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林业科学 ›› 2001, Vol. 37 ›› Issue (6): 34-38.doi: 10.11707/j.1001-7488.20010607

• 论文及研究报告 • 上一篇    下一篇

耐盐基因Bet-A转化小黑杨的研究

杨传平 刘桂丰 梁宏伟 张慧   

  1. 东北林业大学,哈尔滨150040;山东师范大学,济南250014
  • 收稿日期:2001-06-25 修回日期:1900-01-01 出版日期:2001-11-25 发布日期:2001-11-25

STUDY ON THE TRANSFORMATION OF POPULUS SIMONII×P.NIGRA WITH SALT RESISTANCE GENE BET A

Yang Chuanping,Liu Guifeng,Liang Hongwei,Zhang Hui   

  1. Northeast Forestry University Harbin150040;Shandong Teachers University Jinan250014
  • Received:2001-06-25 Revised:1900-01-01 Online:2001-11-25 Published:2001-11-25

摘要:

本研究建立了小黑杨(Populus simonii×P .nigra)组培再生体系,确定小黑杨芽分化最适宜培养基、生根培养基,然后进行卡那霉素敏感性试验。以小黑杨无菌苗叶片为转化受体材料,通过根癌农杆菌(Agrobacterium tumefacien)介导法将外源基因Bet-A(编码胆碱脱氢酶,催化胆碱生成甘氨酸甜菜碱)导入小黑杨(P.simonii×P.nigra)。对获得的11株转化苗进行了PCR扩增检测和斑点杂交检测,其中7株获得了1.7Kb特异性扩增条带和杂交斑点,表现为阳性。从斑点杂交检测的转化苗中选取3株杂交斑点明亮的转化子进行Southern印迹杂交分析,结果证实外源基因已整合到小黑杨基因组中。

关键词: 甘氨酸甜菜碱, Bet-A基因, 遗传转化, 小黑杨

Abstract:

In this study,exogenous gene Bet-A (encoding choline dehydrogenase,synthesizing glycine betaine) was introduced into Populus simonii×P.nigra by Agro-bacterium tumefacien,so as to enhance salt resistance in transformed plant.Firstly,the optimal media of P.simonii×P.nigra for bud differentiation and rooting were established.The sensitivity of the explants to Kanamycin was tested in order to decide a reasonable selective pressure for transformation.Secondly,Bet-A gene was introduced into leaves of asepsis plants of poplar by agrobacterium,and then transformed poplar plants were obtained.PCR and southern blotting analyses showed that Bet-A gene has been integrated into the genome of P.simonii×P.nigra.

Key words: Glycine betaine, Bet-A gene, Genetic transformation, Populus simonii×, P.nigra