烟草打顶对腐胺N- 甲基转移酶基因表达的影响

戚元成1,2, 马雷1, 王菲菲2, 刘卫群1,2,*
河南农业大学1 烟草学院, 2 生命科学学院, 郑州450002

通信作者:刘卫群;E-mail: liuweiqun2004@126.com;Tel: 0371-63555153

摘 要:

水培65 d的烟株进行打顶后, 测定打顶和不打顶烟株上部叶中烟碱含量。分别提取根中总RNA, 以腐胺N-甲基转移酶基因(PMT)特异引物、肌动蛋白基因(actin)为内参作半定量RT-PCR分析; 同时将提取的总RNA转膜, 用地高辛标记PMTRT-PCR产物作探针, 进行Northern杂交分析的结果表明, 打顶后的上部烟叶中烟碱含量和PMT基因表达量显著增加。

关键词:烟草打顶; 腐胺N- 甲基转移酶基因(PMT); 半定量RT-PCR; Northern 杂交

收稿:2009-04-17   修定:2009-06-22

资助:国家烟草专卖局重点项目(110200001011A)。

The Influence of Tobacco (Nicotiana tabacum L.) Topping on the Expression of Putrescine N-methyltransferase Gene

QI Yuan-Cheng1,2, MA Lei1, WANG Fei-Fei2, LIU Wei-Qun1,2,*
1College of Tobacco Science, 2College of Life Science, Henan Agricultural University, Zhengzhou 450002, China

Corresponding author: LIU Wei-Qun; E-mail: liuweiqun2004@126.com; Tel: 0371-63555153

Abstract:

The tobacco (Nicotiana tabacum) plants were topped after water cultured for 65 d and the top leaf nicotine contents of the topped and untopped were examined. Total RNA were extracted respectively from the roots of topped and untopped tobacco plants. Expression analysis of putrescine N-methyltransferase gene (PMT) was carried out by semi-quantitative RT-PCR with the tobacco actin as control and by Northern blotting with the digoxin-labelled PMT RT-PCR product as probe. Results showed that PMT expression and the nicotine content in the top leaf increased after topping.

Key words: tobacco topping; putrescine N-methyltransferase gene (PMT); semi-quantitative RT-PCR; Northern blotting

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