Foundation for Bioproduction: Secretory Stages, Metabolite Profiles and Gene Function of Glandular Trichomes in Cucumber.

Abstract

Glandular trichomes (GTs) are epidermal outgrowths that function as "natural cell factories" for the synthesis of specialized metabolites. Beyond their traditional understanding, GTs on cucumber fruits can form an undesirable trait known as bloom, which negatively affects market value. However, the secretory process, metabolite profiles, and genetic regulation underlying GT development in cucumber remain largely unclear. In this study, we employed scanning electron microscopy (SEM), transmission electron microscopy (TEM), histochemical staining, multi-omics analyses, and liquid chromatography-mass spectrometry (LC-MS) to systematically investigate GT development. The secretory process was classified into four distinct stages via SEM observations: morphogenesis, active metabolism, head sunken, and metabolite release. TEM revealed progressive ultrastructural changes, including increased organelle abundance and expansion of the periplasmic space, which facilitate metabolite transport and release. This process occurs through an autonomous mechanism involving osmiophilic substances and eventual cell rupture. LC-MS analysis identified 744 metabolites belonging to 11 classes, with phenylpropanoids/polyketides-particularly flavonoids-being the most abundant. While metabolite classes are conserved between European greenhouse and North China ecotypes, specific metabolite contents vary significantly. Multi-transcriptome analysis identified 60 candidate genes associated with GT development. Among these, <i>CsaV4_3G003418</i> was functionally validated through virus-induced gene silencing (VIGS) to be involved in early GT development. Collectively, this work elucidates the secretory mechanism and metabolic characteristics of cucumber GTs, providing a foundation for future functional studies and biotechnological applications of secondary metabolites.