Genes on the Y chromosome play important roles in male sex determination and development.The identification of Y-chromosome-specific genes not only provides a theoretical basis for the study of male reproductive devel...Genes on the Y chromosome play important roles in male sex determination and development.The identification of Y-chromosome-specific genes not only provides a theoretical basis for the study of male reproductive development,but also offers genetic control targets for agricultural pests.However,Y-chromosome genes are rarely characterized due to their high repeatability and high heterochromatinization,especially in the oriental fruit fly.In this study,1011 Y-chromosome-specific candidate sequences were screened from 2 to 4 h Bactrocera dorsalis embryo datasets with the chromosome quotient method,6 of which were identified as Y-chromosome-specific sequences by polymerase chain reaction,including typo-gyf,a 19126-bp DNA sequence containing a 575-amino acid open reading frame.Testicular deformation and a significant reduction in sperm number were observed after typo-gyf knockdown with RNA interference in embryos.After typo-gyf knockout with clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9 in the embryonic stage,the sex ratio of the emergent adults was unbalanced,with far more females than males.A genotype analysis of these females with the Y-chromosome gene MoY revealed no sex reversal.Typo-gyf knockout led to the death of XY individuals in the embryonic stage.We conclude that typo-gyf is an essential gene for male survival,and is also involved in testicular development and spermatogenesis.The identification of typo-gyf and its functional verification provide insight into the roles of Y-chromosome genes in male development.展开更多
基金supported by the National Natural Science Foundation of China(no.32220103009)the China Agriculture Research System of MOF and MARA(No.CARS-26)and Hubei Hongshan Laboratory.
文摘Genes on the Y chromosome play important roles in male sex determination and development.The identification of Y-chromosome-specific genes not only provides a theoretical basis for the study of male reproductive development,but also offers genetic control targets for agricultural pests.However,Y-chromosome genes are rarely characterized due to their high repeatability and high heterochromatinization,especially in the oriental fruit fly.In this study,1011 Y-chromosome-specific candidate sequences were screened from 2 to 4 h Bactrocera dorsalis embryo datasets with the chromosome quotient method,6 of which were identified as Y-chromosome-specific sequences by polymerase chain reaction,including typo-gyf,a 19126-bp DNA sequence containing a 575-amino acid open reading frame.Testicular deformation and a significant reduction in sperm number were observed after typo-gyf knockdown with RNA interference in embryos.After typo-gyf knockout with clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9 in the embryonic stage,the sex ratio of the emergent adults was unbalanced,with far more females than males.A genotype analysis of these females with the Y-chromosome gene MoY revealed no sex reversal.Typo-gyf knockout led to the death of XY individuals in the embryonic stage.We conclude that typo-gyf is an essential gene for male survival,and is also involved in testicular development and spermatogenesis.The identification of typo-gyf and its functional verification provide insight into the roles of Y-chromosome genes in male development.
