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        Selection and evaluation of reference genes for quantitative real‐time PCR analysis in lac insect (Kerria lacca)

        Wang Weiwei,Ling Xiaofei,Bashir Nawaz Haider,Lu Qin,Zhang Jinwen,Li Tichu,Chen Hang 한국곤충학회 2022 Entomological Research Vol.52 No.2

        The screening and identification of suitable reference genes are key steps in experiments involving the quantitative real-time polymerase chain reaction. However, the reference genes of Kerria lacca, a lac insect with considerable economic significance, have not been investigated. In this study, we used five statistical algorithms (ΔCt, geNorm, NormFinder, BestKeeper, and RefFinder) to evaluate the stability of eight candidate reference genes (18S rRNA, 28S rRNA, β-ACT, EF1-α, RPL10, RPL13, RPS11,andRPS15) in different developmental stages and tissues of K. lacca and under dsRNA treatment. Our results showed that the most stable reference genes were 28S rRNA and RPS11 for the developmental stages, RPL13 and RPS11 for different tissues, and β-ACT and EF1-α for dsRNA treatments. Although the target genes FA S and 17β-HSD showed consistent expression profiles (alone or in combination) when normalized to the expression levels of the stable genes, those changed when the least stable reference gene (18S rRNA) was used. These results highlight that suitable reference genes should be chosen according to the specific experimental conditions used. The reliability and stability of different reference genes can provide a firm foundation for gene expression analysis in lac insects.

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        Metagenomic analysis suggests unique gut‐microbiota composition and GH family constitution of Macrotermes annandalei

        Cui Yongzhong,Wang Weiwei,Lu Qin,Wu Hai‐Xia,Ling Xiaofei,Liu Juan,Zhang Jinwen,Chen Hang 한국곤충학회 2022 Entomological Research Vol.52 No.3

        Energy supply is a major concern of human survival and development, and that concern demands new sources of renewable clean energy. Cellulose, as a renewable energy material is affected by low activity and high cost of cellulose. Studies of cellulose degrading bacterial microbiota in insect intestine have revealed that these potential microbes can be used as a rich resource of cellulose degrading strains. Here, the present study involves metagenomic sequencing of intestinal microbes of Macrotermes annandalei by Illumina PE150 and a total of 401,190 scaftigs were obtained. Open Reading frame (ORF) species annotation showed that the largest proportion of them belong to the phylum of Bacteroides, followed by Protobacteia, Firmicutes and Mucoromycota. Mapping to the CAZy database indicates that 4,539 genes encoding of glycoside hydrolase belongs to 109 distinct CAZy families. The results provide a theoretical basis for exploring new candidate bacteria for efficient cellulose degradation to obtain high cellulose resource yield.

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