仙人掌苷对荔枝霜疫霉菌的生物活性和蛋白组学分析

doi:10.7668/hbnxb.2011.06.037

摘要/Abstract

摘要： 活性测定表明仙人掌苷对荔枝霜疫霉菌菌丝生长具有良好的抑菌活性,EC50达到173.52μg/mL。为了从蛋白质水平探讨仙人掌苷对荔枝霜疫霉菌的抑菌机制,利用蛋白质双向电泳方法分析仙人掌苷处理前后荔枝霜疫霉菌蛋白质组表达的差异,结合飞行质谱以及数据库检索鉴定差异蛋白。结果发现,仙人掌苷处理后荔枝霜疫霉菌有16个蛋白发生了显著变化,其中11个下调表达蛋白,4个上调表达蛋白,1个诱导表达蛋白。其中鉴定的5种差异蛋白质为:线粒体二羧酸载体蛋白(DIC 1)、半胱氨酸合成酶A、ABC转运蛋白(ABCF-EF-3)、Class III stress response-re-lated ATPase以及假定蛋白BSU31030。功能分析发现这些蛋白在药物胁迫反应、氨基酸与硫代谢、细胞膜药物运输外排、能量代谢等方面发挥重要的作用。这为阐明仙人掌苷对荔枝霜疫霉菌抑制作用的分子机理提供了重要参考。

关键词: 仙人掌苷, 荔枝霜疫霉菌, 抑菌活性, 蛋白组

Abstract: Bioassays showed that cacticin exhibited good antimicrobial activity and EC50was 173． 52 μg /mL． To explore the protein-level antimicrobial mechanism of cacticin in Peronophythora litchi， the proteome of P． litchi mycelium treated with cacticin was analyzed by two-dimensional electrophoresis(2-DE) and MALDI-TOF-MS． Results indicated sixteen protein spots showed significant and reproducible changes after cacticin treatment，among which eleven were down-regulated， four were up-regulated，and one was newly induced by cacticin． Five successfully identified proteins were assigned as mitochondrial dicarboxylate carrier protein(DIC 1),cysteine synthetase A，ATP-binding cassette transporters(ABCF-EF-3),class III stress response-related ATPase and hypothetical protein BSU31030． These proteins were involved in drug stress response，amino acid and sulfur metabolism，membrane transportation and discharge，and energy metabolism． Altogether， the results provided important information on the molecular mechanisms of antimicrobial activity of cacticin against P． litchi．

Key words: Cacticin, Peronophythora litchi, Antimicrobial activity, Proteome

中图分类号:

赖多, 曹永军, 卢梦玲, 徐汉虹. 仙人掌苷对荔枝霜疫霉菌的生物活性和蛋白组学分析[J]. 华北农学报, 2011, 26(6): 196-200. doi: 10.7668/hbnxb.2011.06.037.

LAI Duo, CAO Yong-jun, LU Meng-ling, XU Han-hong. Bioassays and Proteomics Analysis of Peronophythora litchi Treated with Cacticin[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(6): 196-200. doi: 10.7668/hbnxb.2011.06.037.

