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家蚕载脂蛋白-III 直接和通过调控与免疫信号通路相关基因的表达来抑制球孢白僵菌。

Bombyx mori Apolipophorin-III inhibits Beauveria bassiana directly and through regulating expression of genes relevant to immune signaling pathways.

机构信息

School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212100, China.

School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212100, China.

出版信息

J Invertebr Pathol. 2021 Sep;184:107647. doi: 10.1016/j.jip.2021.107647. Epub 2021 Jul 23.

Abstract

Insect Apolipophorin-III is a multifunctional protein and also plays an important role in insect innate immunity. Early transcriptome and proteome studies indicated that the gene expression level of Bombyx mori Apolipophorin-III (BmApoLp-III) in silkworm larvae infected with Beauveria bassiana was significantly up-regulated. In this study, BmApoLp-III gene was cloned, its expression patterns in different larval tissues investigated, the BmApoLp-III protein was successfully expressed with prokaryotic expression system and its antifungal effect was verified. The results showed that the BmApoLp-III gene was expressed in all the tested tissues of the 5th instar larvae infected by B. bassiana, with the highest expression in fat body. The fungistatic zone test showed that the recombinant BmApoLp-III has a significant antifungal effect on B. bassiana. Injecting purified BmApoLp-III to the larvae delayed the onset and death of the infected larvae. Conversely, silencing BmApoLp-III gene by RNAi resulted in early morbidity and death of the infected larvae. At the same time, injecting BmApoLp-III up-regulated the expression of genes including BmβGRP4 and BmMyd88 in the Toll signaling pathway, BmCTL5 and BmHOP in the Jak/STAT signaling pathway, serine proteinase inhibitor BmSerpin5, and antimicrobial peptide BmCecA, but down-regulated the expression of BmTak1 of Imd signaling pathway; while silencing BmApoLp-III gene down-regulated the expression of BmβGRP1 and BmSpaetzle, BmCTL5 and BmHOP, BmSerpin2 and BmSerpin5, BmBAEE and BmPPO2 of relevant pathways and BmCecA, but up-regulated the expression of BmPGRP-Lc and BmTak1 of Imd pathway. These results indicate that the BmApoLp-III could not only directly inhibit B. bassiana, but also participate in regulation of the expression of immune signaling pathway related genes, promote the expression of immune effectors, and indirectly inhibit the reproduction of B. bassiana in the silkworm.

摘要

昆虫载脂蛋白-III 是一种多功能蛋白,在昆虫先天免疫中也发挥着重要作用。早期的转录组和蛋白质组研究表明,感染白僵菌的家蚕幼虫中,Bombyx mori 载脂蛋白-III(BmApoLp-III)的基因表达水平显著上调。本研究克隆了 BmApoLp-III 基因,研究了其在不同幼虫组织中的表达模式,成功地通过原核表达系统表达了 BmApoLp-III 蛋白,并验证了其抗真菌作用。结果表明,BmApoLp-III 基因在家蚕 5 龄幼虫感染白僵菌后所有检测组织中均有表达,在脂肪体中表达量最高。抑菌圈试验表明,重组 BmApoLp-III 对 B. bassiana 具有显著的抗真菌作用。向幼虫体内注射纯化的 BmApoLp-III 可延迟感染幼虫的发病和死亡。相反,通过 RNAi 沉默 BmApoLp-III 基因会导致感染幼虫发病和死亡提前。同时,向幼虫体内注射 BmApoLp-III 可上调 Toll 信号通路中 BmβGRP4 和 BmMyd88 基因、Jak/STAT 信号通路中 BmCTL5 和 BmHOP 基因、丝氨酸蛋白酶抑制剂 BmSerpin5 和抗菌肽 BmCecA 的表达,但下调 Imd 信号通路中 BmTak1 的表达;而沉默 BmApoLp-III 基因则下调了相关通路中 BmβGRP1 和 BmSpaetzle、BmCTL5 和 BmHOP、BmSerpin2 和 BmSerpin5、BmBAEE 和 BmPPO2 以及 BmCecA 的表达,但上调了 Imd 通路中 BmPGRP-Lc 和 BmTak1 的表达。这些结果表明,BmApoLp-III 不仅可以直接抑制 B. bassiana,还可以参与调节免疫信号通路相关基因的表达,促进免疫效应物的表达,从而间接抑制家蚕中 B. bassiana 的繁殖。

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