Abstract

Plant chitinases have been known as pathogenesis-related (PR) proteins, but recent studies suggest that they play functional roles during normal plant growth and development. We previously isolated two cDNA clones encoding endochitinases, EuNOD-CHT1 and -CHT2, from the root nodules of Elaeagnus umbellata. These genes show differential expression patterns, with the EuNOD-CHT1 gene being active in the root nodules and meristems, while EuNOD­CHT2 is preferentially expressed in the infected cells of those nodules. To elucidate the functional roles of these two endochitinases, we have now constitutively expressed each gene in a heterologous plant system, Arabidopsis thaliana. Stable inheritance and expression of the transgenes were confirmed by genomic Southern hybridization and RT-PCR. Our transgenic plants did not differ morphologically from the wild types. However, constitutive expression of EuNOD-CHT1 and -CHT2 in Arabidopsis resulted in increased resistance against a fungal pathogen, Botrytis cinerea, but not against a bacterial agent, Pseudomonas syringae pv. Tomato DC3000. Expression levels were enhanced by both wounding and jasmonic acid treatments (for EuNOD-CHT1), or by jasmonic acid only (for EuNOD-CHT2). These data suggest that EuNOD-CHTl and -CHT2 primarily play defensive roles during root nodule development in E. umbellata.

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참고문헌 (30)

1. Broglie K, Chet I, Holliday M, Cressman R, Biddle P, Knowlton S, Mauvais CJ, Broglie R (1991) Transgenic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254: 1194-1197 
2. Harikrishna K, Jampates-Beale R, Milligan SB, Gasser CS (1996) An endochitinase gene expressed at high levels in the stylar transmitting tissue of tomatoes. Plant Mol Biol 30: 899-911 
3. Koch E, Slusarenko AJ (1990) Fungal pathogens of Arabidopsis thaliana (L.) Heynh. Bot Helv 100: 257-268 
4. Takakura Y, Ito T, Saito H, Inoue T, Komari T, Kuwata S (2000) Flower-predominant expression of a gene encoding a novel class I chitinase in rice (Oryza sativa L.). Plant Mol Biol 42: 883-897 
5. Vance CP (1983) Rhizobium infection and nodulation: A beneficial plant disease? Annu Rev Microbiol 37: 399-424 
6. Graham LS, Sticklen MB (1994) Plant chitinases. Can J Bot 72: 1057-1083 
7. van Hengel AJ, Guzzo F, van Kammen A, de Vries S (1998) Expression pattern of the carrot EP3 endochitinase genes in suspension cultures and in developing seeds. Plant Physiol 117: 43-53 
8. Zhong RQ, Kays SJ, Schroeder Bp, Ye ZH (2002) Mutation of a chitinase-like gene causes ectopic deposition of lignin, aberrant cell shapes, and overproduction of ethylene. Plant Cell 14: 165-179 
9. Kragh KM, Hendricks T, de Jong AJ, Lo Schiavo F, BuCherna N, Hojrup P, Mikkelsen JD, de Vries SC (1996) Characterization of chitinases able to rescue somatic embryos of the temperature-sensitive carrot variant ts11. Plant Mol Biol 31: 631-645 
10. Melchers LS, Stuiver MH (2000) Novel genes for diseaseresistance breeding. Curr Opin Plant Biol 3: 147-152 
11. Clough SJ, Bent AF (1998) Floral dip: A simplified method for Agrobacterium mediated transformation of Arabidopsis thaliana. Plant J 16: 735-743 
12. Samac DA, Shah DM (1994) Effect of chitinase antisense RNA expression on disease susceptibility of Arabidopsis plants. Plant Mol Biol 25: 587-596 
13. Passarinho PA, van Hengel AJ, Fransz PF, de Vries SC (2001) Expression pattern of the Arabidopsis thaliana AtEP3/AtchitlV endochitinase gene. Planta 212: 556-567 
14. Topfer R, Matzeit V, Gronenborn B, Schell J, Steinbiss HH (1987) A set of plant expression vectors for transcriptional and translational fusions. Nucl Acids Res 15: 5890 
15. Wu CT, Leubner-Metger G, Meins F, Bradford K (2001) Class I beta-1, 3-glucanase and chitinase are expressed in the micropylar endosperm of tomato seeds prior to radicle emergence. Plant Physiol 126: 1299-1313 
16. Neuhaus JM, Ahl-Goy P, Hinz U, Flores S, Meins F Jr (1991) High-level expression of a tobacco chitinase gene in Nicotiana sylvestris: Susceptibility of transgenic plants to Cercospora nicotianae infection. Plant Mol Biol 16: 141-151 
17. Gongora CE, Broadway RM (2002) Plant growth and development influenced by transgenic insertion of bacterial chitinolytic enzymes. Mol Breed 9: 123-135 
18. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, Ed 2. Cold Spring Harbor Laboratory Press, New York 
19. Doyle JJ, Doyle JI (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13-15 
20. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15: 473-497 
21. Baron CP, Zambryski C (1995) The plant response in pathogenesis, symbiosis, and wounding: Variations on a common theme? Ann Rev Genet 29: 107-129 
22. de Jong AJ, Cordewener J, Lo Schiavo F, Terzi M, Vandekerckhove J, van Kammen A, de Vries SC (1992) A carrot somatic embryo mutant is rescued by chitinase. Plant Cell 4: 425-433 
23. Hajdukiewicz P, Svab Z, Maliga P (1994) The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25: 989-994 
24. Kim HB, An CS (2002) Differential expression patterns of an acidic chitinase and a basic chitinase in the root nodule of Elaeagnus umbellata. Mol Plant-Microbe Interact 15: 209-215 
25. Whalen MC, Innes RW, Bent AF, Staskawicz BJ (1991) Identification of Pseudomonas syringae pathogens of Arabidopsis and a bacterial locus determining avirulence on both Arabidopsis and soybean. Plant Cell 3: 4959 
26. Passarinho PA, de Vries SC (2002) Arabidopsis chitinases: A genomic survey. In CR Somerville, EM Meyerowitz, eds, The Arabidopsis Book. American Society of Plant Physiologists, Rockville, doi/10.1199/tab.0023. http://www.aspb.org/publications/arabidopsis/ 
27. Hanfrey C, Fife M, Buchanan-Wollaston V (1996) Leaf senescence in Brassica napus: Expression of genes encoding pathogenesis-related proteins. Plant Mol Biol 30: 597-609 
28. Jach G, Gornhardt B, Mundy J, Logemann J, Pinsdorf E, Leah R, Schell J, Maas C (1995) Enhanced quantitative resistance against fungal disease by combinatorial expression of different barley antifungal proteins in transgenic tobacco. Plant J 8: 97-109 
29. Kim HB, Oh CJ, Lee H, An CS (2003) A type-1 chalcone isomerase mRNA is highly expressed in the root nodule of Elaeagnus umbellata. J Plant Biol 46: 263-270  원문보기 상세보기
30. Collinge DB, Kragh KM, Mikkelesen JD, Nielsen KK, Rasmussen U, Vad K(1993) Plantchitinases. Plant J 3: 31-40 

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1. 2006. "" Journal of plant biology = 식물학회지, 49(6): 413~420  원문보기 상세보기