稻瘟菌诱导性水稻Benzoyl-CoA：苯甲酰基转移酶基因的cDNA克隆与鉴定
|
摘要
植物在受到病原物侵染时可以产生一系列复杂的生理生化反应，这些反应大多数是由一系列植物基因的转录活化所介导的，其中包括许多植物的防卫基因及其它一些可能涉及植物与病原物识别，信号转导和防卫基因转录起始等相关的基因。这些基因包括植保素合成途径的关键酶基因。研究这些基因在植物与病原物互作过程中的功能及诱导表达特性，有助于揭示植物-病原物的互作机理；同时，分离这些基因可为植物抗病基因工程提供必要的操作材料。
植保素是植物抵抗外来生物或非生物因素的侵袭而产生的低分子量抗菌活性物质，是植物体内的次生代谢产物，具有侵染可诱导性和体外抗菌性的特点，其在受病原菌侵染的植物体内的诱导合成与积累被认为是重要的抗病机理之一。彭友良(1996)在非亲和性稻瘟菌接种的稻叶中发现两种新的色胺类植保素P1(N-苯甲酰基色胺)和P2(N-肉桂酰基色胺)，根据化学结构推测，苯甲酰类化合物是这两种植保素的前体之一，在非亲和性稻瘟菌侵染的稻叶中可能有苯甲酰转移酶(Benzoyltransferase)的诱导。
在本研究中，作者以一个与其它物种苯甲酰基转移酶基因具有同源性的cDNA片段为探针，筛选非亲和性稻瘟菌侵染诱导的稻叶cDNA文库，获得的了一个苯甲酰基转移酶基因全长cDNA。
苯甲酰基转移酶基因的cDNA全长为1610bp，其中完整的开放阅读框为1335bp(82-1419)，编码一条具有446个氨基酸残基的多肽。而且该多肽含有一个保守的苯甲酰基转移酶催化区HXXXDG，该基因推定的氨基酸序列与来自于大麦。紫杉 康乃馨 长春花的中的苯甲酰基转移酶基因的氨基酸序列一致性高达41.69％。在大肠杆菌中能诱导表达，SDA-PAGE结果显示在74kDa有一差异表达带。并初步测定具有酶的活性。
Northern杂交分析表明：非亲和性稻瘟菌小种能诱导苯甲酰基转移酶转录物的积累；而亲和性稻瘟菌小种不能诱导苯甲酰基转移酶转录物的累积。
以上结果证明：苯甲酰基转移酶与水稻抗稻瘟病相关，能够体外催化合成植保素P1，可以作为抗病基因工程的侯选材料。
Plants show a series of complex physiological and biochemical changes in response to pathogen attack. Most of these responses are mediated by activation of plant genes including various defense genes and those involved in the process of pathogen recognition, signal transduction and initiation of transcription. The functional identification and expressing characterization of plant genes that are induced during interaction with pathogen will be a great help to reveal the mechanism of plant-pathogen interaction; isolation of these genes may provide a good choice for engineering plant disease resistance.
Phytoalexin is utilized by plant to resist biological or none biological invading factors, which is secondary metabolic product. It is an important fungi resistant material that have activity in vitro and can be inducible by invading factors. It is thought to be an important mechanism for its induced synthesis in plant after infected by pathogen. Fan and Peng(1996) isolated two phytoalexin in rice inoculated with incompatible race of Magnaporthe grisea. They accumulate in rice after 48 hours. According to their chemical structure, benzoc acid and cinnamic acid is precursor of this two phytoalexin. The density of JA immediately increase after elicitor., is correspondent with process of phytoalexin induction. It can be inferred that Benzoyltransferase involved in rice resistance reaction to Magnaporthe grisea.
