甘蓝型油菜MAPK1在损伤和病原菌胁迫下的表达模式分析

doi:10.3864/j.issn.0578-1752.2013.20.023

摘要/Abstract

摘要： 【目的】分析BnMAPK1组织特异性表达特征，探究其在损伤和病原菌胁迫下的特征及参加的信号途径。【方法】在电子克隆的基础上，克隆BnMAPK1的启动子序列。利用实时荧光定量PCR（qRT-PCR）检测BnMAPK1的诱导表达特征。【结果】从甘蓝型油菜中克隆了MAPK1的启动子序列，发现它含有多个激素响应元件和逆境胁迫响应元件。还发现植物激素茉莉酸甲酯（MeJA）、水杨酸（SA）、脱落酸（ABA）和过氧化氢（H2O2），及损伤（wound）和核盘菌（sclerotinia sclerotiorum）均能在转录水平上激活BnMAPK1。甘蓝型油菜在损伤和接种核菌盘后，JA信号途径标志基因PDF1.2的表达量被诱导上升，而SA信号途径标志基因PR-1被抑制，推测甘蓝型油菜可能是通过茉莉酸（jasmonic acid，JA）信号途径响应损伤和病原菌入侵。【结论】甘蓝型油菜MAPK1受多种植物激素和胁迫的诱导，并且其启动子中含有多个激素响应元件和逆境胁迫响应元件，可能MAPK1在植物体抵抗外界多种胁迫中起重要作用，并且甘蓝型油菜可能是通过JA信号途径对损伤和病原菌入侵做出响应。

关键词: 甘蓝型油菜 , BnMAPK1 , 损伤 , 病原菌 , 信号途径

Abstract: 【Objective】The objective of this study is to analyze tissue specific expression of BnMAPK1 genes, and explore their characteristics and signaling pathway in wound and pathogenetic fungi stress.【Method】On the basis of silico cloning, the promoters of BnMAPK1 were cloned. BnMAPK1 mRNA expression characteristics after induced were analyzed by Quantitative real-time PCR (qRT-PCR). 【Result】The cloned promoter sequences of BnMAPK1 contain several hormone and stress responsive elements. BnMAPK1 expression could be induced by phytohormones methyl jasmonate (MeJA), salicylic acid (SA), abscisic acid (ABA), hydrogen peroxide (H2O2), wound, and Sclerotinia sclerotiorum. Further experiments showed that PDF1.2 gene was activated but PR-1 was suppressed after wound and S.sclerotinrum treatments, indicate that oilseed rape most likely through jasmonic acid (JA) signaling in response to wound and pathogenetic fungi stress.【Conclusion】BnMAPK1 expression could be induced by several phytohormones and its promoter contains several hormone and stress responsive elements. BnMAPK1 may play an important role in resistance to a variety of stress in plants and oilseed rape might be mediated by JA signaling pathway in response to wound and pathogenetic fungi stress.

Key words: Brassica napus , BnMAPK1 gene , wound , pathogenetic fungi , signaling pathway

陆俊杏, 陆奇丰, 张凯, 柴友荣, 李加纳, 钱伟, 吕俊, 卢坤, 梁颖. 甘蓝型油菜MAPK1在损伤和病原菌胁迫下的表达模式分析[J]. 中国农业科学, 2013, 46(20): 4388-4396.

LU Jun-Xing, LU Qi-Feng, ZHANG Kai, CHAI You-Rong, LI Jia-Na, QIAN Wei, 吕Jun , LU Kun, LIANG Ying. Expression Features of BnMAPK1 in Wound and Pathogenetic Fungi Stress[J]. Scientia Agricultura Sinica, 2013, 46(20): 4388-4396.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2013.20.023

https://www.chinaagrisci.com/CN/Y2013/V46/I20/4388

参考文献

[1]Colcombet J, Hirt H. Arabidopsis MAPKs: A complex signalling network involved in multiple biological processes. Biochemical Journal, 2008, 413: 217-226.

[2]Pitzschke A, Schikora A, Hirt H. MAPK cascade signalling networks in plant defence. Current Opinion in Plant Biology, 2009, 12: 421-426.

[3]Zhang S, Klessig D F. MAPK cascades in plant defense signaling. Trends in Plant Science, 2001, 6: 520-527.

