Cuticle is not only critical for protecting insects from noxious stimuli but is also involved in a variety of metabolic activities.Cuticular proteins(CPs)affect cuticle structure and mechanical properties during insect growth,reproduction,and environmen-tal adaptation.Here,we describe the identification and characterization of a member of the RR-1 subfamily of CPs with an R&R consensus(CPR)in Tribolium castaneum(Tc-CPR69).Although it was previously reported to be highly expressed in the wings,we found that knocking down TcCPR69 by RNA interference(RNAi)did not cause obvi-ous wing abnormalities but markedly disrupted the growth and metamorphosis of beetles with 100％cumulative mortality;additionally,the chitin content of the pharate adult was decreased and the new abdominal cuticle was significantly thinner before molting.Tc-CPR69 showed chitin-binding ability and the expression levels of key genes involved in chitin metabolism(trehalase[TcTRE],chitin synthase[TcCHSA and TcCHSB],and chiti-nase[TcCHT5 and TcCHT10])were also decreased by TcCPR69 knockdown.TcCPR69 gene expression peaked shortly after molting and was increased 2.61 fold at 12 h after 20-hydroxyecdysone(20E)injection.This was reversed by RNAi of the ecdysone-related genes ecdysone receptor(TcECR)and fushi tarazu transcription factor 1(TcFTZ-F1).These results indicate that TcCPR69 is positively regulated by 20E signaling to contribute to cuticle formation and maintain chitin accumulation during the growth and metamor-phosis of beetles.

Jia Xie;Guifang Peng;Miao Wang;Qisheng Zhong;Xiaowen Song;Jingxiu Bi;Jing Tang;Fan Feng;Han Gao;Bin Li

Jiangsu Key Laboratory of Biodiversity and Biotechnology,College of Life Sciences,Nanjing Normal University,Nanjing,China;Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests,Ministry of Education,College of Plant Protection,Hainan University,Haikou,China

中国昆虫科学（英文版）

[1]Jia Xie;Guifang Peng;Miao Wang;Qisheng Zhong;Xiaowen Song;Jingxiu Bi;Jing Tang;Fan Feng;Han Gao;Bin Li-.RR-1 cuticular protein TcCPR69 is required for growth and metamorphosis in Tribolium castaneum)[J].中国昆虫科学（英文版）,2022(06):1612-1628

TcCPR69,Cuticle,CPR69,pharate,TcTRE,TcCHSA,TcCHSB,chiti,TcCHT5,TcCHT10,TcECR,fushi,tarazu,TcFTZ,phosis

RR,cuticular,required,growth,metamorphosis,Tribolium,castaneum,not,only,critical,protecting,insects,from,noxious,stimuli,also,involved,variety,metabolic,activities,Cuticular,proteins,CPs,affect,cuticle,structure,mechanical,properties,during,reproduction,environmen,adaptation,Here,describe,identification,characterization,member,subfamily,consensus,Although,was,previously,reported,highly,expressed,wings,found,that,knocking,by,interference,RNAi,did,cause,obvi,abnormalities,markedly,disrupted,beetles,cumulative,mortality,additionally,chitin,content,adult,decreased,new,abdominal,significantly,thinner,before,molting,showed,binding,ability,expression,levels,key,genes,metabolism,trehalase,synthase,nase,were,knockdown,peaked,shortly,after,increased,fold,hydroxyecdysone,20E,injection,This,reversed,related,receptor,transcription,These,results,indicate,positively,regulated,signaling,contribute,formation,maintain,accumulation

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