Cloning of the gene Lecanicillium psalliotae chitinase Lpchi1 and identification of its potential role in the biocontrol of root-knot nematode Meloidogyne incognita.
The nematophagous fungus Lecanicillium psalliotae (syn. Verticillium psalliotae) is a well-known biocontrol agent. In this study, a chitinase gene Lpchi1 was isolated for the first time from L. psalliotae using degenerate primers and DNA-walking technique. The cloned gene Lpchi1 encoding 423 amino acid residues shares a high degree of homology with other pathogenicity-related chitinases from entomopathogenic and mycoparasitic fungi. The complementary DNA sequence of the mature chitinase was amplified via reverse transcription polymerase chain reaction and expressed well in Pichia pastoris GS115. Through gel filtration, the recombinant chitinase was purified as a protein of ca. 45 kDa with an optimal activity at pH 7.0 and 37.6 degrees C. The purified chitinase LPCHI1 was found degrading chitinous components of eggs of the root-knot nematode Meloidogyne incognita and significantly influence its development. Moreover, our results also demonstrate that the protease Ver112 and the chitinase LPCHI1 from the same fungus interacted on the egg infection.
Laboratory for Conservation and Utilization of Bio-resources, Yunnan University, Kunming 650091, People's Republic of China. kqzhang111@yahoo.com.cn
Zhongwei
Z
Gan
Jinkui
J
Yang
Nan
N
Tao
Lianming
L
Liang
Qili
Q
Mi
Juan
J
Li
Ke-Qin
KQ
Zhang
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002688:Chitinases / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D010063:Ovum / Q000187:drug effects; D010447:Peptide Hydrolases / Q000378:metabolism / Q000494:pharmacology; D010572:Pest Control, Biological; D010802:Phylogeny; D014431:Tylenchoidea / Q000187:drug effects*; D020172:Verticillium / Q000201:enzymology* / Q000235:genetics
D005656:Fungal Proteins; D002688:Chitinases; D010447:Peptide Hydrolases