Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (17): 3537-3546.doi: 10.3864/j.issn.0578-1752.2011.17.006

• PLANT PROTECTION • Previous Articles     Next Articles

Cloning and Bioinformatics Analysis of Chitin Synthase Gene ThChsC from Trichoderma harzianum

JI  Ying, LI  Mei, TIAN  Yun-Long, LIU  Wei-De, NIU  Jing, JIANG  Xi-Liang   

  1. 1. 中国农业科学院植物保护研究所/农业部作物有害生物综合治理综合性重点实验室
    2. 中国农业科学院农业环境与可持续发展研究所
  • Received:2011-01-21 Revised:2011-02-28 Online:2011-09-01 Published:2011-03-16
  • Contact: ying ji E-mail:xljiang@caas.net.cn

Abstract: 【Objective】 The objective of this study is to clone and make bioinformatics analysis of the sequence of chitin synthase gene ThChsC from Trichoderma harzianum. 【Method】 The full length of ThChsC was cloned by polymerase chain reaction (PCR) and genome walking methods, the gene sequence obtained and the putative amino acid sequence were analyzed by bioinformatics software, and the three-dimensional structure of ThChsC was predicted by homology modeling. 【Result】 ThChsC (GenBank accession number HQ419000) coding sequence spans 2 835 bp and is comprised of 4 extrons inserted by 3 introns. The ThChsC contains an open reading frame (ORF) of 2 688 bp capable of encoding a polypeptide of 895 amino acids and shares 80% and 81% identity to Gibberella moniliformis chsB (ACY08039.1) and Colletotrichum graminicola chs2 (AAL23718.1), respectively. Phylogenetic analysis indicates that ThChsC belongs to the type III chitin synthase subfamily. The conserved Chitin_synth_1, Chitin_synth_1N and Chitin_synth_2 domains and three transmembrane domains were found in the predicted ThChsC protein. A conserved DHD domain of glycosyl transferase was found in the three-dimensional structure of ThChsC predicted. 【Conclusion】The chitin synthase gene ThChsC was cloned for the first time from Trichoderma harzianum Th-33. The further study of ThChsC will have an important significance in exploring the mechanism of chitin synthesis of Trichoderma harzianum.

Key words: Trichoderma harzianum, chitin synthase, gene cloning, sequence analysis

[1]Benítez T, Rincón A M, Limón M C, Codón A C. Biocontrol mechanisms of Trichoderma strains. International Microbiology, 2004, 7: 249-260.

[2]冯贻安, 崔志峰. 真菌几丁质合酶的研究进展. 微生物学通报, 2008, 35(2): 267-271.

Feng Y A, Cui Z F. Progress in the studies of fungal chitin synthases. Microbiology, 2008, 35(2): 267-271. (in Chinese)

[3]Silverman S J, Sburlati A, Slater M L, Cabib E. Chitin synthase 2 is essential for septum formation and cell division in Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences of the United States of America, 1988, 85(7): 4735-4739.

[4]Shaw J A, Mol P C, Bowers B, Silverman S J, Valdivieso M H, Durán A, Cabib E. The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle. The Journal of Cell Biology, 1991, 114(1): 111-123.

[5]Bulawa C E. CSD2, CSD3, and CSD4, genes required for chitin synthesis in Saccharomyces cerevisiae: the CSD2 gene product is related to chitin synthases and to developmentally regulated proteins in Rhizobium species and Xenopus laevis. Molecular and Cellular Biology, 1992, 12(4): 1764-1776.

[6]Schmidt M. Survival and cytokinesis of Saccharomyces cerevisiae in the absence of chitin. Microbiology, 2004, 150(10): 3253-3260.

[7]Motoyama T, Fujiwara M, Kojima N, Horiuchi H, Ohta A, Takagi M. The Aspergillus nidulans genes chsA and chsD encode chitin synthases which have redundant functions in conidia formation. Molecular and General Genetic, 1996, 251(2): 442-450.

[8]Ichinomiya M, Hirouchi H, Ohta A. Different functions of the class I and class II chitin synthase genes, chsC and chsA, are revealed by repression of chsB expression in Aspergillus nidulans. Current Genetic, 2002, 42(10): 51-58.

[9]Lee J I, Yu Y M, Rho Y M, Park B C, Chio J H, Park H M, Maeng P J. Differential expression of the chsE gene encoding a chitin synthase of Aspergillus nidulans in response to developmental status and growth conditions. FEMS Microbiology Letters, 2005, 249(6): 121-129.

