Journal
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
Volume 137, Issue -, Pages -Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.ibmb.2021.103624
Keywords
Bombyx mori; Ommochrome; CRISPR; Cas9; Solute carrier transporter
Categories
Funding
- JSPS KAKENHI [17H05047, 20H02997, 26850220, JP20J22954]
- Ministry of Agriculture, Forestry and Fisheries of Japan
- Ibaraki University
- Cooperative Research Grant of the Genome Research for BioResource, NODAI Genome Research Center, Tokyo University of Agriculture
- Grants-in-Aid for Scientific Research [26850220, 20H02997, 17H05047] Funding Source: KAKEN
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The researchers identified a gene called Bmmah in silkworm, the knockout of which results in reddish-brown color in eggs and adult compound eyes. Experimental evidence confirmed that Bmmah is responsible for the b-4 phenotype.
The brown egg 4 (b-4) is a recessive mutant in the silkworm (Bombyx mori), whose egg and adult compound eyes exhibit a reddish-brown color instead of normal purple and black, respectively. By double digest restriction-site associated DNA sequencing (ddRAD-seq) analysis, we narrowed down a region linked to the b-4 phenotype to approximately 1.1 Mb that contains 69 predicted gene models. RNA-seq analysis in a b-4 strain indicated that one of the candidate genes had a different transcription start site, which generates a short open reading frame. We also found that exon skipping was induced in the same gene due to an insertion of a transposable element in other two b-4 mutant strains. This gene encoded a putative amino acid transporter that belongs to the beta-group of solute carrier (SLC) family and is orthologous to Drosophila eye color mutant gene, mahogany (mah). Accordingly, we named this gene Bmmah. We performed CRISPR/Cas9-mediated gene knockout targeting Bmmah. Several adult moths in generation 0 (G0) had totally or partially reddish-brown compound eyes. We also established three Bmmah knockout strains, all of which exhibit reddish-brown eggs and adult compound eyes. Furthermore, eggs from complementation crosses between the b-4 mutants and the Bmmah knockout mutants also exhibited reddish-brown color, which was similar to the b-4 mutant eggs, indicating that Bmmah is responsible for the b-4 phenotypes.
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