Identification of Fungus Lecanicillium attenuatum and Cloning of Its Chitinase Gene LACHI1

doi:10.7668/hbnxb.2014.04.011

Abstract

Abstract: To clone chitinolytic genes from Lecanicillium attenuatum and provide theoretical basis for the inhibition of the chitinases on egg-hatching of root knot and cyst nematode.In this study,an nematode egg-parasitic fungus CGMCC5328 was isolated from Heterodera glycines infecting soybean in Heilongjiang Province.Based on morphological characters and molecular analysis of ITS-rDNA,the strain was identified as L.attenuatum.The chitinase gene LACHI1 from L.attenuatum was cloned using the degenerate PCR primers and RACE techniques.The analysis of the gene LACHI1 and its amino acid sequence was by biology software.We have cloned a chitinase gene LACHI1 from L.attenuatum for the first time.The gene is 1 743 bp in length and contains three putative introns.The ORF of LACHI1 is 1 272 bp in size with encoding protein of 423 aa,molecular mass of 45.9 kDa and pI of 5.90.The chitinase deduced by LACHI1 belongs to glycosyl hydrolase family 18 chitinase.Comparison of the chitinase amino acid sequence with other chitinases from entomopathogenic fungi and nematophagous fungi revealed that the enzymes were highly similar.

Key words: Lecanicillium attenuatum, Chitinase gene, Cloning, Phylogenetic analysis

CLC Number:

ZHAO Yang, CHEN De-xin, WANG Feng-long, HUANG Hua-gang, WU Xia. Identification of Fungus Lecanicillium attenuatum and Cloning of Its Chitinase Gene LACHI1[J]. journal1, 2014, 29(4): 63-70. doi: 10.7668/hbnxb.2014.04.011.

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http://www.hbnxb.net/EN/Y2014/V29/I4/63

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