Details

Autor(en) / Beteiligte

• Wang, Yun
• Xu, Hui‐Qian
• Han, Hong‐Liang
• Chen, Dong
• Jiang, Hongbo
• Smagghe, Guy
• Wang, Jin‐Jun
• Wei, Dong

Titel

CRISPR/Cas9‐mediated knockout of a male accessory glands‐specific gene takeout1 decreases the fecundity of Zeugodacus cucurbitae female

Ist Teil von

• Pest management science, 2024-09, Vol.80 (9), p.4399-4409

Ort / Verlag

Chichester, UK: John Wiley & Sons, Ltd

Erscheinungsjahr

2024

Quelle

Wiley-Blackwell Journals

Beschreibungen/Notizen

• BACKGROUND The melon fly, Zeugodacus cucurbitae (Coquillett), is an invasive Tephritidae pest with robust fertility. The male accessory glands (MAGs) form a vital organ that ensures insect reproductive efficiency. Most of the secreted proteins by MAGs exhibit a male bias expression. Takeout, one of these proteins, is abundantly present in the MAGs of many insects. RESULTS In this study, we identified 32 takeout genes in Z. cucurbitae. The phylogenetic analysis and multiple sequence alignment results showed that Zctakeout1 is the most related homolog to the MAGs‐specific takeout in Tephritidae. The real‐time quantitative PCR results showed that Zctakeout1 was exclusively expressed in the male adult stage, and its expression level gradually increased with the increase in age and then remained stable at the sexually matured stage. The distribution among tissues demonstrated the specific expression of Zctakeout1 in the MAGs, and fluorescence immunohistochemical results confirmed the presence of Zctakeout1 in close proximity to binuclear cells of the mesoderm epidermal MAGs. In continuation, CRISPR/Cas9‐mediated genome editing was employed, resulting in successfully generating a homozygous strain with an +8 bp insertion. The mating experiments with the Zctakeout1−/− males resulted in significant reductions in both the mating rate and egg production of females. CONCLUSION These findings prove that the MAGs‐specific Zctakeout1 is essential in regulating fecundity in female Z. cucurbitae fruit flies. Our data suggests its utilization in future essential insect‐specific gene‐directed sterility insect technique (SIT) by the genetic manipulation to keep these important Tephritidae populations under control. © 2024 Society of Chemical Industry. Zctakeout1 was exclusively expressed in the male adult stage. Immunohistochemistry confirmed the presence of Zctakeout1 in close proximity to binuclear cells in the mesoderm epidermal MAGs. CRISPR/Cas9 was employed to knock out this gene in males, and the mating experiments with the Zctakeout1−/− males resulted in significant reductions in both mating rate and egg production. These findings provide evidence that Zctakeout1 plays an essential role in the regulation of fertility in male Z. cucurbitae fruit flies, suggesting its utilization in future essential insect‐specific gene‐directed SIT for Tephritidae pest control.