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        Molecular cloning, characterization, genomic structure and functional analysis of catalase in Chilo suppressalis

        Jing Xu,Ming-Xing Lu,Dong-lin Huang,Yu-zhou Dua 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.2

        Catalase (CAT), one of the key enzymes involved in protecting organisms from reactive oxygen species (ROS), degrades hydrogen peroxide to oxygen and water. In this study, we isolated the catalase gene (CsCAT) from the striped stemborer, Chilo suppressalis, a serious pest of rice in Asia. Sequence analysis of the genomic DNA indicated that CsCAT contains a 1023 bp intron and a 1524 bp coding sequence. The predicted CsCAT protein contains 507 amino acids and includes the proximal active site (FXRERIPERVVHAKGXGA) and heme-ligand sequence (RLFSYNDTX) that are conserved in catalase enzymes. Phylogenetic analysis indicated that CAT from Lepidopteran moths could be assigned to one well-supported cluster. qRT-PCR indicated that CsCAT was expressed in insect heads, epidermis, fat body, foregut, midgut, hindgut, Malpighian tubules, and hemocytes. Expression of CsCAT was highest in the fat body and hemocytes of diapausing larvae, which suggests a potential important role in insect physiology during diapause period. Expression of CsCAT (measured by qRT-PCR) and enzymatic activity of CsCAT were not significantly altered by temperature stress, which indicates that CsCAT may insensitive to temperature.

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        Identification of qRT-PCR reference genes for Chilo suppressalis (Walker) during parasitism by Cotesia chilonis (Matsumura)

        Zi-Lan Li,Dan-Dan Pan,Ming-Xing Lu,Yu-zhou Dua 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.1

        Quantitative real-time RT-PCR is highly sensitive approach for identifying and studying genes that function in various biochemical and cellular processes. Choosing the proper reference genes is a necessary step in ensuring the accuracy of results obtained with qRT-PCR. Herein, we evaluate the expression stability of nine potential reference genes in Chilo suppressalis parasitized by Cotesia chilonis. Stability was analyzed using the ΔCt method, geNorm, NormFinder, and BestKeeper, and our results show that the combination of TUB, EF1 and NADHD was optimal for normalizing expression. The transcription of target genes Hsp60 and CAT in C. suppressalis during parasitism was used to evaluate reference genes, and the results varied depending on the gene selected for normalization. Therefore, it is imperative to choose the proper reference genes to estimate target gene expression accurately in a given experiment. This study provides insights on gene expression in C. suppressalis and is especially relevant to further experiments that explore the effectiveness of C. chilonis in biological control.

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