Analysis for Intron 1 and 2 of Rice Transformation to Promoter Activity
|School||Nanjing Agricultural College|
|Course||Crop Genetics and Breeding|
|Keywords||Rice Gene expression Intron Darkness inducing|
Rice blast was a fungal diseases and caused by ascomycete Magnaporthe grisea Barr [a sexual generations Pyricularia grisea Saco.] of rice (Herbert,1971), blast and sheath blight and bacterial leaf blight were three main diseases of rice; this disease has occurred almost every cultivation countries and regions of rice, if it had suitable conditions, it’s easy to pop and cause disaster, and even erected; the way to solve this problem is to cultivate disease-resistant varieties, The gene engineering was an efficient methods which transport disease resistance gene into rice varieties, it has a very important theoretical significance and application value in breeding for disease resistance of ricePib gene is the first major blast resistant gene cloned from rice by mapping clone technique and which is belongs to NBS-LRR resistant disease gene family.at the NCBI site,we search the pib gene to find already published pib gene nucleotide sequence, analysis showed that, pib gene 10322bp long,containing the complete structural gene of about 5.4kb,3573bp of ATG and 1000bp upstream section.pib is a 4introns and 5 exons of the split genes,two of the introns is before of the ATG.According to the structural characteristics of Pib gene, we use the 3’deltion of pib promoter to analyze the gene expression in the introns, vectors were each named pNAR1101, pNAR1102 and pNAR1103. And use the Agrobacterium-mediated transformation Nipponbare of rice varieties, a total of 98 transgenic plants were obtained, pNAR1101 had 32 plants, pNAR1102 had 30 plants, pNAR1103 had 36 plants, PCR a showed that:the gene has been integrated into the genome of Nipponbare.By using transgenic rice plants of 3’deletion pib promoter fragment-gus constructions, the relationships between the deletion of introns in pib promoter and activity of pib promoter and its darkness inducing attribute were systematically analyzed. The results from GUS histochemistry analysis revealed that the deletion of the two introns and pib promoter fragments of downstream in its 3’end remained promotion activity and induced activity. with the deletion of introns, the activity of promoter was significantly reduced; the induced activity was siganificantly droped, indicating that the introns can enhance the promotion activity and induced activity,these also verify that the intron components may also be involved in the regulation of induction of components and the regulation of the gene transcription and expression.