The CDNA-AFLP Analysis Of Gene Differential Expression And RNA Editing Of ATP6 Gene Between Cytoplasmic Male Sterile Lines And Their Maintainer Lines In Soybean (Glycine Max (L.) Merrill)

CMS (Cytoplasmic male sterility) 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. In the present paper, the cDNA-AFLP analysis of gene differential expression and RNA editing of atp6 gene were studied between the cytoplasmic male sterile lines NJCMS1A, NJCMS2A and their maintainer lines NJCMS1B, NJCMS2B in soybean (Glycine max(L.)Merrill). The main results were as follows:1. The cDNA-AFLP differential display technology was established in soybean, which methods and steps were improved and optimized referring to those reported on other species. The results showed that the bands of polyacrylamide gel electrophoresis were clear, rich, and ideal. Therefore the established cDNA-AFLP differential display technology could be used in the related studies in soybean.2. The cDNA-AFLP differential display technology was used to study the gene differential expression between the cytoplasmic male sterile lines NJCMS1A, NJCMS2A and their maintainer lines NJCMS1B, NJCMS2B. A differentially expressed fragment named as 2A-2 was isolated from NJCMS2A by cDNA-AFLP. The result of RT-PCR showed that the expression quantity of 2A-2 in the flower buds of NJCMS2A was much higher than that in NJCMS2B. The blast results showed that 2A-2 was high homology with the conservative region of the MADS-box gene. The homology analysis, structure and function prediction of the amino acid sequence showed that the protein encoded by 2A-2 was the transcription factor of the MADS-box gene. The above results suggested that the 2A-2 fragment might be related to the soybean cytoplasmic male sterility.3. The study on the RNA editing of mitochondrial atp6 gene showed that there were less editing sites in the conservative region of atp6-3. The results of the mtDNA sequence comparision showed that only one base difference was found in the conservative region of atp6-3 gene between NJCMS1A and NJCMS1B. In addition, the obvious difference was found on the expression of atp6-1, atp6-2 and atp6-3 between NJCMS1A and NJCMS1B. The relation between the mitochondrial atp6 gene and the soybean cytoplasmic male sterility needed to be further studied.