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Adopting Bacteria in Order to Adapt to Water-How Reed Beetles Colonized the Wetlands (Coleoptera, Chrysomelidae, Donaciinae).

Kleinschmidt B, Kölsch G - Insects (2011)

Bottom Line: Reed beetles are herbivores living on wetland plants, each species being mono- or oligo-phagous.They lay their eggs on the host plant and the larvae live underwater in the sediment attached to its roots.The pupation underwater enabled the reed beetles to permanently colonize the wetlands and to diversify in this habitat underexploited by herbivorous insects (adaptive radiation).

View Article: PubMed Central - PubMed

Affiliation: Zoological Institute, Molecular Evolutionary Biology, University of Hamburg, Martin-Luther-King-Platz 3, 20146 Hamburg, Germany. birgit.kleinschmidt@gmx.net.

ABSTRACT
The present paper reviews the biology of reed beetles (Donaciinae), presents experimental data on the role of specific symbiotic bacteria, and describes a molecular method for the detection of those bacteria. Reed beetles are herbivores living on wetland plants, each species being mono- or oligo-phagous. They lay their eggs on the host plant and the larvae live underwater in the sediment attached to its roots. The larvae pupate there in a water-tight cocoon, which they build using a secretion that is produced by symbiotic bacteria. The bacteria are located in four blind sacs at the foregut of the larvae; in (female) adults they colonize two out of the six Malpighian tubules. Tetracycline treatment of larvae reduced their pupation rate, although the bacteria could not be fully eliminated. When the small amount of bacterial mass attached to eggs was experimentally removed before hatching, symbiont free larvae resulted, showing the external transmission of the bacteria to the offspring. Specific primers were designed to detect the bacteria, and to confirm their absence in manipulated larvae. The pupation underwater enabled the reed beetles to permanently colonize the wetlands and to diversify in this habitat underexploited by herbivorous insects (adaptive radiation).

No MeSH data available.


Related in: MedlinePlus

Specific primers developed for reed beetle symbionts do not yield a PCR product with standard laboratory strains of E. coli (1% agarose gel containing ethidium bromide, UV illumination). Primers used: 1–3: specific for Donacia symbionts; 4–6: specific for Macroplea symbionts; 7–9: general bacterial primers. The samples are: 1, 4, 7: E. coli; 2, 8: D. semicuprea symbiont; 3: negative control (distilled water); 5, 9: Macroplea sp. symbiont; 6: negative control; S: size standard with the fragment size 1,000 bp given (cf. Figure 6).
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f4-insects-02-00540: Specific primers developed for reed beetle symbionts do not yield a PCR product with standard laboratory strains of E. coli (1% agarose gel containing ethidium bromide, UV illumination). Primers used: 1–3: specific for Donacia symbionts; 4–6: specific for Macroplea symbionts; 7–9: general bacterial primers. The samples are: 1, 4, 7: E. coli; 2, 8: D. semicuprea symbiont; 3: negative control (distilled water); 5, 9: Macroplea sp. symbiont; 6: negative control; S: size standard with the fragment size 1,000 bp given (cf. Figure 6).

Mentions: With the molecular tools developed as described above, we were able to specifically detect the symbionts. This specificity was required, because it is not possible to work under sterile conditions that would avoid contamination by, for example, bacteria contained in the digestive tract of the animal. Application of the primers to samples of Escherichia coli did not yield PCR products (Figure 4). All PCR-products sequenced revealed reed beetle symbionts in the BLAST searches conducted. All best hits were sequences of Candidatus Macropleicola, the symbionts of the reed beetle genus Macroplea (accession no. GQ480915 and GQ480918 [39]), and the bacterial symbionts of Donacia semicuprea (accession no. GQ480891, GQ480897, GQ480898, GQ480907, GQ480936; [45]), respectively, consistent with the host species used.


Adopting Bacteria in Order to Adapt to Water-How Reed Beetles Colonized the Wetlands (Coleoptera, Chrysomelidae, Donaciinae).

Kleinschmidt B, Kölsch G - Insects (2011)

Specific primers developed for reed beetle symbionts do not yield a PCR product with standard laboratory strains of E. coli (1% agarose gel containing ethidium bromide, UV illumination). Primers used: 1–3: specific for Donacia symbionts; 4–6: specific for Macroplea symbionts; 7–9: general bacterial primers. The samples are: 1, 4, 7: E. coli; 2, 8: D. semicuprea symbiont; 3: negative control (distilled water); 5, 9: Macroplea sp. symbiont; 6: negative control; S: size standard with the fragment size 1,000 bp given (cf. Figure 6).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4553447&req=5

f4-insects-02-00540: Specific primers developed for reed beetle symbionts do not yield a PCR product with standard laboratory strains of E. coli (1% agarose gel containing ethidium bromide, UV illumination). Primers used: 1–3: specific for Donacia symbionts; 4–6: specific for Macroplea symbionts; 7–9: general bacterial primers. The samples are: 1, 4, 7: E. coli; 2, 8: D. semicuprea symbiont; 3: negative control (distilled water); 5, 9: Macroplea sp. symbiont; 6: negative control; S: size standard with the fragment size 1,000 bp given (cf. Figure 6).
Mentions: With the molecular tools developed as described above, we were able to specifically detect the symbionts. This specificity was required, because it is not possible to work under sterile conditions that would avoid contamination by, for example, bacteria contained in the digestive tract of the animal. Application of the primers to samples of Escherichia coli did not yield PCR products (Figure 4). All PCR-products sequenced revealed reed beetle symbionts in the BLAST searches conducted. All best hits were sequences of Candidatus Macropleicola, the symbionts of the reed beetle genus Macroplea (accession no. GQ480915 and GQ480918 [39]), and the bacterial symbionts of Donacia semicuprea (accession no. GQ480891, GQ480897, GQ480898, GQ480907, GQ480936; [45]), respectively, consistent with the host species used.

Bottom Line: Reed beetles are herbivores living on wetland plants, each species being mono- or oligo-phagous.They lay their eggs on the host plant and the larvae live underwater in the sediment attached to its roots.The pupation underwater enabled the reed beetles to permanently colonize the wetlands and to diversify in this habitat underexploited by herbivorous insects (adaptive radiation).

View Article: PubMed Central - PubMed

Affiliation: Zoological Institute, Molecular Evolutionary Biology, University of Hamburg, Martin-Luther-King-Platz 3, 20146 Hamburg, Germany. birgit.kleinschmidt@gmx.net.

ABSTRACT
The present paper reviews the biology of reed beetles (Donaciinae), presents experimental data on the role of specific symbiotic bacteria, and describes a molecular method for the detection of those bacteria. Reed beetles are herbivores living on wetland plants, each species being mono- or oligo-phagous. They lay their eggs on the host plant and the larvae live underwater in the sediment attached to its roots. The larvae pupate there in a water-tight cocoon, which they build using a secretion that is produced by symbiotic bacteria. The bacteria are located in four blind sacs at the foregut of the larvae; in (female) adults they colonize two out of the six Malpighian tubules. Tetracycline treatment of larvae reduced their pupation rate, although the bacteria could not be fully eliminated. When the small amount of bacterial mass attached to eggs was experimentally removed before hatching, symbiont free larvae resulted, showing the external transmission of the bacteria to the offspring. Specific primers were designed to detect the bacteria, and to confirm their absence in manipulated larvae. The pupation underwater enabled the reed beetles to permanently colonize the wetlands and to diversify in this habitat underexploited by herbivorous insects (adaptive radiation).

No MeSH data available.


Related in: MedlinePlus