Gene Differential Expression and RNA Editing of atp9 between Soybean Cytoplasmic-Nuclear Male Sterile Lines and Their Maintainer Lines
|School||Nanjing Agricultural College|
|Keywords||Soybean [Glycine max (L.) Merr.] Cytoplasmic-nuclear male sterility (CMS) Gene differential expression RNA editing|
Cytoplasmic-nuclear male sterility (CMS) plays an important role in the utilization of crop heterosis. It is of important significance on the theory and practice to study the genetic base and mechanism of CMS. To reveal the molecular mechanism of soybean cytoplasmic-nuclear male sterility, the gene differential expression and RNA editing of atp9 gene between soybean cytoplasmic-nuclear male sterile lines and their maintainer lines were studied in the present paper. The main results were as follows:1. A large number of differential expression bands including special expression bands(TDFs) and high expression bands between soybean cytoplasmic-nuclear male sterile lines NJCMS1A, NJCMS2A and their corresponding maintainer lines NJCMS1B, NJCMS2B were obtained by the cDNA-AFLP technology. The results of TDFs analysis showed that the TDFs from NJCMS1A and NJCMS1B were mainly appeared in the middle and large flower buds, while those from NJCMS2A and NJCMS2B mainly in the small and middle flower buds. Total 55 TDFs were recycled and sequenced. The BLAST analysis showed that the function of the proteins encoded by these 55 TDFs were mainly related to the signal transduction, the electron transport and energy metabolism, the cell wall synthesis and regulation, the protein metabolism, the defense and stress responses. QRT-PCR results showed that the expression of 12 TDFs had spatial and temporal differences.2. A special expression fragment TDFs25 was isolated from the large flower buds of NJCMS1B by cDNA-AFLP. The blast results showed that TDFs25 was high homology with the conservative region of the DUF620 gene, which was an unknown functional protein. The full-length cDNA of DUF620 was 1308bp and encoded 436 amino acids. The semi-quantitative and quantitative PCR results showed that the DUF620 gene expression in the large flower buds of NJCMSIA and NJCMS2A were significantly lower than those of NJCMS1B and NJCMS2B. It was inferred that DUF620 gene might be related to soybean cytoplasmic-nuclear male sterility. 3. The sequence analysis showed that the cDNA sequence of atp9 from NJCMS1A was the same as that from NJCMS1B, while the cDNA sequence of atp9 from NJCMS2A was different from that of NJCMS2B. The protein trans-membrane structure prediction showed that the atp9 protein structure of NJCMS2A was obviously different from that of NJCMS2B. The relation between atp9 and soybean cytoplasmic-nuclear male sterility needed to be further studied.