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CRISPR/Cas9 介导的 LW-opsin 基因敲除降低了小菜蛾的趋光性。

CRISPR/Cas9-mediated knockout of LW-opsin reduces the efficiency of phototaxis in the diamondback moth Plutella xylostella.

机构信息

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.

出版信息

Pest Manag Sci. 2021 Jul;77(7):3519-3528. doi: 10.1002/ps.6405. Epub 2021 May 12.

Abstract

BACKGROUND

Opsins are crucial for animal vision. The identity and function of opsins in Plutella xylostella remain unknown. The aim of the research is to confirm which opsin gene(s) contribute to phototaxis of P. xylostella.

RESULTS

LW-opsin, BL-opsin and UV-opsin, were identified in the P. xylostella genome. LW-opsin was more highly expressed than the other two opsin genes, and all three genes were specifically expressed in the head. Three P. xylostella strains, LW-13 with a 13-bp deletion in LW-opsin, BL + 2 with a 2-bp insertion in BL-opsin, and UV-29 with a 5-bp insertion and a 34-bp deletion in UV-opsin, were established from the strain G88 using the CRISPR/Cas9 system. Among the three opsin-knockout strains, only male and female LW-13 exhibited weaker phototaxis to lights of different wavelengths and white light than G88 at 2.5 lx due to defective locomotion, and LW-13 was defective to sense white, green and infrared lights. The locomotion of LW-13 was reduced compared with G88 at 2.5, 10, 20, 60, 80, 100, and 200 lx under the green light, but the locomotion of LW-13 female was recovered at 80, 100 and 200 lx. The defective phototaxis to the green light of male LW-13 was not affected by light intensity, while the defective phototaxis to the green light of female LW-13 was recovered at 10, 20, 60, 80, 100, and 200 lx.

CONCLUSION

LW-opsin is involved in light sensing and locomotion of P. xylostella, providing a potential target gene for controlling the pest. © 2021 Society of Chemical Industry.

摘要

背景

视蛋白对视动物的视觉至关重要。小菜蛾的视蛋白的身份和功能尚不清楚。本研究旨在确定哪些视蛋白基因参与小菜蛾的趋光性。

结果

在小菜蛾基因组中鉴定出 LW-opsin、BL-opsin 和 UV-opsin。LW-opsin 的表达水平高于其他两种视蛋白基因,并且这三种基因都在头部特异性表达。通过 CRISPR/Cas9 系统从小菜蛾 G88 菌株中建立了三种小菜蛾品系:LW-13 品系在 LW-opsin 中缺失 13 个碱基对,BL+2 品系在 BL-opsin 中插入 2 个碱基对,UV-29 品系在 UV-opsin 中插入 5 个碱基对和缺失 34 个碱基对。在这三种视蛋白敲除品系中,只有雄性和雌性 LW-13 在 2.5 lx 时对不同波长的光和白光的趋光性比 G88 弱,这是由于运动能力受损,并且 LW-13 无法感知白光、绿光和红外线。在绿光下,LW-13 的运动能力与 G88 相比,在 2.5、10、20、60、80、100 和 200 lx 时降低,但 LW-13 雌虫在 80、100 和 200 lx 时恢复。雄性 LW-13 对绿光的趋光性缺陷不受光强度的影响,而雌性 LW-13 对绿光的趋光性缺陷在 10、20、60、80、100 和 200 lx 时恢复。

结论

LW-opsin 参与了小菜蛾的光感和运动,为控制害虫提供了一个潜在的靶基因。 © 2021 化学工业协会。

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