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

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 u...

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Main Authors: Yongfeng Wang, Meijuan Shi, Jiameng Yang, Lu Ma, Xuedong Chen, Meng Xu, Ruji Peng, Guang Wang, Zhonghua Pan, Yanghu Sima, Shiqing Xu
Format: Article
Language:English
Published: MDPI AG 2023-02-01
Series:Insects
Subjects:
Online Access:https://www.mdpi.com/2075-4450/14/3/245
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author Yongfeng Wang
Meijuan Shi
Jiameng Yang
Lu Ma
Xuedong Chen
Meng Xu
Ruji Peng
Guang Wang
Zhonghua Pan
Yanghu Sima
Shiqing Xu
author_facet Yongfeng Wang
Meijuan Shi
Jiameng Yang
Lu Ma
Xuedong Chen
Meng Xu
Ruji Peng
Guang Wang
Zhonghua Pan
Yanghu Sima
Shiqing Xu
author_sort Yongfeng Wang
collection DOAJ
description 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 <i>Ser3</i>, 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 (<i>Ser3</i><sup>+/+</sup>). 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 <i>PPAE</i>, <i>SP21</i> 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<sub>2</sub>O<sub>2</sub>) 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.
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spelling doaj.art-d3aff474c97c46c9b3eb1893da22fadb2023-11-17T11:45:56ZengMDPI AGInsects2075-44502023-02-0114324510.3390/insects14030245Sericin <i>Ser3</i> Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in SilkwormYongfeng Wang0Meijuan Shi1Jiameng Yang2Lu Ma3Xuedong Chen4Meng Xu5Ruji Peng6Guang Wang7Zhonghua Pan8Yanghu Sima9Shiqing Xu10School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaSchool of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou 215123, ChinaThe 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 <i>Ser3</i>, 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 (<i>Ser3</i><sup>+/+</sup>). 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 <i>PPAE</i>, <i>SP21</i> 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<sub>2</sub>O<sub>2</sub>) 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.https://www.mdpi.com/2075-4450/14/3/245<i>Bombyx mori</i>silk gland transgenicimmune melanization responsemelanin synthesis
spellingShingle Yongfeng Wang
Meijuan Shi
Jiameng Yang
Lu Ma
Xuedong Chen
Meng Xu
Ruji Peng
Guang Wang
Zhonghua Pan
Yanghu Sima
Shiqing Xu
Sericin <i>Ser3</i> Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm
Insects
<i>Bombyx mori</i>
silk gland transgenic
immune melanization response
melanin synthesis
title Sericin <i>Ser3</i> Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm
title_full Sericin <i>Ser3</i> Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm
title_fullStr Sericin <i>Ser3</i> Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm
title_full_unstemmed Sericin <i>Ser3</i> Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm
title_short Sericin <i>Ser3</i> Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm
title_sort sericin i ser3 i ectopic expressed in posterior silk gland affects hemolymph immune melanization response via reducing melanin synthesis in silkworm
topic <i>Bombyx mori</i>
silk gland transgenic
immune melanization response
melanin synthesis
url https://www.mdpi.com/2075-4450/14/3/245
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