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作物学报 ›› 2012, Vol. 38 ›› Issue (07): 1232-1239.doi: 10.3724/SP.J.1006.2012.01232

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

烟草细胞质雄性不育系及其保持系的花蕾差异蛋白质分析

祁建民1,**，马红勃1,2,**，徐建堂1，陈美霞1,3，周东新4，王涛5，陈顺辉6   

  1. 1 福建农林大学生命科学学院 / 作物遗传育种与综合利用教育部重点实验室, 福建福州 350002; 2 南京农业大学作物遗传与种质创新国家重点实验室, 江苏南京 210095; 3 宁德师范学院生物工程系, 福建宁德 352100; 4 福建省龙岩市烟草科学研究所, 福建龙岩 364000; 5 福建省南平市烟草公司, 福建南平 353200; 6 福建省福州市烟草科学研究所, 福建福州 350003
  • 收稿日期:2011-11-17 修回日期:2012-02-22 出版日期:2012-07-12 网络出版日期:2012-04-06
  • 基金资助:

    本研究由中国烟草总公司重点项目(110201002006)资助

Proteomic Analysis of Bud Differentiation between Cytoplasmic Male-Sterile Line and Maintainer in Tobacco

QI Jian-Min1,**,MA Hong-Bo1,2,**,XU Jian-Tang1,CHEN Mei-Xia1,3,ZHOU Dong-Xin4,WANG Tao5,CHEN Shun-Hui6   

  1. 1 Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2 National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China; 3 Ningde Normal University Biological Engineering, Ningde 352100, China; 4 Longyan Tobacco Branch Company, Longyan 364000, China; 5 Nanping Tobacco Company, Nanping 353200 China; 6 Fuzhou Institute of Tobacco Science of Fujian Province, Fuzhou 350003, China?
  • Received:2011-11-17 Revised:2012-02-22 Published:2012-07-12 Published online:2012-04-06

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被引次数

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摘要： 为探讨烟草雄性不育的分子遗传机理，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14.4~97.6 kD、等电点4~7线性范围内，检测到约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，分别被鉴定为核酮糖-1,5-二磷酸羧化酶/加氧酶、多酚氧化酶、Patatin homolog蛋白、PSI 9 kD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。

关键词: 烟草, 细胞质雄性不育, 花蕾, 差异蛋白质, 双向凝胶电泳

Abstract: To explorethe moleculargenetic mechanismsof cytoplasmic male sterility in tobacco,we studied the differential protein between the cytoplasmic male-sterile line and its maintainer. Immobilized pH gradient two-dimensional gel electrophoresis (2-DE) technique 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-1,5-bisphosphate carboxylase/oxygenase, polyphenol oxidase, patatin homolog, 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.

Key words: Tobacco, Cytoplasmic male sterility, Bud, Differential protein, 2-DE

[1]Yang T-Z(杨铁钊), Ding Y-L(丁永乐), Liu R-X(刘仁祥), Wu J(吴军), Chen X-P(陈学平), Luo Y-G(罗义贵), Cui H(崔红). Tobacco Breeding (烟草育种学). Beijing: China Agriculture Press, 2003. pp 114–132 (in Chinese)

[2]Liu Z-S(刘忠松), Guan C-Y(官春云), Chen S-Y(陈社员). Theory and Practice of Plant Male-sterility (植物雄性不育机理的研究及应用). Beijing: China Agriculture Press, 2001. pp 105–110 (in Chinese)

[3]Liu Z-D(刘祖洞), Jiang S-H(江绍慧). Genetics (遗传学). Beijing: Higher Education Press, 1979. p 197 (in Chinese)

[4]Li J-G(李继耕). Chloroplast Molecular Genetics in Higher Plants (高等植物叶绿体分子遗传学). Beijing: Beijing Agricultural University Press, 1987. pp 33–158 (in Chinese)

[5]Zhang X-W(张学文), Wang B(王斌). A new discovered dsRNA associated with cytoplasmic male-sterile rice. Acta Genet Sin (遗传学报), 1990, 17(4): 289–293 (in Chinese with English abstract)

[6]Zhu Y-G(朱英国), Li R-Q(利容千), Wang M-Q(王明全). Biology About Rice Male-Sterile (水稻雄性不育生物学). Wuhan: Wuhan University Press, 2000. pp 100–123 (in Chinese)

[7]Wen L(文李), Liu G(刘盖), Zhang Z-J(张再君), Tao J(陶钧), Wan C-X(万翠香), Li S-Q(李绍清), Zhu Y-G(朱英国). Preliminary proteomics analysis of the total proteins of HL type cytoplasmic male sterility rice anther. Hereditas (遗传), 2006, 28(3): 311–316 (in Chinese with English abstract)

