Identification And Analysis Of Transgenic Rice Lines HD3's

Rice (Oryza sativa) is a gramineous monocot plants, is the world’s major foodcrops. By Agrobacterium-mediated gene transfer to the cry1C*Rice Songjing9, getTransgenic rice varieties HD3. This study of Transgenic rice varieties HD3be Bastaresistance screening, PCR detection, Southern blot detection, identification of indoorinsect resistance, RT-PCR detection and identification of Bt protein detection, choosea target gene PCR-positive exogenous gene is a low-copy integration, higher levelsof Bt gene transcription and translation, and strong resistance to stem borer transgenicrice lines.The main results are as follows:1. By Agrobacterium tumefaciens EHA105(pBar-cry1c*)-mediatedtransformation of rice varieties Songjing the9th to the bar Basta resistant gene andcry1c*gene PCR-positive plants, normal growth and development as a principle,after five generations of selection nurture successful7HD3transgenic rice lines.2. Transgenic rice lines HD3the Bt gene RT-PCR assay showed that differentstrains of mRNA expression of leaves highest HD3-7, the lowest HD3-3; stem sheathhighest HD3-7, the lowest the HD3-1; Infloresence highest HD3-7, the lowest HD3-4.3. Transgenic rice lines HD3Bt protein content were identified, the results showdifferent different strains of Bt protein content, leaf highest HD3-7, the lowest HD3-3;Infloresence highest HD3-7, the lowest HD3-4; highest stem sheath is HD3-7, thelowest HD3-1.4. Transgenic rice lines HD3conducted Southern blot hybridization identified,results showed HD3-1, HD3-2, HD3-3, HD3-4, HD3-5and HD3-7and other six linesare single-copy integration.5. Transgenic rice lines HD3indoor insect resistance identification results show that, in descending order of insect resistance for HD3-7, HD3-3, HD3-5, HD3-4,HD3-6, HD3-2and HD3-1. Combined expression of mRNA and protein expression ofBt results analysis, the higher the level of gene transcription insect cry1C*The moreBt protein expression, the stronger anti-borer resistance.