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Liu Quan-quan,Zhang Chen,Zhou Jin-cheng,Dong Qian-jin,Huo Liang-xiao,Dong Hui 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.2
Ploidy diversity provides valuable scientific information, thus making the detection technique of ploidy important. However, traditional methods of cytological observation and flow cytometry are either laborious or expensive. We here report a simple and rapid, effective and economical quantitative PCR (qPCR) approach to determine the ploidy of a parasitoid species Trichogramma dendrolimi Matsumura, an economically important biocontrol agent. We applied a mitochondrial gene cytochrome oxidase (COI) and a nuclear gene forkhead to evaluate the mitochondrial number per nuclear copy in a thelytokous Wolbachia-infected strain of T. dendrolimi and its bisexual uninfected counterparts. The 2 −ΔCq values calculated from C q values which resulted from qPCR experiments were significantly larger in haploid males than that in diploid females. Haploid males possessed about 2.69 times mitochondrial number per nuclear copy as diploid females. Not a single significant difference was found between diploid females from thelytokous and bisexual strains. Based on the differences in relative mitochondrial content, we were allowed to distinguish between haploid males and diploid females. Moreover, the number of mitochondria significantly decreased with higher ploidy level but was not affected by Wolbachiainfection. Our study supplied an available tool to investigate the ploidy diversity in sex determination of T. dendrolimi and thelytokous manipulation of Wolbachia, which is the crucial step to further study their underlying mechanisms. This will in turn contribute to the biocontrol efficiency by enhancing the female production and hence the parasitism rate.