Wu Taoyan, Zhao Ya, Wang Zhenying, Song Qisheng, Wang Zengxia, Xu Qiuwen, Wang Yingjuan, Wang Libao, Zhang Yiqiang, Feng Congjing
Department of Plant Protection, College of Horticulture and Plant Protection, Yangzhou University, Yangzhou, Jiangsu Province 225009, China.
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Dev Comp Immunol. 2018 Feb;79:31-43. doi: 10.1016/j.dci.2017.10.004. Epub 2017 Oct 12.
Pattern recognition receptors (PRRs) are biosensor proteins that bind to non-self pathogen associated molecular patterns (PAMPs). β-1,3-glucan recognition proteins (βGRPs) play an essential role in immune recognition and signaling pathway of insect innate immunity. Here, we report the cloning and characterization of cDNA of OfβGRP3 from Ostrinia furnacalis larvae. The OfβGRP3 contains 1455 bp open reading frame, encoding a predicted 484 amino acid residue protein. In hemocytes, the expression levels of OfβGRP3 in Escherichia coli-challenged group were higher than those of Bacillus subtilis-challenged group at 2, 4, 8, 10 and 12 h post injection (HPI). In fat body, OfβGRP3 expression in both B. subtilis and E. coli-challenged group was significantly higher than that in untreated group from 4 to 10 HPI, and then the expression continuously dropped from 12 to 36 HPI. The OfβGRP3 expression in laminarin-injected group was higher than that in lipopolysaccharides (LPS)-injected group in various test tissues from 4 to 24 HPI. The LT of E. coli-infected OfβGRP3-RNAi larvae (1.0 days) was significantly lower compared with that of E. coli infected wild-type larvae (3.0 days) (p < 0.01). Only 10.2% Sephadex G50 beads (degree 3) were completely melanized in the larvae inoculated with OfβGRP3 dsRNA, as compared to 48.8% in control larvae (p < 0.01). A notable reduction in the PO activity and IEARase activity in hemolymph was also detected in the OfβGRP3 knockdown larvae. Our study demonstrates that OfβGRP3 is one of PRR members involved the PPO-activating system in O. furnacalis larvae.
模式识别受体(PRRs)是一类与非自身病原体相关分子模式(PAMPs)结合的生物传感器蛋白。β-1,3-葡聚糖识别蛋白(βGRPs)在昆虫先天免疫的免疫识别和信号通路中发挥着重要作用。在此,我们报道了亚洲玉米螟幼虫OfβGRP3 cDNA的克隆与特征分析。OfβGRP3含有1455 bp的开放阅读框,编码一个预测的484个氨基酸残基的蛋白质。在血细胞中,注射大肠杆菌后2、4、8、10和12小时(HPI),大肠杆菌攻击组中OfβGRP3的表达水平高于枯草芽孢杆菌攻击组。在脂肪体中,枯草芽孢杆菌和大肠杆菌攻击组中OfβGRP3的表达在4至10 HPI时均显著高于未处理组,然后在12至36 HPI时表达持续下降。在4至24 HPI的各个测试组织中,注射海带多糖组中OfβGRP3的表达高于注射脂多糖(LPS)组。与大肠杆菌感染的野生型幼虫(3.0天)相比,大肠杆菌感染的OfβGRP3-RNAi幼虫的LT(1.0天)显著降低(p < 0.01)。接种OfβGRP3 dsRNA的幼虫中,只有10.2%的Sephadex G50珠(3级)完全黑化,而对照幼虫中为48.8%(p < 0.01)。在OfβGRP3敲低的幼虫中,还检测到血淋巴中PO活性和IEARase活性显著降低。我们的研究表明,OfβGRP3是参与亚洲玉米螟幼虫PPO激活系统的PRR成员之一。