The Identification Of Rice T-DNA Insertional Mutants And The Preliminary Study On A Heading-delayed Mutant Line

Rice is the staple food for more than one third of the world population. Because of its small genome size, relatively well developed genetic maps and easily to be transferred, rice is a model plant for functional genomics research. With the completion of rice (Oryza sativa) genome sequencing, large information will be available which may play important roles in rice functional genomics. Obtaining a mutant library by insertion is a powerful strategy for the research of rice functional genomics.The aim of this thesis was to generate a population of T-DNA insertional mutant, made identification and analysis of important T-DNA insertional mutants, which supplied technique and material to the research of rice functional genomics. The major results obtained were follows:1. On the basis of the collection of T-DNA tag lines which was established, lots of mutants with distinct phenotype variation were found in Ti generation, including dwarfing mutant, heading-delayed mutant, few tiller, curling leaf, dissilient hull and grain shape etc.2. Some rice mutated lines resulted from T-DNA insertion have been screened through genetic analysis between To and Ti, together with the molecular biology proofs (PCR analysis). A set up of the efficient technology of high-throughput thermal asymmetric interlaced PCR (TAIL-PCR) was established.3. A Heading-delayed mutant line named M29 was characterized, which indicated that it was co-segregated with Basta resistance. Flanking sequence at T-DNA insertion site of the tagging line was obtained by tail-pcr. Homologous comparison was conducted by BLASTn and BLASTx with the data in GenBank. The chromosome location of flanking sequences and possible encoding proteins were presumed.4. The whole sequence of gene lh (temporary name) of the mutant was analyzed and presumed in the bioinformatics pipeline against the TIGR Rice Pseudomolecules (virtual contigs), at the same the whole fragment of the gene had been obtained through LA-PCR, and it was about 5Kb. The further cloning the lh and construction of the plasmid vector were in progress.