Identification of Two Diamondback Moth Parasitoids, Diadegma fenestrale and Diadegma semiclausum, Using LAMP for Application in Biological Control

The diamondback moth, Plutella xylostella L., is a lepidopteran pest that damages various vegetable plants belonging to the genus Brassica worldwide. Various biological controls, such as parasitoid wasps, have been used control this pest. Among these, Diadegma semiclausum and Diadegmafenestrale are widely globally. In field-based research, investigation of population dynamics parasitoids rate parasitism within very important. However, achieving profundity in research difficult when morphologically similar species coexist field. morphological characteristics D. fenestrale similar, they both parasitize P. larvae. Therefore, accurately identify these species, study, we developed molecular diagnostic method by using loop-mediated isothermal amplification (LAMP). mitochondrial genome partial nucleotide sequences, including ITS region semiclausum, were analyzed for use diagnosis markers. results showed homology was 94%, due excessively low D loop, but actual higher than particularly coding region. species-specific primers LAMP designed based on encoding COX3, optimal reaction condition four (F3, B3, FIP, BIP) 63 °C 35 min. A primer capable classifying ITS2 region, 40 Under conditions upon addition loop LB, efficiency increased, time shortened more 5 limit concentration up 10 pg under conditions; therefore, it possible detect even concentrations. For assay with DNA-releasing technique, without DNA extraction process, incubating tissue sample or homogenized whole body at 95 case fenestrale, diagnose can be variety ways determine whether has parasitized process field mass breeding, distinguish parasitic This LAMP-based applied xylostella.