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 。
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.\%
出处
《林业科学》
EI
CAS
CSCD
北大核心
2001年第6期34-38,共5页
Scientia Silvae Sinicae
基金
黑龙江省杰出青年基金课题
国家"973"项目 (G19990 160 0 3)
国家转基因专项 (J0 0 B 0 0 1)资助
