TRANSCRIPTION FACTOR Bmsage PLAYS A CRUCIAL ROLE IN SILK GLAND GENERATION IN SILKWORM, Bombyx mori

Salivary gland secretion is altered in Drosophila embryos with loss of function of the sage gene. Saliva has a reduced volume and an increased electron density according to transmission electron microscopy, resulting in regions of tube dilation and constriction with intermittent tube closure. Howeve...

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Bibliographic Details
Published in:Archives of insect biochemistry and physiology 2015-10, Vol.90 (2), p.59-69
Main Authors: Xin, Hu‐hu, Zhang, Deng‐pan, Chen, Rui‐ting, Cai, Zi‐zheng, Lu, Yan, Liang, Shuang, Miao, Yun‐gen
Format: Article
Language:English
Subjects:
Animals
Bmsage
Bombyx - embryology
Bombyx - genetics
Bombyx - physiology
Bombyx mori
Clustered Regularly Interspaced Short Palindromic Repeats
development
Drosophila
Embryo, Nonmammalian
embryogenesis
Exocrine Glands - embryology
Exocrine Glands - physiology
Female
fibroins
genes
Genomes
germ cells
homozygosity
Insect Proteins - genetics
Insect Proteins - metabolism
Larva - genetics
Larva - growth & development
larvae
mutagenesis
mutants
Mutation
open reading frames
phenotype
progeny
sage
saliva
salivary glands
sampling
secretion
silk
silk gland
silk glands
silkworm
silkworms
transcription factor
Transcription factors
Transcription Factors - genetics
Transcription Factors - metabolism
transmission electron microscopy
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Summary:Salivary gland secretion is altered in Drosophila embryos with loss of function of the sage gene. Saliva has a reduced volume and an increased electron density according to transmission electron microscopy, resulting in regions of tube dilation and constriction with intermittent tube closure. However, the precise functions of Bmsage in silkworm (Bombyx mori) are unknown, although its sequence had been deposited in SilkDB. From this, Bmsage is inferred to be a transcription factor that regulates the synthesis of silk fibroin and interacts with another silk gland‐specific transcription factor, namely, silk gland factor‐1. In this study, we introduced a germline mutation of Bmsage using the Cas9/sgRNA system, a genome‐editing technology, resulting in deletion of Bmsage from the genome of B. mori. Of the 15 tested samples, seven displayed alterations at the target site. The mutagenesis efficiency was about 46.7% and there were no obvious off‐target effects. In the screened homozygous mutants, silk glands developed poorly and the middle and posterior silk glands (MSG and PSG) were absent, which was significantly different from the wild type. The offspring of G₀ mosaic silkworms had indel mutations causing 2‐ or 9‐bp deletions at the target site, but exhibited the same abnormal silk gland structure. Mutant larvae containing different open‐reading frames of Bmsage had the same silk gland phenotype. This illustrated that the mutant phenotype was due to Bmsage knockout. We conclude that Bmsage participates in embryonic development of the silk gland.
ISSN:0739-4462
1520-6327
DOI:10.1002/arch.21244