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作物学报 ›› 2010, Vol. 36 ›› Issue (12): 2091-2098.doi: 10.3724/SP.J.1006.2010.02091

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

含CBS结构域的小麦TaCDCP1基因的克隆及其表达分析

王晓敏1,冯浩1,孙燕飞1,刘博1,王晓杰1,徐亮胜1,于秀梅1,魏国荣1,黄丽丽1,康振生1,2,*   

  1. 1 西北农林科技大学植物保护学院，陕西杨凌 712100；2 西北农林科技大学陕西省农业分子生物学重点实验室，陕西杨凌 712100
  • 收稿日期:2010-05-24 修回日期:2010-08-03 出版日期:2010-12-12 网络出版日期:2010-10-22
  • 通讯作者: 康振生,E-mail:kangzs@nwsuaf.edu.cn
  • 基金资助:

    本研究由国家自然科学基金重点项目(30930064)，现代农业产业技术体系建设专项资金和高等学校学科创新引智计划资助项目(B07049)资助。

Cloning and Expression Analysis of a CBS Domain Containing Protein Gene TaCDCP1 from Wheat

WANG Xiao-Min1,FENG Hao1,SUN Yan-Fei1,LIU Bo1,WANG Xiao-Jie1,XU Liang-Sheng1,YU Xiu-Mei1,WEI Guo-Rong1,HUANG Li-Li1,KANG Zhen-Sheng1,2,*   

  1. 1 College of Plant Protection, Northwest A&F University, Yangling 712100, China; 2 Shaanxi Provincial Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, Yangling 712100, China
  • Received:2010-05-24 Revised:2010-08-03 Published:2010-12-12 Published online:2010-10-22
  • Contact: 康振生,E-mail:kangzs@nwsuaf.edu.cn

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摘要： 利用电子克隆和RT-PCR技术，在本实验室前期构建的非亲和互作的SSH文库基础上，从条锈菌侵染的小麦水原11叶片中首次分离出一个含CBS结构域蛋白的基因，暂命名为TaCDCP1 (Triticum aestivum CBS domain containing protein 1)。TaCDCP1包含一个完整的654 bp的开放阅读框，编码217个氨基酸。推测编码的蛋白具有两个CBS保守结构域，不含跨膜区且无信号肽，定位在叶绿体基质内；经过同源比对，小麦TaCDCP1氨基酸序列与大麦、水稻和玉米等的同源序列的相似性较高；该基因表达量在小麦叶中显著高于在根和茎中；在小麦与条锈菌的非亲和、亲和组合中，TaCDCP1基因均受到条锈菌诱导，分别在接种后18 h和96 h达到表达高峰，非亲和组合表达量在侵染前期(接种后18~48 h)高于亲和组合，而在侵染后期(接种后96~120 h)低于亲和组合；外源植物激素脱落酸诱导该基因上调表达，苄基腺嘌呤，乙烯，赤霉素，茉莉酸甲酯和水杨酸处理后其表达量在不同程度上受到抑制；TaCDCP1在低温和干旱条件下表达量上升，在机械伤害和高盐处理下表达量无明显差异。表明TaCDCP1可能通过脱落酸等信号途径参与小麦对条锈菌的防御反应，同时参与低温和干旱环境下的信号转导途径。这些结果对于明确CBS结构域的功能以及CBS结构域蛋白尤其是TaCDCP1在小麦与条锈菌互作中的作用奠定了基础。

