Mechanized hand duster pollination maintains key fruit traits and mogroside V biosynthesis in Siraitia grosvenorii

• Hand duster pollination (HDP) improves pollination efficiency in S. grosvenorii . • HDP could replace hand supplementary pollination (HSP) in S. grosvenorii . • HDP does not alter mogroside V synthesis and content. Siraitia grosvenorii is a high-value crop prized for its fruits that contain mogrosides, particularly mogroside V, a natural zero-calorie sweetener with substantial health benefits. However, its cultivation is constrained by complete reliance on labor-intensive artificial pollination, due to its dioecious nature and inability to utilize on wind or insect pollination. This study assessed the feasibility of mechanized hand duster pollination (HDP) as an alternative to hand supplementary pollination (HSP) in S. grosvenorii , combining agronomic and molecular analyses. Results showed that HDP achieved a fruit set rate comparable to that of HSP, alongside an identical final mogroside V content, despite slight transient decreases in fruit size and weight. Transcriptomic analyses revealed HDP induced early stress responses but did not alter core mogroside biosynthetic pathways; WGCNA further confirmed that gene co-expression networks associated with fruit quality, specifically mogroside V accumulation, are independent of pollination method. Thus, HDP emerges as a labor-saving, efficient alternative to HSP, directly facilitating sustainable commercial cultivation of S. grosvenorii without compromising the yield of its key bioactive compound.