Abstract
A new basic chitinase gene, designated RC24, was isolated from a rice genomic library. The predicted RC24 protein contains 322 amino acid residues and exhibits 68% to 95% amino acid identity with known class I rice chitinases. RC24 protein expressed in Escherichia coli exhibited chitinase activity and strongly inhibited bacterial growth. Two transcription start sites of the RC24 gene were mapped by primer extension analysis of both rice native RNA and in vitro transcribed RNA using a RC24 promoter/GUS (β-glucuronidase) gene fusion as a template. The 5′-franking region of RC24 contained several putative stress-responsive cis-acting elements. A basal level of RC24 transcripts was detected in rice root and stem tissues, but not in leaf tissues. RC24 transcripts rapidly accumulated within 1 h after fungal elicitor treatment of suspension-cultured cells, and the levels continued to increase for at least 9 h. RC24 transcript accumulation was also observed in intact leaf tissues upon wounding. Transgenic rice plants containing the RC24/GUS gene fusion further confirmed that the RC24 gene showed a tissue-specific expression pattern and that transcription of the RC24 promoter was sensitively and rapidly activated by wounding.
Similar content being viewed by others
References
1.Anuratha CS, Huang JK, Pingali A, Muthukrishnan S: Isolation and characterization of a chitinase and its cDNA clone from rice. J Plant Biochem Biotechnol 1: 5–10 (1992).
Ausubel FM, Brent R, Kington RZ, Moore DD, Seideman JG, Smith JA, Struhl K. Current Protocols in Molecular Biology. Wiley, New York (1987).
Boller T, Gehri A, Mauch F, Vögeli U: Chitinase in bean leaves: induction by ethylene, purification, properties and possible function. Planta 157: 22–31 (1983).
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).
Broekaert WF, Parijs JV, Allen AK, Peumans WJ: Comparison of some molecular, enzymatic and antifungal properties of chitinases from thorn-apple, tobacco and wheat. Physiol Mol Plant Path 33: 319–331 (1988).
Broglie K, Chet I, Holliday M, Cressman R, Biddle P, Knowlton S, Mauvais CJ, Broglie R: Transgenic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254: 1194–1197 (1991).
Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159 (1987).
Chory J, Peto C, Feinbaum R, Pratt L, Ausubel F: Arabidopsis thaliana mutant that develops as a light-grown plant in the absence of light. Cell 58: 991–999 (1989).
Church GM, Gilbert W: Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995 (1984).
Collinge DB, Kragh KM, Mikkelsen JD, Nielsen KK, Rasmussen U, Vad K: Plant chitinases. Plant J 3: 31–40 (1993).
Del Sal G, Manfioletti G, Schneider C: A common miniscale preparation of template DNA from phagemids, phages or plasmids suitable for sequencing. Biotechniques 7: 514–520 (1989).
Dixon RA, Lamb CJ: Molecular communication in plant: microbia pathogen interactions. Annu Rev Plant Physiol Plant Mol Biol 41: 339–367 (1990).
Dixon RA, Harrison MJ, Lamb CJ: Early events in the activation of plant defense responses. Annu Rev Phytopath 32: 479–501 (1994).
Huang JK, Wen L, Swegle M, Tran HC, Thin TH, Naylor HM, Muthukrishnan S, Reeck GR: Nucleotide sequence of a rice genomic clone that encodes a class I endochitinase. Plant Mol Biol 16: 479–480 (1991).
Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 287–305 (1987).
Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).
Jones KA, Kadonaga JT, Rosenfield PJ, Keller JT, Tjian R. A cellular DNA-binding protein that activates eukaryotic transcription and DNA replication. Cell 48: 79–89 (1987).
Kim YK, Baek JM, Park HY, Choi YD, Kim SI: Isolation and characterization of cDNA clones encoding class I chitinase in suspension cultures of rice cell. Biosci Biotechnol Biochem 58: 1164–1166 (1994).
Li L, Qu R, Kochko A, Fauquet C, Beachy RN: Anuimproved rice transformation system using the biolistic method. Plant Cell Rep 12: 250–255 (1993).
Lois R, Dietrich A, Hahlbrock K, Schulz W: A phenylalanine ammonialyase gene from parsley: structure, regulation and identification of elicitor and light responsive cis-acting elements. EMBO J 8: 1641–1648 (1989).
Mauch F, Mauch-Mani B, Boller T: Antifugal hydrolases in pea tissue. II. Inhibition of fungal growth by combinations of chitinase and β-1,3-glucanase. Plant Physiol 88: 936–942 (1988).
Molano J, Duran A, Cabib E: A rapid and sensitive assay for chitinase using tritiated chitin. Anal Biochem 83: 648–656 (1977).
Nishizawa Y, Hibi T: Rice chitinase gene: cDNA cloning and stress-induced expression. Plant Sci 76: 211–218 (1991).
Nishizawa Y, Kishimoto N, Saito A, Hibi T: Sequence variation, differential expression and chromosomal location of rice chitinase genes. Mol Gen Genet 241: 1–10 (1993).
Roby D, Broglie K, Cressman R, Biddle P, Chet N, Broglie R: Activation of a bean chitinase promoter in transgenic tobacco plants by phytopathogenic fungi. Plant Cell 2: 999–1007 (1990).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Sharp JK, Valent B, Albersheim P: Purification and partial characterization of a β-glucan fragment that elicits phytoalexin accumulation in soybean. J Biol Chem 259: 11312–11320 (1972).
Schlumbaum A, Mauch F, Vögeli U, Boller T: Plant chitinases are potent inhibitors of fungal growth. Nature 324: 365–367 (1986).
Sierra F: A Laboratory Guide to In Vitro transcription. BioMethod, vol 2. Birkhauser, Boston (1990).
Studidier FW, Rosenberg AH, Dunn JJ, Dubendortt JW: Use of T7 RNA polymerase to direct expression of cloned genes. Meth Enzymol 185: 60–89 (1990).
Zhu Q, Lamb CJ: Isolation and characterization of a rice gene encoding a basic chitinase. Mol Gen Genet 226: 289–296 (1991).
Zhu Q, Maher EA, Masoud S, Dixon RA, Lamb CJ: Enhanced protection against fungal attack by constitutive co-expression of chitinase and glucanase genes in transgenic tobacco. Bio/technology 12: 807–812 (1994).
Zhu Q, Chappell J, Hedrick SA, Lamb CJ: Accurate in-vitro transcription from circularized plasmid templates by plant whole cell extracts. Plant J 7: 1021–1030 (1995).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Xu, Y., Zhu, Q., Panbangred, W. et al. Regulation, expression and function of a new basic chitinase gene in rice (Oryza sativa L.). Plant Mol Biol 30, 387–401 (1996). https://doi.org/10.1007/BF00049319
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00049319