[1]曾维英,杨守萍,盖钧镒,等.大豆质核互作雄性不育系NJCMS1A和其保持系的不同器官蛋白质组比较[J].大豆科学,2008,27(01):8-14.[doi:10.11861/j.issn.1000-9841.2008.01.0008]
　ZENG Wei-ying,YANG Shou-ping,GAI Jun-yi,et al.Comparative Proteome Analysis of Different Organs between Cytoplasmicnuclear Malesterile Line NJCMS1A and Its Maintainer in Soybeans[J].Soybean Science,2008,27(01):8-14.[doi:10.11861/j.issn.1000-9841.2008.01.0008]
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大豆质核互作雄性不育系NJCMS1A和其保持系的不同器官蛋白质组比较

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第27卷 期数: 2008年01期 页码: 8-14 栏目: 出版日期: 2008-02-25
Title:
Comparative Proteome Analysis of Different Organs between Cytoplasmicnuclear Malesterile Line NJCMS1A and Its Maintainer in Soybeans
作者:
曾维英1; 2; 杨守萍1; 盖钧镒1; 喻德跃1
1 南京农业大学大豆研究所，国家大豆改良中心，作物遗传与种质创新国家重点实验室，江苏南京210095；
2广西农业科学院玉米研究所，广西南宁530227
Author(s):
ZENG Wei-ying1; 2; YANG Shou-ping1; GAI Jun-yi1; and Yu De-yue1
1 Nanjing Agricultural University，Soybean Research Institute，National Center for Soybean Improvement，National Key Laboratory of Crop Genetics and Germplasm Enhancement，Nanjing 210095，Jiangsu；
2 Maize Research Institute of Guangxi Academy of Agricultural Sciences，Nanning 530227，Guangxi，China
关键词:
大豆; 质核互作雄性不育; 不同器官; 蛋白质组; 双向凝胶电泳
Keywords:
Soybean; Cytoplasmic-nuclear male-sterility; Different organs; Protome; Two-dimensional gel electrophoresis (2-DE)
DOI:
10.11861/j.issn.1000-9841.2008.01.0008
文献标志码:
A
摘要:
质核互作雄性不育在杂种优势利用中起着重要作用，探讨质核互作雄性不育发生的机理具有重要的理论和实践意义。本研究开展大豆质核互作雄性不育系NJCMS1A与其保持系NJCMS1B的不同器官蛋白质组比较分析。以大豆质核互作雄性不育系NJCMS1A和其保持系NJCMS1B的种子、叶片和花药为材料，采用双向凝胶电泳技术对其蛋白质进行分离，考马斯亮蓝染色，获得重复性好的蛋白质双向电泳图谱，利用PDQuest 软件处理分析，寻找差异表达蛋白。结果发现不育系NJCMS1A与其保持系NJCMS1B的种子2-DE图谱间存在7个差异表达蛋白点，其中3个在NJCMS1A中表达而在NJCMS1B中缺失，3个在NJCMS1A中缺失而在NJCMS1B中表达，1个在NJCMS1A中表达量明显增强；苗期叶片2-DE图谱间基本一致，没有差异表达蛋白点；单核小孢子期花药2-DE图谱间有9个差异表达蛋白点，其中3个在NJCMS1A中表达而在NJCMS1B中缺失，6个在NJCMS1A中缺失而在NJCMS1B中表达；二胞花粉期花药2-DE图谱间有24个差异表达蛋白点，其中10个在NJCMS1A中表达而在NJCMS1B中缺失，12个在NJCMS1A中缺失而在NJCMS1B中表达，2个在NJCMS1B中表达量明显增强。两系花药间存在较多差异表达蛋白点，种子间仅有少量差异表达蛋白点，而苗期叶片间基本没有差异表达蛋白点，说明不育基因表达具有时空性和器官特异性，与育性有关的蛋白主要在花药中表达。
Abstract:
Cytoplasmicn-uclear male sterility plays an important role in the production of hybrid seeds.The knowledge of the mechanism of the cytoplasmicn-uclear male sterility is essential to hybrid seed production.The present paper was aimed at the comparative proteome analysis of different organs between the cytoplasmic-nuclear male-sterile line NJCMS1A and its maintainer NJCMS1B in soybeans.The materials were the seeds，young leaves and anthers from the male-sterile line NJCMS1A and its maintainer NJCMS1B.Two-dimensional gel electrophoresis (2-DE) technique was used to separate the protein spots and the gels were stained with Coomassie Blue G-250.The obtained 2-DE maps were pretty consistent among replications.The difference between the protein maps of NJCMS1A and those of NJCMS1B was analyzed with the PDQuest image software.Seven protein spots were found differentially expressed between NJCMS1A and NJCMS1B seeds.Among these，3 protein spots were present in the seed protein map of NJCMS1A but absent in that of NJCMS1B，and 3 protein spots present in that of NJCMS1B but absent in that of NJCMS1A，another protein spot was up-regulated significantly in the map of NJCMS1A in comparison with that of NJCMS1B.Nine protein spots were found differentially expressed between NJCMS1A and NJCMS1B anthers at uninucleate microspore stage.Among them，3 protein spots were present in the anther protein map of NJCMS1A but absent in that of NJCMS1B，and 6 protein spots present in that of NJCMS1B but absent in that of NJCMS1A.Twenty-four protein spots were found differentially expressed between NJCMS1A and NJCMS1B anthers at binucleate pollen stage.Among them，10 protein spots were present in the anther protein map of NJCMS1A but absent in that of NJCMS1B，and 12 protein spots present in that of NJCMS1B but absent in that of NJCMS1A，another two protein spots were up-regulated significantly in the map of NJCMS1B in comparison with that of NJCMS1A.No protein spot was found differentially expressed between NJCMS1A and NJCMS1B leaves.The above results showed that there were lots of difference between the 2-DE maps of anthers of NJCMS1A and those of NJCMS1B，a little difference between the 2-DE maps of seeds of NJCMS1A and those of NJCMS1B，but no difference between the 2-DE maps of leaves of NJCMS1A and those of NJCMS1B.It was inferred that the expression of the male sterile genes was spacially and periodly，and the anther was the major organ in which the protein related to the male sterility was expressed.