The author characterized one full-length cDNA. Sequencing result showed that the full-length cDNA which consists of 1610bp has an intact ORF (82-1419) that encodes an 446-amino-acid protein. BLAST search indicated that the deduced amino acid sequence shares 41.69% identity with that transferase of from Hordeum, Taxus chinensis,.Catharanthus, Diabnthus caryophyllus respectively. Then there is a different band with 74kDa in the SDA-PAGE , after indued with IPTG .the result of expressing Benzoyltransferase in E. coli showed Benzoyltransferase has the transferase activity. Benzoyltransferase transcript in rice leaves can be induced by blast fungus infection.
植保素是植物抵抗外来生物或非生物因素的侵袭而产生的低分子量抗菌活性物质，是植物体内的次生代谢产物，具有侵染可诱导性和体外抗菌性的特点，其在受病原菌侵染的植物体内的诱导合成与积累被认为是重要的抗病机理之一。彭友良(1996)在非亲和性稻瘟菌接种的稻叶中发现两种新的色胺类植保素P1(N-苯甲酰基色胺)和P2(N-肉桂酰基色胺)，根据化学结构推测，苯甲酰类化合物是这两种植保素的前体之一，在非亲和性稻瘟菌侵染的稻叶中可能有苯甲酰转移酶(Benzoyltransferase)的诱导。
在本研究中，作者以一个与其它物种苯甲酰基转移酶基因具有同源性的cDNA片段为探针，筛选非亲和性稻瘟菌侵染诱导的稻叶cDNA文库，获得的了一个苯甲酰基转移酶基因全长cDNA。
苯甲酰基转移酶基因的cDNA全长为1610bp，其中完整的开放阅读框为1335bp(82-1419)，编码一条具有446个氨基酸残基的多肽。而且该多肽含有一个保守的苯甲酰基转移酶催化区HXXXDG，该基因推定的氨基酸序列与来自于大麦。紫杉 康乃馨 长春花的中的苯甲酰基转移酶基因的氨基酸序列一致性高达41.69％。在大肠杆菌中能诱导表达，SDA-PAGE结果显示在74kDa有一差异表达带。并初步测定具有酶的活性。
Northern杂交分析表明：非亲和性稻瘟菌小种能诱导苯甲酰基转移酶转录物的积累；而亲和性稻瘟菌小种不能诱导苯甲酰基转移酶转录物的累积。
以上结果证明：苯甲酰基转移酶与水稻抗稻瘟病相关，能够体外催化合成植保素P1，可以作为抗病基因工程的侯选材料。
Plants show a series of complex physiological and biochemical changes in response to pathogen attack. Most of these responses are mediated by activation of plant genes including various defense genes and those involved in the process of pathogen recognition, signal transduction and initiation of transcription. The functional identification and expressing characterization of plant genes that are induced during interaction with pathogen will be a great help to reveal the mechanism of plant-pathogen interaction; isolation of these genes may provide a good choice for engineering plant disease resistance.
Phytoalexin is utilized by plant to resist biological or none biological invading factors, which is secondary metabolic product. It is an important fungi resistant material that have activity in vitro and can be inducible by invading factors. It is thought to be an important mechanism for its induced synthesis in plant after infected by pathogen. Fan and Peng(1996) isolated two phytoalexin in rice inoculated with incompatible race of Magnaporthe grisea. They accumulate in rice after 48 hours. According to their chemical structure, benzoc acid and cinnamic acid is precursor of this two phytoalexin. The density of JA immediately increase after elicitor., is correspondent with process of phytoalexin induction. It can be inferred that Benzoyltransferase involved in rice resistance reaction to Magnaporthe grisea.
The author characterized one full-length cDNA. Sequencing result showed that the full-length cDNA which consists of 1610bp has an intact ORF (82-1419) that encodes an 446-amino-acid protein. BLAST search indicated that the deduced amino acid sequence shares 41.69% identity with that transferase of from Hordeum, Taxus chinensis,.Catharanthus, Diabnthus caryophyllus respectively. Then there is a different band with 74kDa in the SDA-PAGE , after indued with IPTG .the result of expressing Benzoyltransferase in E. coli showed Benzoyltransferase has the transferase activity. Benzoyltransferase transcript in rice leaves can be induced by blast fungus infection.
