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Identification of mutations involved in the requirement of potassium for growth of typical Melissococcus plutonius strains.

ABSTRACT: Melissococcus plutonius is a fastidious honeybee pathogen, and the addition of KH(2)PO(4) to culture medium is required for its growth. Using genome sequences and a newly developed vector, we showed that mutations in genes encoding Na(+)/H(+) antiporter and cation-transporting ATPase are involved in the potassium requirement for growth.

SUBMITTER: Takamatsu D

PROVIDER: S-EPMC3675957 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature

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Identification of mutations involved in the requirement of potassium for growth of typical Melissococcus plutonius strains.

Takamatsu Daisuke D Arai Rie R Miyoshi-Akiyama Tohru T Okumura Kayo K Okura Masatoshi M Kirikae Teruo T Kojima Atsuko A Osaki Makoto M

Applied and environmental microbiology 20130412 12

Melissococcus plutonius is a fastidious honeybee pathogen, and the addition of KH(2)PO(4) to culture medium is required for its growth. Using genome sequences and a newly developed vector, we showed that mutations in genes encoding Na(+)/H(+) antiporter and cation-transporting ATPase are involved in the potassium requirement for growth. ...[more]

PMID: 23584776

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Development of duplex PCR assay for detection and differentiation of typical and atypical Melissococcus plutonius strains.

Project description:Melissococcus plutonius is the causative agent of an important honeybee disease, European foulbrood (EFB). In addition to M. plutonius strains with typical characteristics (typical M. plutonius), we recently reported the presence of atypical M. plutonius, which are phenotypically and genetically distinguished from typical M. plutonius. Because typical and atypical M. plutonius may have different pathogenic mechanisms, differentiation of these two types is very important for diagnosis and more effective control of EFB. In this study, therefore, a duplex PCR assay was developed to detect and differentiate typical and atypical M. plutonius rapidly and easily. On the basis of the results of comparative genomic analyses, we selected Na(+)/H(+) antiporter gene and Fur family transcriptional regulator gene as targets for detection of typical and atypical strains, respectively, by PCR. Under optimized conditions, the duplex PCR system using the designed primers successfully detected and differentiated all typical and atypical M. plutonius strain/isolates tested, while no product was generated from any other bacterial strains/isolates used in this study, including those isolated from healthy honeybee larval guts. Detection limits of the PCR were 50 copies of chromosome/reaction for both types, and it could detect typical and atypical M. plutonius directly from diseased honeybee larvae. Moreover, the duplex PCR diagnosed mixed infections with both M. plutonius types more precisely than standard culture methods. These results indicate that the duplex PCR assay developed in this study is extremely useful for precise diagnosis and epidemiological study of EFB.

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