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耐盐基因Bet-A转化小黑杨的研究
STUDY ON THE TRANSFORMATION OF POPULUS SIMONII×P.NIGRA WITH SALT RESISTANCE GENE BET A
【摘要】 本研究建立了小黑杨 (Populussimonii×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(Agrobacteriumtumefacien)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
【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;
- 【文献出处】 林业科学 ,Scientia Silvae Sinicae , 编辑部邮箱 ,2001年06期
- 【分类号】S722.3
- 【被引频次】140
- 【下载频次】388