Rice Empty Sterile 131 (cry1c *) Molecular Detection And Insect Resistance Evaluation

Rice is one of the main staple food, about half of the world population (most of them are Asian) lived on it. The area of rice planting in China is about a quarter of the total rice planting area in the world, and is one third of crop planting area in China, so high and stable rice yield is important for securing the food safety of China and the world. Rice yield can be affected by many factors, among which stem borer is one of the major cause of rice yield loss. At present, rice pest is chiefly controlled by chemical pesticides, the use of which usually can cause pesticides contaminant, environmental pollution and increase of rice production costs. Biotic method can come over these problems. Insecticidal genes from other species(such as Bacillus thuringienesis) can be introduced into the rice genome of Heilongjiang province by transgenic method to culture transgenic insect-resistant rice varieties, which is of significant meaning for rice production of Heilongjiang province.Insecticidal gene was introduced into Kongyul31, which is the major rice varieties of Heilongjiang province, by Agrobacterium mediated transformation by key laboratory of molecular biology of Heilongjiang University. And series of cry1C* transgenic Kongyul31 plantlets were got. In this study, these transgenic plantlets were analyzed in molecular levels and insect-resistant evaluation was also done. The main results in this study are as follows:1. PCR analysis of To, T1, T4 generations of crylC* transgenic Kongyu131 plantlets showed there were PCR positive plantlets in all of the four generations, so cry1C* gene had integrated into the genome of Kongyu 131 and could inherit stably.2. To confirm the expression of bar gene, T1 generation plantlets were undergone PPT-resistance assay in the lab, and herbicide basta were sprayed over T4 generation plantlets in the field. By these tests PPT-resistant plantlets and basta-resistant plantlets were separately got in T1 and T4 generation, which meant bar gene had integrated into the genome of Kongyu131, and could express and inherit stably in different generations. And it also could indirectly prove that cry1C*gene which was closely linked with bar gene expressed stably in different generations.3. Bioassay in the laboratory showed that T4 generation of cry1C* transgenic Kongyu 131 were highly resistant to striped stem borer, the mortality of the striped stem borer larvae was nearly 100% after fed by 5 days.4. Southern blot showed that most of insect-resistant T4 generation plantlets contained single copy. These single copy transgenic plantlets could be used to culture stem borer-resistant transgenic rice varieties of Heilongjiang province.5. Field evaluation showed that transgenic plantlets can resist pest damage well, and there was no great agronomic traits changing.