Research On Transgenic Rapeseed (Brassica Napus) With Gastrodia Antifungal Protein Gene By Agrobacterium Tumefaciens Mediated Method

Rape (Brassica napus L. ) is a kind of very important economic crop and oil plant in our country. Its importance was significantly increased during the past several years because of the successful breeding of 'double low' rape. But all the regions that plants rape were threatened by the illness that caused by fungi. The yield and quality of rape were greatly decreased by the illness, especially the double-low lines which were more sensitive because of low content of thioglycoside.Gastrodiae ( Gastrodia elata B1. ) is a kind of plant that can symbiose with fungi, because it contains GAFP (Gastrodia Antifungal Protein) in the out layer of its corm. Wang Yiqin et al. (2000) have cloned the GAFP gene successfully, and proved that the prokaryotic expression product of this gene has the repressive activity to fungi.In the present research, Brasscia napus L. double-low cultivar westar was used as experiment material, the hypocotyls were used as explants and were transformed by GAFP gene via Agrobacterium mediated transformation system. Transgenic plants were obtained after co-culture, antibiotic screen and root regeneration. The transgenic plants were detected with PCR, genomic Southern blot and RT-PCR. The results suggest that foreign GAFP gene has already been combined into the rape genome and were expressed. The factors that influence the frequency of Agrobacterium mediated transformation was also discussed.