Cloning And Expression Analysis Of Ogucms Related Genes Of Brassica Oleracea

During the research of OguCMS expression pattern of anther in cabbage, we got four down- regulated expression EST sequences and one up-regulated expression EST sequence, The full length cDNA of these five OugCMS related genes were cloned in silicon by using these ESTs as querying probes. Their encoded protein sequence and putative functions were discussed. Spatial and temperal expression characteristics were analyzed by using RT-PCR and quantitative RT-PCR by using different plant organs and flower buds at different developmental stages of Brassica oleracea.Main conclusions are listed as follows:1. BoBHLH1 is a new member of bHLH transcription factors family related to OguCMS. The results showed that a 309 bp ORF and a 102 amino acids peptide were coded by this gene, located the gene in mitochondria by sub-cellular localization. It has 87% identity with AtBHLH151 in Arabidopsis, has an obviously domain of bHLH. Expression of BoBHLH1 in different organs and different development periods in cabbage anther were elucidated by using RT-PCR and Real time RT-PCR. BoBHLH1 is preferentially expressed in cabbage anther and have two expression peaks in early and late stages of anther development.2. BoMYB1 is a MYB transcription factor family member in the cabbage, the length of cDNA is 751bp, containing a 699bp ORF, encoding 232 amino acids. Located in the mitochondria, the gene has 83% identity with AtMYB21 of R2R3MYB transcription family in Arabidopsis chromosome 1. The gene is predominantly expressed in anther, have a peak in late stage of anther development.3. BoMF1 is a function-unknown gene; the full length of genomic DNA is 1128bp, containing introns, the length of cDNA is 586bp, containing a 447bp ORF, and encoding 148 amino acids, located in the mitochondria. The function of gene needs further study. By RT-PCR and quantitative PCR analysis showed that the gene is predominantly expressed in anther, have a peak in late stage of anther development.4. BoMF2 is an up-regulated genes in cabbage, the length of cDNA is 696bp, containing a 624bp ORF, encoding 207 amino acids, located in the nucleus. Its Protein has 78% identity with At2g42940, is a putative DNA binding proteins, and BoMF2 function is a function-unknown gene. The gene is predominantly expressed in anther; the expression has a peak the S2 stage of anther development, and then decreased in late stage of anther development.5. BoMF3 may be involved in the regulation of caspase activity, the length of cDNA is 1156bp, contains a 1032bpORF, encoding 343 amino acids, located in the extracellular. The gene has 78% identity with At1g06260. BoMF3 predominantly expressed in petals, the expression reached a peak in the S4 period, and then decreased in late stage of anther development.