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While deposition of hydrocarbons on insect cuticles as a key measure for water conservation has been extensively investigated, we know little about other mechanisms for preventing water loss in insects. Here, we report two fatty acid synthetic genes that are independent of hydrocarbon production but crucial for water retention in the German cockroach <jats:italic>Blattella germanica<\/jats:italic> (L.). First, an integument enriched fatty acid elongase gene (<jats:italic>BgElo1<\/jats:italic>) was identified as a critical gene for desiccation resistance in <jats:italic>B. germanica<\/jats:italic>; however, knockdown of <jats:italic>BgElo1<\/jats:italic> surprisingly failed to cause a decline in cuticular lipids. In addition, RNA interference (RNAi)\u2010knockdown of an upstream fatty acid synthase gene (<jats:italic>BgFas3<\/jats:italic>) showed a similar phenotype, and transmission electron microscopy analysis revealed that <jats:italic>BgFas3<\/jats:italic>\u2010 or <jats:italic>BgElo1<\/jats:italic>\u2010RNAi did not affect cuticle architecture. Bodyweight loss test showed that repression of <jats:italic>BgFas3<\/jats:italic> and <jats:italic>BgElo1<\/jats:italic> significantly increased the weight loss rate, but the difference disappeared when the respiration was closed by freeze killing the cockroaches. A water immersion test was performed, and we found that <jats:italic>BgFas3<\/jats:italic>\u2010 and <jats:italic>BgElo1<\/jats:italic>\u2010RNAi made it difficult for cockroaches to recover from drowning, which was supported by the upregulation of hypoxia\u2010related genes after a 10\u2010h recovery from drowning. Moreover, a dyeing assay with water\u2010soluble Eosin Y showed that this was caused by the entry of water into the respiratory system. Our research suggests that <jats:italic>BgFas3<\/jats:italic> and <jats:italic>BgElo1<\/jats:italic> are required for both inward and outward waterproofing of the respiratory system. This study benefits the understanding of water retention mechanisms in insects.<\/jats:p>","DOI":"10.1111\/1744-7917.12900","type":"journal-article","created":{"date-parts":[[2021,2,8]],"date-time":"2021-02-08T04:37:55Z","timestamp":1612759075000},"page":"33-50","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Two putative fatty acid synthetic genes of <i>BgFas3<\/i> and <i>BgElo1<\/i> are responsible for respiratory waterproofing in <i>Blattella germanica<\/i>"],"prefix":"10.1111","volume":"29","author":[{"given":"Xiao\u2010Jin","family":"Pei","sequence":"first","affiliation":[{"name":"State Key Laboratory of Crop Stress Biology for Arid Areas and Key Laboratory of Integrated Pest Management on Crops in Northwestern Loess Plateau, Ministry of Agriculture Northwest A&amp;F University  Yangling Shaanxi 712100 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