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在后部丝腺中异位表达的丝胶蛋白通过减少家蚕黑色素合成影响血淋巴免疫黑化反应。

Sericin Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm.

作者信息

Wang Yongfeng, Shi Meijuan, Yang Jiameng, Ma Lu, Chen Xuedong, Xu Meng, Peng Ruji, Wang Guang, Pan Zhonghua, Sima Yanghu, Xu Shiqing

机构信息

School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, China.

Institute of Agricultural Biotechnology & Ecology (IABE), Soochow University, Suzhou 215123, China.

出版信息

Insects. 2023 Feb 28;14(3):245. doi: 10.3390/insects14030245.

Abstract

The transgenesis of silkworms is an important way to innovate genetic resources and silk function. However, the silk-gland (SG) of transgenic silkworms, which is the most concerned target tissue of sericulture, often suffers from low vitality, stunting and other problems, and the reasons are still unknown. This study trans engineered recombinant , a middle silk gland (MSG) specific expression gene, in the posterior silk gland (PSG) of the silkworm, and studied hemolymph immune melanization response changes in mutant pure line SER (). The results showed that although the mutant had normal vitality, the melanin content and phenoloxidase (PO) activity in hemolymph related to humoral immunity were significantly reduced, and caused significantly slower blood melanization and weaker sterilization ability. The mechanism investigation showed that the mRNA levels and enzymatic activities of phenylalanine hydroxylase (PAH), tyrosine hydroxylase (TH) and dopamine decarboxylase (DDC) in the melanin synthesis pathway in mutant hemolymph, as well as the transcription levels of the , and serpins genes in the serine protease cascade were significantly affected. Moreover, the total antioxidant capacity, superoxide anion inhibition capacity and catalase (CAT) level related to the redox metabolic capacity of hemolymph were significantly increased, while the activities of superoxide dismutase (SOD) and glutathione reductase (GR), as well as the levels of hydrogen peroxide (HO) and glutathione (GSH), were significantly decreased. In conclusion, the anabolism of melanin in the hemolymph of PSG transgenic silkworm SER was inhibited, while the basic response level of oxidative stress was increased, and the hemolymph immune melanization response was decreased. The results will significantly improve the safe assessment and development of genetically modified organisms.

摘要

家蚕转基因是创新遗传资源和蚕丝功能的重要途径。然而,转基因家蚕的丝腺作为养蚕业最关注的靶组织,常出现活力低下、发育不良等问题,其原因尚不清楚。本研究在家蚕后部丝腺中过表达了一个中部丝腺特异性表达基因,并研究了突变纯系SER的血淋巴免疫黑化反应变化。结果表明,虽然突变体活力正常,但与体液免疫相关的血淋巴中黑色素含量和酚氧化酶(PO)活性显著降低,导致血液黑化明显减慢,杀菌能力减弱。机制研究表明,突变体血淋巴中黑色素合成途径中苯丙氨酸羟化酶(PAH)、酪氨酸羟化酶(TH)和多巴胺脱羧酶(DDC)的mRNA水平和酶活性,以及丝氨酸蛋白酶级联反应中serpin、serpin和serpin基因的转录水平均受到显著影响。此外,与血淋巴氧化还原代谢能力相关的总抗氧化能力、超氧阴离子抑制能力和过氧化氢酶(CAT)水平显著升高,而超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GR)的活性以及过氧化氢(HO)和谷胱甘肽(GSH)的水平显著降低。综上所述,后部丝腺转基因家蚕SER血淋巴中黑色素的合成代谢受到抑制,而氧化应激的基础反应水平升高,血淋巴免疫黑化反应降低。该结果将显著提高转基因生物的安全评估和开发水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/82bc/10051610/3d709c14fae1/insects-14-00245-g001.jpg

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