Isolation and characterization of a cold-induced nonculturable suppression mutant of Vibrio vulnificus.
The viable but nonculturable (VBNC) suppression mutant formed platable cells at low temperature stress after inoculation in artificial seawater (ASW). Suppression subtractive hybridization was used to identify differentially expressed genes among cDNAs of the VBNC suppression mutant and the wild-type Vibrio vulnificus strain. Glutathione S-transferase was identified as a responsive gene of the VBNC suppression mutant in our assay, and was highly expressed from the VBNC suppression mutant at low temperature stress. Culturability tests revealed that the wild-type cells were sensitive to oxidative stress in the hydrogen peroxide (H(2)O(2)) and to 1-chloro-2,4-dinitrobenzene (CDNB) compared with the VBNC suppression mutant cells. Adding glutathione showed that many wild-type V. vulnificus cells maintained culturability in cold ASW. These results suggest that non-nutritional growth inhibitors, such as peroxide that accumulates at low temperatures, influence VBNC in V. vulnificus cells.
Abe
Akihisa
A
Nippon Suisan Kaisha, Ltd., Food Safety Research Center, 559-6 Kitano-Machi, Hachioji, Tokyo 192-0906, Japan. aki-abe@nissui.co.jp
Ohashi
Eiji
E
Ren
Huifeng
H
Hayashi
Tetsuhito
T
Endo
Hideaki
H
D003080:Cold Temperature; D004137:Dinitrochlorobenzene / Q000494:pharmacology; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D006146:Guanidines / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D009154:Mutation; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D041261:Vibrio vulnificus / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D014871:Water Microbiology
D004137:Dinitrochlorobenzene; D006146:Guanidines; D012333:RNA, Messenger; C082988:N-methyl-N'-nitroguanidine; D006861:Hydrogen Peroxide; D005982:Glutathione Transferase