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Spiroplasma lampyridicola sp. nov., from the Firefly Beetle Photuris pennsylvanicus

Clauzell Stevens¹, Ah Yin Tang¹, Edward Jenkins^1,, Rickey L. Goins¹, Joseph G. Tully², David L. Rose², Meghnad Konai³, David L. Williamson⁴, Patricia Carle⁵, Joseph M. Bové⁵, Kevin J. Hackett³, Frank E. French⁶, Jimmy Wedincamp⁶, Roberta B. Henegar⁷ and Robert F. Whitcomb^1,*

¹Department of Agricultural Sciences and Department of Microbiology, School of Veterinary Medicine, Tuskegee University, Tuskegee, Alabama 36088
²Mycoplasma Section, Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, Frederick Cancer Research Facility, Frederick, Maryland 21702
³Insect Biocontrol Laboratory, Vegetable Laboratory, USDA Agricultural Research Service, BARCW, Beltsville, Maryland 20705
⁷Vegetable Laboratory, USDA Agricultural Research Service, BARCW, Beltsville, Maryland 20705
⁴Department of Anatomical Sciences, State University of New York, Stony Brook, New York 11794
⁵Laboratoire de Biologie Cellulaire et Moléculaire, Institut Nationale de Recherche Agronomique, 33883 Villenave d'Ornon Cedex, France
⁶Department of Biology, Georgia Southern University, Statesboro, Georgia 30460

^* Corresponding author. Mailing address: Vegetable Laboratory, GH3-1, Bldg. 010, Range 2, BARCW, Beltsville, MD 20705. Phone: (301) 504-8339. Fax: (301) 504-6017.

ABSTRACT

Spiroplasma strain PUP-1^Twas isolated from the gut fluids of a firefly beetle (Photuris pennsylvanicus) collected in Maryland. Cells of the strain were shown by dark-field microscopy to be helical, motile filaments. Ultrastructural examination by electron microscopy revealed filamentous cells bounded by a single cytoplasmic membrane and no evidence of a cell wall. The cells were not sensitive to 500 U of penicillin per ml and grew under aerobic conditions in M1D, SP-4, and M-2 broth formulations, as well as in conventional mycoplasma medium. The doubling times at 15, 20, 25, and 30°C were 83.1, 32.0, 14.9, and 9.8 h, respectively. Suboptimal growth occurred at 37°C, and no growth was apparent in cultures maintained at 10 or 40°C. The organism required cholesterol for growth and produced acid from glucose, fructose, and trehalose; arginine and urea were not hydrolyzed. The results of previous serological analyses of strain PUP-1^Tindicated that the organism was not related to the then currently established Spiroplasma species or group representatives, and the organism was classified as the representative of group XIX. Additional testing of strain PUP-1^Tagainst recently recognized Spiroplasma species or group representatives by both metabolism inhibition and deformation tests confirmed the unique serological status of the organism. The guanine-plus-cytosine content of the DNA was 26 ± 1 mol%, and the genome size was 1,375 kbp. These values clearly differentiate strain PUP-1^Tfrom group XXI strain W115, with which it cross-reacted reciprocally at a low level in deformation and metabolism inhibition tests. We propose that strain PUP-1 (= ATCC 43206) should be recognized as the type strain of a new species, Spiroplasma lampyridicola.

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