Rice Bel Gene: Cloning, Expression Pattern Analysis And Its Transformation To Tabacco

Bentazon lethality/sensitivity of Rice mutant 8077S is controlled by a pair of recessive gene bel, it became a hotspot because of its useful in two-line hybrid rice breeding. In this research, we confirmed that Bel is located in the same position as SNP 151, using mutant analysis and bioinformatics methods. We then cloned Bel gene and surveyed its expression pattern via semi-quantity RT-PCR. Then it was constructed to plant expression vector and made transgenic tobacco analysis. The main result described as below:1. In the basis of former study of SSR and SNP genetic mapping, we made gene prediction and bioinformatics analysis of bel mapping region. We conclude that SNP151 locus is what bel gene locus; it was mutated by a single base deletion.2. We cloned Bel gene, constructed its coding sequence to pMD18-T vector and sequencing it, NCBI blast find that it's a member of cytochrome P450 gene family named CYP81A6, compose by two exon and encodes a protein with 513 amino acid residues.3. Semi-quantity RT-PCR analysis proved that Bel is bentazon inducible and express highest on 12h after bentazon application. It also showed tissue expression difference with the highest mRNA acculation in leaf tissue.4. We subcloned bel into plant expression vector pKYL71, and transform pKYL71-Bel to GV3101, transfect it with tobacco leaf dishes. Transgenic tobacco was proved by PCR, bentazon treatment reveal that transgenic tobacco acquired bentazon resistance.