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《Chinese Journal of Biotechnology》 1991-01
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Insect Resistance of Transgenic Tohacco Plants Expressing δ-endotoxin Gene of Bacillus thuringiensis

Tian Yingchuan;Qin Xiaofeng;Xu Bingyin;Li Taiyuan;Fang Rongxiang;Mang Keqiang;Li Wengu;Fu Wenjun;Li Yiping;Zhang Shufang;Xie Qiangjang Institute of Microbiology, Academia Sinica, Beijing Institute of Entomology, Academia Sinica, Shanghai Institute of Zoology, Academia Sinica Beijing  
The original B.thuringiensis δ-endotoxin (Bt toxin)gene clones TH12and TH48 contain two different classes of homologous genes, the 5.3kbclass and 6.6kb class respectively. Bt toxin genes of both classes modj-fied at the 5'-end and truncated at the 3′-end can still be expressed toproduce the entomocidal, truncated toxin proteins in E.coli. The modi-fied Bt toxin genes were inserted into the plant binary expression vec-tor pBin437 (a derivative plasmid of pBin 19) and were transferedinto tobacco genome by disarmed Ti plasmid pAL 4404 gene transfersystem. Southern blot and DNA slot blot analysis indicate that the Bttoxin genes have been integrated into tobacco genome at a copy numberof 1--5. Northern blot analysis of poly A~+ RNAs from progenies ofthe transgenic plants revealed that Bt toxin genes of both 5.3 and 6.6kb classes were expressed in transgenic plants, though the transcriptswere degraded to RNAs of lower molecular weights. In entomocidal test,5 plants from the progeny of 5.3 kb class gene-transformed SR1 tobaccoplants and 3 plants from that of 6.6kb class gene-transformed plantswere found to be toxic to the testing larvea of H. assulta. In compari-sion with the control, mortality of the insects feeded on transgenicplants reached 40--50% and the growth of the survived insects was re-markably inhibited. These results indicated that the modified Bt genes ofthe 5.3kb and 6.6kb classes were expressed in transgenic plants andcould confer the transgenic plants a new character of insect resistance.
【Fund】: 国家科委和国际科学文化中心的世界实验室(WLICSC 日内瓦 洛桑)资助项目
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