参考文献/References:

[1]Bradford M M.1976.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding.Analytical Biochemisty，72(1):248-254

[2]Ding D R，Gai J Y，Cui Z L，and Qiu J X.2002.Development of a cytoplasmic-nuclear male-sterile line of soybean.Euphytica，124:85-91
[3]Fan J M.2003. Studies on cyto-morphological and cytochemical features of cytoplasmic-nuclear male-sterile lines of soybeans[Glycine max(L.)Merrill].Thesis for M S，Agronomy College of Nanjing Agricultural University，Supervisor:Yang S P (凡军民.2003.大豆质核互作雄性不育系的细胞形态学和细胞化学特征的研究.硕士学位论文，南京农业大学农学院，导师:杨守萍)
[4]Gai J Y， ed.2006.Crop breeding (Species)，2nd-edn.China Agriculture Press，China，Beijing，pp.223（盖钧镒，著.2006.作物育种学各论.中国农业出版社，中国，北京，pp.223)
[5]Gai J Y， Ding D R，Cui Z L，and Qiu J X.1999.Development and performance of the cytoplasmic nuclear male sterile line NJCMS1A of soybean.Scientia Agricultura Sinica，32(5)：23-27（盖钧镒，丁德荣，崔章林，邱家驯.1999.大豆质核互作雄性不育系NJCMS1A的选育及其特性.中国农业科学，32(5)：23-27)
[6]Gallardo K，Job C，Groot S P C，Puype M，Demol H，Vandekerckhove J，and Job D.2001.Proteomic analysis of Arabidopsis seed germination and priming.Plant Physiology，126：835-848
[7]Men S Z， and Liu B L.2001.Studies on protein composition of different organs of wheat plant under control of a male sterile gene(MS2).Acta Agronomica Sinica，27(1)：117-122 (门淑珍，刘博林.2001.小麦显性雄性核不育材料后代不育株与可育株不同器官的蛋白质比较研究.作物学报，27(1):117-122)
[8]Shao J Z， and Gu Y.2006.Analysis of flag leaf proteins of cross rice by two-dimensional electrophoresis.Journal of Hubei Normal University(Natural Science)，26(1)：1-5 (邵锦震，顾勇.2006.水稻剑叶蛋白质的双向电泳分析.湖北师范学院学报(自然科学版)，26(1)：1-5)
[9]Teng X Y， and Chen X H.1996.Comparative studies on the constitution of protein between t-type cytoplasmic male sterile wheat and its maintainer.Acta Agronomica Sinica，22(3)：264-270（滕晓月，陈雪晖.1996.小麦T型细胞质雄性不育系和保持系蛋白质的比较研究.作物学报，22(3)：264-270）
[10]Wei L，Ding Y，Hu Y J，and Yu J H.2002.Analysis of leaf proteins of zidao male-sterile line by 2D-PAGE.Acta Genetica Sinica，29(8)：696-699（魏磊，丁毅，胡耀军，余金洪.2002.紫稻细胞质雄性不育系叶片全蛋白双向电泳分析.遗传学报，29(8)：696-699)
[11]Wen L，Liu G， Zhang Z J，Tao J，Tao J，Wan C X，Li S Q，and Zhu Y G.2006.Preliminary proteomics analysis of the total proteins of HL type cytoplasmic male sterility rice anther.Hereditas，28(3):311-316 (文李，刘盖，张再君，陶钧，万翠香，李绍清，朱英国.2006.红莲型水稻细胞质雄性不育花药蛋白质组学初步分析.遗传，28(3)：311-316)
[12]Xu X Y， Zheng R，Li C M，Gai J Y，and Yu D Y.2006.Differential proteomic analysis of seed germination in soybean.Progress in Biochemistry and Biophysics，33(11)：1106-1112（徐晓燕，郑蕊，李春梅，盖钧镒，喻德跃.2006.大豆种子萌发过程中的差异蛋白质组研究.生物化学与生物物理进展，33(11)：1106-1112)
[13]Yang S P， Duan M P，Meng Q C，Qiu J，Fan J M，Zhao T J，Yu D Y，and Gai J Y.2007.Inheritance and gene tagging of male fertility restoration of cytoplasmic-nuclear male-sterile line NJCMS1A in soybean.Plant Breeding，126：302-305
[14]Zheng R，and Yu D Y.2005.Improvement of two-dimensional electrophoresis of soybean seed protein for proteome analysis.Soybean Science，24(3)：166-170（郑蕊，喻德跃.2005.适于蛋白质组研究的大豆种子蛋白双向电泳技术的改进.大豆科学，24(3)：166-170)

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备注/Memo

基金项目：国家重点基础研究发展计划项目(2002CB111304，2004CB7206，2006CB101708)；国家自然科学基金项目(30490250，30671314)；国家高技术研究发展计划项目(2002AA211052，2006AA100104)；长江学者和创新团队发展计划资助项目(PCSIRT)

作者简介：曾维英（1981-），女，硕士研究生，研究方向为植物遗传育种。
通讯作者（Corresponding author）：杨守萍，副教授，博士。Tel：025-84396463；E-mail: spyang@njau.edu.cn 喻德跃，教授，博士。Tel：025-84396410；E-mail:dyyu@njau.edu.cn

更新日期/Last Update: 2014-10-08