[8]Wang S-G(王世刚), Wang Y(王宇), Tang Y(唐咏), Xu G(徐光), Xu M(许明), Wei Y-T(魏毓棠). Leaf proteomic analysis of pepper cytoplasmic male sterility and maintainer lines during flowering. China Veg (中国蔬菜), 2007, (2): 13–16 (in Chinese with English abstract)

[9]Zeng W-Y(曾维英), Yang S-P(杨守萍), Gai J-Y(盖钧镒), Yu D-Y(喻德跃). Proteomic study of anther differentiation between cytoplasmic-nuclear male-sterile line NJCMS1A and its maintainer in soybean [Glycine max (L) Merr.]. Sci Agric Sin (中国农业科学), 2007, 40(12): 2679–2687 (in Chinese with English abstract)

[10]Zhang W-J(张维佳), Wang Y(王艳), Li X-M(李雪梅), Fan B-L(范宝莉), Wang Z-Y(王振英), Peng Y-K(彭永康), Xie C-J(解超杰), Liu Z-Y(刘志勇), Sun Q-X(孙其信). The changes in chloroplast proteome caused by different nuclear background in cytoplasmic male-sterile (CMS) of wheat. J Mol Cell Biol (分子细胞学报), 2007, 40(1): 84–89 (in Chinese with English abstract)

[11]Leipner J, Horn R. Nuclear and cytoplasmic differences in the mitochondrial respiration and protein expression of CMS and maintainer lines of sunflower. Euphytica, 2002, 123: 411–419

[12]Zhang W-J(张维佳), Li C-Z(李纯正), Huang H-Q(黄海泉), Wang Z-Y(王振英), Peng Y-K(彭永康). Different proteins in mitochondrial proteome of T-type maize cytoplasmic male-sterile line and its maintainer line. J Mol Cell Biol (分子细胞生物学报), 2007, 40(6): 410–418 (in Chinese with English abstract)

[13]Xu K, Cao M J, Zhu Y G, Pan G T, Rong T Z. Differential expression of mitochondrial proteins between C-type cytoplasmic male sterility line C48-2 and its maintainer line in maize. Acta Agron Sin (作物学报), 2008, 34(2): 232–237 (in Chinese with English abstract)

[14]Liu Q-Y(刘齐元), Liu F-H(刘飞虎), Huang H-Q(黄海泉), He K-X(何宽信), Zhu X-W(朱肖文). Equilibrium relationship between tobacco cytoplasmic male sterility and bud endogenous hormone. Tob Sci Technol (烟草科技), 2006, (5): 51–53 (in Chinese with English abstract)

[15]Chen W(陈伟), Huang C-M(黄春梅), Lü L-X(吕柳新). The improved method of two-dimensional electrophores is of protein in recalcitrant plant litchi. J Fujian Agric Univ (福建农业大学学报), 2001, 30(1): 123–126 (in Chinese with English abstract)

[16]Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem, 1976, 72: 248–254

[17]Liu Q-Y(刘齐元), Liu F-H(刘飞虎), He K-X(何宽信), Zhu X-W(朱肖文), Li L-X(李立新). Preliminary study on the relationship between cytoplasmic male sterility and the contents of flower bud endogenous hormone in tobacco. Acta Agric Boreali Sin (华北农学院), 2006, 21(5): 79–82 (in Chinese with English abstract)

[18]Li X-P(李杏普), Lan S-Q(兰素缺), Liu Y-P(刘玉平). Studies on pigment and its related physio-biochemical properties of blue or purple grain wheat. Acta Agron Sin (作物学报), 2003, 29(1): 157–158 (in Chinese with English abstract)

[19]Racusen D. Lipid acylhydrolase of patatin. Can J Bot, 1984, 62: 1640–1644

[20]Li C-H(李灿辉), Wang J(王军), Long W-B(龙维彪). The development on patatin tuber protein in potato. Chin Potato J (马铃薯杂志), 1998, 12(3): 179–186 (in Chinese)

[21]Wei J(魏捷), Yu H(余辉), Li L-B(李良璧), Kuang T-Y(匡廷云), Wang J-S(王居硕), Gong Y-D(公衍道), Zhao N-M(赵南明). The study on the photodamage of pigments and proteins of the photosystem I particles of Spinach oleracea L. Chin Sci Bull (科学通报), 2000, 45(6): 612–617 (in Chinese with English abstract)
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