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利用RAPD分子标记对番茄杂交种纯度的鉴定研究
引用本文:李丽,郑晓鹰,E.Klocke.利用RAPD分子标记对番茄杂交种纯度的鉴定研究[J].广西植物,2003,23(2):149-154,148.
作者姓名:李丽  郑晓鹰  E.Klocke
作者单位:1. 北京农林科学院蔬菜研究中心,北京,100089
2. 联邦育种研究中心园艺作物研究所,德国
基金项目:中德合作研究项目资助(99 02)
摘    要:应用RAPD(RandomlyamplifiedpolymorphicDNA)分子标记对番茄京丹1号和毛粉802的F1代杂交种纯度进行鉴定的实验研究。该项研究使用了10个碱基的单随机引物和10个碱基的双随机引物进行扩增。在60个单引物扩增反应中获得7个京丹1号父本特有的核酸标记片段。但在14个双随机引物对京丹1号和毛粉802杂交组合的扩增反应中获得了7个京丹1号F1代杂交种特有的核酸标记片段和5个毛粉802父本特有的标记带。实验结果显示,双引物的扩增反应对鉴定双亲亲缘关系极近的杂交种纯度较单引物扩增反应更有效。其中,京丹1号的14个标记片段在北京蔬菜研究中心,种子纯度检测室又进行了重复扩增实验。实验结果为87%的RAPD标记可以在使用不同的PCR仪和不同来源的Taq酶的实验条件下得到。RAPD分子标记技术对鉴定双亲亲缘关系极近的杂交种纯度是真实可靠的。

关 键 词:RAPD  单引物  双引物  京丹1号  毛粉802  杂交种纯度

Identification of hybrid purity of tomato (Lyco-persicon esculentum) using RAPD markers
E.Klocke.Identification of hybrid purity of tomato (Lyco-persicon esculentum) using RAPD markers[J].Guihaia,2003,23(2):149-154,148.
Authors:EKlocke
Abstract:To determine the genetic purity of two commercial tomato (Lycopersicon esculentum) hybrids,Jingdan No.1 and Maofen 802,randomly amplified polymorphic DNA (RAPD) markers were applied to discriminate hybrids from their parental inbred lines. Single primer and two-primer RAPD reactions were assayed. Sixty single primers were applied to discriminate the Jingdan No.1 and Maofen 802 hybrids and their parental inbred lines.Six of them generated polymorphic bands that were different between Jingdan No.1 and its female line. But no informative products produced between Maofen 802 and its parental lines. Jingdan No.1 and Maofen 802 were screened with 14 two-primer reactions. Three two-primer reactions showed polymorphic bands discriminated Jingdan No.1 hybrid from its parental inbred lines and Maofen 802 hybrid from its female inbred lines. The result demonstrate that two-primer RAPD reactions are more effective than the single primer reactions. Six single primer reactions and three two-primer X04-K13, K13-G12, X04-K11 reactions were repeated in National engineering research center for vegetable, Seed purity testing laboratory. 87% paternal-specific markers and hybrid-specific markers in the first experiment, which discriminated Jingdan No.1 hybrid from his parental were generated on another PCR machine, MJ Research PTC-200 and with a different source Taq polymerase purchased from Katara company.
Keywords:Lycopersicon esculentum  RAPD  two primer  hybrid purity  
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