Abstract

The silk gland synthesizes and secretes a large amount of protein and stores liquid silk protein at an extremely high concentration. Interestingly, silk proteins and serine protease inhibitors are orderly arranged in the silk gland lumen and cocoon shells. Silk fiber formation and the spinning mechanism have not been fully elucidated. Therefore, we conducted a comparative transcriptome analysis of seven segments of the single silk gland to characterize internal changes in the silk gland during the 5th instar of mature larvae. In total, 3121 differentially expressed genes were identified in the seven segments. Genes highly expressed in the middle silk gland (MSG) were mainly involved in unsaturated fatty acid biosynthesis, fatty acid metabolism, apoptosis—fly, and lysosome pathways, whereas genes highly expressed in the posterior silk gland (PSG) were mainly involved in ribosome, proteasome, citrate cycle, and glycolysis/gluconeogenesis pathways. Thus, the MSG and PSG differ greatly in energy source use and function. Further, 773 gradually upregulated genes (from PSG to MSG) were involved in energy metabolism, silk protein synthesis, and secretion, suggesting that these genes play an important role in silk fiber formation. Our findings provide insights into the mechanism of silk protein synthesis and transport and silk fiber formation.

Highlights

  • Silk is one of the most mysterious and attractive materials in nature and has been widely used in biomedicine, soft-tissue engineering, biosensing, textiles, cosmetics, and other fields

  • It was very interesting to find genes related to lipid metabolism and lipid transport to be highly expressed in middle silk gland (MSG)

  • Numerous genes involved in ribosome, biosynthesis of amino acids, and tyrosine metabolism pathways were identified in the posterior silk gland (PSG), which means protein synthesis is still occurring at large scale in the PSG during larval maturation

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Summary

Introduction

Silk is one of the most mysterious and attractive materials in nature and has been widely used in biomedicine, soft-tissue engineering, biosensing, textiles, cosmetics, and other fields. Sericins are soluble glue proteins that coat and cement the silk fibers, and mainly include sericin 1 (Ser1, ~400 kDa), sericin 2 (Ser2, ~230 kDa and 120 kDa), and sericin 3 (Ser3, ~250 kDa) [9,10] They are secreted by different segments of the middle silk gland (MSG). The mixture moves forward to the anterior silk gland (ASG) and spins out through the ASG and spinneret, accompanied by structural conformational changes This protein secretion process leads to the orderly composition of silk fibroins and sericins in silk fibers. SGF-2 is a multisubunit activator complex containing Awh, Ldb, and Lcaf, which control PSG-specific expression of silk fibroin genes [23]. This is consistent with the previously reported gradient distribution of serine protease inhibitors in the cocoon shell, which enables silkworm pupae to resist infection [18]

The MSG is Functionally More Divergent than the PSG
Discussion
Silk Gland Dissection and Sample Collection
RNA Preparation and Transcriptome Sequencing
Sequencing Data Processing
Findings
Differential Expression Analysis
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