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Sterile insect technique (SIT) has emerged as an environmentally sustainable pest management strategy. However, discovery of molecular targets applicable for SIT implementation in\n                      <jats:italic>Z.\u2009tau<\/jats:italic>\n                      still constitutes a significant research gap.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>RESULTS<\/jats:title>\n                    <jats:p>\n                      We conducted comparative transcriptome analysis of four male tissues (midgut, Malpighian tubules, fat body, and testis), identifying 9653 differentially expressed genes (DEGs) with predominant testis enrichment. Bioinformatics screening revealed 3020 testis\u2010specific highly expressed genes showing significant functional enrichment in cytoplasmic translation, cytosolic ribosome assembly, and oxidoreductase activity. Quantitative real\u2010time PCR (qRT\u2010PCR) assay was utilized to confirm 10 testis\u2010specific genes, including two serine\/threonine kinases (\n                      <jats:italic>ZtTSSK1<\/jats:italic>\n                      and\n                      <jats:italic>ZtTSSK3<\/jats:italic>\n                      ) which were significantly enriched in spermatid development. Fluorescence\n                      <jats:italic>in situ<\/jats:italic>\n                      hybridization (FISH) localized the two genes specifically to the transformation zone of\n                      <jats:italic>Z.\u2009tau<\/jats:italic>\n                      testis. Functional characterization via RNAi bioassay demonstrated that suppression of\n                      <jats:italic>ZtTSSK<\/jats:italic>\n                      s expression levels reduced spermatozoa number and impaired male fertility.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>CONCLUSION<\/jats:title>\n                    <jats:p>\n                      These results establish\n                      <jats:italic>ZtTSSK<\/jats:italic>\n                      s as crucial regulators of male fertility in\n                      <jats:italic>Z.\u2009tau<\/jats:italic>\n                      and identify them as potential molecular targets for developing SIT\u2010based interventions against this economically significant pest. \u00a9 2026 Society of Chemical Industry.\n                    <\/jats:p>\n                  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