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酚氧化酶激活机制对亚洲玉米螟对苏云金芽孢杆菌杀虫蛋白抗性的贡献。

Contribution of phenoloxidase activation mechanism to Bt insecticidal protein resistance in Asian corn borer.

机构信息

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, PR China.

Institute of Plant Protection, Gansu Academy of Agricultural Sciences, Lanzhou 730070, PR China.

出版信息

Int J Biol Macromol. 2020 Jun 15;153:88-99. doi: 10.1016/j.ijbiomac.2020.03.003. Epub 2020 Mar 3.

Abstract

Phenoloxidase (PO) is a crucial enzyme in the Arthropods melanization process, in which synthesized melanin rapidly acts at the site of injury and infection. In this study, we observed significant changes in humoral and cellular responses after exposing susceptible and resistant strains to a sub-lethal concentration of Cry1Ah toxin. Based on STRING v 11.0 computational protein-protein interaction analysis, we selected seven immune genes namely Prophenoloxidase PPO1b, PP03, Serpin-3, Serpin-5, Beta-1,3-glucan recognition protein, Immulectin-3 and Serine protease SP105 reported in Asian corn borer. Quantitative real-time PCR gene expression studies showed Cry1Ah resistant strain had higher expression of PPO1b, PP03, Serpin-3, Beta-1,3-glucan recognition protein, Immulectin-3 and Serine protease SP105 genes in midgut and hemocyte samples. This study also investigated and found that the level of prophenoloxidation (proPO) activity in Cry1Ah resistant strains was significantly higher than susceptible strains. Cry1Ah toxin significantly increased the resistant strain's immune responses, the difference was observed through assays of bacterial agglutination and phagocytosis. Additionally, immune response induced by Cry1Ah toxin influences the microbiome composition associated with the host system. These parameters seem to explain the contribution of PO/PO regulating proteins render the host to resist the Cry1Ah toxin.

摘要

酚氧化酶(PO)是节肢动物黑化过程中的关键酶,在该过程中合成的黑色素迅速作用于损伤和感染部位。在这项研究中,我们观察到易感和抗性品系暴露于亚致死浓度 Cry1Ah 毒素后,体液和细胞反应发生了显著变化。基于 STRING v 11.0 计算的蛋白质-蛋白质相互作用分析,我们选择了 7 种免疫基因,即亚洲玉米螟中的原酚氧化酶 PPO1b、PP03、丝氨酸蛋白酶抑制剂-3、丝氨酸蛋白酶抑制剂-5、β-1,3-葡聚糖识别蛋白、免疫球蛋白-3 和丝氨酸蛋白酶 SP105。定量实时 PCR 基因表达研究表明,Cry1Ah 抗性品系中 PPO1b、PP03、丝氨酸蛋白酶抑制剂-3、β-1,3-葡聚糖识别蛋白、免疫球蛋白-3 和丝氨酸蛋白酶 SP105 基因在中肠和血细胞样本中的表达更高。本研究还发现,Cry1Ah 抗性品系的原酚氧化酶(proPO)活性水平明显高于易感品系。Cry1Ah 毒素显著增加了抗性品系的免疫反应,通过细菌凝集和吞噬作用的测定可以观察到这种差异。此外,Cry1Ah 毒素诱导的免疫反应会影响与宿主系统相关的微生物组组成。这些参数似乎解释了 PO/PO 调节蛋白的作用,使宿主能够抵抗 Cry1Ah 毒素。

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