The method of detecting DNA from sweet potato weevils, Cylas formicarius deteriorated in trap
2019
Kikukawa, K. (Naha Plant Protection Station, Okinawa (Japan)) | Arakaki, S. | Miyazaki, I. | Adachi, H. | Ishige, Y.
The sweet potato weevil, Cylas formicarius (Fabricius) (Coleoptera: Brentidae), is a serious pest of sweet potatoes. The sterile insect technique (SIT) is used for eradicating C. formicarius in Okinawa, Japan. In the eradication programs using SIT, marking methods are essential to distinguish sterile weevils from wild weevils; however, current methods have certain shortcomings and alternative methods have been in need of development. Previous research revealed a genetic difference between the population in the Ryukyu Islands and Ogasawara Islands, suggesting a possible use of the populations in Ogasawara as sterile insects in the Ryukyu Islands. For the introduction of this genetic marking method, we investigated the effective procedure to detect DNA from C. formicarius bodies that deteriorated after being left in harsh environmental conditions in the field trap for a few weeks. We could increase the detection rate of DNA extracted from deteriorated body from 47% in the conventional method to 100% by using Ampdirect Plus (Ampdirect) as the PCR amplification reagent to suppress the influence of PCR inhibitors. This result shows that DNA can be detected from C. formicarius bodies that have deteriorated in the field trap using Ampdirect.
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