[4]Jonak C, Okresz L, Bogre L, Hirt H. Complexity, cross talk and integration of plant MAP kinase signalling. Current Opinion in Plant Biology, 2002, 5: 415-424.

[5]Leon J, Rojo E, Sanchez-Serrano J J. Wound signalling in plants. Journal of Experimental Botany, 2001, 52: 1-9.

[6]Seo S, Okamoto M, Seto H, Ishizuka K, Sano H, Ohashi Y. Tobacco MAP kinase: A possible mediator in wound signal transduction pathways. Science, 1995, 270: 1988-1992.

[7]Seo S, Sano H, Ohashi Y. Jasmonate-based wound signal transduction requires activation of WIPK, a tobacco mitogen-activated protein kinase. The Plant Cell, 1999, 11: 289-298.

[8]Penninckx I A, Eggermont K, Terras F R, Thomma B P, De Samblanx G W, Buchala A, Metraux, J P, Manners J M, Broekaert W F. Pathogen-induced systemic activation of a plant defensin gene in Arabidopsis follows a salicylic acid-independent pathway. The Plant Cell, 1996, 8: 2309-2323.

[9]Thomma B P, Eggermont K, Penninckx I A, Mauch-Mani B, Vogelsang R, Cammue B P, Broekaert W F. Separate jasmonate-dependent and salicylate-dependent defense-response pathways in Arabidopsis are essential for resistance to distinct microbial pathogens. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95: 15107-15111.

[10]Feys B J, Parker J E. Interplay of signaling pathways in plant disease resistance. Trends in Genetics, 2000, 16: 449-455.

[11]尹恒, 杨金丽, 李曙光, 赵小明, 白雪芳, 马小军, 杜昱光. 受壳寡糖诱导的油菜MAPK基因的克隆与分析. 作物学报, 2008, 34(5): 743-747.

Yin H, Yang J L, Li S G, Zhao X M, Bai X F, Ma X J, Du Y G. Cloning and analysis of BnMPK4, a novel MAP kinase gene induced by oligochitosan in Brassica napus. Acta Agronomica Sinica, 2008, 34(5): 743-747. (in Chinese)

[12]Wang Z, Mao H, Dong C, Ji R, Cai L, Fu H, Liu, S. Overexpression of Brassica napus MPK4 enhances resistance to Sclerotinia sclerotiorum in oilseed rape. Molecular Plant-Microbe Interactions, 2009, 22: 235-244.

[13]Ortiz-Masia D, Perez-Amador M A, Carbonell J, Marcote M J. Diverse stress signals activate the C1 subgroup MAP kinases of Arabidopsis. Febs Letters, 2007, 581: 1834-1840.

[14]Umezawa T, Sugiyama N, Takahashi F, Anderson J C, Ishihama Y, Peck S C, Shinozaki K. Genetics and phosphoproteomics reveal a protein phosphorylation network in the abscisic acid signaling pathway in Arabidopsis thaliana. Science Signaling, 2013, 6: 1-13.

[15]Takemoto D, Hardham A R, Jones D A. Differences in cell death induction by phytophthora elicitins are determined by signal components downstream of MAP kinase kinase in different species of Nicotiana and cultivars of Brassica rapa and Raphanus sativus. Plant Physiology, 2005, 138: 1491-1504.

[16]Liu S Y, Liu R H, Latunde-Dada A O, Cools H J, Foster S J, Huang Y J, Fitt B D L. Comparison of Leptosphaeria biglobosa-induced and chemically induced systemic resistance to L-maculans in Brassica napus. Chinese Science Bulletin, 2007, 52: 1053-1062.

[17]Chen X, Truksa M, Shah S, Weselake R J. A survey of quantitative real-time polymerase chain reaction internal reference genes for expression studies in Brassica napus. Analytical Biochemistry, 2010, 405: 138-140.

[18]Bustin S A, Benes V, Garson J A, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl M W, Shipley G L, Vandesompele J, Wittwer C T. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry, 2009, 55: 611-622.

[19]Rojo E, Solano R, Sanchez-Serrano J J. Interactions between signaling compounds involved in plant defense. Journal of Plant Growth Regulation, 2003, 22: 82-98.

[20]Zhang S, Klessig D F. Salicylic acid activates a 48-kD MAP kinase in tobacco. The Plant Cell, 1997, 9: 809-824.

