This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Konai, M. Articles by Whitcomb, R. F. Search for Related Content PubMed PubMed Citation Articles by Konai, M. Articles by Whitcomb, R. F. Agricola Articles by Konai, M. Articles by Whitcomb, R. F.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Sep 1996, 3453-3458, Vol 62, No. 9
Copyright © 1996, American Society for Microbiology

Improved Cultivation Systems for Isolation of the Colorado Potato Beetle Spiroplasma

M Konai, KJ Hackett, DL Williamson, JJ Lipa, JD Pollack, GE Gasparich, EA Clark, DC Vacek and RF Whitcomb
USDA Agricultural Research Service, Insect Biocontrol Laboratory, Beltsville Agricultural Research Center, Beltsville, Maryland 20705; Department of Anatomical Sciences, State University of New York, Stony Brook, New York 11794; Department of Biological Control and Quarantine, Institute of Plant Protection, Poznan, Poland; Department of Medical Microbiology and Immunology, Ohio State University, Columbus, Ohio 43210; and USDA Animal and Plant Health Inspection Service, Mission Biological Control Center, Mission, Texas 78573

In North America, the Colorado potato beetle, Leptinotarsa decemlineata, is often infected with the host-specific, gut-inhabiting Colorado potato beetle spiroplasma (CPBS). CPBS is apparently a commensal, but it may be useful in biocontrol if it can be transformed to express an insect-lethal gene. Difficulty in cultivating the organism, however, has hindered the development of a suitable transformation system. In this study, we eliminated the need for coculturing CPBS with insect cells. CPBS was reliably isolated with the BBL Anaerobic GasPak Jar system (low redox, enhanced CO(inf2)), which was easier to use and less expensive than insect cell coculture methods. A further advantage is a reduction in contaminating insect cell components. Use of anaerobiosis should facilitate early-passage screening of isolates for extrachromosomal elements, for use in gene vector constructs. The unique spiral (decreasing amplitude of coils) morphology of CPBS was preserved by anaerobiosis. The use of low-pH (6.0 to 6.5) media allowed aerobic adaptation of CPBS to M1D and SP-4 broth media. These formulations permitted the first cultivation of CPBS on solid media, an accomplishment that will simplify the selection of molecular transformants. Potato beetles collected at four sites in Poland yielded CPBS strains similar to those previously obtained from populations in North America.

This article has been cited by other articles:

• Oduori, M. A., Lipa, J. J., Gasparich, G. E. (2005). Spiroplasma leucomae sp. nov., isolated in Poland from white satin moth (Leucoma salicis L.) larvae. Int. J. Syst. Evol. Microbiol. 55: 2447-2450 [Abstract] [Full Text]  
• Razin, S., Yogev, D., Naot, Y. (1998). Molecular Biology and Pathogenicity of Mycoplasmas. Microbiol. Mol. Biol. Rev. 62: 1094-1156 [Abstract] [Full Text]