引文
[1] 王金生，1999，《分子植物病理学》。中国农业出版社。
[2] 卢圣栋主编，1999，《现代分子生物学实验技术》。中国协和医科大学出版社
[3] 范军，1999，中国农业大学博士学位论文
[4] 郑文杰，2000，中国农业大学博士学位论文
[5] 吴春霞，2002，中国农业大学博士学位论文
[6] 秦庆明 2002，中国农业大学博士学位论文
[7] 李晖 2002，中国农业大学博士学位论文
[8] 范军 彭友良 一种新的水稻植保素的诱导，纯化及特性研究植物病理学报 1996。26(3)：217-221
[9] 范军 罗招庆 彭友良 一种新的水稻植保素的纯化及其性状分析
[10] Kevin Walker and Rodney Croteau, Taxol biosynthesis: Molecular cloning of a benzoyl-CoA: taxane 2α-O-benzoyltransferase cDNA from Taxus and functional expression in Escherichia coli, PNAS, Dec 2000; 97: 13591-13596.
[11] Kevin Walker, Robert Long, and Rodney Croteau, The final acylation step in Taxol biosynthesis: Cloning of the taxoid Cl3-side-chain N-benzoyltransferase from Taxus, PNAS, Jul 2002; 99: 9166-9171
[12] D.L.Nandi, S.V.Lucas, and L. T. Webster, Jr.Benzoyl-coenzyme A: glycine N-acyltransferase and phenylacetyl-coenzyme A: glycine N-acyltransferase from bovine liver mitochondria. Purification and characterization. J. Biol. Chem., Aug 1979; 254:7230-7237.
[13] Mary Jo Farmer, Pierre Czernic, Anthony Michael, and Jonathan Negrei Identification and characterization of cDNA clones encoding hydroxycinnamoyl-CoA: tyramine N-hydroxycinnamoyltransferase from tobacco.Eur.J.Biochem., Aug 1999; 263:686-694.
[14] Laurent Hoffmann, Stéphane Maury, Francoise Martz, Pierrette Geoffroy, and Michel Legrand. Purification, Cloning, and Properties of an Acyltransferase Controlling Shikimate and Quinate Ester Intermediates in Phenylpropanoid Metabolism. J. Biol. Chem., Jan 2003; 278:95-103.
[15] Kevin Walker, Shingo Fujisaki, Robert Long, and Rodney Croteau
[16] Molecular cloning and heteroiogous expression of the C-13 phenylpropanoid side chain-CoA acyltransferase that functions in Taxol biosynthesis PNAS, Oct 2002; 99:12715-12720
[17] Kim Burhenne, Brian K. Kristensen, and Sren K. Rasmussen. A New Class of N-Hydroxycinnamoyltransferases. purification, cloning, and expression of a barley agmatine coumaroyltransferase (ec 2.3.1.64).3. Biol. Chem., Apr 2003; 278:13919-13927.
[18] John C. D'Auria, Feng Chen, and Eran Pichersky. Characterization of an Acyltransferase Capable of Synthesizing Benzylbenzoate and Other Volatile Esters in Flowers and Damaged Leaves of
Clarkia breweri. Plant Physiology, Sep 2002; 130: 466-476.
[19] Farmer MJ, Czernic P, Michael A, Negrel J. Identification and characterization of cDNA clones encoding hydroxycinnamoyl-CoA: tyramine N-hydroxycinnamoyltransferase from tobacco.. Eur J Biochem. 1999 Aug; 263(3): 686-94.
[20] Yang Q, Grimmig B, Matern U.Anthranilate N-hydroxycinnamoyl/benzoyltransferase gene from carnation: rapid elicitation of transcription and promoter analysis. Plant Mol Biol. 1998 Dec; 38(6): 1201-14.