[21]Gomi K, Ogawa D, Katou S, Kamada H, Nakajima N, Saji H, Soyano T, Sasabe M, Machida Y, Mitsuhara I, Ohashi Y, Seo S. A mitogen-activated protein kinase NtMPK4 activated by SIPKK is required for jasmonic acid signaling and involved in ozone tolerance via stomatal movement in tobacco. Plant and Cell Physiology, 2005, 46: 1902-1914.

[22]Romeis T, Piedras P, Zhang S, Klessig D F, Hirt H, Jones J D. Rapid Avr9- and Cf-9 -dependent activation of MAP kinases in tobacco cell cultures and leaves: Convergence of resistance gene, elicitor, wound, and salicylate responses. The Plant Cell, 1999, 11: 273-287.

[23]Laudert D, Weiler E W. Allene oxide synthase: A major control point in Arabidopsis thaliana octadecanoid signalling. The Plant Journal, 1998, 15: 675-684.

[24]Harms K, Ramirez I I, Pena-Cortes H. Inhibition of wound-induced accumulation of allene oxide synthase transcripts in flax leaves by aspirin and salicylic acid. Plant Physiology, 1998, 118: 1057-1065.

[25]Clarke J D, Volko S M, Ledford H, Ausubel F M, Dong X. Roles of salicylic acid, jasmonic acid, and ethylene in cpr-induced resistance in Arabidopsis. The Plant Cell, 2000, 12: 2175-2190.

[26]Rojo E, Titarenko E, Leon J, Berger S, Vancanneyt G, Sanchez-Serrano J J. Reversible protein phosphorylation regulates jasmonic acid-dependent and -independent wound signal transduction pathways in Arabidopsis thaliana. The Plant Journal, 1998, 13: 153-165.

[27]Baudouin E, Meskiene I, Hirt H. Short communication: Unsaturated fatty acids inhibit MP2C, a protein phosphatase 2C involved in the wound-induced MAP kinase pathway regulation. The Plant Journal, 1999, 20: 343-348.

[28]Reymond P, Weber H, Damond M, Farmer E E. Differential gene expression in response to mechanical wounding and insect feeding in Arabidopsis. The Plant Cell, 2000, 12: 707-720.

[29]Perez-Amador M A, Leon J, Green P J, Carbonell J. Induction of the arginine decarboxylase ADC2 gene provides evidence for the involvement of polyamines in the wound response in Arabidopsis. Plant Physiology, 2002, 130: 1454-1463.

[30]Guan L M, Zhao J, Scandalios J G. Cis-elements and trans-factors that regulate expression of the maize Cat1 antioxidant gene in response to ABA and osmotic stress: H2O2 is the likely intermediary signaling molecule for the response. The Plant Journal, 2000, 22: 87-95.

[31]Orozco-Cardenas M L, Narvaez-Vasquez J, Ryan C A. Hydrogen peroxide acts as a second messenger for the induction of defense genes in tomato plants in response to wounding, systemin, and methyl jasmonate. The Plant Cell, 2001, 13: 179-191.

[32]Ortiz-Masia D, Perez-Amador M A, Carbonell P, Aniento F, Carbonell J, Marcote M J. Characterization of PsMPK2, the first C1 subgroup MAP kinase from pea (Pisum sativum L.). Planta, 2008, 227: 1333-1342.

[33]Blanco F A, Zanetti M E, Casalongué C A, Daleo G R. Molecular characterization of a potato MAP kinase transcriptionally regulated by multiple environmental stresses. Plant Physiology and Biochemistry, 2006, 44: 315-322.

[34]Wang X J, Zhu S Y, Lu Y F, Zhao R, Xin Q, Wang X F, Zhang D P. Two coupled components of the mitogen-activated protein kinase cascade MdMPK1 and MdMKK1 fromfrom apple function in ABA signal transduction. Plant and Cell Physiology, 2010, l51: 754-766.

[35]Petersen M, Brodersen P, Naested H, Andreasson E, Lindhart U, Johansen B, Nielsen H B, Lacy M, Austin M J, Parker J E, Sharma S B, Klessig D F, Martienssen R, Mattsson O, Jensen A B, Mundy J. Arabidopsis MAP kinase 4 negatively regulates systemic acquired resistance. Cell, 2000, 103: 1111-1120.

[36]Nakagami H, Pitzschke A, Hirt H. Emerging MAP kinase pathways in plant stress signalling. Trends in Plant Science, 2005, 10: 339-346.