Experimental & Applied Acarology

, Volume 39, Issue 3–4, pp 257–271 | Cite as

Cardinium symbionts induce haploid thelytoky in most clones of three closely related Brevipalpus species

  • Thomas V. M. GrootEmail author
  • Johannes A. J. Breeuwer


Bacterial symbionts that manipulate the reproduction of their host to increase their own transmission are widespread. Most of these bacteria are Wolbachia, but recently a new bacterium, named Cardinium, was discovered that is capable of the same manipulations. In the host species Brevipalpus phoenicis (Acari: Tenuipalpidae) this bacterium induces thelytoky by feminizing unfertilized haploid eggs. The related species B. obovatus and B. californicus are thelytokous too, suggesting that they reproduce in the same remarkable way as B. phoenicis. Here we investigated the mode of thelytokous reproduction in these three species. Isofemale lines were created of all three species and 19 lines were selected based on variation in mitochondrial COI sequences. All B. phoenicis and B. californicus lines (10 and 4 lines, respectively) produced males under laboratory conditions up to 6.7%. In contrast, males were absent from all B. obovatus lines (5 lines). Additional experiments with two B. phoenicis isofemale lines showed that males can be produced by very young females only, while older females produce daughters exclusively. For most lines it was shown that they are indeed feminized by a bacterium as treatment with antibiotics resulted in increased numbers of males up to 13.5%. Amplification and identification of specific gyrB sequences confirmed that those lines were infected with Cardinium. Three out of the five B. obovatus lines did not produce males after treatments with antibiotics, nor did they contain Cardinium or any other bacterium that might induce thelytoky. In these lines thelytoky is probably a genetic property of the mite itself. Despite the different causes of thelytoky, flow cytometry revealed that all 19 lines were haploid. Finally, the taxonomic inferences based on the mitochondrial COI sequences were incongruent with the classical taxonomy based on morphology, suggesting that a taxonomic revision of this group is necessary.


Brevipalpus phoenicis Brevipalpus obovatus Brevipalpus californicus Acari Tenuipalpidae Cardinium Thelytoky Spanandric males 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



We thank Dr. G. de Moraes and N.C. Mesa for morphologically identifying the mite species. We also thank the people at the Burger’s Bush tropical greenhouse for allowing us to collect mites and their help in doing so. The advise by M. Stift and R. Bregman on the flow cytometry analysis was much appreciated. We thank Prof. S. Menken for his remarks on an earlier draft in this manuscript. This study was supported by the Netherlands Foundation for the Advancement of Tropical Research (WOTRO), grant number W89–141.


