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Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (05): 788-793.doi: 10.3724/SP.J.1006.2010.00788

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Transgenic of Ramie with Synthetic CryIA+CpTI Gene by Agrobacterium tumefaciens-Mediated

 MA Xiong-Feng1,3,YU Chun-Meng1,TANG Shou-Wei1,GUO San-Dui2,ZHANG Dui2,WANG Yan-Zhou1,SHU Ai-Guo1,SHU Si-Yuan1,XIONG He-Beng1*   

  1. 1Institute of Bast Fiber Crops,Chinese Academy of Agricultural Sciences/Key Laboratory of Genetic Improvement & Engineering Microbiology for Bast Fiber Crops,Changsha 420105,China;2Biotechnology Research Center,Chinese Academy of Agricultural Sciences,Beijing 100081 China;3Institute of Cotton Research,Chinese Academy of Agricultural Sciences,Anyang 455000,China
  • Received:2009-08-26 Revised:2009-12-21 Online:2010-05-12 Published:2010-03-15
  • Contact: XIONG He-Ping,E-mail:ramiexhp@2118.cn;Tel:0731-88998500 E-mail:ibfc-maxif@163.com

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Abstract:

Ramie [Boehmeria nivea L. (Guad.)] is a perennial herbaceous plant of the urticaceae family. Ramie affected by many insect pests above and below the ground during the growing season. The pests damage has become an important factor affecting ramie yield and quality in China. Application of chemical insecticides not only is less effective and costly, but also causes environmental pollution. Development of a genetic transformation system to introduce desired genes (e.g. toxin genes) into ramie could improve its agromonic traits such as resistance to insects. The study was conducted to transfer CryIA gene and CpTI gene into the ramie variety Zhongzhu 1. The CryIA gene encodes Bt toxin protein from Bacillus thuringiensis toxinof Vitis pseudoreticulata; while, the CpTI gene encodes a cowpea trypsin inhibitor produced by Vigna unguiculata. Synthesized CryIA geneand CpTI gene were inserted into plant expression vectorpGBI4ABC whichcarried high efficient bivalent insect resistant genes. With high frequency of adventitious shoot induction and plant regeneration, ramie hypocotyls of the variety Zhongzhu 1 were transformed with Agrobacterium tumefaciens strain LB4404 harboring the plant expression vector pGBI4ABC, and then shoot buds formed from calli induced from hypocotyls were selected with the mixed solution of 50 mg L–1 Kanamycin and 300 mg L–1 Cefotaxime. The result showed that 32 Kanamycin-resistant plants were obtained, and therein 26 plants were positive in PCR test, indicating that 80% of the Kanamycin-resistant plants contain CryIA gene and CpTI gene. Southern blot and GUS analysis also verified that the foreign genes of CryIA and CpTI were successfully integrated into the genome of the transgenic plants and expressed.

Key words: Ramie, Insect resistant genes, Agrobacterium tumefaciens, Transegenetic plants

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