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豌豆蚜防控的关键RNAi靶标基因筛选与分析

引用本文：宗诗轩,李涛,黄清,程保平,赵弘巍.豌豆蚜防控的关键RNAi靶标基因筛选与分析.植物保护学报,2023,50(5):1280-1288

DOI：10.13802/j.cnki.zwbhxb.2023.2022062

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作者	单位	E-mail
宗诗轩	南京农业大学植物保护学院, 南京 210095
李涛	南京农业大学植物保护学院, 南京 210095
黄清	南京农业大学植物保护学院, 南京 210095
程保平	广东省农业科学院植物保护研究所, 广东省植物保护新技术重点实验室, 农业农村部华南果蔬绿色防控重点实验室, 广州 510640	chengbaoping@gdaas.cn
赵弘巍	南京农业大学植物保护学院, 南京 210095	hzhao@njau.edu.cn

中文摘要:为筛选防控豌豆蚜Acyrthosiphon pisum的关键RNA干扰（RNA interference，RNAi）靶标基因，通过构建精氨酸激酶（arginine kinase，AK）、超氧化物歧化酶（superoxide dismutase，SOD）和几丁质合成酶（chitin synthase，CHS）这3个与豌豆蚜生长发育相关基因的dsRNA，于室内检测dsAK、dsSOD和dsCHS处理后对其致死效果，并比较4种dsRNA递送方式对其致死效果的影响，筛选获得防治效果明显的dsRNA及其递送方式。结果显示，处理48~168 h后，不同基因dsRNA处理组豌豆蚜的死亡率比阴性对照组高2.19倍~4.39倍，其中dsAK处理组豌豆蚜的死亡率为27.7%~91.1%，显著高于dsSOD、dsCHS处理组及阴性对照组。使用dsAK通过人工饲喂法和叶片注射饲喂法处理豌豆蚜，48 h即可出现显著致死效果。dsAK通过人工饲喂方法处理72 h后，豌豆蚜AK基因表达量比空白对照组降低43.2%，同时可抑制豌豆蚜的生长发育，使其体长比阴性对照组减少了5.9%。表明dsAK对豌豆蚜有较好的致死效果，在蚜虫的绿色防控方面有很好的应用前景。

中文关键词:豌豆蚜 RNA干扰精氨酸激酶超氧化物歧化酶几丁质合成酶 dsRNA 柑橘

Screening and analysis of the pivotal RNAi target genes for the control of pea aphid Acyrthosiphon pisum

Author Name	Affiliation	E-mail
Zong Shixuan	College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China
Li Tao	College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China
Huang Qing	College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China
Cheng Baoping	Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China, Ministry of Agriculture and Rural Affairs Guangdong Provincial Key Laboratory of High Technology for Plant Protection Plant Protection Research Institute, GuangdongAcademy ofAgricultural Sciences, Guangzhou 510640, Guangdong Provice, China	chengbaoping@gdaas.cn
Zhao Hongwei	College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China	hzhao@njau.edu.cn

Abstract:To screen for the key RNA interference (RNAi) target genes for controlling pea aphid Acyrthosiphon pisum, the dsRNAs targeting three genes associated with the growth and development of A. pisum, namely arginine kinase (AK), superoxide dismutase (SOD), and chitin synthase (CHS), were constructed. By comparing the lethal effects of different dsRNA (dsAK, dsSOD, and dsCHS) treatments on A. pisum, and the effects of four different dsRNA delivery methods on the lethal effects of aphids, an optimized A. pisum control method was proposed. The results showed that after 48-168 h, the mortality rate of aphids in the treatment group was 2.19-4.39 times higher compared to the negative control group. The lethal rate of dsAK to aphids was 27.7%-91.13%, significantly higher than that those of dsSOD, dsCHS treatment groups and the negative control group. Significant mortality effects on A. pisum could be observed within 48 h after dsAK treatment through artificial diet feeding or feeding with leaves injected with dsRNA solution. At 72 h after treatment with dsAK through artificial diet feeding, the expression of AK gene in A. pisum was reduced by 43.2% compared with blank control, while it also suppressed the growth and development of aphids, resulting in a 5.9% decrease in body length compared to the negative control. The results indicated that dsAK exhibited a significant lethal effect on pea aphids, showing promising prospects for its application in environment-friendly aphid prevention and control.

keywords:Acyrthosiphon pisum RNAi arginine kinase superoxide dismutase chitin synthase dsRNA citrus

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