关键词: 小麦, 条锈菌, CBS结构域, 非生物胁迫, 基因表达

Abstract: To elucidate the defense response of wheat(Triticum aestivum L.) to Puccinia striiformis f. sp. tritici (Pst), we constructed the incompatible interaction SSH cDNA library of wheat(cv. Suwon 11) leaves infected by Pst CYR23. A total of 652 unigenes were identified and 424 genes were annotated. On the basis of previous study, according to cDNA sequence LWSRP2502 (Genbank accession No. EV254338), a full-length sequence of the CBS domain containing protein gene, tentatively designated as TaCDCP1 (Triticum aestivum CBS domain containing protein 1), was isolated and characterized from wheat leaves infected by Pst through in silico cloning and reverse transcription PCR (RT-PCR) approaches.The open reading frame of TaCDCP1 was 654 bp in length andpredicted to encode 217 amino acids protein which contained two conserved cystathionine beta-synthase (CBS) domains and was without transmembrane domain or signal peptide sequence. The deduced protein was predicted existing in chloroplast stroma. The amino acid sequence of TaCDCP1 shares 92%, 72%, and 63% identify withthe homologs in barley (Hordeum vulgare) , rice (Oryza sativa) and maize (Zea mays), respectively. The TaCDCP1 gene was highly expressed in leaves than in roots and stems. Challenged by Pst, TaCDCP1 was induced by this fungus in both incompatible and compatible interactions, with the maximal expression at 18 h post inoculation (hpi) and 24 hpi, respectively. Its transcript accumulation was much higher in the incompatible interaction than in the compatible interaction at the early stage of infection (18–48 hpi), but much lower at the late stage (96–120 hpi). The expression of TaCDCP1 wasalso up-regulated after treated by phytohormones such as abscisic acid (ABA), and down-regulated by benzyladenine, ethylene, gibberellins, methyl jasmonate and salicylic acid to a certain degree. And it was obviously up-regulated by various abiotic stresses, such as low temperature and drought. However, mechanical wound and high salinity stress could not induce the expression of TaCDCP1. These results suggest that TaCDCP1 is probably involved in the disease resistance and defense response in wheat to Pst through ABA and ethylene pathways, and also participate in the signal transmission pathways under low temperature, and drought conditions.

Key words: Wheat, Stripe rust fungus, CBS domain, Abiotic stresses, Gene expression

