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Although the functional importance of genes in EPFs' pathogenesis have been extensively studied, the precise ultrastructural mechanism of the fungal infection, particularly penetration of the host insect cuticles, is not well understood.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>In this study, we investigated the morphology and ultrastructure of the larval cuticle of the red flour beetle, <jats:italic>Tribolium castaneum<\/jats:italic>, after treatment with <jats:italic>B. bassiana<\/jats:italic> ERL1170 expressing an enhanced green fluorescent protein (<jats:italic>Bb<\/jats:italic>\u2010eGFP). The <jats:italic>Bb<\/jats:italic>\u2010eGFP showed high virulence against the larvae, with approximately 90% mortality at 48\u2009h after treatment (HAT) and 100% at 72 HAT under our infection conditions. In these larvae, the regions of the body wall with flexible cuticles, such as the ventral and ventrolateral thorax and abdomen, became darkly melanized, but there was little to no melanization in the rigid dorsal cuticular structures. Confocal microscopy and transmission electron microscopy (TEM) indicated that germinated conidia on the surface of the larval cuticle were evident at 6 HAT, which formed penetration pegs and began to penetrate the several cuticle layers\/laminae by 12 HAT. The penetration pegs then developed invading hyphae, some of which passed through the cuticle and reached the epidermal cells by 24 HAT. The larval cuticle was aggressively and extensively disrupted by 48 HAT, and a number of outgrowing hyphae were observed at 72 HAT.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSIONS<\/jats:title><jats:p>Our results indicate that <jats:italic>Bb<\/jats:italic>\u2010eGFP is capable of infection and penetrating <jats:italic>T. castaneum<\/jats:italic> larvae shortly after inoculation (~24 HAT) at the body regions with apparently flexible and membranous cuticles, such as the ventral intersegmental regions and the ventrolateral pleura. This study provides details on the histopathogenesis of the host cuticle by infection and penetration of EPFs, which can facilitate the management of insect pests. \u00a9 2022 Society of Chemical Industry.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ps.6962","type":"journal-article","created":{"date-parts":[[2022,5,5]],"date-time":"2022-05-05T01:55:31Z","timestamp":1651715731000},"page":"3356-3364","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Ultrastructural analysis of beetle larva cuticles during infection with the entomopathogenic fungus, <i>Beauveria bassiana<\/i>"],"prefix":"10.1002","volume":"78","author":[{"given":"Sehyeon","family":"Baek","sequence":"first","affiliation":[{"name":"Department of Agricultural Biology Jeonbuk National University  Jeonju South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mi Young","family":"Noh","sequence":"additional","affiliation":[{"name":"Department 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