Role of miR-276-3p in regulation of chitin metabolism in Aedes albopictus larvae

doi:10.11853/j.issn.1003.8280.2023.05.001

Abstract

Abstract: Objective To investigate the role of miR-276-3p in the chitin metabolism of Aedes albopictus larvae, so as to provide a new target for the control of Ae. albopictus using chitin metabolism-related miRNAs.Methods miR-276-3p-specific mimic agomir and inhibitor antagomir chitosan nanoparticles were synthesized to overexpress and suppress the expression of miR-276-3p in Ae. albopictus larvae through feeding, respectively. The relative expression of miR-276-3p in larvae was measured by reverse transcription quantitative real-time PCR. The chitin levels of larvae and pupae were determined using biochemical assay. The survival rate and pupation rate of larvae and the eclosion rate of pupae were calculated. Two-way ANOVA was used for mirR-276-3p expression differences analysis, T-test was used for chitin content comparison between different groups, and the log-rank test and Mantel-Cox test were used for survival rate difference analysis.Results miR-276-3p agomir chitosan nanoparticles effectively overexpressed miR-276-3p in Ae. albopictus larvae, the expression on the 1st and 3rd days increased by 73.96% (F=932.401, P<0.001) and 16.95% (F=114.551, P=0.047) respectively, while miR-276-3p antagomir chitosan nanoparticles significantly suppressed the expression of miR-276-3p in Ae. albopictus larvae, the expression on the 1st and 5th days decreased by 13.42% (F=293.632, P<0.001) and 1.49% (F=69.542, P=0.014) respectively. The overexpression of miR-276-3p increased the chitin content by 25.34% (t=85.838, P=0.019). The survival rate decreased significantly, and the difference between the experimental group and the control group on the 8th day was statistically significant (χ²=77.490, P=0.020), and the pupation rate was 70.00%, which was 90.00% and lower than that of the control group. The suppression of miR-276-3p reduced chitin levels in the larvae by 40.06% (t=71.811, P=0.028) and pupae by 46.82% (t=92.147, P=0.011) of Ae. albopictus.Conclusions miR-276-3p has significant effects on the chitin content,survival rate,and pupation rate of Ae. albopictus larvae. This study provides basic data for mosquito control by using miRNAs to regulate chitin metabolism in Ae. albopictus larvae.

Key words: Aedes albopictus, Agomir, Antagomir, miR-276-3p, Feed

摘要： 目的探讨miR-276-3p在白纹伊蚊幼虫几丁质代谢中的作用,为实现利用几丁质代谢相关miRNA防控白纹伊蚊提供新的靶标。方法合成miR-276-3p特异性模拟物agomir和抑制剂antagomir的壳聚糖纳米颗粒,通过饲喂法过表达或抑制白纹伊蚊幼虫体内miR-276-3p的表达。利用反转录实时荧光定量PCR（RT-qPCR）检测幼蚊体内miR-276-3p的相对表达量,利用生化分析法测定幼蚊及蛹的几丁质含量变化,并统计幼蚊存活率、化蛹率及蛹的羽化率。表达量差异比较使用two-way方差分析,几丁质含量两组间比较使用t检验,存活率差异分析采用乘积极限法的log-rank检验和Mantel-Cox检验。结果miR-276-3p agomir壳聚糖纳米颗粒可有效过表达白纹伊蚊幼虫体内miR-276-3p的表达量,第1和第3天后表达量较对照分别上调73.96%（F=932.401,P<0.001）、16.95%（F=114.551,P=0.047）；miR-276-3p antagomir壳聚糖纳米颗粒可有效抑制miR-276-3p的表达,第1和第5天后表达量较对照分别下调13.42%（F=293.632,P<0.001）和1.49%（F=69.542,P=0.014）；过表达miR-276-3p导致幼虫几丁质含量上升了25.34%（t=85.838,P=0.019）,幼虫存活率下降,第8天较对照组差异有统计学意义（χ²=77.490,P=0.020）,化蛹率为70.00%,低于对照组的90.00%；抑制miR-276-3p表达导致白纹伊蚊幼虫与蛹的几丁质含量分别下降了40.06%（t=71.811,P=0.028）和46.82%（t=92.147,P=0.011）。结论miR-276-3p对于白纹伊蚊幼虫的几丁质含量、存活率及化蛹率存在显著影响,相关研究结果为利用miRNAs调控白纹伊蚊幼虫的几丁质代谢,进而实现蚊虫防控提供了基础资料。

关键词: 白纹伊蚊, Agomir, Antagomir, miR-276-3p, 饲喂

CLC Number:

R384.1
Q965

FU Jing-wen, LIU Wen-juan, ZHANG Zhong, ZHANG Rui-ling. Role of miR-276-3p in regulation of chitin metabolism in Aedes albopictus larvae[J]. Chinese Journal of Vector Biology and Control, 2023, 34(5): 589-595.

傅菁文, 刘文娟, 张忠, 张瑞玲. miR-276-3p在白纹伊蚊幼虫几丁质代谢调控中的作用研究[J]. 中国媒介生物学及控制杂志, 2023, 34(5): 589-595.

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