{"title":"BTB-ZF转录因子的破坏导致家蚕幼虫体中的雌性不育和黑化","authors":"Kenta Tomihara , Takashi Kiuchi","doi":"10.1016/j.ibmb.2023.103982","DOIUrl":null,"url":null,"abstract":"<div><p>The <em>dilute black</em> (<em>bd</em><span>) of the silkworm </span><span><em>Bombyx </em><em>mori</em></span><span> is a recessive mutant that produces a grayish-black color in the larval integument, instead of the characteristic white color found in wild-type larvae. In addition, eggs produced by </span><em>bd</em> females are sterile due to a deficiency in the micropylar apparatus. We identified candidate genes responsible for the <em>bd</em> phenotype using publicly available RNA-seq data. One of these candidate genes was homologous to the <em>maternal gene required for meiosis</em> (<span><em>mamo</em></span>) of <span><em>Drosophila melanogaster</em></span>, which encodes a broad-complex, tramtrack, and bric-à-brac-zinc finger (BTB-ZF) transcription factor essential for female fertility. In three independent <em>bd</em> strains, the expression of the <em>B. mori mamo</em> (<em>Bmmamo</em>) was downregulated in the larval integument. Using a CRISPR/Cas9-mediated knockout strategy, we found that <em>Bmmamo</em><span> knockout mutants exhibit a grayish-black color in the larval integument and female infertility. Moreover, larvae obtained from the complementation cross between </span><em>bd/+</em> mutants and heterozygous knockouts for the <em>Bmmamo</em> also exhibited a grayish-black color, indicating that <em>Bmmamo</em> is responsible for the <em>bd</em><span> phenotype. Gene expression analysis using </span><em>Bmmamo</em><span> knockout mutants suggested that the BmMamo protein suppresses the expression of melanin synthesis genes. Previous comparative genome analysis revealed that the </span><em>Bmmamo</em> was selected during silkworm domestication, and we found that <em>Bmmamo</em> expression in the larval integument is higher in <em>B. mori</em><span> than in the wild silkworm </span><em>B. mandarina</em>, suggesting that the <em>Bmmamo</em><span> is involved in domestication-associated pigmentation changes of the silkworm.</span></p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":3.2000,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Disruption of a BTB-ZF transcription factor causes female sterility and melanization in the larval body of the silkworm, Bombyx mori\",\"authors\":\"Kenta Tomihara , Takashi Kiuchi\",\"doi\":\"10.1016/j.ibmb.2023.103982\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>The <em>dilute black</em> (<em>bd</em><span>) of the silkworm </span><span><em>Bombyx </em><em>mori</em></span><span> is a recessive mutant that produces a grayish-black color in the larval integument, instead of the characteristic white color found in wild-type larvae. In addition, eggs produced by </span><em>bd</em> females are sterile due to a deficiency in the micropylar apparatus. We identified candidate genes responsible for the <em>bd</em> phenotype using publicly available RNA-seq data. One of these candidate genes was homologous to the <em>maternal gene required for meiosis</em> (<span><em>mamo</em></span>) of <span><em>Drosophila melanogaster</em></span>, which encodes a broad-complex, tramtrack, and bric-à-brac-zinc finger (BTB-ZF) transcription factor essential for female fertility. In three independent <em>bd</em> strains, the expression of the <em>B. mori mamo</em> (<em>Bmmamo</em>) was downregulated in the larval integument. Using a CRISPR/Cas9-mediated knockout strategy, we found that <em>Bmmamo</em><span> knockout mutants exhibit a grayish-black color in the larval integument and female infertility. Moreover, larvae obtained from the complementation cross between </span><em>bd/+</em> mutants and heterozygous knockouts for the <em>Bmmamo</em> also exhibited a grayish-black color, indicating that <em>Bmmamo</em> is responsible for the <em>bd</em><span> phenotype. Gene expression analysis using </span><em>Bmmamo</em><span> knockout mutants suggested that the BmMamo protein suppresses the expression of melanin synthesis genes. Previous comparative genome analysis revealed that the </span><em>Bmmamo</em> was selected during silkworm domestication, and we found that <em>Bmmamo</em> expression in the larval integument is higher in <em>B. mori</em><span> than in the wild silkworm </span><em>B. mandarina</em>, suggesting that the <em>Bmmamo</em><span> is involved in domestication-associated pigmentation changes of the silkworm.</span></p></div>\",\"PeriodicalId\":330,\"journal\":{\"name\":\"Insect Biochemistry and Molecular Biology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.2000,\"publicationDate\":\"2023-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Insect Biochemistry and Molecular Biology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0965174823000760\",\"RegionNum\":2,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Insect Biochemistry and Molecular Biology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0965174823000760","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Disruption of a BTB-ZF transcription factor causes female sterility and melanization in the larval body of the silkworm, Bombyx mori
The dilute black (bd) of the silkworm Bombyx mori is a recessive mutant that produces a grayish-black color in the larval integument, instead of the characteristic white color found in wild-type larvae. In addition, eggs produced by bd females are sterile due to a deficiency in the micropylar apparatus. We identified candidate genes responsible for the bd phenotype using publicly available RNA-seq data. One of these candidate genes was homologous to the maternal gene required for meiosis (mamo) of Drosophila melanogaster, which encodes a broad-complex, tramtrack, and bric-à-brac-zinc finger (BTB-ZF) transcription factor essential for female fertility. In three independent bd strains, the expression of the B. mori mamo (Bmmamo) was downregulated in the larval integument. Using a CRISPR/Cas9-mediated knockout strategy, we found that Bmmamo knockout mutants exhibit a grayish-black color in the larval integument and female infertility. Moreover, larvae obtained from the complementation cross between bd/+ mutants and heterozygous knockouts for the Bmmamo also exhibited a grayish-black color, indicating that Bmmamo is responsible for the bd phenotype. Gene expression analysis using Bmmamo knockout mutants suggested that the BmMamo protein suppresses the expression of melanin synthesis genes. Previous comparative genome analysis revealed that the Bmmamo was selected during silkworm domestication, and we found that Bmmamo expression in the larval integument is higher in B. mori than in the wild silkworm B. mandarina, suggesting that the Bmmamo is involved in domestication-associated pigmentation changes of the silkworm.
期刊介绍:
This international journal publishes original contributions and mini-reviews in the fields of insect biochemistry and insect molecular biology. Main areas of interest are neurochemistry, hormone and pheromone biochemistry, enzymes and metabolism, hormone action and gene regulation, gene characterization and structure, pharmacology, immunology and cell and tissue culture. Papers on the biochemistry and molecular biology of other groups of arthropods are published if of general interest to the readership. Technique papers will be considered for publication if they significantly advance the field of insect biochemistry and molecular biology in the opinion of the Editors and Editorial Board.