外源基因在苹果植株中稳定性研究

以常规离体继代培养的转豇豆胰蛋白酶抑制剂（Cowpea trypsin inhibitor,CpTI）基因苹果（Malusdomestica Borkh.）（包括皇家嘎拉、王林、乔纳金和富士等60个株系）组培苗为试材,应用卡那霉素筛选、PCR扩增及RT-PCR扩增对其外源基因的稳定性进行研究。结果表明,在60个常规继代培养6～8年的转CpTI基因苹果转化株系中均可检测出CpTI特异基因片断;除2个转化株系在50 mg.L-1卡那霉素浓度下出现花叶现象,表现缺乏卡那霉素抗性外,其他株系在含相同浓度卡那霉素的培养基中生长正常。RT-PCR检测结果表明,转入的外源CpTI基因在43个株系中得到了较高...

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Bibliographic Details
Published in东北农业大学学报 Vol. 42; no. 7; pp. 97 - 101
Main Author 周瑞金 杜国强 师校欣
Format Journal Article
LanguageChinese
Published 河南科技学院园林学院,河南新乡,453003%河北农业大学园艺学院,河北保定,071001 2011
Subjects
nptⅡ基因
稳定性
继代培养
苹果
豇豆胰蛋白酶抑制剂基因
nptⅡ基因
苹果
继代培养
稳定性
豇豆胰蛋白酶抑制剂基因
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ISSN1005-9369
DOI10.3969/j.issn.1005-9369.2011.07.018

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Summary:以常规离体继代培养的转豇豆胰蛋白酶抑制剂（Cowpea trypsin inhibitor,CpTI）基因苹果（Malusdomestica Borkh.）（包括皇家嘎拉、王林、乔纳金和富士等60个株系）组培苗为试材,应用卡那霉素筛选、PCR扩增及RT-PCR扩增对其外源基因的稳定性进行研究。结果表明,在60个常规继代培养6～8年的转CpTI基因苹果转化株系中均可检测出CpTI特异基因片断;除2个转化株系在50 mg.L-1卡那霉素浓度下出现花叶现象,表现缺乏卡那霉素抗性外,其他株系在含相同浓度卡那霉素的培养基中生长正常。RT-PCR检测结果表明,转入的外源CpTI基因在43个株系中得到了较高水平的表达,在15个株系中表达强度低,2个株系未检测到特异表达条带。
Bibliography:ZHOU Ruijin,DU Guoqiang,SHI Xiaoxin（1.College of Landscape Architecture,Henan Institute of Science and Technology,Xinxiang Henan 453003,China;2.College of Horticulture,Agricultural University of Hebei,Baoding Hebei 071001,China）
23-1391/S
apple; nptⅡ gene; CpTI gene; subculture; stability
Using the transgenic lines of apple（Malus domestica Borkh.） carrying exogenous cowpea trypsin inhibitor（CpTI） gene including Royal Gala,Orin,Jonagold and Fuji as the test materials,the stability of exogenous gene in transgenic lines conserved for 6-8 years by subculture at normal temperature were studied by PCR analysis techniques,RT-PCR analysis and Kanamycin resistance evaluation.PCR analysis results showed that all transgenic lines could generate the expectable band.The leaves of two lines presented yellow when the plantlets cultured on the medium with 50 mg·L-1 Kanamycin,which indicated the lack of the Kanamycin resistance,and the others could grow normally.Testing of the expression of exogenous CpTI by RT-PCR analysis in 6
ISSN:1005-9369
DOI:10.3969/j.issn.1005-9369.2011.07.018