| 甘蓝型油菜抗菌核病近等基因系和感病亲本蛋白差异的初步研究 |
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| 引用本文： | 皇甫海燕,官春云.甘蓝型油菜抗菌核病近等基因系和感病亲本蛋白差异的初步研究[J].中国农业科学,2010,43(10):2000-2007. |
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| 作者姓名： | 皇甫海燕 官春云 |
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| 作者单位： | (湖南农业大学农学院/国家油料改良中心湖南分中心); |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 【目的】建立适合于提取油菜叶片全蛋白的三氯乙酸/丙酮沉淀法,为今后油菜蛋白质组学的研究提供参考,明确菌核菌诱导抗、感菌核病油菜后的蛋白差异表达情况,为寻找抗菌核病基因提供线索。【方法】以油菜抗菌核病近等基因系98C40(抗原来自湘油15)和感菌核病亲本98C40为材料,在其生长的苗期阶段,接种菌核菌72h之后,分别提取其叶片全蛋白。运用双向电泳技术、PD-quest软件、质谱技术对其进行差异蛋白质组分析。【结果】检测到抗病98C40(抗原来自湘油15)与感病亲本98C40之间有4个差异蛋白点,蛋白点a,线粒体ATP合酶F1β亚基(mitochondrial F1ATP synthase beta subunit);蛋白点c,磷酸甘油酸激酶(phosphoglycerate kinas);蛋白点d,噻唑生物合成酶(THI1);这3个蛋白只在抗病98C40(抗原来自湘油15)上表达,而在感病亲本98C40上不表达。蛋白点b,1,5-二磷酸核酮糖羧化酶/加氧酶(ribulose-1,5-bisphosphate carboxylase/oxygenase);按5倍差异表达量计算,在抗病98C40(抗原来自湘油15)上高表达,而在感病亲本98C40上低表达。【结论】这4个蛋白都是油菜代谢相关蛋白,它们可能在油菜抗菌核病的反应中发挥作用。
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| 关 键 词： | 油菜 菌核病 蛋白质组学 近等基因系 双向电泳 质谱鉴定 |
| 收稿时间： | 2009-12-01; |
A Preliminary Analysis of Differential Proteins Between Brassica napus Resistance to Sclerotinia sclerotiorum Near-isogenic Lines and Susceptible Parent |
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| HUANGFU Hai-yan,GUAN Chun-yun.A Preliminary Analysis of Differential Proteins Between Brassica napus Resistance to Sclerotinia sclerotiorum Near-isogenic Lines and Susceptible Parent[J].Scientia Agricultura Sinica,2010,43(10):2000-2007. |
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| Authors: | HUANGFU Hai-yan GUAN Chun-yun |
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| Affiliation: | HUANGFU Hai-yan,GUAN Chun-yun (Department of Agronomy,Hunan Agricultural University/the Branch Center of Hunan,The National Improvement Center of Oilcrop,Changsha 410128) |
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| Abstract: | Objective] A trichoroacetic acid/acetone method was established for extracting the full leaf-proteins of rapeseed which provided the reference for the rapeseed proteomics research in the future. The difference in protein expression was identified after the resistance and susceptibility to Sclerotinia sclerotiorum rapeseed were induced by Sclerotinia sclerotiorum. It offered the clues to searching for the resistance to Sclerotinia sclerotiorum genes. Method] Near-isogonic lines 98C40 (the antigen derived from Xiangyou 15) is resistant to Sclerotinia sclerotiorum and the parent 98C40 is susceptible to Sclerotinia sclerotiorum which were inoculated with Sclerotinia sclerotiorum during their seedling stage. Seventy-two hours after inoculation, full leaf-proteins of them were extracted respectively. Two-dimensional eloctrophoresis (2-DE), PD-quest software and mass technology were used in differential proteomies analysis. Result] Significant difference of 4 protein spots were observed between them. Protein spot a is mitoehondrial F1 ATP synthase beta subunit, spot c is phosphoglycerate kinase and spot d is thiazole biosynthetic enzymes (THI1),which express only in 98C40 (the antigen derived from Xiangyou 15) not expressed in parent 98C40. Protein spot b is ribuiose-1,5-bisphosphate earboxylase/oxygenase expressed highly in 98C40 (the antigen derived from Xiangyou 15) and lowly in parent 98C40 based on 5 times difference of expression. Conclusion] The four proteins are all closely related to the rapeseed metabolism.They may play a role in resistancc to Sclerotinia sclerotiorum. |
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| Keywords: | rapeseed Sclerotinia sclerotiorum proteomics neat-isogenic lines two-dimensional electrophoresis mass spectrometry |
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