Identification of reference genes for normalization of gene expression in Liposcelis entomophila (Psocoptera: Liposcelididae)

Involvement of Methoprene‐tolerant and Krüppel homolog 1 in juvenile hormone‐mediated vitellogenesis of female Liposcelis entomophila (End.) (Psocoptera: Liposcelididae)

“…The β‐actin gene of L. entomophila (GenBank accession no. MT603494.1) was used as a reference gene to normalize the target gene expression and to correct for sample‐to‐sample variation Specific qPCR primers were designed using the NCBI profile blast ( http://www.ncbi.nlm.nih.gov/tools/primer-blast ) for the experiment (Table 1 ) (S. Y. Miao et al, 2021 ). The reaction was performed in a 10 μl volume containing 0.4 μl of cDNA, 5 μl of 2 × TB Green™ Premix Ex Taq II (Tli RNaseH Plus) (Takara), 0.3 μl of each primer (10 mM), and 4 μl of RNase‐free H 2 O and tested on a LightCycler 96 instrument (Roche Diagnostics, Basel) using the following protocol: 95°C for 2 min (for predenaturation), 40 cycles of 95°C for 15 s, and 60°C for 30 s (for fragment amplification and signal collection, respectively), and 60°C–95°C with 0.5°C increasing per 5 s (for dissociation and melting curve analysis on the specific amplification).…”

“…The RNAi bioassay comprised a treatment group ( dsLeMet ‐fed or dsLeKrh1 ‐fed) and a control group ( dsGFP ‐fed). RNAi was performed as described (S. Y. Miao et al, 2021 ; Wei et al, 2020 ) and was further optimized as follows: (1) For each group, dsRNA dissolved in 200 μl of nuclease‐free water was added to a glass bottle (25 ml) containing 10 mg of sieved artificial diet. The final concentration of dsRNAs was 200 μg per mg of artificial diet.…”

“…RNAi was performed as described (S. Y. Miao et al, 2021;Wei et al, 2020) and was further optimized as follows:…”

“…This phenomenon has also been found in the chemical control of N. lugens (Wu, Ge, et al, 2020 ). Our previous study has shown that JH III significantly induced the expression of the vitellogenin ( Vg ) gene and affected the oviposition of L. entomophila (S. Y. Miao et al, 2021 ). Insect reproduction has been a focus of research in pest control, but relevant information is limited in L. entomophila .…”