Sericin Ser3 Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm

SIMPLE SUMMARY: Silkworms have remarkable protein synthesis ability, which is favored by biomaterials and biomedicine researchers. The silk gland is the most concerned target tissue of sericulture. Many studies have focused on introducing exogenous functional fibrin genes into the silkworm genome, and have obtained biomedical proteins with good biological activity and multifunctional silk fibers with special properties using the silk gland bioreactor. However, the silk gland of transgenic silkworms often suffers from low vitality, stunting and other problems, and the reasons are still unknown; the biosafety of transgenic silkworms is also unknown. Therefore, we studied the effect of the recombinant Ser3 ectopic gene expressed in the posterior silk gland on hemolymph melanization and metabolism in silkworms. The results showed that although the mutant had normal vitality in normal environments, hemolymph metabolism and immunity were significantly affected. Therefore, the results have positive significance to promote the safe assessment and development of genetically modified organisms. 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 Ser3, 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 (Ser3(+/+)). 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 PPAE, SP21 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 (H(2)O(2)) 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.