Inducible overexpression of cecropin B decreases the susceptibility of the transgenic silkworm, Bombyx mori (Lepidoptera: Bombycidae), to bacteria

IF 1.3 4区 农林科学 Q2 ENTOMOLOGY Applied Entomology and Zoology Pub Date : 2022-11-16 DOI:10.1007/s13355-022-00807-4
Rasalkar Sandhya Yashwant, Dyna Susan Thomas, Chitra Manoharan, Upendra Nongthomba, Vankadara Sivaprasad, Ravikumar Gopalapillai
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Abstract

Silkworms and other insects confer innate immunity by expressing antimicrobial peptides (AMPs) through the induction of Toll and IMD pathways. Cecropin B, an AMP from Bombyx mori L. (Lepidoptera: Bombycidae), has a broad range of antibacterial activity against both Gram-positive and -negative bacteria. Bacterial flacherie is one of the severe diseases in silkworms. In an attempt to develop silkworm strains expressing antibacterial properties, a transgenic vector, piggyBac overexpressing the cecropin B gene, was constructed under its promoter. The vector had GFP under the control of the elongation factor (ELFα) promoter as a marker for screening transgenic silkworms. Transgenic silkworms were generated by microinjecting the piggyBac vector along with the helper vector into the silkworm eggs. The mRNA level of cecropin B in the fat body of transgenic lines was higher than the non-transgenic lines in response to Escherichia coli and Staphylococcus aureus infections. The overexpression of cecropin B was confirmed by Western blot analysis. Transgenic lines' mortality was significantly decreased compared to non-transgenic lines’ post-bacterial infections. These results imply that overexpressing an endogenous AMP gene can enhance the resistance of silkworms in response to bacterial infections.

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cecropin B的诱导过表达降低了转基因家蚕(鳞翅目:家蚕科)对细菌的易感性
家蚕和其他昆虫通过诱导Toll和IMD通路表达抗菌肽(AMPs)来赋予先天免疫。Cecropin B是家蚕(鳞翅目:家蚕科)的一种AMP,对革兰氏阳性和阴性细菌都有广泛的抗菌活性。细菌性绒毛病是家蚕的严重病害之一。为了培育具有抗菌特性的家蚕菌株,在其启动子下构建了过表达天蚕素B基因的转基因载体piggyBac。该载体以伸长因子(ELFα)启动子控制的GFP作为筛选转基因家蚕的标记。将piggyBac载体与辅助载体一起微注射到家蚕卵中,获得了转基因家蚕。对大肠杆菌和金黄色葡萄球菌感染的反应中,转基因品系脂肪体中cecropin B mRNA水平高于非转基因品系。Western blot分析证实cecropin B过表达。与非转基因品系相比,转基因品系的细菌感染后死亡率显著降低。这些结果表明,过表达内源性AMP基因可以增强家蚕对细菌感染的抗性。
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来源期刊
CiteScore
2.70
自引率
7.70%
发文量
37
审稿时长
6-12 weeks
期刊介绍: Applied Entomology and Zoology publishes articles concerned with applied entomology, applied zoology, agricultural chemicals and pest control in English. Contributions of a basic and fundamental nature may be accepted at the discretion of the Editor. Manuscripts of original research papers, technical notes and reviews are accepted for consideration. No manuscript that has been published elsewhere will be accepted for publication.
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