Cloning And Expression Analysis Of The Male Sterile Related Genes In Chinese Cabbage

Chinese cabbage stamen development is regulated by the genes fine control, deeply analysis to the genes expression patterns, help to create male sterile line, solve the problem of hybrid seed production methods and utilization of heterosis. Using Illumina high-throughput sequencing technology, we do transcriptome sequencing analysis for fertile plant and sterile plant bud of Chinese cabbage male sterile influenza a dual system. According to the data has gotten, in the experiment, we choose 10 differentially expressed and function unknown genes and have them on cloning and expression analysis, the main results is as follows:1. In the experiment, the ID for the 10 genes are Bra020916, Bra008228, Bra025386, Bra023621, Bra040503, Bra005946, Bra024610, Bra020864, Bra008396 and Bra010803, we do sequence feature analyses to these differentially expressed genes, obtained the base sequences of genes, amino acid sequence, isoelectric point and molecular weight, and get the protein type, the modification locus of each protein and their positions. Besides Bra040503, Bra008396 is secreted protein, other are all non secretory proteins.2. Through real-time fluorescence quantitative RT-PCR for the different parts of the fertile and sterile material, it is concluded that the highly expression of those related genes in the fertile plant;According to the results of the analysis in different organs of fertile plants, the gene expression in floral organ significantly higher than the expression in root, stem and leaf area; According to the results of the analysis on various levels fertile bud,Bra020916, Bra008228, Bra025386,Bra023621, Bra040503 these five genes expression quantity is higher in the late flower bud;Bra005946, Bra024610,Bra020864, Bra008396 these four genes work in the middle of the flower bud;Bra010803 gene plays a main role in the early flower bud.3. We selected four genes to Bra020916, Bra00822, Bra008396 and Bra020864 do cloning and carrier construction in the 10 genes we studied, the carrier of successful build into Arabidopsis and do function verification, do PCR verification and GUS staining on the transgenic plants, it is proved that purpose gene has been successfully transferred.