Selection of available suicide vectors for gene mutagenesis using chiA (a chitinase encoding gene) as a new reporter and primary functional analysis of chiA in Lysobacter enzymogenes strain OH11.
Here, three different suicide vectors were evaluated for the possibility of performing gene mutagenesis in strain OH11 using the chiA gene (accession number: DQ888611) as a new reporter. Suicide vector pEX18GM was selected, and it was successfully applied for disruption and in-frame deletions in the chiA gene in strain OH11, which was confirmed by PCR amplification and Southern hybridization. The chiA-deletion mutant OH11-3 did not have the ability to produce chitinase on chitine selection medium. Interestingly, the chiA-deletion mutants displayed wild-type antimicrobial activity against Saccharomyces cerevisiae, Magnaporthe grisea, Phytophthora                         capsici, Rhizoctonia solani, Sclerotinia sclerotiorum and Pythium ultimum. Our data suggest that chitinase might not be a unique lytic enzyme in controlling S. cerevisiae, M. grisea, P. capsici, and P. ultimum. R. solani, S. sclerotiorum. Also, suicide vector pEX18GM might be explored as a potential tool for gene deletions in L. enzymogenes, which will facilitate the molecular study of mechanisms of biological control in L. enzymogenes.
College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China.
Guoliang
G
Qian
Yansheng
Y
Wang
Dongyu
D
Qian
Jiaqin
J
Fan
Baishi
B
Hu
Fengquan
F
Liu
D001426:Bacterial Proteins / Q000235:genetics*; D015139:Blotting, Southern; D056915:DNA Copy Number Variations; D005822:Genetic Vectors / Q000235:genetics*; D053528:Lysobacter / Q000235:genetics*; D016296:Mutagenesis / Q000235:genetics / Q000502:physiology*
D001426:Bacterial Proteins