  1. Bensasson D, Zhang D-X, Hartl DL, Hewitt GM (2001) Mitochondrial pseudogenes: evolution’s misplaced witnesses. Trends Ecol Evol 16:314–321PubMedCrossRefGoogle Scholar
  2. Breeuwer JAJ, Werren JH (1993) Cytoplasmic incompatibility and bacterial density in Nasonia vitripennis. Genetics 135:565–574PubMedGoogle Scholar
  3. Carson HL, Chang LS, Lyttle TW (1982) Decay of female sexual behavior under parthenogenesis. Science 218:68–70PubMedCrossRefGoogle Scholar
  4. Chigira A, Miura K (2005) Detection of ‘Candidatus Cardinium’ bacteria from the haploid host Brevipalpus californicus (Acari: Tenuipalpidae) and effect on the host. Exp Appl Acarol 37:107–116PubMedCrossRefGoogle Scholar
  5. Dedeine F, Bandi C, Bouletreau M and Kramer LH (2003) Insights into Wolbachia obligatory symbiosis. In: Bourtzis K, Miller TA (eds) Insect Symbiosis. CRC Press, Florida, pp 267–282Google Scholar
  6. Diaz PI, Chalmers NI, Rickard AH, Kong C, Milburn CL, Palmer Jr RJ, Kolenrander PE (2006) Molecular characterization of subject-specific oral microflora during initial colonization of enamel. Appl Environ Microbiol 72:2837–2848PubMedCrossRefGoogle Scholar
  7. Doyle JJ (1991) DNA protocols for plants. In: Hewitt G, Johnson AWB, Young JPW (eds) Molecular techniques in taxonomy, vol 57, pp. 283–293Google Scholar
  8. Groot TVM, Janssen A, Pallini A, Breeuwer JAJ (2005) Adaptation in the asexual false spider mite Brevipalpus phoenicis: evidence for frozen niche variation. Exp Appl Acarol 36:165–176PubMedCrossRefGoogle Scholar
  9. Haramoto FH (1969) Biology and control of Brevipalpus phoenicis (Geijskes) (Acarina: Tenuipalpidae). Tech Bull Hawaii Agricul Exp Stat 68:1–63Google Scholar
  10. Helle W, Bolland HR, Heitmans WRB (1980) Chromosomes and types of parthenogenesis in the false spider mites (Acari: Tenuipalpidae). Genetica 54:45–50CrossRefGoogle Scholar
  11. Hoy MA, Jeyaprakash A (2004) Microbial diversity in the predatory mite Metaseiulus occidentalis (Acari: Phytoseiidae) and its prey, Tetranychus urticae (Acari: Tertranychidae). Biol Control 32:427–441CrossRefGoogle Scholar
  12. Hunter MS, Perlman SJ, Kelly SE (2003) A bacterial symbiont in the Bacteriodetes induces cytoplasmic incompatibility in the parasitoid wasp Encarsia pergandiella. Proc Roy Soc ser B 270:2185–2190CrossRefGoogle Scholar
  13. Hurst GDD, Jiggins FM (2005) Problems with mitochondrial DNA as marker in population, phylogeographic and phylogenetic studies: the effects of inherited symbionts. Proc Roy Soc ser B 272:1525–1534CrossRefGoogle Scholar
  14. Hurst GDD, Jiggins FM, Majerus MEN (2003) Inherited microorganisms that selectively kill male hosts: the hidden players of insect evolution? In: Bourtzis K, Miller DJ (eds) Insect symbiosis. CRC Press, Florida, pp 177–198Google Scholar
  15. Johnston JS, Ross LD, Beani L, Hughes DP, Kathirithamby J (2004) Tiny genomes and endoreduplication in Strepsiptera. Insect Mol Biol 13:581–585PubMedCrossRefGoogle Scholar
  16. Kennedy JS (1995) Functional ecology of the false spider mite, Brevipalpus phoenicis (Geijskes). Dissertation, Université Catholique de Louvain, Louvain-la-NeuveGoogle Scholar
  17. Kondo N, Nikoh N, Ljichi N, Shimada M, Fukatsu T (2002) Genome fragment of Wolbachia endosymbiont transferred to X chromosome of host insect. Proc Nat Acad Sci USA 99:14280–14285PubMedCrossRefGoogle Scholar
  18. Muller HJ (1949) The Darwinian and modern conceptions of natural selection. Proc Am Philos Soc 93:459–470PubMedGoogle Scholar
  19. Nagesha Chandra BK, Channabasavanna GP (1974) Biology of guave scarlet mite, Brevipalpus phoenicis (Geijskes) (Acarina: Tenuipalpidae). Proc. 4th Int Cong Acarol pp 167–176Google Scholar
  20. Navajas M, Gutierrez J, Lagnel J, Boursot P (1996) Mitochondrial cytochrome oxidase I in tetranychid mites: a comparison between molecular phylogeny and changes of morphological and life history traits. Bull Entomol Res 86:407–417CrossRefGoogle Scholar