http://www.hbnxb.net/CN/Y2011/V26/I6/196

参考文献

[1] 岑贞陆，黄思良. 一种高效分离荔枝霜疫霉病菌的新方法[J]. 植物保护， 2008， 34(6): 137-138.
[2] 陶挺燕，何凡，范鸿雁，等. 几种杀菌剂对荔枝霜疫霉病的室内毒力测定[J]. 农药， 2010， 49(1): 72-73.
[3] Kim K S，Lee S，Lee Y S， et al. Anti-oxidant activities of the extracts from the herbs of Artemisia apiacea[J]. J Ethnopharmacol， 2003， 85: 69-72.
[4] 孙言伟，姜颖，贺福初. 差异蛋白质组学的研究进展[J]. 生命科学， 2005， 17(2): 137-140.
[5] 潘怡欧，刘琳琳，张炬红，等. 茉莉酸诱导的拟南芥叶片蛋白质组分析[J]. 华北农学报，2010，25(2): 35-39.
[6] Fernández-Acero F J， Jorge I，Calvo E， et al. Proteomic analysis of phytopathogenic fungus Botrytis cinerea as a potential tool for identifying pathogenicity factors， therapeutic targets and for basic research[J]. Arch Microbiol， 2007， 187: 207-215.
[7] Kim S T，Yu S，Kim S G， et al. Proteome analysis of rice blast fungus(Magnaporthe grisea) proteome during appressorium formation[J]. Proteomics，2004，4(11): 3579 -3587.
[8] 谢娟，黄明星，卢代华，等. 水稻稻瘟菌抗性相关蛋白的双向电泳分析[J]. 中国生物工程杂志，2006，26(12): 92-98.
[9] 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.
[10] Campo S，Carrascal M，Coca M，et al. The defence response of germinating maize embryos against fungal in fection: a proteomics approach[J]. Proteomics， 2004，4:383-396.
[11] Chan Z L，Wang Q，Xu X B， et al. Functions of defenserelated proteins and dehydrogenasesin resistance response induced by salicylic acid in sweet cherry fruits at different maturity stages[J]. Proteomics，2008，8: 4791 -4807.
[12] Candiano G，Bruschi M，Musante L， et al. Blue silver: a very sensitive colloidal Coomassie G-250 staining for proteome analysis[J]. Electrophoresis， 2004， 25: 1327- 1333.
[13] Dahl S G，Sylte I，Ranvna A W. Structures and models of transporter proteins[J]. J Pharmacol Exp Ther，2004， 309(3): 853-860.
[14] 谢浩，李其昌，郭小明. 膜转运蛋白的功能性超表达和纯化[J]. 中国生物化学与分子生物学报， 2007，23(12): 1051-1058.
[15] Lancar-Benba J，Foucher B，Saint-Macary M. Characterization， purification and properties of the yeast mitochondrial dicarboxylate carrier(Saccharomyces cerevisiae)[J]. Biochimie， 1996， 78: 195-200.
[16] Stergiopoulos I，Zwiers L H，De Waard M A. Secretion of natural and synthetic compounds from filamentous fungi by membrane transporters of the ATP-binding cassette and major facilitator superfamily[J]. Eur J Plant Pathol， 2002， 108: 719-734.
[17] Mamnun Y M，Schller C，Kuchler K. Expression regulation of the yeast PDR5 ATP-binding cassette(ABC) transporter sugGests a role in cellular detoxification during the exponential growth phase[J]. FEBS Lett， 2004， 559(1-3): 111-117.
[18] Hanekop N，Zaitseva J， Jenewein S， et al. Molecular insights into the mechanism of ATP-hydrolysis by the NBD of the ABC-transporter HlyB[J]. FEBS Lett，2006，580 (4): 1036-1041.
[19] Theodoulou F L，Holdsworth M，Baker A. Peroxisomal ABC transporters[J]. FEBS Lett，2006，580: 1139-1155.
[20] 王华丙，张振义，包锐，等. ABC 转运蛋白的结构与转运机制[J]. 生命的化学， 2007， 27(3): 208-210.
[21] 乌艳红，米福贵，李志明，等. 紫花苜蓿铝激活苹果酸转运蛋白基因的克隆与亚细胞定位[J]. 华北农学报， 2011， 26(1): 72-77.
[22] 孟成生，李爱丽，王省芬，等. 小麦脂质转运蛋白cDNA结构及相应肽链构造特征的分析[J]. 华北农学报， 2007， 22(2): 1-5.
[23] 闫洪波. 两个类ADP-葡萄糖转运蛋白基因的克隆和特征[J]. 河南农业科学， 2009(2): 50-53.
[24] 田双起，秦广雍，黄新，等. 拟南芥跨液泡膜磷转运蛋白基因突变体的初步筛选[J]. 河南农业科学，2008(4): 45-47.
[25] 杨泽峰，张丹，敖雁，等. 植物蔗糖转运基因家族功能分化的生物信息学分析[J]. 安徽农业科学，2009， 37(29): 14061-14065.
[26] Skov J，Lemmens M，Geise H. Role of Fusarium culmorum ABC transporter(FcABC1) during infection of wheat and barley[J]. Physiol Mol Plant Path，2002，108(7): 719-734.
[27] Hayashi K，Schoonbeek H，Sugiura H，et al. Multidrug resistance in Botrytis cinerea associated with decreased accumulation of the azole fungicide oxpoconazole and increased transcription of the ABC transporter gene BcatrD[J]. Pestic Biochem Physiol， 2001， 70: 168-179.
[28] Hayashi K，Schoonbeek H，De Waard M A. Expression of the ABC transporter BcatrD from Botrytis cinerea reduces sensitivity to sterol demethylation inhibitor fungicides[J]. Pestic Biochem Physiol， 2002， 73: 110-121.
[29] Hamamoto H，Hasegawa K，Nakaune R， et al. The role of the ABC transporter gene PMR1 in demethylation inhibitor resistance in Penicillium digitatum[J]. Pestic Biochem Physiol， 2001， 70: 19-26.

选择文件类型/文献管理软件名称

选择包含的内容