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A single ortholog of teashirt and tiptop regulates larval pigmentation and adult appendage patterning in Bombyx mori.
Insect Biochemistry and Molecular Biology ( IF 3.2 ) Pub Date : 2020-03-31 , DOI: 10.1016/j.ibmb.2020.103369
Ru Zhang 1 , Zhongjie Zhang 2 , Yongping Huang 3 , Airong Qian 4 , Anjiang Tan 2
Affiliation  

Two paralogous genes, teashirt (tsh) and tiptop (tio), encode zinc-finger transcription factors and play important roles in insect growth and development. In the fruit fly, Drosophila melanogaster, tsh promotes trunk segmental identities and contributes to the patterning of other tissues during the embryonic stage. During the adult stage, tsh contributes to the specification and patterning of appendages, including the leg, wing and eye. While tio acts redundantly with tsh, flies lacking tio function are viable without deleterious phenotypes. This gene pair is present in the genomes of all Drosophila species but only as a single homologue in several other insect species. In Oncopeltus fasciatus and Tribolium castaneum, tsh/tio has been functionally characterized as specifying the identity of the leg during the adult stage. However, in lepidopteran insects which include large numbers of pests in agriculture and forestry, as well as the major silk producer silkworm Bombyx mori, the biological functions of tsh/tio are still poorly understood. In the current study, we performed functional analysis of tsh/tio by using both CRISPR/Cas9-mediated mutagenesis and transposon-mediated ectopic expression in B. mori. The results show that loss of tsh/tio function affected pigmentation during the larval stage and appendage pattering during the adult stage. RNA-seq analysis and subsequent q-RT-PCR analysis revealed that depletion of tsh/tio significantly elevated the expression of the kynurenine 3-monooxygenase gene, as well as melanin synthase-related genes during the larval stage. Furthermore, ubiquitous ectopic expression of tsh/tio induces developmental retardation and eventually larval lethality. These data reveal evolutionarily conserved functions of tsh/tio in controlling adult appendage patterning, as well as the novel function of regulating larval pigmentation in B. mori, providing novel insights into how tsh/tio regulates insect growth and development.

中文翻译:

单一的直立的茶衬衫和顶部装饰物可调节家蚕中的幼虫色素沉着和成年肢体的形态。

茶衬衫(tsh)和尖顶(tio)这两个同源基因编码锌指转录因子,并在昆虫的生长发育中起重要作用。在果蝇中,果蝇(trosophila melanogaster,tsh)促进了树干的节段身份,并有助于在胚胎期形成其他组织的图案。在成年阶段,tsh有助于附件(包括腿,翼和眼睛)的规格和样式。虽然tio与tsh冗余发挥作用,但缺乏tio功能的果蝇没有有害表型是可行的。该基因对存在于所有果蝇物种的基因组中,但在其他几种昆虫物种中仅作为单个同源物存在。在筋膜Oncopeltus fasciatus和Tribolium castaneum中,tsh / tio在功能上已被指定为在成年阶段确定腿的身份。然而,在鳞翅目昆虫中,其中包括农业和林业中的大量害虫,以及主要的蚕桑生产商蚕Bombyx mori,对tsh / tio的生物学功能仍知之甚少。在当前的研究中,我们通过使用CRISPR / Cas9介导的诱变和转座子介导的异位表达来进行tsh / tio的功能分析。结果表明,tsh / tio功能的丧失会影响幼虫阶段的色素沉着和成年阶段的附肢拍打。RNA-seq分析和随后的q-RT-PCR分析表明,tsh / tio的消耗在幼虫期显着提高了犬尿氨酸3-单加氧酶基因以及黑色素合酶相关基因的表达。此外,tsh / tio的普遍异位表达诱导发育迟缓,最终导致幼虫致死率。这些数据揭示了tsh / tio在控制成年附肢图案方面的进化保守功能,以及调节B. mori中幼虫色素沉着的新功能,从而为tsh / tio如何调节昆虫的生长和发育提供了新颖的见解。
更新日期:2020-03-31
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