Yashwant Rasalkar Sandhya, Thomas Dyna Susan, Manoharan Chitra, Roy Gourab, Kunjupillai Vijayan, Mishra Rakesh Kumar, Nongthomba Upendra, Gopalapillai Ravikumar
Seri-Biotech Research Laboratory, Central Silk Board, Kodathi, Bengaluru, 560 030, India.
Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bengaluru, 560 012, India.
Mol Biotechnol. 2022 Jun;64(6):711-724. doi: 10.1007/s12033-021-00438-0. Epub 2022 Jan 31.
The sericulture industry faces substantial economic losses due to severe pathogenic infections caused by fungi, viruses, and bacteria. The development of transgenic silkworms against specific pathogens has been shown to enhance disease resistance against a particular infection. A single gene or its products that can confer protection against multiple pathogens is required. In an attempt to develop silkworms with enhanced immunity against multiple pathogens, we generated transgenic silkworm lines with an overexpressed NF-kB transcription factor, Relish 1, under two different promoters. Separately, a potent anti-fungal gene, Drosomycin, was also expressed in transgenic silkworms. Both Relish 1 and Drosomycin transgenic silkworms had single copy genomic integration, and their mRNA expression levels were highly increased after infection with silkworm pathogens. The overexpression of the Relish 1 in transgenic silkworms resulted in the upregulation of several defense-related genes, Cecropin B, Attacin, and Lebocin, and showed enhanced resistance to Nosema bombycis (microsporidian fungus), Nucleopolyhedrovirus (BmNPV), and bacteria. The Drosomycin expressing transgenic silkworms showed elevated resistance to N. bombycis and bacteria. These findings demonstrate the role of Relish 1 in long-lasting protection against multiple pathogens in silkworms. Further, the successful introduction of a foreign gene, Drosomycin, also led to improved disease resistance in silkworms.
由于真菌、病毒和细菌引起的严重致病性感染,养蚕业面临着巨大的经济损失。已证明开发针对特定病原体的转基因蚕可增强对特定感染的抗病能力。需要一种能够抵御多种病原体的单一基因或其产物。为了培育对多种病原体具有增强免疫力的蚕,我们在两种不同的启动子下生成了过表达NF-kB转录因子Relish 1的转基因蚕品系。另外,一种强效抗真菌基因Drosomycin也在转基因蚕中表达。Relish 1和Drosomycin转基因蚕均具有单拷贝基因组整合,并且在感染家蚕病原体后它们的mRNA表达水平大幅提高。转基因蚕中Relish 1的过表达导致几种防御相关基因(天蚕素B、攻击素和勒博辛)的上调,并显示出对家蚕微孢子虫(微孢子虫真菌)、核型多角体病毒(BmNPV)和细菌的抗性增强。表达Drosomycin的转基因蚕对家蚕微孢子虫和细菌的抗性增强。这些发现证明了Relish 1在蚕对多种病原体的长期保护中的作用。此外,成功导入外源基因Drosomycin也提高了蚕的抗病能力。