Florida Entomologist (2008) 91, 494-497

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Jennifer M. Zaspel and Marjorie A. Hoy (2008)
Comparison of short-term preservation and assay methods for the molecular detection of Wolbachia in the Mediterranean flour moth Ephestia kuehniella
Florida Entomologist 91 (3), 494-497
Abstract: Our results are consistent with previous studies (Fukatsu 1999) suggesting acetone and 95% EtOH are adequate short-term specimen preservation methods for detecting endosymbiotic bacterial DNA within an insect host. Although not statistically significant, an apparent reduction in Wolbachia wspA copy number and HF PCR band intensity of the symbiont DNA isolated from specimens stored in 95% EtOH at room temperature was observed (Fig. 1A, Table 1). Averages in the total DNA concentration and purity of the stored specimens at week 101 were not significantly different among replicates from the same treatment nor were they significantly different between treatments. Regression analyses revealed an increase in total DNA concentration over time for each treatment, and a decrease in absorbance over time. Additionally, a reduction in band intensity was observed for specimens stored in 95% EtOH at room temperature at the last sample date (week 101), suggesting longterm storage in 95% EtOH at room temperature may not be an optimal storage method because it may damage the DNA or inhibit the PCR. The results reported here are specific to E. kuehniella, but they indicate that when Wolbachia infection density in the host is high and the DNA extraction method is consistent, both standard and HF PCR are adequate for detecting Wolbachia after a 2-yr storage period in both acetone and 95% EtOH.
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Database assignments for author(s): Marjorie A. Hoy

Research topic(s) for pests/diseases/weeds:
molecular biology - genes
Research topic(s) for beneficials or antagonists:
molecular biology - genes

Pest and/or beneficial records:

Beneficial Pest/Disease/Weed Crop/Product Country Quarant.

Ephestia kuehniella
Wolbachia (genus - entomopathogens)