Studies On Production Of Herbicide-resistant Rice Plants Mediated By Agrobacterium Tumefaciens

Rice is the most important food crop in the world and has become one of the model crop for the research of molecular biology. Exogenous Bar gene and atzA gene were introduced into rice restoring lines and maintaining lines respectively. The main results are as follows:1. Two japonica varieties were used for the genetic transformation (Oryza saliva L.) by using Agrobacterium tumefaciences EHA105 harboring binary vector containing Bar gene and selectable marker gene of HPT. 77 and 55 transgenic rice lines were obtained respectively. PCR analysis showed that Bar gene had been integrated into the rice genome. The segregation of HPT gene in most of T1 progeny corresponded to the Mendelian ratio.2 . An Agrobacterium binary vector carrying chlorohydrolase gene was successfully constructed based on pCambia1301. The stragety of construction plant binary vector is as follows: first, the target gene atzA has been prepared by digestion with EcoRI/Hindâ…¢, then the fragment can be ligated with vector pBluescript SK(+/-) cut with the two same enzyme. The recombinant plasmid pBS-SK-atzA was digested by EcoRI/Hindâ…¢, which can separate the 1.5kb atzA gene fragment ; Second, plasmid pBS-SK-atzA and pCambiaBOl were digested by EcoRI/Hindâ…¢ at the same time, then ligating the two fragment into a new plasmid vector p1301-atzA.3 . The target gene was successfully introduced into mature embryos of a Japonica rice cultivar Jindaol07 by Agrobacterium-mediated transformation. 249 transgenic plants were obtained. The exogenous atzA gene in those transgenic plants that expressing resistance to herbicide atrazine was confirmed with PCR and PCR-Southern blot analysis .4. The results of herbicide resistance of transgenic rice showed that most of transgenic rice plants have behaved common herbicide resistance to atrazine , but the expression level of herbicide resistance is different.