Preliminary Study Of The Resistance To Rice Blast And Sheath Blight Of Transgenic Rice With Luffin-a、luffin-b Gene From The Seeds Of Luffa Cylindrical

Rice is one of the most important food crop in the world, while because of the fungus diseases caused by Rice blast disease and the Rice Sheath Blight seriously affect the high and stable yield. The traditional way by chemical pesticide prevention not only has some bad effect on environment, but also dose bad to farmers and consumers’health. It is the most economic and effective method to develop disease-resistant varieties. It is a efficient way to gain disease-resistant varieties by transgenic technology with disease-resistant genes constantly discovered and cloned. Some experiment has proved that the rice sucrose synthase gene promoter pRSUSl is a tissue specificity promoter which could drive exogenous genes high expression in the roots、stems、leave, but did not in seed. Luffin is one of the members of ribosome-inactivating protein, it is a toxic protein than can inhibit the protein synthesis in the cell ribosome by a specific target of the large rRNA, leads to cell death. It has a broad resistance to the virus、fungi and insects. In this research,we select the rice sucrose synthase gene promoter pRSUS1and Luffin a、Luffin b gene which have safety genetically modified safety, then construct two expression vectors, one is pRSUS1link up with Luffin a, the other is pRSUS1link up with Luffin b for the Agrobacterium-mediated transform to a kind of rice tp309、We use the PCR and RT-PCR technology to detect the genetically modified plants which are selected by hygromycin, and identify whether these plants have resistance to rice blast and sheath blight or not and carry on agronomic traits of the survey. Through this way, we wish we can screen out not only has the disease resistance and do not affect the agronomic characters of genetically modified rice loss. So far, the main study results obtained are as follows:traditional1. The expression vector pRSUSl-Luffin a、pRSUS1-Luffin b were transformed into TP309by Agrobacterium tumefaciens mediated transformation.11transgenic plants containing pRSUS1-Luffin a and24transgenic plants which containing pRSUS1-Luffin b are screened from T0generation by PCR.2.The T1transgenic plants generation of pRSUSl-Luffin a were analysed by PCR、RT-PCR and resistance to sheath blight, the results show that8lines of transgenic plants were coincident with Free separation principle of3:1ratio of Mendel’s Law, RT-PCR results were positive, seven lines of transgenic plants have a certain resistance to sheath blight compared with TP309. The T2transgenic plants generation of pRSUS1-Luffin a were analysed by PCR、RT-PCR and resistance to sheath blight, the results show that4lines of transgenic plants have a certain resistance to sheath blight compared with TP309and RT-PCR results were positive,A1-3/1-7transgenic plant was inoculated rice blast, the resistance to rice blast is at3level.410of transgenic plants of pRSUS1-Luffin b were detected the expression of target genes by the PT-PCR technology and results are positive5. The Tl transgenic plants generation of pRSUS1-Luffin b were analysed by PCR、RT-PCR and resistance to sheath blight, the results show that5lines of transgenic plants were positive by PCR and RT-PCR, five lines of transgenic plants have a certain resistance to sheath blight compared with TP309. B4-2and B6-4transgenic plant was inoculated rice blast, the resistance to rice blast is at3level6. A1-3/1-7and B6-4transgenic plants were analysed of general resistance to rice blast, results show that transgenic plants have a certain resistance to rice blast kinds of10,11,13,21,44,47compared with TP309.7. The T2pRSUS1-Luffin a and T1pRSUS1-Luffin b generation transgenic plants were investigated with the several major agronomic traits, results show that the plant height of TP309are lower than that of pRSUS1-Luffin a-、pRSUS1-Luffin b-and transgenic plants as well as the tillers of TP309,but others agronomic traits were similar, so we consider agronomic traits were not changed.