Identification Of Traits Of HD 1 And HD 3 Lines And Assessment Of Soil Environmental Safety Of Transgenic Insect-resistant Early Indica Rice

Rice?Oryza sativa L.?is native to tropical Asia,which is an annual gramineous plant with 24 chromosomes.Heilongjiang is one of the major rice producing provinces in China,but also suffers from a serious agricultural pest problem that costing millions a year.Traditional pest prevention and control methods rely mainly on chemical pesticides,but because of its widespread use over many years,not only destroyed the ecological environment,developed pests resistance,seriously the chemicals pose a ut also cause a threat to human and animal health.The conventional breeding is not only a long cycle,and also it is difficult to develop excellent insect-resistant varieties.Fortunately,with the development of molecular biological technology,the emergence of the Bt gene of rice has become an effective way to solve this problem.At present,rice Kongyu 131 which is the largest japonica rice varieties in China has the largest planting area in Heilongjiang province.Songjing 9 with high yield and quality is also widely cultivated.In this study,the transgenic Bt gene Kongyu 131 and Songjing 9 were used as test materials to coduct depth and comprehensive molecular traits,identification of agronomic traits and assessment of soil safety.The results are as follows:1,Through Basta resistance screening,test strip test and conventional PCR detection,through the screening of T8^T10 generations,successful identification of turn Bt gene Kongyu 131 and Songjing 9,HD₁?Kongyu 131?cry1C*/bar??and HD₃?Songjing 9?cry1C*/bar??line.2,The expression level of target gene mRNA and the transcriptional level of Bt gene in the transgenic rice HD1 and HD3 heading stages,seedling tissues and brown rice showed that:the expression of mRNA in the same growth phase of the same tissue of the different strains was different:HD1-1>HD1-2,HD3-1>HD3-2;and there were obvious difference in the same growth phase of the different tissue of the same strains.There were significantly difference in the cry1C*protein expression in the different growth phase of the same tissue of the same strains,while that were significantly difference in the same growth phase of the different tissue of the same strains;and there were difference in the cry1C*protein expression in the samet growth phase of the same tissue of the different strains:HD₁-1>HD₁-2,HD₃-1>HD₃-2;the cry1C*protein expression in the different tissues in the growth phase,ripening stage are the lowest.3,Transgenic gene rice HD₁ and HD₃ were genetic stability in every generations,and were single copy inserted into the rice genome,the insertion site of exogenous gene was intergenic region,the exogenous gene of HD1-1 was inserted into chromosome 3,and HD3-1 was inserted into chromosome 8.4,Compared with CK,there was no significant difference in agronomic traits and rice quality in the HD₁ and HD₃ lines with cry1C*gene,HD₁-1 and HD₃-1 are better.5,Compared with the cultivation of CK soil,there was no significant difference in the number of bacteria,fungi and actinomycetes in root and intertidal field soil which planting cry1C*transgenic rice HD1 and HD3,and found that there were no cry1C*gene transferred to soil microorganism.