[21] Yang Q, Reinhard K, Schiltz E, Matern UCharacterization and heterologous expression of hydroxycinnamoyl/benzoyl-CoA: anthranilate N-hydroxycinnamoyl/benzoyitransferase from elicited cell cultures of carnation, Dianthus caryophyllus L. Plant Mol Biol. 1997 Dec; 35(6): 777-89.
[22] Reinhard K, Matern U The biosynthesis of phytoalexins in Dianthus caryophyllus L.cell cultures: induction of benzoyl-CoA: anthranilate N-benzoyltransferase activity. Arch Biochem Biophys. 1989 Nov 15; 275(1): 295-301
[23] Nandi DL, Lucas SV, Webster LT Jr Benzoyl-coenzyme A: glycine N-acyltransferase and phenylacetyl-coenzyme A: glycine N-acyltransferase from bovine liver mitochondria. Purification and characterization. J Biol Chem. 1979 Aug 10; 254(15): 7230-7.
[24] Walker K, Long R, Croteau R Taxol biosynthetic genes. Phytochemistry. 2001 Sep; 58(1): 1-7. Review
[25] Walker K, Croteau R. Taxol biosynthesis: molecular cloning of a benzoyl-CoA: taxane 2alpha-O-benzoyltransferase cDNA from taxus and functional expression in Escherichia coli. Proc Natl Acad Sci U S A. 2000 Dec 5; 97(25): 13591-
[26] Uknes S, Mauch-Mani B, Moyer M, et al., 1992, Acquired resistance in Arabidopsis. Plant Cell, 4: 645-656
[27] yang Y, Shan J and Klessing DF, 1997, Signal perception and transduction in plant defense response, Genes & Development, 11: 1621-1639
[28] Yamada K, Mori H, Yamaki S, 1999, Identification and cDNA cloning of a protein abundantly expressed during apple fruit development. Plant Cell Physiol., 40(2): 198-204
[29] Waterhouse PM, Wang MB, Lough T, 2001, Gene silencing as an adaptive defence against viruses. Nature, 411: 834-842
[30] Ward ER, Uknes SJ, Williams SC, et al., 1991, Coordinate gene activity in response to agents that induced systemic acquired resistance. Plant Cell, 3: 1085-1094
[31] Uknes S, MauchoMani B, Moyer M, et al., 1992, Acquired resistance in Arabidopsis. Plant Cell, 4: 645-656
[32] Uknes S, Winter AM, Delaney, et al., 1993, Biological induction of systemic acquired resistance in Arabidopsis. Mol. Plant-Microbe Interact., 6: 692-698
[33] Uknes S, Mauch-Mani B, Moyer M, et al., 1992, Acquired resistance in Arabidopsis. Plant Cell, 4:
645-656
[34] Staskawicz B J, Ausubel FM, Baker B J, et al., 1995, Molecular genetics of plant disease resistance. Science, 268: 661-667
[35] Seo S, Sano H, Ohashi Y, et al., 1997, Jasmonic acid in wound signal transduction pathways. Physiol. Plant, 101: 740-745
[36] Sano H, Seo S, Koizumi N, et al., 1996, Regulation by cytokinins of endogenous levels of jasmonic and salicylic acids in mechanically wounded tobacco plants. Plant Cell Physiol., 37: 762-769
[37] Ryan CA, 1990, Protease inhibitors in plants: Gene for improving defense against insects and pathogens. Annu. Rev. Phytopathol., 28: 425-449
[38] Reymond P and Farmer EE, 1998, Jasmonate and salicylate as global signals for defense gene expression. Curr. Opin. Plant Biol., 1: 404-411
[39] Peng YL, 1994, A molecular strategy to manipulate rice blast disease resistance. In Integrated Resource Management for Sustainable Agriculture. Ed. by Shi and Cheng. Beijing Agricultural University Press: 450-453
[40] Leon J, Rojo E and Sanchez-Serrano J, 2001, Wound signalling in plants. J. Exp. Bot., 52: 1-9
[41] Koiwai A, Noguchi M, Tamaki E, 1971, Change in the amino acid composition of tobacco cells in suspension culture. Phytochemistry, 10: 561-565
[42] Bart P. H. J. Thomma, et al., 1998. Separate jasmonate-dependent and salicylate-dependent defense-response pathways in Arabidopsis are essential for resistance to distinct microbial pathogens PNAS, 95: 15107-151119.
