摘要
【目的】西花蓟马Frankliniella occidentalis (Pergande)是一种重要的外来入侵害虫,其繁殖能力强、寄主范围广、抗药性强,给我国蔬菜、花卉等经济作物造成了严重危害。本研究旨在评估膜饲喂和显微注射dsActin对西花蓟马Actin基因的沉默效率,以期为蓟马类等小型昆虫基因功能的研究提供方法和依据。【方法】通过膜饲喂和显微注射的方法将体外合成的dsActin导入西花蓟马二龄若虫体内,用RT-qPCR方法检测Actin基因的mRNA表达;通过单头饲养方法观察统计西花蓟马成虫体长、成虫翅膀或胸腹部畸形率及死亡率。【结果】dsActin膜饲喂后24、48、72 h,Actin基因的相对表达量分别为对照组的97%、91%和98%;通过显微注射将dsActin注入西花蓟马体腔,在注射后24、48、72 h,Actin基因的表达量分别为对照组的68%、56%和53%。注射dsActin的西花蓟马在第24~120 h死亡率为44%~98%,显著高于ds GFP对照组;此外,dsActin组个体体长仅为ds GFP对照组的90%,且翅膀或胸腹部出现畸形率为41%。【结论】显微注射dsActin能显著沉默西花蓟马Actin基因mRNA水平的表达,并引起西花蓟马个体不正常发育和死亡,从而建立蓟马类小型昆虫RNAi体系。
【Objective】 Application of ds Actin to control the proliferating, multiple-hosting, insecticide-resistant invasive western flower thrips on vegetables and ornamental crops in China was investigated.【Method】 Synthetic dsActin or ds GFP(as control) was introduced into the 2 nd-instar nymphs of Frankliniella occidentalis(Pergande) by either membrane-feeding or microinjection. The treated thrips were collected for mRNA expression detection by RT-qPCR at 24, 48 and 72 h after the treatment. Effects of 0.5 μg·μL-1 dsRNA on the body length, wing or thorax malformation, and mortality of the insects were monitored in an individual culture experiment conducted in the laboratory.【Result】 The relative expression of Actin mRNA in the thrips was similar to that of control 24, 48 and 72 h after membrane-feeding. But the expressions in the injection group decreased to 68% in 24 h, 56% in 48 h, and 53% in 72 h. Significantly higher mortality rate(44%-98% in 24-120 h), shorter body length(90% of control), and higher bodily deformation rate(41%) were observed in the dsRNA-treated thrips than control.【Conclusion】 The Actin mRNA expression in F. occidentalis was significantly suppressed by ds Actin injection at the test level to induce malformations or death in the insects. It suggested that an in vivo RNA interference method was made available for studying the gene functions and control of thrips.
作者
李恒
周知恩
陈勇
陆承聪
张祥琴
王谅
陈艺欣
田厚军
林硕
张洁
游泳
魏辉
LI Heng;ZHOUZhi-en;CHEN Yong;LU Cheng-cong;ZHANG Xiang-qin;WANGLiangu;CHENYi-xin;TIAN Hou-jun;LIN Shuo;ZHANG Jie;YOU Yong;WEI Hui(State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;Institute of Plant Protection,Fujian Academy of Agricultural Sciences/Fujian Key Laboratory far Monitoring and Integrated Management of Crop Pests/Fuzhou Scientific Observing and Experimental Station of Crop Pests,Ministry ofAgriculture and Rural Affairs,Fuzhou,Fujian 350003,China;Biotechnology and Germplasm Resource Institute,Yunnan Academy of Agricultural Sciences/Yunnan Provincial Key Laboratory of Agricultural Biotechnology/Key Laboratory far Gene Resources and Germplasm Innovation of Southwest Cropst Ministry of Agriculture and Rural Affairs,Kunming,Yunnan 650223,China)
出处
《福建农业学报》
CAS
CSCD
北大核心
2019年第10期1179-1184,共6页
Fujian Journal of Agricultural Sciences
基金
国家重点研发计划项目（2018YFD0201102、2018YFD0201209）
国家自然科学基金项目（31871936、31560499）
福建省科技重大专项（2017NZ0003-1-4）
福建省科技计划公益类专项（2017R1025-9、2018R1025-7、2019R1024-2、2019R1024-3）
福建省农业科学院科技项目（STIT2017-3-2、AGY2018-5、AC2017-6、YC2019003）
云南省应用基础研究计划面上项目（2016FB063）
云南省应用基础研究计划重点项目（2018FA020）
云南省中青年学术技术带头人后备人才项目(2015HB081)
关键词
西花蓟马
RNA干扰
膜饲喂法
显微注射法
Frankliniella occidentalis
RNA interference
membrane-feeding
microinjection
