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Applied and Environmental Microbiology, March 2007, p. 1569-1575, Vol. 73, No. 5
0099-2240/07/$08.00+0     doi:10.1128/AEM.00981-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Effect of Superoxide Dismutase Gene Inactivation on Virulence of Pseudomonas aeruginosa PAO1 toward the Silkworm, Bombyx mori{triangledown}

Kazuhiro Iiyama,1* Yuuka Chieda,1,3 Jae Man Lee,2 Takahiro Kusakabe,2 Chisa Yasunaga-Aoki,1 and Susumu Shimizu1

Laboratory of Insect Pathology and Microbial Control, Institute of Biological Control, Faculty of Agriculture, Graduate School, Kyushu University, Fukuoka 812-8581, Japan,1 Laboratory of Silkworm Science, Faculty of Agriculture, Graduate School, Kyushu University, Fukuoka 812-8581, Japan,2 Japan Society for the Promotion of Science, Tokyo, Japan3

Received 26 April 2006/ Accepted 30 December 2006

To investigate the role of superoxide dismutase (SOD) in virulence against the silkworm, Bombyx mori, mutants of Pseudomonas aeruginosa PAO1 lacking manganese-SOD (PAO1sodM), iron-SOD (PAO1sodB), or both (PAO1sodMB) were generated. The mutants were injected into the hemocoel of B. mori. The virulence decreased in the order PAO1 = PAO1sodM > PAO1sodB > PAO1sodMB. In particular, PAO1sodMB was avirulent at a dose of 105 cells or less. The sod double mutant PAO1sodMB was then complemented with either pSodM or pSodB in trans. In both the complemented strains, the virulence was partially restored. Of the two plasmids, pSodB contributed more to the virulence of P. aeruginosa against B. mori. The results of growth in B. mori hemolymph broth and microscopic analysis suggested that a longer lag phase and superoxide sensitivity correlated with decreased virulence in sod mutants. In conclusion, the SODs are required for full virulence of P. aeruginosa against B. mori and Fe-SOD is more important than Mn-SOD in the infection process.

* Corresponding author. Mailing address: Laboratory of Insect Pathology and Microbial Control, Institute of Biological, Faculty of Agriculture, Graduate School, Kyushu University, Fukuoka 812-8581, Japan. Phone: 81 92 642 3033. Fax: 81 92 642 4421. E-mail: iiyama{at}grt.kyushu-u.ac.jp.

{triangledown} Published ahead of print on 12 January 2007.

Applied and Environmental Microbiology, March 2007, p. 1569-1575, Vol. 73, No. 5
0099-2240/07/$08.00+0     doi:10.1128/AEM.00981-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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