Function analysis and characterisation of a novel chitinase, MdCht9, in Musca domestica

Insect chitinases have been proposed as potential targets for pest control. In this work, a novel group IV chitinase gene, MdCht9, from Musca domestica was found to have multiple functions in the physiological activity, including chitin regulation, development and antifungal immunity. The MdCht9 gen...

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Bibliographic Details
Published in:Insect molecular biology 2024-04, Vol.33 (2), p.157-172
Main Authors: Zhao, Wen‐Jing, Li, Yan, Jiao, Zhen‐Long, Su, Pei‐Pei, Yang, Long‐Bing, Sun, Chao‐Qin, Xiu, Jiang‐Fan, Shang, Xiao‐Li, Guo, Guo
Format: Article
Language:English
Subjects:
Animals
anti‐microbial activity
Biological activity
Chitin
Chitin - metabolism
Chitinase
Chitinases - metabolism
E coli
Function analysis
Fungicides
Gene expression
Houseflies - genetics
Houseflies - metabolism
Insects
Larva
Larvae
Lethality
Microbial activity
Microorganisms
Musca domestica
Pest control
Pests
Phylogeny
Proteins
Recombinant Proteins - genetics
RNA-mediated interference
Salivary gland
Salivary glands
transcriptome
Transcriptomes
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Summary:Insect chitinases have been proposed as potential targets for pest control. In this work, a novel group IV chitinase gene, MdCht9, from Musca domestica was found to have multiple functions in the physiological activity, including chitin regulation, development and antifungal immunity. The MdCht9 gene was cloned and sequenced, its phylogeny was analysed and its expression was determined in normal and 20E treated larvae. Subsequently, RNA interference (RNAi)‐mediated MdCht9 knockdown was performed, followed by biochemical assays, morphological observations and transcriptome analysis. Finally, the recombinant protein MdCht9 (rMdCht9) was purified and tested for anti‐microbial activity and enzyme characteristics. The results showed that MdCht9 consists of three domains, highly expressed in a larval salivary gland. RNAi silencing of MdCht9 resulted in significant down‐regulation of chitin content and expression of 15 chitin‐binding protein (CBP) genes, implying a new insight that MdCht9 might regulate chitin content by influencing the expression of CBPs. In addition, more than half of the lethality and partial wing deformity appeared due to the dsMdCht9 treatment. In addition, the rMdCht9 exhibited anti‐microbial activity towards Candida albicans (fungus) but not towards Escherichia coli (G−) or Staphylococcus aureus (G+). Our work expands on previous studies of chitinase while providing a potential target for pest management. MdCht9 consists of three domains, highly expressed in larval salivary gland. MdCht9 might regulate the chitin content by influencing the expression of chitin‐binding proteins as well as degrading chitin. MdCht9 played multiple roles in housefly development, digestion and immunity processes.
ISSN:0962-1075
1365-2583
DOI:10.1111/imb.12887