State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, China.
Joint International Research Laboratory of Ecological Pest Control, Ministry of Education, Fuzhou, China.
Insect Sci. 2023 Aug;30(4):1035-1045. doi: 10.1111/1744-7917.13150. Epub 2022 Dec 26.
Cryptochrome 1 (CRY1) functions as a light-responsive photoreceptor, which is crucial for circadian rhythms. The identity and function of CRY1 in Plutella xylostella remain unknown. In this study, cry1 was cloned and identified in P. xylostella. Then, a cry1-knockout strain (Cry1-KO) of P. xylostella with a 2-bp deletion was established from the strain Geneva 88 (G88) using the CRISPR/Cas9 technology. No daily temporal oscillation of cry1 was observed in G88 and Cry1-KO, and cry1 mean daily transcription of Cry1-KO was lower than that of G88. Both G88 and Cry1-KO demonstrated rhythmic locomotion under the light/dark condition with Cry1-KO being more active than G88 in the daytime, whereas Cry1-KO completely lost rhythmicity under constant darkness. The developmental period of pre-adult of Cry1-KO was longer than that of G88; the lifespan of the Cry1-KO male adult was shorter than that of G88; the fecundity of Cry1-KO was lower than that of G88; and Cry1-KO showed lower intrinsic rate of increase (r), net reproduction rate (R ), finite increase rate (λ), and longer mean generation time (T) than G88. Our results indicate that cry1 is involved in the regulation of locomotor circadian rhythm and development in P. xylostella, providing a potential target gene for controlling the pest and a basis for further investigation on circadian rhythms in lepidopterans.
CRY1（隐花色素 1）作为一种光响应光受体，对昼夜节律至关重要。CRY1 在小菜蛾中的身份和功能尚不清楚。本研究在小菜蛾中克隆并鉴定了 cry1。然后，使用 CRISPR/Cas9 技术从小麦蛾品系 Geneva 88（G88）中建立了一个带有 2 个碱基缺失的 cry1 敲除株系（Cry1-KO）。在 G88 和 Cry1-KO 中未观察到 cry1 的每日时间振荡，并且 Cry1-KO 的 cry1 平均每日转录水平低于 G88。在光/暗条件下，G88 和 Cry1-KO 均表现出节律性运动，但 Cry1-KO 在白天比 G88 更活跃，而 Cry1-KO 在持续黑暗下完全失去节律性。Cry1-KO 的预成虫发育期长于 G88；Cry1-KO 雄性成虫的寿命短于 G88；Cry1-KO 的繁殖力低于 G88；Cry1-KO 的内禀增长率（r）、净生殖率（R）、有限增长率（λ）和平均世代时间（T）均低于 G88。我们的结果表明，cry1 参与了小菜蛾运动昼夜节律和发育的调节，为控制害虫提供了一个潜在的靶基因，并为进一步研究鳞翅目昼夜节律奠定了基础。