  21. Oomen PA (1982) Studies on population dynamics of the scarlet mites, Brevipalpus phoenicis, a pest of tea in Indonesia. Mededelingen Landbouwhogeschool Wageningen 82:1–82Google Scholar
  22. Palmer JD, Adams KL, Cho Y, Parkinson CL, Qui Y-L, Song K (2000) Dynamic evolution of plant mitochondrial genomes: mobile genes and introns and highly variable mutation rates. Proc Nat Acad Sci USA 97:6960–6966PubMedCrossRefGoogle Scholar
  23. Pijnacker LP, Ferwerda MA, Bolland HR, Helle W (1980) Haploid female parthenogenesis in the false spider mite Brevipalpus obovatus (Acari: Tenuipalpida). Genetica 51:211–214CrossRefGoogle Scholar
  24. Pijnacker LP, Ferwerda MA, Helle W (1981) Cytological investigations on the female and male reproductive system of the parthenogenetic privet mite Brevipalpus obovatus Donnadieu (Phytoptipalpidae, Acari). Acarologia 22:157–163Google Scholar
  25. Razoux Schultz L (1961) Enkele notities over de oranje mijt, Brevipalpus phoenicis Geijskes, op thee in Indonesie. Mededelingen landbouwhogeschool Gent 26:1694–1702Google Scholar
  26. Rigaud T, Juchault P, Mocquard J-P (1997) The evolution of sex determination in isopod crustaceans. BioEssays 19:409–416CrossRefGoogle Scholar
  27. Rigaud T, Pennings PS, Juchault P (2001) Wolbachia bacteria effects after experimental interspecific transfer in terrestrial isopods. J Invertebr Pathol 77:251–257PubMedCrossRefGoogle Scholar
  28. Sambrook J, Frisch EF, Maniatis T (1989) Molecular cloning. Cold Spring Harbor Press, Cold Spring Harbor, New YorkGoogle Scholar
  29. Siegel S, Castellan NJ Jr (1988) Nonparametric statistics for the behavioral sciences, McCraw-Hill, New YorkGoogle Scholar
  30. Stouthamer R, Breeuwer JAJ, Hurst GDD (1999) Wolbachia pipientis: microbial manipulator of arthropod reproduction. Ann Rev Microbiol 53:71–102CrossRefGoogle Scholar
  31. Swofford DL (1998) PAUP*, phylogenetic analysis using parsimony (*and other methods). Sinauer Assoc., SunderlandGoogle Scholar
  32. Thompson JD, Higins DG, Gibson TJ (1994) CLUSTAL W: improving the senitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedCrossRefGoogle Scholar
  33. Weeks AR, Marec F, Breeuwer JAJ (2001) A mite species that consists entirely of haploid females. Science 292:2479–2482PubMedCrossRefGoogle Scholar
  34. Weeks AR, Velten R, Stouthamer R (2003) Incidence of a new sex ratio distorting endosymbiotic among arthropods. Proc Roy Soc ser B 270:1857–1865CrossRefGoogle Scholar
  35. Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703PubMedGoogle Scholar
  36. Welbourn WC, Ochoa R, Kane EC, Erbe EF (2003) Morphological observations on Brevipalpus phoenicis (Acari: Tenuipalpidae) including comparisons with B. californicus and B. obovatus. Exp Appl Acarol 30:107–133PubMedCrossRefGoogle Scholar
  37. Werren JH, O’Neill SL (1997) The evolution of heritable symbionts. In: O’Neill SL, Hoffmann AA, Werren JH (eds) Influential passengers; inherited microorganisms and arthropod reproduction. Oxford University Press, Oxford, pp 1–41Google Scholar
  38. Zchori-Fein E, Perlman SJ (2004) Distribution of the bacterial symbiont Cardinium in arthropods. Mol Ecol 13:2009–2016PubMedCrossRefGoogle Scholar
  39. Zchori-Fein E, Perlman SJ, Kelly SE, Katzir N, Hunter MS (2004) Characterization of a Bacteriodetes symbiont in Encarsia wasps (Hymenoptera: Aphelinidae): proposal of ‘Candidatus Cardinium hertigii’. Int J Syst Evol Microbiol 54:961–968PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Thomas V. M. Groot
    • 1
    Email author
  • Johannes A. J. Breeuwer
    • 1
  1. 1.Section Evolutionary Biology, Institute for Biodiversity and Ecosystem DynamicsUniversity of AmsterdamAmsterdamThe Netherlands

Personalised recommendations