The Screening Of Rice T-DNA-inserted Mutants And The Research Of Floral Development Mutant And The Gene Clone Of Heading-date-related Mutant

Rice became a model plant of functional genomics research, because of the staple food, the smaller genome, well developed genetic maps and being transferred easily. With the completion of sequencing of the rice genome, the post-genome era is coming. Namely, post-genome is functional genome and proteome. In order to research the functional genome of rice, researchers created a mass of rice mutants by various methods, and especially created many T-DNA inserted mutants, which contained the specific T-DNA as a tag of isolating the specific gene.In this study, we screened, identified and analysis of the library of rice T-DNA inserted mutants, for ZH11 as receptor. The main results are following:1 The traits of transferred plants of ZH11 by transferring T-DNA are investigated and screened. 1071 stable lines of phenotype variant are obtained and 173 lines show T-DNA insertion through testing the 450 lines of these by PCR with the specific primers of Bar gene. The types of mutants includes heading date, height, leaf color, leaf shape, the number of tillers and so on.2 TAIL-PCR and adaptor-mediated nested PCR are applied to amplify the flanking sequences of the T-DNA insertion in the study. By compare with the efficiency of them, the result shows the efficiency of adaptor-mediated nested PCR is higher of that of TAIL-PCR.3 By screeing of the mutants of T-DNA-inserted ZH11, the mutants with late heading date is obtained. Further more the mutant show smaller weight per kilograin and shorted root. Genetic analysis show the relative traits are controlled by a pair of genes and codominant. By sequencing and aligment of the flanking aequence obtained by adaptor-mediated nest PCR, it is found that T-DNA inserts into the last exon of the gene, the gene is a putative early flowering gene, and named Early Flowering 3(EF3). The gene locates in Chrl, and encode a putative monocopper oxidase precursor. By cpmpare the rice genome with the sequence and function of the relative cloned genes, EF3 is thought be a novel gene controlling rice heading date. In order to test the function of EF3, the function complement vector and two RNAi vectors are constructed. The testing of the function is doing.4 A mutagenesis rice mutant L2 layer aberrant differentiation ladl have the differentiation of L2 layers of male and female reproductive organs. Genetic analysis indicated ladl was a single recessive gene. By microscopic observation, we discovered the ladl mutant had one or two regular layers in the anther wall and at least seven archespores or archespore-like cells in the ovules in stage Ov2. Because of seed setting by being pollinated with the pollen of wild type, at least one MMC can undergo meiosis and produce functional megaspores. By compared with nzz/spl mutants in Arabidopsis and mpslmutant in rice and microscopic observation, we purpose that LAD1 gene should be an allele or upstream gene of MSP1 and regulate the development and differentiation of L2 layers in anthers and ovules by cell communication between cell layers.