白粉菌侵染<em>Pm21</em>近等基因系的早期蛋白组差异研究

doi:10.3969/j.issn.1000-7091.2012.01.002

摘要/Abstract

摘要： 分析接种小麦白粉菌E09后Pm21近等基因系及其亲本叶片的总蛋白质差异,从蛋白质组调控角度揭示Pm21基因抗白粉病的抗病机理。以Pm21近等基因系及其亲本百农3217为试材,显微观察接种后5 d内的叶片表面细胞与白粉菌孢子表型差异;采用非离子去污剂提取总蛋白质,运用双向电泳、质谱分析与数据库检索技术,比较Pm21近等基因系及其亲本接菌第4天后的叶片总蛋白质差异。超显微和显微观察结果表明,12 h后百农3217叶细胞表面与Pm21近等基因系相比褶皱明显;在接菌第2天后,在Pm21近等基因系白粉菌孢子生长明显慢于其亲本表面孢子,且孢子侵入个数较少,不萌发现象较多;在第4天后Pm21近等基因系被侵染叶片细胞出现过敏性反应;通过MALDI-TOF-MS/MS质谱测序和MASCOT软件序列分析,注释了15个表达上调蛋白点;功能分类表明,差异蛋白质分别参与叶绿体光合呼吸作用(40%) 、放氧作用(13%) 、代谢物合成(20%) 、抗病、动力蛋白、运输等生理功能。蛋白质组水平发现,光合呼吸蛋白和叶谷氨酰胺合成酶在Pm21近等基因系感病3～4 h后就大量表达,可能是导致叶片明显变绿原因;叶谷氨酰胺合成酶的增加与光合呼吸蛋白的增加密不可分;光合呼吸蛋白和抗坏血酸过氧化物酶的上调表达为植物体在高H2O2浓度下正常生长提供了保障。另外,β-1,3-葡聚糖苷酶的过量表达也是Pm21基因前期抵御白粉菌侵染有效物质。钾离子通道蛋白可能在抗病防御过程中起到了信号转导的作用。

关键词: 白粉菌, 近等基因系, Pm21, 显微观察, 双向电泳, 蛋白质组

Abstract: The purpose of this project was to study inoculated by E09 in wheat leaves to confer durable and broad spectrum resistance to wheat powdery mildew through proteomic analysis. To carry out the cytological observations of Wheat(Triticum aestivum) cultivars Bainong 3217 and near-isogenic Bainong 3217/Pm21(BC₇F₆) plants were inoculated with E09 powdery mildew after 5 days. Total proteins were extracted by nonionic detergent, were separated by two-dimensional gel electrophoresis(2-DE) 3 replicates and analysed through MALDI-TOF-MS/MS mass spectrometry. Microscopic observation results showed that Bainong 3217 leaves cell surface folds after 12hr, and spores growing significantly inhibited on leave surface of near-isogenic Bainong 3217/Pm21(BC₇F₆) after 2 days inoculation. Besides, hypersensitive reaction was found at 4th day post inoculation. And 15 of up-regulated proteins were identified homologous to known proteins in various databases by MALDI-TOF-MS/MS and MASCOT analysis. The predicted functions of the 5 proteins were related to Chloroplasts photosynthesis(40%), oxygen releasing(13%), Metabolites synthesis(20%), resistance, transcription. One result of leaves deepening green may cause by Enzyme Rubisco and Glutamine synthetase leaf isozyme proteins increased. These proteins increased after about 3 or 4 hours infection. Enzyme Rubisco and Ascorbate peroxidase expressed were up-regulated to protect Calvin cycle in high concentration of H₂O₂. In addition, g lucan endo-1, 3-beta-D-glucosidase precursor induced were up-regulated to suppress further development of haustoria. The potassium channel protein may play an important role in early disease-resistance signal transduction in Pm21.

Key words: Powdery mildew, Near-isogenic lines, Pm21, Microscopy, 2-D electrophoresis, Proteomics

中图分类号:

Q78
S435

李达, 李强, 关西贞, 张卫东, 田纪春. 白粉菌侵染Pm21近等基因系的早期蛋白组差异研究[J]. 华北农学报, 2012, 27(1): 5-11. doi: 10.3969/j.issn.1000-7091.2012.01.002.

