Identification Of Cry1C * And Cry2A * Genes In Super Japonica Rice

Rice was vital for people, because it was the main food for above two billions people all over the world. However, insect damage was the major enemy for rice, which could make bad influence on its production and quality. According to related reports, insect damage could cause huge economic losses in every year. The employment of traditional chemical synthesized pesticides could lead to environmental pollution and impair human health. To breed insectinsect-resistant rice varieties is the most economical and effective method. The genes resisting Lepidoptera damage are not found in cultivated rice and related species. Therefore, it was meaningful for people to use transgenic Technology to solve insect damage problem, especially Borer damage.In this work, T3 transgenic super japonica rice Songjing9 lines, crylC*and cry2A* being transformed into them, were detected by Basta resistance analysis, PCR, insect bioassay, Southern blot, RT-PCR, Bt protein quantitative analysis and agronomic traits investigation. Some transgenic super japonica rice lines were selected out. They had positive PCR results, low copy integration foreign gene, high expression and translation levels of BT gene, strong resistance to Rice Stem Borers, and they had no significant difference about agronomic traits with the control. The main results of this study included:1. Basta resistance screening of 106 T3 transgenic super japonica rice lines,98 lines of them showing the Basta resistance, the resistance rate as high as 92.45%.4 kinds of different transgenic materials, TlC(Songjing9(cry1C*)), T2A(Songjing9(cry2A*)), T1C/2A(Songjing9 (cry1C*/2A*)) and TBar(Songjing9(Bar)), were obtained by PCR screening of Basta-resistant.2. The insect bioassay in the laboratory of the selected 3 transgenic super japonica rice lines showed that the insect resistance of T1C/2A were the highest, TIC were in the second position, T2A were the lowest The insect-resistant level of different lines in T1C/2A、T1C and T2A were different. T1C/2A-2 was the strongest in T1C/2A; T1C-1 was the strongest in T1C; T2A-9 was the strongest in T2A.3. The copy numbers of Bt gene and bar in transgenic super japonica rice lines were dectected by Southern blot. The result showed that the lines had higher insect resistance were 1-2 copies of Bt gene integration. Through RT-PCR of Bt gene, the transcription of the different transgenic lines were different. Among TIC, T1C-1 was the highest;Among T2A,T2A-9 was the highest; Among T1C/2A, two Bt gene transcripition of T1C/2A-2 were highest. Through the Bt protein quantitative detection, the result was almost the same with mRNA express level of Bt gene. The expression of Bt protein in leaves of T1C-1 and T2A-9 were highest among their own materials, respectively. The expression of Bt protein of T1C/2A-2 were highest in co-transformation material. By the combination of the insect bioassay in the laboratory and gene expression detection, it was found that the lines with good insect resistance had relatively high BT gene expression.4. By agronomic traits investigation,6 transgenic super japonica rice lines, T1C-1, T1C-4, T2A-2, T2A-7, T2A-9 and T1C/2A-2, were selected. They had good insect resistance, low copy integration and high expression of BT gene, and no significant difference with control.