摘要
为探讨烟草雄性不育的分子遗传机制,对烟草细胞质雄性不育系和保持系进行差异蛋白质组学研究。采用固相pH梯度SDS-PAGE双向电泳,对烟草细胞质雄性不育系MSK326及其保持系K326雄蕊原基分化期、四分体时期及单核时期花蕾蛋白质进行分离,经考马斯亮蓝染色,获得了分辨率和重复性较好的双向电泳图谱。使用PDQuest软件分析蛋白质图谱,利用基质辅助激光解吸电离飞行时间串联质谱分析,然后用Mascot软件对NCBInr数据库搜索。在分子量14.4～97.6kD、等电点4～7线性范围内,检测到365个蛋白点,其中7个蛋白质点在保持系中出现,在不育系中缺失,被鉴定为核酮糖-1,5-二磷酸羧化酶/加氧酶、多酚氧化酶、Patatin homolog蛋白、PSI9kD蛋白4类蛋白,推测不育系MSK326雄性不育性可能与营养代谢紊乱、间接抑制雄性器官发育、养分供给不足、免疫能力低和能量转移受到抑制等有关。
To explore the molecular genetic mechanisms of cytoplasmic male sterility （CMS） in tobacco, we studied the differen- tial proteins between the CMS line and its maintainer. Immobilized pH gradient two-dimensional gel electrophoresis （2-DE） tech- nique was used to separate the protein spots while gels were stained with the Coomassie Blue G-250. The difference between protein maps of bud from cytoplasmic male-sterile line MSK326 and maintainer line K326 was analyzed with PDQuest image software. Matrix-assisted laser-adsorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） technique was used to obtain the peptide mass of differentially expressed protein spots. The Mascot software was used to search the protein database NCBInr to identify those spots interested. A total of 365 protein spots were detected within Mr 14.4-97.6 kD and pH 4-7. Seven protein spots appeared in the protein map of maintainer line K326 but absent in that of CMS line MSK326, which were identified as ribulose-l,5-bisphosphate carboxylase/oxygenase, polyphenol oxidase, patatin homolog, and PSI 9 kD protein. It was inferred that the male sterility of MSK326 might be related to energy metabolism turbulence, abnormality in male organ development, deficient nutrition supply and immunity, and inhibition of energy transfer.
出处
《作物学报》
CAS
CSCD
北大核心
2012年第7期1232-1239,共8页
Acta Agronomica Sinica
基金
中国烟草总公司重点项目(110201002006)资助
