 765 bp in I-2 gene. Compared with the sequence of I-2 gene, th