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作物学报 ›› 2009, Vol. 35 ›› Issue (5): 786-794.doi: 10.3724/SP.J.1006.2009.00786

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

白粉菌(Erysiphe graminis)侵染诱导表达的小麦糖苷水解酶基因TaGlc2的克隆与鉴定

Ramesh N PUDAKE**,辛明明**,尹玉静**,解超杰,倪中福,孙其信*   

  1. 中国农业大学农业生物技术国家重点实验室/农业部作物基因组学与遗传改良重点开放实验室/北京市作物遗传改良重点实验室/教育部作物杂种优势研究及利用重点实验室,北京100193
  • 收稿日期:2008-10-07 修回日期:2009-02-17 出版日期:2009-05-12 网络出版日期:2009-03-20
  • 通讯作者: 孙其信

Cloning and Characterization of a Novel Wheat Glycoside Hydrolase Gene TaGlc2 Induced by Powdery Mildew Pathogen(Erysiphe graminis) Infection

Ramesh N PUDAKE**,XIN Ming-Ming**,YIN Yu-Jing1,XIE Chao-Jie,NI Zhong-Fu,SUN Qi-Xin*   

  1. State Key Laboratory for Agrobiotechnology/Key Laboratory of Crop Heterosis and Utilization,Ministry of Education/Key Laboratory of Crop Genomics and Genetic Improvement,Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University,Beijing 100193,China
  • Received:2008-10-07 Revised:2009-02-17 Published:2009-05-12 Published online:2009-03-20
  • Contact: SUN Qi-Xin
  • Supported by:

    The work was supported by the National High Technology Research and Development Program of China(2006AA10A104),Fok Ying-Tung Education Foundation(94021),and Natural Science Foundation of Beijing, China(6061003 and 30871528)

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摘要：

真菌病害是世界小麦生产中的最重要的病害之一。目前，在小麦中已经鉴定出一批小麦病菌侵染诱导基因，包括病程相关基因和抗真菌水解酶基因（葡聚糖酶基因和几丁质酶基因）。最近的研究表明，植物1,3-β-葡聚糖酶参与对真菌侵染的防卫。本研究从小麦cDNA文库中克隆出一个编码小麦1,3-β-葡聚糖酶的新基因，命名为 TaGlc2。该基因编码的氨基酸序列与糖苷水解酶基因家族17高度同源。采用实时定量PCR分析方法对TaGlc2基因在白粉菌侵染(Erysiphe graminis)后小麦叶片中的表达模式进行研究，发现TaGlc2基因在白粉菌侵染6 h后表达明显增强，至24 h达到峰值，说明该基因受白粉菌侵染诱导表达。还获得了TaGlc2基因5'上游调控区序列，发现存在与病菌侵染响应有关的顺式元件。小麦TaGlc2基因在小麦白粉病抗性上可能具有重要作用。

关键词: 1,3-ß-葡聚糖酶, 白粉菌, 真菌侵染, 基因表达, 小麦

Abstract:

Fungal diseases cause serious yield losses of wheat worldwide. Up to date, numerous genes involved in the wheat-pathogen response have been identified. These include pathogenesis related (PR) genes and antifungal hydrolases such as glucanase and chitinase genes. Recently, there has been increasing number of studies providing evidence of the potential involvement of 1,3-β-glucanase in defense against fungal infection. In this study we identified a cDNA encoding a 1,3-β-glucanase, designated TaGlc2, from wheat cDNA library. The deduced peptide sequence of TaGlc2 is similar to a glycoside hydrolase family 17. Using real time PCR, the expression pattern of TaGlc2 in wheat seedlings inoculated with powdery mildew pathogen (Erysiphe graminis) was determined. The results showed that TaGlc2 is inducible in response to fungal infection. The 5' genomic region of TaGlc2 was isolated and it contains some cis-elements which are reported to be involved in pathogenesis response.

Key words: 1,3-ß-glucanase, Powdery mildew, Fungal inoculation, Gene expression, Wheat


[1] Wessels J G H, Sietsma J H. Fungal cell wall: a survey. Tanner W, Loewus F A, eds. Encyclopedia of plant physiology, new series, plant carbohydrates II, vol. 13B. Berlin: Springer, 1981. pp 352–394

[2] V?geli-Lange R, Fründt C, Hart C M, Beffa R, Nagy F, Meins F Jr. Evidence for a role of β-1,3-glucanase in dicot seed germination. Plant J, 1994, 5: 273–278

[3] Worrall D, Hird DL, Hodge R, Paul W, Draper J, Scott R. Premature dissolution of the microsporocyte callose wall causes male sterility in transgenic tobacco. Plant Cell, 1992, 4: 759–771

[4] Ori N, Sessa G, Lotan T, Himmelhoch S, Fluhr R. A major stylar matrix polypeptide (sp41) is a member of the pathogenesis-related protein superclass. EMBO J, 1990, 9: 3429–3436

[5] Roulin S, Buchala A J. The induction of 1,3-β-glucanases and other enzymes in groundnut leaves infected with Cercospora arachidicola. Physiol Mol Plant Path, 1995, 46: 471–489

[6] Anguelova-merhar V S, Van der Westhuizen A J, Pretorius Z A. β-1,3-glucanase and chitinase activities and the resistance response of wheat to leaf rust. J Phytopath, 2001, 149: 381–384

