The Impact of Pollinating Fig Wasps' Entry on Fig Development and the Hormonal Regulation of Sex Differentiation in Ficus hispida

Ficus trees (Moraceae) play a vital role in sustaining the stability of tropical and subtropical rainforests. The obligate mutualism between Ficus species and their pollinating fig wasps renders them an exemplary model for investigating insect-plant coevolution. In this study, we employed Ficus hispida Linn. f., an ecologically significant fig species in tropical rainforests, to conduct a wasp-introduction controlled experiment in the field. This method enabled us to precisely delineate the developmental stages of figs. We collected samples at specific intervals and examined the impact of pollinating fig wasp entry on the hormonal metabolism of male and female figs using liquid chromatography-tandem mass spectrometry analysis. The findings demonstrate that pollinator entry significantly decreases fig abscission. Moreover, it substantially altered the developmental indices of the figs. Unpollinated figs exhibit elevated levels of abscisic acid (ABA), which increases the likelihood of fig abortion and reduces the probability of pollinator entry into senescent figs. Following pollinator entry, indole-3-acetic acid (IAA) levels rise in both male and female figs. Male figs show higher concentrations of 1-aminocyclopropane-1-carboxylic acid (ACC), jasmonic acid (JA), and salicylic acid (SA), whereas these changes are less pronounced in female figs. Additionally, pollinated male figs display increased levels of cytokinins (CKs) and other hormones compared to female figs, suggesting a coordinated hormonal response to the stress induced by pollinator oviposition and gall development. Our findings suggest that the entry of pollinators likely triggers the transition from the female to the interfloral phase, with hormonal regulation playing a crucial role in the reproductive dimorphism of figs. This research can offer novel insights into the mechanisms underlying fig-wasp mutualism.