Identification of testis development-related genes by combining Iso-Seq and RNA-Seq in .

(Walker) is an invasive pest. An effective method to control this pest is the sterile insect technique (SIT). To better apply this technique, it is necessary to understand testis development progression. Differentially expressed genes (DEGs) during testis development were analyzed by PacBio Iso-Seq and RNA-seq. RNA-Seq library of testes on day 1, 6, and 11 post eclosion were constructed. We identified 755 and 865 differentially expressed genes in the comparisons of T6 (testes on day 6) vs. T1 and T11 vs. T1, respectively. The KEGG pathway analysis showed that the DEGs were significantly enriched in retinol metabolism, vitamin B6 metabolism, and ascorbate and aldarate metabolism pathways. Knockdown of -like (-like), and , the representative gene in each of the above 3 pathways, reduced the hatching rate of offspring. In addition, we identified 107 spermatogenesis-related orthologous genes in , of which exhibited significantly up-regulated expression throughout testis development, and the knockdown of this gene reduced offspring hatching rate. Our data indicated that -like, , , and genes were related to testis development, and they were conserved in tephritid species. These results suggested that this gene might have the same function in tephritid. The findings provide an insight into testis development and spermatogenesis in tephritid species.