LI Da, LI Qiang, GUAN Xi-zhen, ZHANG Wei-dong, TIAN Ji-chun. Proteomic Analyses of the Early Pm21 Wheat Near Isogenic Lines and Their Parents Affected by Powdery Mildew[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(1): 5-11. doi: 10.3969/j.issn.1000-7091.2012.01.002.

http://www.hbnxb.net/CN/Y2012/V27/I1/5

参考文献

[1] 肖世和,庄巧生.中国小麦品种改良及系谱分析[M] 北京:中国农业出版社,2003:469-487.
[2] 李洪杰,王晓鸣,宋凤景,等.中国小麦品种对白粉病的抗性反应与抗病基因检测[J] 作物学报,2011,37(6):943-954
[3] 牛吉山.小麦抗白粉病遗传育种研究[M] 北京:中国农业科学技术出版社,2007:21-25.
[4] Qi L L,Chen P D,Liu D J,et al.Identification,mapping and application of polymorphic DNA associated with resistance gene Pm21 of wheat[J] Genome,1996,39:191-197.
[5] 齐莉莉,陈佩度,刘大钧,等.小麦白粉病新抗原--基因Pm21[J] 作物学报,1995,21(3):257-262.
[6] 刘金元,陶文静,刘大钧,等.小麦簇毛麦易位系6VS/6AL 中6VS 的遗传传递及其所携Pm21 基因的遗传稳定性分析[J] 植物学报,1999,41(10):1058-1060.
[7] 孙晓丽,周荣华,贾继增.白粉菌诱导的小麦叶片程序性细胞死亡[J] 扬州大学学报:农业与生命科学版,2006,27(2):21-25.
[8] 郭兴启,温孚江,朱汉城.烟草感染马铃薯Y 病毒(PVY) 后光合作用的变化规律[J] 浙江大学学报:农业与生命科学版,2000,26(1):75-78.
[9] Li Q,Chen X M,Li D,et al.Differences in Protein Expression and Ultrastructure between Two Wheat Near Isogenic Lines Affected by Powdery Mildew[J] Russian Journal of Plant Physiology,2011,58(4):686-695.
[10] 肖明刚.玉米疯顶病病原菌检测和病害防治技术研究[D] 北京:中国农业科学院,2004:20-21.
[11] Andrej S,Matthias W,Ole V,et al.Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide Gels[J] European Molecular Biology Laboratory(EMBL) Anal Chem,1996,68(5):850 858.
[12] Marco T,Kogel K H,Huckelhoven R.Superoxide and hydrogen peroxide play different roles in the nonhost interaction of barley and wheat with inappropriate formae speciales of Blumeria graminis[J] Mol Plant-Microbe Interact,2004,17:304-312
[13] Li A L,Wang M L,Zhou R H,et al.Comparative analysis of early H₂O₂ accumulation in compatible and incompatible wheat powdery mildew interactions[J] Plant Pathol,2005,54:308-316.
[14] Bienert G P,Mφller A L B,Kristiansen A,et al.Specific aquaporins facilitate the diffusion of hydrogen peroxide across membranes[J] The Journal of Biological Chemistry,2007,282(2):1183-1192.
[15] Cao A Z,Xing L P,Wang X Y,et al.Serine/threonine kinase gene Stpk-V,a key member of powdery mildew resistance gene Pm21,confers powdery mildew resistance in wheat[C]//Proceedings of the National Academy of Sciences of the United States of America(PNAS).2011,1016981108.
[16] Niu J S,Yu L,Ma Z Q.Molecular clonging and characterization of a Beclin-1 like gene in wheat[J] Journal of Plant Physiology and Molecular Biology,2002b,28:275-280.
[17] Oliveira I C,Brears T,Knight T J,et al.Overexpression of cytosolic glutamine synthetase.Relation to nitrogen,light,and photorespiration[J] Plant Physiol,2002,129:1170-1180.
[18] Tucker M R,Paech N A,Willemse M T,et al.Dynamics of callose deposition and beta-1,3-glucanase expression during reproductive events in sexual and apomictic Hieracium[J] Planta,2001,212(4):487-498.
[19] 于振,李倩,赵建叶,江帆,等.栽培小麦Brock 和京411 感染白粉菌后蛋白质组的变化[J] 作物学报,2009,35(11):2064-2072.
[20] Lesser M P.Oxidative stress in marine environments:biochemistry and physiological ecology[J] Annu Rev Physiol,2006,68:253-278.
[21] 孙卫红,王伟青,孟庆伟.植物抗坏血酸过氧化物酶的作用机制、酶学及分子特性[J] 植物生理学通讯,2005,41(2):143-147.
[22] Najami N,Janda T,Barriah W,et al.Ascorbate peroxidase gene family in tomato:its identification and characterization[J] Mol Genet Genomics,2008,279:171-182.
[23] Nakano Y,Asada K.Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts[J] Plant Cell Physiol,1981,22:867.
[24] 曹宛虹.作为叶绿体H₂O₂ 分解系统关键酶的抗坏血酸过氧化物酶[J] 植物生理学通讯,1994,30(6):452-458.
[25] 李强,张卫东,田纪春.小麦抗白粉病基因Pm21 抗病差异的蛋白质组学研究[J] 中国农业科学,2009,42(8):2778-2783.
[26] Amrutha R N,Sekhar P N,Varshney R K,et al.Genomewide analysis and identification of genes related to potassium transporter families in rice(Oryza sativa L.)[J] Plant Science,2007,172:708-721.

