Mutations in a β-group of solute carrier gene are responsible for egg and eye coloration of the brown egg 4 (b-4) mutant in the silkworm, Bombyx mori
Mutations in a β-group of solute carrier gene are responsible for egg and eye coloration of the brown egg 4 (b-4) mutant in the silkworm, Bombyx mori
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溶质载体基因 β 组的突变导致家蚕 Bombyx mori 中棕色卵 4 (b-4) 突变体的卵和眼睛颜色
DOI：
10.1016/j.ibmb.2021.103624
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发表时间：
2021
影响因子：
3.8
通讯作者：
Kiuchi Takashi
中科院分区：
文献类型：
--
作者：
Tomihara Kenta;Satta Katsuya;Matsuzaki Shohei;Yoshitake Kazutoshi;Yamamoto Kimiko;Uchiyama Hironobu;Yajima Shunsuke;Futahashi Ryo;Katsuma Susumu;Osanai-Futahashi Mizuko;Kiuchi Takashi
褐卵4号（B-4）是家蚕的一个隐性突变体，其卵和成虫复眼的颜色分别由正常的紫色和黑色变为红褐色。通过双酶切限制性位点相关DNA测序（ddRAD-seq）分析，我们将与b-4表型相关的区域缩小到约1.1 Mb，其中包含69个预测的基因模型。对ab-4株的RNA-seq分析表明，其中一个候选基因具有不同的转录起始位点，产生了一个短的开放阅读框架。我们还发现，外显子跳跃诱导在同一个基因，由于在其他两个b-4突变株的转座因子的插入。该基因编码一种氨基酸转运蛋白，属于β-组溶质转运蛋白（SLC）家族，与果蝇眼色突变基因mah同源。因此，我们将该基因命名为Bmmah。我们进行了CRISPR/Cas9介导的基因敲除靶向Bmmah。第0代（G 0）有几只成虫复眼全部或部分呈红褐色。我们还建立了三个Bmmah敲除菌株，所有这些菌株都表现出红棕色的卵和成虫复眼。此外，b-4突变体与Bmmah基因敲除突变体互补杂交所得的卵也呈现红褐色，与b-4突变体的卵相似，表明Bmmah基因是造成b-4表型的原因。
Thebrown 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 theb-4phenotype to approximately 1.1 Mb that contains 69 predicted gene models. RNA-seq analysis in ab-4strain 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 twob-4mutant strains. This gene encoded a putative amino acid transporter that belongs to the β-group of solute carrier (SLC) family and is orthologous toDrosophilaeye color mutant gene,mahogany(mah). Accordingly, we named this geneBmmah. We performed CRISPR/Cas9-mediated gene knockout targetingBmmah. Several adult moths in generation 0 (G0) had totally or partially reddish-brown compound eyes. We also established threeBmmahknockout strains, all of which exhibit reddish-brown eggs and adult compound eyes. Furthermore, eggs from complementation crosses between theb-4mutants and theBmmahknockout mutants also exhibited reddish-brown color, which was similar to theb-4mutant eggs, indicating thatBmmahis responsible for theb-4phenotypes.