[43] Adam T., et al., 1989. Consequence of O~(2-) generation during a bacterially induced hypersensitive reaction in tobacco: deterioration of membrane lipids. Physiol. Mol. Plant Pathoio. 34: 13-26.
[44] Antoniw J.F. et al., 1980. Comparision of three pathogensis-related proteins from plants of two cultivars of tobacco infected with TMV. J. Gen. Virol. 47: 79.
[45] Beardmore, J., et al., Cellular lignification as a factor in the hypersensitive resistance of wheat to stem rust, Physiol. Plant Pathol., 22, 209, 1983.
[46] Btischges R. et al., 1997. The barley Mlo gene: a novel control element of plant pathogen resistance. Cell. 88: 695-705.
[47] Cohen Y, et al., 1993. Local and systemic protection against phytophthora infestans induced in potato and tomato plants by jasmonic acid and jasmonie methyl ester.
[48] Darvill A.G. and Albersheim P., 1984. Phytoalexions and their Elicitors-A defense against microbial infection in plants. Ann. Rev. Plant Physiol. 35: 242-275.
[49] Dempsey DA., et al., 1998. Engirmering disease and pest resistance in plants Trends Microbiol. 6: 54-61.
[50] Despres C., et al., 1995. The activation of the potato PR 10a gene requires the phosphorylation of the nuclear factor PBF-1. Plant Cell. 7: 589-598.
[51] Ebel A., et al., 1998. Early events in the eliciation of plant defese Planta. 86: 335-348
[52] Flor HH. 1971. Curent status of gene-for-gene concept Annu. Rev. Phytopathol. 9: 275-296.
[53] Gaspar, T., et al., Peroxidases: A Survey of Their Biochemical and Physiological Roles in Higher Plants, University of Geneva Press, Geneva, 1982.
[54] Hulbert Scot H., et al., 2001. Resistance gene complexes: Evolution and utilization. Annual review of phytopathology. 39:285-312.
[55] Legrand M., Kauffmann and Geoffroy P., 1987. Biological function of pathogenesis-related proteins: for tobacco pathogenesis-related proteins are chitinases. Proc. Natl. Acad. Sci. USA. 84: 6750-675
[56] Liong Xiong, et al., 2001. Identification of defence-related rice genes by suppression subtractive hybridization and differential screening. MPMI. Vol.14, No.5, 685-692.
[57] Luo G Z, An G, Wu R. 1994. A PCR differential screening method for rapid isolation of clones from a cDNA library. Biotechniques. 16(4): 672-675.
[58] Rasmussen, J.B., et al., cDNA cloning and systemic expression of acidic peroxidases associated with systemic acquired resistance to disease in cucumber, Physiol. Mol. Plant Pathol., 46, 389, 1995.
[59] Sambrook J, et al., 1989. Molecular cloning Cold spring harbor laboratory press. 304-491
[60] Song Wen-Yuan, et al., 1995. A receptor kinase-like protein encoded by the rice disease resistance gene Xa21. Science, 270: 1804-1806.
[61] Thomma BPHT, et al., 1998. Separate jasmonate-dependent and salicylate-dependent defense-response pathway in Arabidopsis are essential for resistance to distinct microbial pathogens. Proc. Natl.Acad.Sci.USA. 95:1507-1511.
[62] Xie DX, et al. 1988. Coll: an Arabidopsis gene reqired for Jasm onate-requlated defence and fertility. Science. 1091-1094.
