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Positional cloning of a Bombyx pink-eyed white egg locus reveals the major role of cardinal in ommochrome synthesis

机译:Bombyx粉红眼白卵基因座的位置克隆揭示了红衣主教在全色素合成中的主要作用

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摘要

Ommochromes are major insect pigments involved in coloration of compound eyes, eggs, epidermis and wings. In the silkworm Bombyx mori, adult compound eyes and eggs contain a mixture of the ommochrome pigments such as ommin and xanthommatin. Here, we identified the gene involved in ommochrome biosynthesis by positional cloning of B. mori egg and eye color mutant pink-eyed white egg (pe). The recessive homozygote of pe has bright red eyes and white or pale pink eggs instead of a normal dark coloration due to the decrease of dark ommochrome pigments. By genetic linkage analysis, we narrowed down the pe-linked region to ~258 kb, containing 17 predicted genes. RNA sequencing analyses showed that the expression of one candidate gene, the ortholog of Drosophila haem peroxidase cardinal, coincided with egg pigmentation timing, similar to other ommochrome-related genes such as Bm-scarlet and Bm-re. In two pe strains, a common missense mutation was found within a conserved motif of B. mori cardinal homolog (Bm-cardinal). RNA interference-mediated knockdown and transcription activator-like effector nuclease (TALEN)-mediated knockout of the Bm-cardinal gene produced the same phenotype as pe in terms of egg, adult eye and larval epidermis coloration. A complementation test of the pe mutant with the TALEN-mediated Bm-cardinal-deficient strain showed that the mutant phenotype could not be rescued, indicating that Bm-cardinal is responsible for pe. Moreover, knockdown of the cardinal homolog in Tribolium castaneum also induced red compound eyes. Our results indicate that cardinal plays a major role in ommochrome synthesis of holometabolous insects.
机译:色素是涉及复眼,卵,表皮和翅膀着色的主要昆虫色素。在家蚕中,成年复眼和卵含有诸如ommin和xanthommatin等全色素的混合物。在这里,我们通过定位克隆桑蚕双歧杆菌卵和眼睛颜色突变的粉红色眼白卵(pe)来鉴定参与全色素生物合成的基因。 pe的隐性纯合子具有鲜红的眼睛和白色或淡粉红色的鸡蛋,而不是正常的深色,这是因为深色的全色色素减少了。通过遗传连锁分析,我们将pe连接区域缩小到〜258 tokb,其中包含17个预测基因。 RNA测序分析表明,一种候选基因果蝇血红素过氧化物酶的直向同源基因的表达与卵的色素沉着时间相吻合,类似于其他与色素相关的基因,例如Bm-scarlet和Bm-re。在两个pe菌株中,在桑蚕双胞胎主要同系物(Bm-cardinal)的保守基序中发现了常见的错义突变。 RNA干扰介导的敲除和转录激活子样效应核酸酶(TALEN)介导的Bm-主要基因的敲除产生了与pe相同的表型,包括蛋,成年眼和幼虫表皮的着色。用TALEN介导的 Bm-基数缺陷菌株对pe突变体进行的互补试验表明,该突变体的表型无法挽救,表明 Bm-基数是造成这种情况的原因。 pe 。此外,在 Tribolium castaneum 主要同源物的敲除也诱导了红色复眼。我们的结果表明,基数在全代谢昆虫的全色素合成中起主要作用。

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