Asymmetrical Interactions between Wolbachia and Spiroplasma Endosymbionts Coexisting in the Same Insect Host

ABSTRACT We investigated the interactions between the endosymbionts Wolbachia pipientis strain wMel and Spiroplasma sp. strain NSRO coinfecting the host insect Drosophila melanogaster. By making use of antibiotic therapy, temperature stress, and hemolymph microinjection, we established the following strains in the same host genetic background: the SW strain, infected with both Spiroplasma and Wolbachia; the S strain, infected with Spiroplasma only; and the W strain, infected with Wolbachia only. The infection dynamics of the symbionts in these strains were monitored by quantitative PCR during host development. The infection densities of Spiroplasma exhibited no significant differences between the SW and S strains throughout the developmental course. In contrast, the infection densities of Wolbachia were significantly lower in the SW strain than in the W strain at the pupal and young adult stages. These results indicated that the interactions between the coinfecting symbionts were asymmetrical, i.e., Spiroplasma organisms negatively affected the population of Wolbachia organisms, while Wolbachia organisms did not influence the population of Spiroplasma organisms. In the host body, the symbionts exhibited their own tissue tropisms: among the tissues examined, Spiroplasma was the most abundant in the ovaries, while Wolbachia showed the highest density in Malpighian tubules. Strikingly, basically no Wolbachia organisms were detected in hemolymph, the principal location of Spiroplasma. These results suggest that different host tissues act as distinct microhabitats for the symbionts and that the lytic process in host metamorphosis might be involved in the asymmetrical interactions between the coinfecting symbionts.

[1]  T. Fukatsu,et al.  Tissue-specific infection dynamics of male-killing and nonmale-killing spiroplasmas in Drosophila melanogaster. , 2006, FEMS microbiology ecology.

[2]  M. Shimada,et al.  Infection density of Wolbachia endosymbiont affected by co-infection and host genotype , 2005, Biology Letters.

[3]  W. Miller,et al.  Evidence for a Global Wolbachia Replacement in Drosophila melanogaster , 2005, Current Biology.

[4]  J. Cande,et al.  Widespread Prevalence of Wolbachia in Laboratory Stocks and the Implications for Drosophila Research , 2005, Genetics.

[5]  T. Fukatsu,et al.  Rickettsia Symbiont in the Pea Aphid Acyrthosiphon pisum: Novel Cellular Tropism, Effect on Host Fitness, and Interaction with the Essential Symbiont Buchnera , 2005, Applied and Environmental Microbiology.

[6]  L. B. Klaczko,et al.  Male‐killing Spiroplasma naturally infecting Drosophila melanogaster , 2005, Insect molecular biology.

[7]  J. Jaenike,et al.  EXPRESSION AND MODULATION OF EMBRYONIC MALE‐KILLING IN DROSOPHILA INNUBILA: OPPORTUNITIES FOR MULTILEVEL SELECTION , 2005, Evolution; international journal of organic evolution.

[8]  F. Vavre,et al.  Multiple infections and diversity of cytoplasmic incompatibility in a haplodiploid species , 2005, Heredity.

[9]  K. Bourtzis,et al.  Heads or Tails: Host-Parasite Interactions in the Drosophila-Wolbachia System , 2004, Applied and Environmental Microbiology.

[10]  F. Vavre,et al.  Virulence, Multiple Infections and Regulation of Symbiotic Population in the Wolbachia-Asobara tabida Symbiosis , 2004, Genetics.

[11]  S. Naitza,et al.  Antimicrobial defences in Drosophila: the story so far. , 2004, Molecular immunology.

[12]  T. Fukatsu,et al.  Occurrence of Chaperonin 60 and Chaperonin 10 in primary and secondary bacterial symbionts of aphids: Implications for the evolution of an endosymbiotic system in aphids , 1993, Journal of Molecular Evolution.

[13]  T. Fukatsu,et al.  Changing partners in an obligate symbiosis: a facultative endosymbiont can compensate for loss of the essential endosymbiont Buchnera in an aphid , 2003, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[14]  F. Vavre,et al.  Strain‐specific regulation of intracellular Wolbachia density in multiply infected insects , 2003, Molecular ecology.

[15]  T. Fukatsu,et al.  Diversity of Wolbachia Endosymbionts in Heteropteran Bugs , 2003, Applied and Environmental Microbiology.

[16]  N. Moran,et al.  Side‐stepping secondary symbionts: widespread horizontal transfer across and beyond the Aphidoidea , 2003, Molecular ecology.

[17]  T. Fukatsu,et al.  Population Dynamics of Male-Killing and Non-Male-Killing Spiroplasmas in Drosophila melanogaster , 2003, Applied and Environmental Microbiology.

[18]  T. Fukatsu,et al.  Hidden from the host: Spiroplasma bacteria infecting Drosophila do not cause an immune response, but are suppressed by ectopic immune activation , 2003, Insect molecular biology.

[19]  H. Ishikawa,et al.  Regulation of Wolbachia Density in the Mediterranean Flour Moth, Ephestia kuehniella, and the Almond Moth, Cadra cautella , 2003, Zoological science.