[63] Suzuki K, et al., 1995. Studies on elicitor-signal transduction leading to differential expression of defense genes in cultured tobacco cells. Plant cell. Physiol. 36: 282-289.
[2] 卢圣栋主编，1999，《现代分子生物学实验技术》。中国协和医科大学出版社
[3] 范军，1999，中国农业大学博士学位论文
[4] 郑文杰，2000，中国农业大学博士学位论文
[5] 吴春霞，2002，中国农业大学博士学位论文
[6] 秦庆明 2002，中国农业大学博士学位论文
[7] 李晖 2002，中国农业大学博士学位论文
[8] 范军 彭友良 一种新的水稻植保素的诱导，纯化及特性研究植物病理学报 1996。26(3)：217-221
[9] 范军 罗招庆 彭友良 一种新的水稻植保素的纯化及其性状分析
[10] Kevin Walker and Rodney Croteau, Taxol biosynthesis: Molecular cloning of a benzoyl-CoA: taxane 2α-O-benzoyltransferase cDNA from Taxus and functional expression in Escherichia coli, PNAS, Dec 2000; 97: 13591-13596.
[11] Kevin Walker, Robert Long, and Rodney Croteau, The final acylation step in Taxol biosynthesis: Cloning of the taxoid Cl3-side-chain N-benzoyltransferase from Taxus, PNAS, Jul 2002; 99: 9166-9171
[12] D.L.Nandi, S.V.Lucas, and L. T. Webster, Jr.Benzoyl-coenzyme A: glycine N-acyltransferase and phenylacetyl-coenzyme A: glycine N-acyltransferase from bovine liver mitochondria. Purification and characterization. J. Biol. Chem., Aug 1979; 254:7230-7237.
[13] Mary Jo Farmer, Pierre Czernic, Anthony Michael, and Jonathan Negrei Identification and characterization of cDNA clones encoding hydroxycinnamoyl-CoA: tyramine N-hydroxycinnamoyltransferase from tobacco.Eur.J.Biochem., Aug 1999; 263:686-694.
[14] Laurent Hoffmann, Stéphane Maury, Francoise Martz, Pierrette Geoffroy, and Michel Legrand. Purification, Cloning, and Properties of an Acyltransferase Controlling Shikimate and Quinate Ester Intermediates in Phenylpropanoid Metabolism. J. Biol. Chem., Jan 2003; 278:95-103.
[15] Kevin Walker, Shingo Fujisaki, Robert Long, and Rodney Croteau
[16] Molecular cloning and heteroiogous expression of the C-13 phenylpropanoid side chain-CoA acyltransferase that functions in Taxol biosynthesis PNAS, Oct 2002; 99:12715-12720
[17] Kim Burhenne, Brian K. Kristensen, and Sren K. Rasmussen. A New Class of N-Hydroxycinnamoyltransferases. purification, cloning, and expression of a barley agmatine coumaroyltransferase (ec 2.3.1.64).3. Biol. Chem., Apr 2003; 278:13919-13927.
[18] John C. D'Auria, Feng Chen, and Eran Pichersky. Characterization of an Acyltransferase Capable of Synthesizing Benzylbenzoate and Other Volatile Esters in Flowers and Damaged Leaves of
Clarkia breweri. Plant Physiology, Sep 2002; 130: 466-476.
[19] Farmer MJ, Czernic P, Michael A, Negrel J. Identification and characterization of cDNA clones encoding hydroxycinnamoyl-CoA: tyramine N-hydroxycinnamoyltransferase from tobacco.. Eur J Biochem. 1999 Aug; 263(3): 686-94.
[20] Yang Q, Grimmig B, Matern U.Anthranilate N-hydroxycinnamoyl/benzoyltransferase gene from carnation: rapid elicitation of transcription and promoter analysis. Plant Mol Biol. 1998 Dec; 38(6): 1201-14.