[1]Chen X M, Line R F. Inheritance of stripe rust (yellow rust) resistance in the wheat cultivar Carstens V. Euphytica, 1993, 71: 107–113
[2]Chen X M. Epidemiology and control of stripe rust (Puccinia striiformis f. sp. tritici) on wheat. Can J Plant Pathol, 2005, 27: 314–337
[3]Li Z-Q(李振岐), Zeng S-M(曾士迈). Stripe Rust in China (中国小麦锈病). Beijing: China Agriculture Press, 2002. pp 2–3 (in Chinese)
[4]Bateman A. The structure of a domain common to archaebacteria and the homocystinuria disease protein. Trends Biochem Sci, 1997, 22: 12–13
[5]Hemant R K, Anil K S, Sudhir K S, Sneh L S P, Ashwani P. Genome wide expression analysis of CBS domain containing proteins in Arabidopsis thaliana (L.) Heynh and Oryza sativa L. reveals their developmental and stress regulation. BMC Genomics, 2009, 10: 200
[6]Woods A, Cheung P C F, Smith F C, Davison M D, Scott J, Beri R K, Carling D. Characterization of AMP-activated protein kinase and subunits. J Biol Chem, 1996, 271: 10282–10290
[7]Sintchak M D, Fleming M A, Futer O, Raybuck S A, Chambers S P, Caron P R, Murcko M A, Wilson K P. Structure and mechanism of inosine monophosphate dehydrogenase in complex with the immunosuppressant mycophenolic acid. Cell, 1996, 85: 921–930
[8]Schmidt-Rose T, Jentsch T J. Reconstitution of functional voltage-gated chloride channels from complementary fragments of CLC-1. J Biol Chem, 1997, 272: 20515–20521
[9]Shan X Y, Kruger W D. Correction of disease-causing CBS mutations in yeast. Nat Genet, 1998, 19: 91–93
[10]Ignoul S, Eggermont J. CBS domains: structure, function, and pathology in human proteins. Am J Physiol-Cell Physiol, 2005, 289: 1369–1378
[11]Wang X L, Ren X, Zhu L L, He G C. OsBi1, a rice gene, encodes a novel protein with a CBS-like domain and its expression is induced in responses to herbivore feeding. Plant Sci, 2004, 166: 1581–1588
[12]Manickavelu A, Kawaura K, Oishi K, Shin-I T, Kohara Y, Yahiaoui N, Keller B, Suzuki A, Yano K, Ogihara Y. Comparative gene expression analysis of susceptible and resistant near-isogenic lines in common wheat infected by Puccinia triticina. DNA Res, 2010, DOI: 10.1093/dnares/dsq009
[13]Yu X-M(于秀梅). Construction of SSH cDNA library of Wheat Leaves Induced by Puccinia striiformis and Its ESTs Analysis. PhD Dissertation of Northwest A&F University, 2006. pp 62–66 (in Chinese with English abstract)
[14]Kang Z-S(康振生), Li Z-Q(李振岐). Discovery of pathogenic isolates of stripe rust on cultivar Lovrin 10 at normal temperature. J Northwest Agric Coll (西北农林科技大学学报), 1984, 12(4): 18–28 (in Chinese with English abstract)
[15]Zhang H B, Zhang D B, Chen J, Yang Y H, Huang Z J, Huang D F, Wang X C, Huang R F. Tomato stress-responsive factor TSRF1 interacts with ethylene responsive element GCC box and regulates pathogen resistance to Ralstonia solanacearum. Plant Mol Biol, 2004, 55: 825–834
[16]Sato K, Shin-I T, Seki M, Shinozaki K, Yoshida H, Takeda K, Yamazaki Y, Conte M, Kohara Y. Development of 5006 full-length cDNAs in barley: a tool for accessing cereal genomics resources. DNA Res, 2009, 16: 81–89
[17]Alexandrov N N, Brover V V, Freidin S, Troukhan M E, Tatarinova T V, Zhang H, Swaller T J, Lu Y P, Bouck J, Flavell R B. Insights into corn genes derived from large-scale cDNA sequencing. Plant Mol Biol, 2009, 69: 179–194
[18]Yu X M, Yu X D, Qu Z P, Huang X J, Guo J, Han Q M, Zhao J, Huang L, Kang Z S. Cloning of a putative hypersensitive induced reaction gene from wheat infected by stripe rust fungus. Gene, 2008, 407: 193–198
[19]Yu G X, Braun E, Wise R P. Rds and Rih mediate hypersensitive cell death independent of gene-for-gene resistance to the oat crown rust pathogen Puccinia coronata f. sp. avenae. Mol Plant Microbe Interact, 2001, 14: 1376–1383
[20]Hand J-D(韩建东), Cao Y-Y(曹远银), Yao P(姚平). Hypersensitive response and activity dynamics of defense enzymes induced by elicitor(s) from wheat-stem rust interaction systerm. Acta Agr Boreali-Sinica (华北农学报), 2009, 24(1): 79–82 (in Chinese with English abstract)
[21]Wang C F, Huang L L, Buchenauer H, Han Q M, Zhang H C, Kang Z S. Histochemical studies on the accumulation of reactive oxygen species (O2- and H2O2) in the incompatible and compatible interaction of wheat-Puccinia striiformis f. sp. tritici. Physiol Mol Plant Pathol, 2007, 71: 230–239
[22]Kang Z S, Huang L L, Buchenauer H. Ultrastructural changes and localization of lignin and callose in compatible and incompatible interactions between wheat and Puccinia striiformis. J Plant Dis Protect, 2002, 109: 25–37
[23]Zhu Q, Dröge-Laser W, Dixon R A, Lamb C. Transcriptional activation of plant defense genes. Curr Opin Genet Dev, 1996, 6: 624–630
[24]Glazebrook J. Genes controlling expression of defense responses in Arabidopsis. Curr Opin Plant Biol, 1999, 2: 280–286
[25]Glazebrook J. Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens. Annu Rev Phytopathol, 2005, 43: 205–227
[26]Koornneef A, Pieterse C M J. Crosstalk in defense signaling. Plant Physiol, 2008, 146: 839–844
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