[7] Cheong Y H, Kim C Y, Chun H J, Moon B C, Park H C, Kim K J, Lee S, Han C, Lee S Y, Cho M J. Molecular cloning of a soybean class III β-1,3-glucanase gene that is regulated both developmentally and in response to pathogen infection. Plant Sci, 2000, 154: 71–81

[8] Shi Y, Zhang Y, Shih D S. Cloning and expression analysis of two β-1,3-glucanase genes from Strawberry. J Plant Physiol, 2006, 163: 956–967

[9] Zhao T J, Zhao S Y, Chen H M, Zhao Q Z, Hu Z M, Hou B K, Xia G M. Transgenic wheat progeny resistant to powdery mildew generated by Agrobacterium inoculum to the basal portion of wheat seedling. Plant Cell Rep, 2006, 25: 1119–1204

[10] Mackintosh C A, Lewis J, Radmer L E, Shin S, Heinen S J, Smith L A, Wyckoff M N, Macky R D, Evans C K, Kravchenko S, Baldridge G D, Zeyen R J, Muehlbauer G J. Overexpression of defense response genes in transgenic wheat enhances resistance to Fusarium head blight. Plant Cell Rep, 2007, 26: 479–488

[11] Clarke B C, Moran L B, Appels R. DNA analyses in wheat breeding. Genome, 1989, 32: 334–339

[12] Bouzidi M F, Franchel J, Tao Q, Stormo K, Mraz A, Nicolas P, Mouzeyar S. A sunflower BAC library suitable for PCR screening and physical mapping of targeted genomic regions. Theor Appl Genet, 2006, 113: 81–89

[13] Altschul S F, Madden T L, Sch?ffer A A, Zhang J, Zhang Z, Miller W, Lipman D J. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl Acids Res, 1997, 25: 3389–3402

[14] Jeanmougin F, Thompson J D, Gouy M, Higgins D G, Gibson T J. Multiple sequence alignment with Clustal X. Trends Biochem Sci, 1998, 23: 403–405

[15] Thompson J D, Higgins D G, Gibson T J. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions- specific gap penalties and weight matrix choice. Nucl Acids Res, 1994, 22: 4673–4680

[16] Felsenstein J. PHYLIP (Phylogeny Inference Package) version 3.6. Department of Genome Sciences, University of Washington, Seattle, 2005

[17] Marchler-Bauer A, Anderson J B, Cherukuri P F, DeWeese-Scott C, Geer L Y, Gwadz M, He S, Hurwitz D I, Jackson J D, Ke Z, Lanczycki C J, Liebert C A, Liu C, Lu F, Marchler G H, Mullokandov M, Shoemaker B A, Simonyan V, Song J S, Thiessen P A, Yamashita R A, Yin J J, Bryant S H. CDD: a Conserved Domain Database for protein classification. Nucl Acids Res, 2005, 33(D): 192–196

[18] Nakai K, Horton P. PSORT: a program for detecting the sorting signals of proteins and predicting their subcellular localization. Trends Biochem Sci, 1999, 24: 34–35

[19] Bendtsen J D, Nielsen H, Heijne G, Brunak S. Improved prediction of signal peptides: SignalP 3.0. J Mol Biol, 2004, 340: 783–795

[20] Li W L, Faris J D, Muthukrishnan S, Liu D J, Chen P D, Gill B S. Isolation and characterization of novel cDNA clones of acidic chitinases and β-1,3-glucanases from wheat spikes infected by Fusarium graminearum. Theor Appl Genet, 2001, 102: 353–362

[21] Muradov A, Petrasovits L, Davidson A, Scott K J. A cDNA clone for a pathogenesis-related protein 1 from barley. Plant Mol Biol, 1993, 23: 439–442

[22] Higa-Nishiyama A, Ohsato S, Banno S, Woo S H, Fujimura M, Yamaguchi I, Kimura M. Cloning and characterization of six highly similar endo-1,3-β-glucanase genes in hexaploid wheat. Plant Physiol Biochem, 2006, 11: 1361–1368

[23] Lai D M, H?j P B, Fincher G B. Purification and characterization of (1→3, 1→4)-β-glucan endohydrolases from germinated wheat (Triticum aestivum). Plant Mol Biol, 1993, 22: 847–859

[24] Cruz-Ortega R, Cushman J C, Ownby J D. cDNA clones encoding 1,3-β-glucanase and a fimbrin-like cytoskeletal protein are induced by Al toxicity in wheat roots. Plant Physiol, 1997, 114: 1453–1460

[25] Henrissat B, Davies G J. Glycoside hydrolases and glycosyltransferases. Families, modules, and implications for genomics. Plant Physiol, 2000, 124: 1515–1519

[26] Ray S, Anderson J M, Urmeev F I, Goodwin S B. Rapid induction of a protein disulfide isomerase and defense-related genes in wheat in response to the hemibiotrophic fungal pathogen Mycosphaerella graminicola. Plant Mol Biol, 2003, 53: 741–754

[27] Cohen L, Eyal Z. The histology of processes associated with the infection of resistant and susceptible wheat cultivars with Septoria tritici. Plant Path, 1993, 42: 737–743
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