[21] Yang Q, Reinhard K, Schiltz E, Matern UCharacterization and heterologous expression of hydroxycinnamoyl/benzoyl-CoA: anthranilate N-hydroxycinnamoyl/benzoyitransferase from elicited cell cultures of carnation, Dianthus caryophyllus L. Plant Mol Biol. 1997 Dec; 35(6): 777-89.
[22] Reinhard K, Matern U The biosynthesis of phytoalexins in Dianthus caryophyllus L.cell cultures: induction of benzoyl-CoA: anthranilate N-benzoyltransferase activity. Arch Biochem Biophys. 1989 Nov 15; 275(1): 295-301
[23] Nandi DL, Lucas SV, Webster LT Jr Benzoyl-coenzyme A: glycine N-acyltransferase and phenylacetyl-coenzyme A: glycine N-acyltransferase from bovine liver mitochondria. Purification and characterization. J Biol Chem. 1979 Aug 10; 254(15): 7230-7.
[24] Walker K, Long R, Croteau R Taxol biosynthetic genes. Phytochemistry. 2001 Sep; 58(1): 1-7. Review
[25] Walker K, Croteau R. Taxol biosynthesis: molecular cloning of a benzoyl-CoA: taxane 2alpha-O-benzoyltransferase cDNA from taxus and functional expression in Escherichia coli. Proc Natl Acad Sci U S A. 2000 Dec 5; 97(25): 13591-
[26] Uknes S, Mauch-Mani B, Moyer M, et al., 1992, Acquired resistance in Arabidopsis. Plant Cell, 4: 645-656
[27] yang Y, Shan J and Klessing DF, 1997, Signal perception and transduction in plant defense response, Genes & Development, 11: 1621-1639
[28] Yamada K, Mori H, Yamaki S, 1999, Identification and cDNA cloning of a protein abundantly expressed during apple fruit development. Plant Cell Physiol., 40(2): 198-204
[29] Waterhouse PM, Wang MB, Lough T, 2001, Gene silencing as an adaptive defence against viruses. Nature, 411: 834-842
[30] Ward ER, Uknes SJ, Williams SC, et al., 1991, Coordinate gene activity in response to agents that induced systemic acquired resistance. Plant Cell, 3: 1085-1094
[31] Uknes S, MauchoMani B, Moyer M, et al., 1992, Acquired resistance in Arabidopsis. Plant Cell, 4: 645-656
[32] Uknes S, Winter AM, Delaney, et al., 1993, Biological induction of systemic acquired resistance in Arabidopsis. Mol. Plant-Microbe Interact., 6: 692-698
[33] Uknes S, Mauch-Mani B, Moyer M, et al., 1992, Acquired resistance in Arabidopsis. Plant Cell, 4:
645-656
[34] Staskawicz B J, Ausubel FM, Baker B J, et al., 1995, Molecular genetics of plant disease resistance. Science, 268: 661-667
[35] Seo S, Sano H, Ohashi Y, et al., 1997, Jasmonic acid in wound signal transduction pathways. Physiol. Plant, 101: 740-745
[36] Sano H, Seo S, Koizumi N, et al., 1996, Regulation by cytokinins of endogenous levels of jasmonic and salicylic acids in mechanically wounded tobacco plants. Plant Cell Physiol., 37: 762-769
[37] Ryan CA, 1990, Protease inhibitors in plants: Gene for improving defense against insects and pathogens. Annu. Rev. Phytopathol., 28: 425-449
[38] Reymond P and Farmer EE, 1998, Jasmonate and salicylate as global signals for defense gene expression. Curr. Opin. Plant Biol., 1: 404-411
[39] Peng YL, 1994, A molecular strategy to manipulate rice blast disease resistance. In Integrated Resource Management for Sustainable Agriculture. Ed. by Shi and Cheng. Beijing Agricultural University Press: 450-453
[40] Leon J, Rojo E and Sanchez-Serrano J, 2001, Wound signalling in plants. J. Exp. Bot., 52: 1-9
[41] Koiwai A, Noguchi M, Tamaki E, 1971, Change in the amino acid composition of tobacco cells in suspension culture. Phytochemistry, 10: 561-565
[42] Bart P. H. J. Thomma, et al., 1998. Separate jasmonate-dependent and salicylate-dependent defense-response pathways in Arabidopsis are essential for resistance to distinct microbial pathogens PNAS, 95: 15107-151119.
