Li Ling, Zhang Wen-Bing, Shan Yan-Min, Zhang Zhuo-Ran, Pang Bao-Ping
Research Center for Grassland Entomology, Inner Mongolia Agricultural University, Hohhot, China.
Inner Mongolia Forestry and Grassland Pest Control and Quarantine Station, Hohhot, China.
Front Physiol. 2021 Jun 9;12:678698. doi: 10.3389/fphys.2021.678698. eCollection 2021.
Odorant-binding proteins (OBPs) and chemosensory proteins (CSPs) play a fundamental role in insect olfaction. (Joannis) is a new pest with outbreak status in the Inner Mongolia grasslands, northern China. In this study, six olfactory protein genes (, , , , , and ) were cloned by RACE and expressed by constructing a prokaryotic expression system. Their binding affinities to 13 compounds from host volatiles () were determined by fluorescence-binding assay. In order to further explore the olfactory functions of and , RNA interference (RNAi) and electroantennogram (EAG) experiments were conducted. Ligand-binding assays showed that the binding properties of the six recombinant proteins to the tested volatiles were different. GdauOBP6, GdauOBP15, GdauCSP4, and GdauCSP5 could bind several tested ligands of host plants. It was suspected that GdauOBP6, GdauOBP15, GdauCSP4, and GdauCSP5 were related to the host location in We also found that there were different EAG responses between males and females when the and genes were silenced by RNAi. The EAG response of females to 2-hexenal was significantly decreased in dsRNA-OBP15-injected treatment compared to the control, and the dsRNA-CSP5-treated females significantly reduced EAG response to eight tested host volatiles (1,3-dithiane, 2-hexenal, methyl benzoate, dimethyl trisulfide, myrcene, hexanal, 1,3,5-cycloheptatriene, and p-xylene). However, the EAG response had no significant difference in males. Both GdauOBP15 and GdauCSP5 may have different functions between males and females in and may play more important roles in females searching for host plants.
气味结合蛋白(OBPs)和化学感受蛋白(CSPs)在昆虫嗅觉中起着重要作用。(乔阿尼斯)是中国北方内蒙古草原上具有爆发态势的一种新害虫。在本研究中,通过RACE克隆了六个嗅觉蛋白基因(,,,,,和),并构建原核表达系统进行表达。通过荧光结合测定法确定了它们与寄主挥发物中的13种化合物()的结合亲和力。为了进一步探究和的嗅觉功能,进行了RNA干扰(RNAi)和触角电位(EAG)实验。配体结合测定表明,六种重组蛋白与测试挥发物的结合特性不同。GdauOBP6、GdauOBP15、GdauCSP4和GdauCSP5可以结合几种寄主植物的测试配体。推测GdauOBP6、GdauOBP15、GdauCSP4和GdauCSP5与(某种昆虫)寻找寄主有关。我们还发现,当通过RNAi沉默和基因时,雌雄虫之间存在不同的EAG反应。与对照相比,注射dsRNA-OBP15处理的雌虫对2-己烯醛的EAG反应显著降低,dsRNA-CSP5处理的雌虫对八种测试寄主挥发物(1,3-二硫戊环、2-己烯醛、苯甲酸甲酯、二甲基三硫醚、月桂烯、己醛、1,3,5-环庚三烯和对二甲苯)的EAG反应显著降低。然而,雄虫的EAG反应没有显著差异。GdauOBP15和GdauCSP5在(某种昆虫)的雌雄虫之间可能具有不同功能,并且可能在雌虫寻找寄主植物中发挥更重要的作用。