Overexpression of a chitinase gene PbChia1 from Plasmodiophora brassicae
increases broad spectrum disease resistance
Abstract
Chitinase plays an important role in plant resistance against chitin
containing pathogens by degrading chitin. Clubroot, caused by
Plasmodiophora brassicae, is a major disease in cruciferous crops
worldwide, including vegetable and rapeseed. Chitin is the main
component of P. brassicae spore cell walls, chitinases can
enhance the plant’s defense system as they act on chitin. However, the
function of chitinases in P. brassicae has not reported. Here, we
found that chitin is the functional component of P. brassicae by
wheat germ agglutinin staining and commercial chitinase treatment
. Chitin extracted from resting spores of P. brassicae was
characterized by infrared spectroscopy, X-ray diffraction and thermo
gravimetry analysis.The chitinase PbChia1 was identified by chitin
pull-down assays combined with LC-MS/MS method. We also showed that
PbChia1 was a typical secreted chitinase, could bind to chitin and
showed chitinase activity in vitro, significantly degrade the
number of resting spores of P. brassicae and significantly
relieve the severity of clubroot symptom, the biocontrol effect was
61.29%. Overexpression of PbChia1 in Arabidopsis thaliana
resulted in susceptibility to P. brassicae, increase host
survival rate and seed yields, and transgenic plants enhanced in
chitin-triggered reactive oxygen species burst, MAPK activation and
expression of immune-related genes. PbChia1 transgenic plants
showed resistance to other pathogens, such as biotrophic bacteria
Pst DC3000, necrotrophic fungi Sclerotinia sclerotiorum
1980 and Rhizoctonia solani. The findings indicate that
chitinases PbChia1 is a candidate gene with potential application
for broad spectrum disease resistance in breeding plants to prevent
clubroot.