[43] Adam T., et al., 1989. Consequence of O~(2-) generation during a bacterially induced hypersensitive reaction in tobacco: deterioration of membrane lipids. Physiol. Mol. Plant Pathoio. 34: 13-26.
[44] Antoniw J.F. et al., 1980. Comparision of three pathogensis-related proteins from plants of two cultivars of tobacco infected with TMV. J. Gen. Virol. 47: 79.
[45] Beardmore, J., et al., Cellular lignification as a factor in the hypersensitive resistance of wheat to stem rust, Physiol. Plant Pathol., 22, 209, 1983.
[46] Btischges R. et al., 1997. The barley Mlo gene: a novel control element of plant pathogen resistance. Cell. 88: 695-705.
[47] Cohen Y, et al., 1993. Local and systemic protection against phytophthora infestans induced in potato and tomato plants by jasmonic acid and jasmonie methyl ester.
[48] Darvill A.G. and Albersheim P., 1984. Phytoalexions and their Elicitors-A defense against microbial infection in plants. Ann. Rev. Plant Physiol. 35: 242-275.
[49] Dempsey DA., et al., 1998. Engirmering disease and pest resistance in plants Trends Microbiol. 6: 54-61.
[50] Despres C., et al., 1995. The activation of the potato PR 10a gene requires the phosphorylation of the nuclear factor PBF-1. Plant Cell. 7: 589-598.
[51] Ebel A., et al., 1998. Early events in the eliciation of plant defese Planta. 86: 335-348
[52] Flor HH. 1971. Curent status of gene-for-gene concept Annu. Rev. Phytopathol. 9: 275-296.
[53] Gaspar, T., et al., Peroxidases: A Survey of Their Biochemical and Physiological Roles in Higher Plants, University of Geneva Press, Geneva, 1982.
[54] Hulbert Scot H., et al., 2001. Resistance gene complexes: Evolution and utilization. Annual review of phytopathology. 39:285-312.
[55] Legrand M., Kauffmann and Geoffroy P., 1987. Biological function of pathogenesis-related proteins: for tobacco pathogenesis-related proteins are chitinases. Proc. Natl. Acad. Sci. USA. 84: 6750-675
[56] Liong Xiong, et al., 2001. Identification of defence-related rice genes by suppression subtractive hybridization and differential screening. MPMI. Vol.14, No.5, 685-692.
[57] Luo G Z, An G, Wu R. 1994. A PCR differential screening method for rapid isolation of clones from a cDNA library. Biotechniques. 16(4): 672-675.
[58] Rasmussen, J.B., et al., cDNA cloning and systemic expression of acidic peroxidases associated with systemic acquired resistance to disease in cucumber, Physiol. Mol. Plant Pathol., 46, 389, 1995.
[59] Sambrook J, et al., 1989. Molecular cloning Cold spring harbor laboratory press. 304-491
[60] Song Wen-Yuan, et al., 1995. A receptor kinase-like protein encoded by the rice disease resistance gene Xa21. Science, 270: 1804-1806.
[61] Thomma BPHT, et al., 1998. Separate jasmonate-dependent and salicylate-dependent defense-response pathway in Arabidopsis are essential for resistance to distinct microbial pathogens. Proc. Natl.Acad.Sci.USA. 95:1507-1511.
[62] Xie DX, et al. 1988. Coll: an Arabidopsis gene reqired for Jasm onate-requlated defence and fertility. Science. 1091-1094.
[63] Suzuki K, et al., 1995. Studies on elicitor-signal transduction leading to differential expression of defense genes in cultured tobacco cells. Plant cell. Physiol. 36: 282-289.