title: Cloning and characterization of chitinases from interior spruce and lodgepole pine.
abstract: Chitinases have been implicated in the defence of conifers against insects and pathogens. cDNA for six chitinases were cloned from interior spruce (Picea glauca x engelmannii) and four from lodgepole pine (Pinus contorta). The cloned interior spruce chitinases were annotated class I PgeChia1-1 and PgeChia1-2, class II PgeChia2-1, class IV PgeChia4-1, and class VII PgeChia7-1 and PgeChia7-2; lodgepole pine chitinases were annotated class I PcChia1-1, class IV PcChia4-1, and class VII PcChia7-1 and PcChia7-2. Chitinases were expressed in Escherichia coli with maltose-binding-protein tags and soluble proteins purified. Functional characterization demonstrated chitinolytic activity for the three class I chitinases PgeChia1-1, PgeChia1-2 and PcChia1-1. Transcript analysis established strong induction of most of the tested chitinases, including all three class I chitinases, in interior spruce and lodgepole pine in response to inoculation with bark beetle associated fungi (Leptographium abietinum and Grosmannia clavigera) and in interior spruce in response to weevil (Pissodes strobi) feeding. Evidence of chitinolytic activity and inducibility by fungal and insect attack support the involvement of these chitinases in conifer defense.
authors.affiliation: Michael Smith Laboratories, University of British Columbia, 312-2185 East Mall, Vancouver, British Columbia V6T 1Z4, Canada; Department of Botany, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada.
authors.forename: N
authors.initials: N
authors.lastname: Kolosova
authors.affiliation: Department of Wood Science, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada.
authors.forename: C
authors.initials: C
authors.lastname: Breuil
authors.affiliation: Michael Smith Laboratories, University of British Columbia, 312-2185 East Mall, Vancouver, British Columbia V6T 1Z4, Canada; Department of Botany, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada; Department of Forest and Conservation Sciences, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada. Electronic address: bohlmann@msl.ubc.ca.
authors.forename: J
authors.initials: J
authors.lastname: Bohlmann
mesh_terms: D000818:Animals; D020134:Catalytic Domain; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D004790:Enzyme Induction; D004926:Escherichia coli / Q000235:genetics; D006868:Hydrolysis; D008969:Molecular Sequence Data; D055155:Ophiostomatales / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity; D028222:Picea / Q000201:enzymology*; D028223:Pinus / Q000201:enzymology*; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology / Q000469:parasitology / Q000517:prevention & control; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic; D048889:Weevils / Q000254:growth & development
chemical_list: D010940:Plant Proteins; D002686:Chitin; D002688:Chitinases
keywords: Chitinases; Conifer defence; Coniferales; Dendroctonus ponderosae; Grossmannia clavigera; Interior spruce; Leptographium abietinum; Lodgepole pine; Mountain pine beetle; Picea glauca x engelmannii; Pinaceae; Pinus contorta; Pissodes strobi; Plant–insect and plant–pathogen interactions; White pine weevil