[20]  Marjorie A. Hoy Insect Symbiosis , 2003 .

[21]  M. Strand,et al.  Insect hemocytes and their role in immunity. , 2002, Insect biochemistry and molecular biology.

[22]  M. Shimada,et al.  Internal Spatiotemporal Population Dynamics of Infection with Three Wolbachia Strains in the Adzuki Bean Beetle, Callosobruchus chinensis (Coleoptera: Bruchidae) , 2002, Applied and Environmental Microbiology.

[23]  P. Gullan,et al.  Secondary (γ-Proteobacteria) Endosymbionts Infect the Primary (β-Proteobacteria) Endosymbionts of Mealybugs Multiple Times and Coevolve with Their Hosts , 2002, Applied and Environmental Microbiology.

[24]  N. Moran,et al.  Estimating Population Size and Transmission Bottlenecks in Maternally Transmitted Endosymbiotic Bacteria , 2002, Microbial Ecology.

[25]  M. Shimada,et al.  Prevailing triple infection with Wolbachia in Callosobruchus chinensis (Coleoptera: Bruchidae) , 2002, Molecular ecology.

[26]  U. Theopold,et al.  The coagulation of insect hemolymph , 2002, Cellular and Molecular Life Sciences CMLS.

[27]  P. Gullan,et al.  Secondary (gamma-Proteobacteria) endosymbionts infect the primary (beta-Proteobacteria) endosymbionts of mealybugs multiple times and coevolve with their hosts. , 2002, Applied and environmental microbiology.

[28]  P. Pennings,et al.  Wolbachia bacteria effects after experimental interspecific transfers in terrestrial isopods. , 2001, Journal of invertebrate pathology.

[29]  Q. Zhang,et al.  Infection density of Wolbachia and incompatibility level in two planthopper species, Laodelphax striatellus and Sogatella furcifera. , 2001, Insect biochemistry and molecular biology.

[30]  T. Fukatsu,et al.  Spiroplasma Symbiont of the Pea Aphid, Acyrthosiphon pisum (Insecta: Homoptera) , 2001, Applied and Environmental Microbiology.

[31]  K. Bourtzis,et al.  Wolbachia neither induces nor suppresses transcripts encoding antimicrobial peptides , 2000, Insect molecular biology.

[32]  N. Moran,et al.  Secondary Endosymbionts of Psyllids Have Been Acquired Multiple Times , 2000, Current Microbiology.

[33]  H. Ishikawa,et al.  Wolbachia infection and cytoplasmic incompatibility in the cricket Teleogryllus taiwanemma. , 2000, The Journal of experimental biology.

[34]  H. Ishikawa,et al.  Genomic copy number of intracellular bacterial symbionts of aphids varies in response to developmental stage and morph of their host. , 2000, Insect biochemistry and molecular biology.

[35]  T. Fukatsu Acetone preservation: a practical technique for molecular analysis , 1999, Molecular ecology.

[36]  H. Ishikawa,et al.  Intracellular Bacterial Symbionts of Aphids Possess Many Genomic Copies per Bacterium , 1999, Journal of Molecular Evolution.

[37]  F. Rousset,et al.  Wolbachia infections are distributed throughout insect somatic and germ line tissues. , 1999, Insect biochemistry and molecular biology.

[38]  G. Hurst,et al.  Wolbachia pipientis: microbial manipulator of arthropod reproduction. , 1999, Annual review of microbiology.

[39]  T. Fukatsu,et al.  Specific detection of intracellular symbiotic bacteria of aphids by oligonucleotide-probed in situ hybridization , 1998 .

[40]  F. Rousset,et al.  Phylogeny and PCR–based classification of Wolbachia strains using wsp gene sequences , 1998, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[41]  A. Hoffmann,et al.  Population dynamics of the Wolbachia infection causing cytoplasmic incompatibility in Drosophila melanogaster. , 1998, Genetics.

[42]  J. Werren Biology of Wolbachia. , 2003, Annual review of entomology.

[43]  John H. Werren,et al.  Influential passengers: inherited microorganisms and arthropod reproduction , 1997 .

[44]  G. Markakis,et al.  Wolbachia infection and cytoplasmic incompatibility in Drosophila species. , 1996, Genetics.

[45]  A. Hoffmann,et al.  Cytoplasmic incompatibility in Australian populations of Drosophila melanogaster. , 1994, Genetics.

[46]  F. Rousset,et al.  Widespread occurence of the proteobacteria Wolbachia and partial cytoplasmic incompatibility in Drosophila melanogaster , 1994 .

[47]  T. Hays,et al.  A fertility region on the Y chromosome of Drosophila melanogaster encodes a dynein microtubule motor. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[48]  D. Williamson,et al.  6 – SEX RATIO ORGANISMS (SPIROPLASMAS) OF Drosophila , 1979 .

[49]  P. Buchner Endosymbiosis of Animals with Plant Microorganisms , 1965 .

[50]  B. Sakaguchi,et al.  Distribution of "sex-ratio" agent in tissues of Drosophila willistoni. , 1961, Genetics.

[51]  B. Sakaguchi,et al.  Nature of "Sex-ratio" Agent in Drosophila , 1961, Science.