选择文件类型/文献管理软件名称

选择包含的内容

白粉菌侵染Pm21近等基因系的早期蛋白组差异研究

Proteomic Analyses of the Early Pm21 Wheat Near Isogenic Lines and Their Parents Affected by Powdery Mildew

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	高素红, 葛长虹, 周国娜, 姚淑娟, 路常宽, 高宝嘉. 绿盲蝽取食诱导赤霞珠冬芽全蛋白双向电泳体系的建立与优化[J]. 华北农学报, 2022, 37(1): 181-187.
[2]	田志刚, 张树伟, 陈芳, 常利芳, 贾举庆, 张晓军, 李欣. 响应白粉菌的小麦NBS基因筛选[J]. 华北农学报, 2021, 36(6): 189-194.
[3]	李丹, 赵存鹏, 刘素恩, 王凯辉, 王兆晓, 刘旭, 郭宝生, 耿军义. 黄萎病菌胁迫下陆地棉冀2658根部蛋白质组研究[J]. 华北农学报, 2020, 35(S1): 322-331.
[4]	李永红, 周语潮, 田启航, 常瑞丰, 刘国俭, 陈湖, 王召元. 桃树枝条皮层响应低温的特异蛋白质2-DE分析[J]. 华北农学报, 2020, 35(S1): 102-108.
[5]	唐秀英, 王会民, 龙起樟, 黄永兰, 芦明, 万建林. 低温胁迫对水稻苗期根系影响的蛋白质组学研究[J]. 华北农学报, 2019, 34(6): 82-88.
[6]	顾倩, 丁维维, 蒋明月, 苏晓帅, 李小娟, 肖凯. *过表达小麦锌指型转录因子基因TaZAT8烟草根系差异蛋白的鉴定*[J]. 华北农学报, 2019, 34(5): 45-51.
[7]	王鹏, 王军节, 李贞彪. 适用于蛋白质组分析的粉红单端孢菌蛋白提取方法的建立[J]. 华北农学报, 2019, 34(3): 209-216.
[8]	张辉, 白晨, 王华忠, 张惠忠, 李晓东, 付增娟, 赵尚敏, 鄂圆圆, 张自强, 王良, 张必周. 甜菜丛根病抗性差异性状及蛋白胶点鉴定比较研究[J]. 华北农学报, 2018, 33(5): 52-59.
[9]	王晓丹, 肖钢, 常涛, 张振乾, 官春云, 邬贤梦. 高油酸油菜脂肪酸代谢的蛋白质组学与转录组学关联分析[J]. 华北农学报, 2017, 32(6): 31-36.
[10]	张辉, 白晨, 王华忠, 张惠忠, 李晓东, 付增娟, 赵尚敏, 鄂圆圆, 张自强, 王良. 甜菜双向电泳体系条件的优化[J]. 华北农学报, 2017, 32(5): 112-116.
[11]	张晋龙, 刘义, 渠云芳, 康娜娜, 杜海烨, 黄晋玲. 棉花不同茸毛性状近等基因系的选育与生理生化特性研究[J]. 华北农学报, 2017, 32(3): 137-142.
[12]	边立娜, 范宝莉, 孙德岭, 彭永康. 花椰菜细胞质雄性不育(CMS)与可育花粉母细胞不同发育期的比较蛋白质组研究[J]. 华北农学报, 2016, 31(S1): 101-105.
[13]	刘辉, 査日扬, 马永周, 叶玉秀. H₂S处理下对小麦根蛋白质组表达谱分析的研究[J]. 华北农学报, 2016, 31(S1): 328-334.
[14]	秦海霞, 侯俊峰, 丁会纳, 夏先春, 何中虎, Morris F Craig, 王晨阳, 马冬云. Puroindoline b位点近等基因系对小麦籽粒淀粉含量和组分及其特性的影响[J]. 华北农学报, 2016, 31(6): 39-43.
[15]	杜欢, 马彤彤, 侯晓梦, 张颖, 白志英, 李存东. 20对大麦株高近等基因系农艺与产量性状差异及相关性分析[J]. 华北农学报, 2016, 31(5): 114-121.

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

白粉菌侵染Pm21近等基因系的早期蛋白组差异研究

Proteomic Analyses of the Early Pm21 Wheat Near Isogenic Lines and Their Parents Affected by Powdery Mildew

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价