RESUMO
The noa gene is an essential gene encoding a very long chain fatty acid elongase. In this study, we cloned the noa gene of Bactrocera dorsalis, which encodes a protein sharing 84.50% identity to the NOA in Drosophila melanogaster. The expression profiles indicated that the transcriptional level of noa was high at the egg stage and in the testis tissue. The results showed that noa expression was up-regulated after Listeria monocytogenes, Staphylococcus aureus and Escherichia coli infection. Silencing of noa would influence the expression of immune related genes, including MyD88 and defensin in the Toll pathway and relish and diptericin in the Imd pathway. Moreover, infection with L. monocytogenes and S. aureus after feeding ds-noa, the expression of MyD88 and defensin down-regulated significantly in ds-noa group compared with in ds-egfp group, indicating that noa interference influenced the activation of the Toll pathway. Meanwhile, infection with L. monocytogenes and E. coli, which activated the Imd pathway, do not cause increase of the mRNA levels of relish and diptericin in ds-noa group as severely as in ds-egfp treatment, indicating that the Imd pathway was also repressed after silences of noa.
Assuntos
Infecções Bacterianas/imunologia , Proteínas de Insetos/metabolismo , Óxido Nítrico Sintase/metabolismo , Tephritidae/imunologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Sequência de Aminoácidos , Animais , Antígenos de Diferenciação/genética , Antígenos de Diferenciação/metabolismo , Defensinas/genética , Defensinas/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/imunologia , Regulação da Expressão Gênica/genética , Imunidade Inata/genética , Proteínas de Insetos/genética , Masculino , Dados de Sequência Molecular , Óxido Nítrico Sintase/genética , RNA Interferente Pequeno/genética , Receptores Imunológicos/genética , Receptores Imunológicos/metabolismo , Homologia de Sequência de Aminoácidos , Transdução de Sinais , Receptores Toll-Like/metabolismoRESUMO
In the male reproductive system of insects, the male accessory glands and ejaculatory duct (MAG/ED) are important organs and their primary function is to enhance the fertility of spermatozoa. Proteins secreted by the MAG/ED are also known to induce post-mating changes and immunity responses in the female insect. To understand the gene expression profile in the MAG/ED of the oriental fruit fly Bactrocera dorsalis (Hendel), that is an important pest in fruits, we performed an Illumina-based deep sequencing of mRNA. This yielded 54,577,630 clean reads corresponding to 4.91Gb total nucleotides that were assembled and clustered to 30,669 unigenes (average 645bp). Among them, 20,419 unigenes were functionally annotated to known proteins/peptides in Gene Orthology, Clusters of Orthologous Groups, Kyoto Encyclopedia of Genes and Genomes pathway databases. Typically, many genes were involved in immunity and these included microbial recognition proteins and antimicrobial peptides. Subsequently, the inducible expression of these immunity-related genes was confirmed by qRT-PCR analysis when insects were challenged with immunity-inducible factors, suggesting their function in guaranteeing fertilization success. Besides, we identified some important reproductive genes such as juvenile hormone- and ecdysteroid-related genes in this de novo assembly. In conclusion, this transcriptomic sequencing of B. dorsalis MAG/ED provides insights to facilitate further functional research of reproduction, immunity and molecular evolution of reproductive proteins in this important agricultural pest.