Differential Proteomic Studies Of Cytoplasmic-Nuclear Male-Sterile Lines And Their Maintainers In Soybean (Glycine Max(L.)Merrill)

Cytoplasmic-nuclear male sterility is a mojor tool in the hybrid seed production for utilization of heterosis in crops. The knowledge of the mechanism of the cytoplasmic-nuclear male-sterility is essential to hybrid seed production. Few studies have been done on the proteomics of the male sterility in crops, especially no relevant paper has been reported in soybean so far. The soybean cytoplasmic-nuclear male-sterile lines NJCMS1A and NJCMS2A were developed through consecutive backcross procedures in the National Center for Soybean Improvement, Nanjing Agricultural University. The present paper was aimed at the differential proteomic studies of the different organs between NJCMS1A, NJCMS2A and their maintainers NJCMSIB, NJCMS2B respectively. The results were as follows:1.The proteomic approach was used to detect the differentially expressed proteins of the different organs between the male-sterile lines NJCMS 1 A, NJCMS2A and their maintainers NJCMS IB, NJCMS2B respectively. The results showed that the differences were investigated on the 2-DE maps of anthers and seeds, but not found on those of leaves between NJCMS1A, NJCMS2A and NJCMS1B, NJCMS2B respectively. The above results showed that the expression of the proteins related to the male sterility were spacially and periodly.2.The matrix-assisted laser-adsorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technique was used to obtain the peptide mass fingerprinting of the differentially expressed proteins in anthers between NJCMS1A and NJCMS1B. In the anthers at uninucleate microspore stage, 7 differentially expressed protein spots were identified. In the anthers at binucleate pollen stage, 16 differentially expressed protein spots were identified. According to the literature, the functions, especially those related to the male sterility of the major differentially expressed proteins, including 1-aminocyclopropane-1-carboxylate oxidase (ACC oxidase), ACC synthase 2, cysteine proteinase, vacuolar H~+-ATPase A subunit, MADS box protein, starch branching enzyme, and UDP-glucose pyrophosphorylase etc. were reviewed and discussed. It was inferred that the male sterility of NJCMS1A might be related to energy metabolism turbulence, the programmed cell death (PCD), ethylene excessive synthesis, starch synthesis suffocation and the abnormal function of the flower developmental gene.3.The matrix-assisted laser-adsorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technique was used to obtain the peptide mass fingerprinting of the differentially expressed proteins in anthers between NJCMS2A and NJCMS2B. In the anthers at uninucleate microspore stage, 4 differentially expressed protein spots were identified. In the anthers at binucleate pollen stage, 14 differentially expressed protein spots were identified. According to the literature, the functions, especially those related to the male sterility of the major differentially expressed proteins, including Heat shock 22 kDa protein, cysteine proteinase, vacuolar H+-ATPase A subunit, MADS box protein, and starch branching enzyme were reviewed and discussed. It was inferred that the male sterility of NJCMS2A might be related to energy metabolism turbulence, the programmed cell death (PCD), starch synthesis suffocation and the abnormal function of the flower developmental gene.4.To synthesize the results of NJCMS1A and NJCMS2A, it was found that the MADS box protein and starch branching enzyme didn't appear in the anthers at both uninucleate microspore and binucleate pollen stages of NJCMS1A and NJCMS2A, but appeared in those of NJCMS1B and NJCMS2B; cysteine proteinase, vacuolar H~+-ATPase A subunit, adenosine/AMP deaminase, AIG1-like protein, oligouridylate binding protein, cullin, beta-amyrin synthase and hypothetical protein MtrDRAFT_AC146570g8vl appeared in the anthers at binucleate pollen stages of NJCMS1A and NJCMS2A, but didn't appear in those of NJCMS1B and NJCMS2B. These proteins might be related to the male sterility of NJCMS1A and NJCMS2A stably. It was suggested to clone the genes of the above proteins and study their functions, especially those related to the male sterility.