Molecular Identification And Insect Resistance Evaluation Of Transgenic Aphid Resistant Rice Line HD4

In this work,HD4 transgenic super japonica rice Songjing9 families were detected by Basta resistance analysis,PCR,GICA,insect bioassay,Southern blot,RT-PCR,ELISA.They had positive PCR results,well Objective expression,low copy integration foreign gene,high expression and translation levels of Bt gene,strong resistance to Rice Stem Borers.The main results of this study included:1.Transgenic super japonica rice Songjing9 lines(pBar-cry2A*)was based on cry2A*being transformed into them based on agrobacterium EHA105(pBar-cry2A*)and(pBar-cry1C*)mixed.This study begin from transforming 135 different independent rice plants of resistant callus which harboring the cry2A*gene and bar gene.The principle is based on the positive testing of Basta,positive PCR testing of cry2A*and bar gene,positive GICA testing of cry2A*gene,normal growth and development,by selecting T0,T1?T4 generation,and nurtured 49 transgenic rice families(bar-cry2A*)named the HD4 families.2.The RT-PCR of Bt gene of the selected 8 HD4 families showed that the expression level of HD4-8 and HD4-1 were the highest,HD4-5 was the lowest.Eight families shows different into expression levels of the Bt protein,and the highest is HD4-8 and HD4-1,the lowest is HD4-5.According to the strength of the trend caused by the different organs of the Bt protein content between the blades>Inflorescence>stem sheath>brown rice.3.The copy numbers of Bt gene and bar in transgenic super japonica rice lines were detected by Southern blot.The result showed that the lines had higher insect resistance were 1-3 copies of Bt gene integration.4.The insect bioassay in the laboratory of the selected 8 HD4 rice families showed that the insect resistance of HD4-8 were the highest,HD4-2 and HD4-5 were the lowest.Compared to RT-PCR results and Bt protein quantitative analysis results found that,the high Bt protein quantitative analysis results in good and the insect resistance results in better.