[10]Takeshita N, Yamashita S, Ohta A, Hirouchi H. Aspergillus nidulans class V and VI chitin synthases CsmA and CsmB, each with a myosin motor-like domain, perform compensatory functions that are essential for hyphal tip growth. Molecular Microbiology, 2006, 59(5): 1380-1394.

[11]Ichinomiya M, Motoyama T, Fujiwara M, Takagi M, Hirouchi H, Ohta A. Repression of chsB expression reveals the functional importance of class IV chitin synthase gene chsD in hyphal growth and conidiation of Aspergillus nidulans. Microbiology, 2002, 148: 1335-1347.

[12]Au-Young J, Robbins P W. Isolation of a chitin synthase gene (CHS1) from Candida albicans by expression in Saccharomyces cerevisiae. Molecular Microbiology, 1990, 4(2): 197-207.

[13]Sudoh M, Watanabe M, Mio T, Nagahashi S, Yamada-Okabe H, Takagi M, Arisawa M. Isolation of canCHSlA, a variant gene of Candida albicans chitin synthase. Microbiology, 1995, 141: 2673-2679.

[14]Sudoh M, Tatsuno K, Ono N, Ohta A, Chibana H, Yamada-Okabe H, Arisawa M. The Candida albicans CHS4 gene complements a Saccharomyces cerevisiae skt5/chs4 mutation and is involved in chitin biosynthesis. Microbiology, 1999, 145: 1613-1622.

[15]Munro C A, Winter K, Buchan A, Henry K, Becker J M, Brown A J  P, Bulawa C E, Gow N A R. Chs1 of Candida albicans is an essential chitin synthase required for synthesis of the septum and for cell integrity. Molecular Microbiology, 2001, 39(5): 1414-1426.

[16]Munro C A, Whitton R K, Hughes H B, Rella M, Selvaggini S, Gow N A R. CHS8-a fourth chitin synthase gene of Candida albicans contributes to in vitro chitin synthase activity, but is dispensable for growth. Fungal Genetics and Biology, 2003, 40(2): 146-158.

[17]Mellndo E, Aufauvre-Brown A, Gow N A R, Holden D W. The Aspergillus fumigates chsC and chsG genes encode Class III chitin synthase with different functions. Molecular Microbiology, 1996, 20(3): 667-679.

[18]Mellado E, Dubreucq G, Mol P, Sarfati J, Paris S, Diaquin M, Holden D W, Rodriguez-Tudela J L, Latgé J P. Cell wall biogenesis in a double chitin synthase mutant (chsG-/chsE-) of Aspergillus fumigatus. Fungal Genetics and Biology, 2003, 38(1): 98-109.

[19]Choquer M, Boccara M, Goncalves I R, Soulié M C, Vidal-Cros A. Survey of the Botrytis cinerea chitin synthase multigenic family through the analysis of six euascomycetes genomes. European Journal of Biochemistry, 2004, 271(11): 2153-2164.

[20]Wang Z, Szaniszlo P J. WdCHS3, a gene that encodes a class III chitin synthase in Wangiella (Exophiala) dermatitidis, is expressed differentially under stress conditions. Journal of Bacteriology, 2000, 182(4): 874-881.

[21]Lee J I, Choi J H, Park B C, Park Y H, Lee M Y, Park H M, Maeng P J. Differential expression of the chitin synthase genes of Aspergillus nidulans, chsA, chsB, and chsC, in response to developmental status and environmental factors. Fungal Genetics and Biology, 2004, 41(6): 635-646.

[22]Penttilä M, Nevalainenb H, Rättö M, Salminen E, Knowles J. A versatile transformation system for the cellulolytic filamentous fungus Trichoderma ressei. Gene, 1987, 61(2): 155-164.

[23]Vriend G. WHAT IF: A molecular modeling and drug design program. Journal of Molecular Graphics, 1990, 8(1): 52-56.

[24]Fritz T A, Hurley J H, Trinh L B, Shiloach J, Tabak L A. The beginnings of mucin biosynthesis: The crystal structure of UDP-GalNAc: polypeptide α-N-acetylgalactosaminyltaransferase-T1. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(43): 15307-15312.

[25]李秀峰, 庄佩君, 唐振华. 真菌中的几丁质合成酶, 世界农药, 2000, 22(5): 33-38.

Li X F, Zhuang P J, Tang Z H. The chitin synthases of fungal. World Pesticide, 2000, 22(5): 33-38. (in Chinese)
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