CRISPR/Cas9-mediated tyrosine hydroxylase knockout in Ectropis grisescens results in defects in the melanization of the integument, excluding sclerotized appendages

文献类型： 外文期刊

第一作者： Li, Jia-li

作者： Li, Jia-li;Yuan, Ting-ting;Cai, Xiao-ming;Luo, Zong-xiu;Bian, Lei;Xiu, Chun-li;Fu, Nan-xia;Chen, Zong-mao;Li, Zhao-qun;Li, Jia-li;Liu, Nai-yong;Yuan, Ting-ting

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关键词： tyrosine hydroxylase; CRISPR; Cas9; sclerotization; pigmentation; melanin pathway

期刊名称：ENTOMOLOGIA GENERALIS （ 影响因子：6.608； 五年影响因子：4.803 ）

ISSN： 0171-8177

年卷期： 2022 年 42 卷 6 期

页码：

收录情况： SCI

摘要： Pigmentation is a variable trait of insects that help in adapting to the external natural environment. It plays an important role in concealment, mimicry, sex selection signals, and immune responses. Ectropis grisescens is the main lepidopteran pest in the tea gardens of China. The use of body color to simulate tea branches is a key characteristic of E. grisescens larvae that aids in predator evasion. However, no study has reported melanization in E. grisescens. In the pres-ent study, the E. grisescens tyrosine hydroxylase (EgTH) gene was cloned and characterized. The phylogenetic tree and stage expression profiles revealed that EgTH may be involved in the melanin pathway. CRISPR/Cas9-mediated targeted mutagenesis of EgTH was conducted to understand its functional role in pigmentation; it was revealed that the intensity of the black pigment systemically and proportionally decreased in the cuticle of the mutant larvae but did not disappear. Importantly, the mutation had no effect on the pigmentation of sclerotized tissues including the crochets, spiracles, claws, ocelli and mouth parts. This study provides the first evidence that CRISPR/Cas9-mediated EgTHknockout induces marked body pigmentation defects that can be visualized in the larvae. It was also demonstrated that another gene or pathway with a function similar to that of EgTH could be present. Additionally, EgTH might be not involved in the pigmentation of sclero-tized tissues in E. grisescens. Further, EgTH mutations do not lead to embryonic death in insect species. Thus, the present study provides new insights into the function of insect TH genes.

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