Tueux, G.; Pouilly, N.; Bernigaud-Samatan, J.; Blanchet, N.; Boniface, M.-C.; Catrice, O.; CARRERE, S.; Gouzy, J.; Jacquemot, M.-P.; Lauber, E.; Legendre, A.; Moreau, S.; Moroldo, M.; Roldan, A.; Carlier, A.; Langlade, N.

Nectar is a hub for plant-pollinator interactions, yet gene-level causal links between plant genetic variation, pollinator foraging and nectar microbial assembly remain poorly resolved. Using near-isogenic lines, innovative field time-lapse monitoring of pollinator visits and long-read amplicon sequencing of nectar microbiota, we show that a natural single-nucleotide variant at a cell-wall invertase gene (HaCWINV2) controls sunflower nectar chemistry and influences both pollinators and microbes. Plants homozygous for a loss-of-function HaCWINV2 allele produce sucrose-rich nectar, resulting in fewer bee visits under field conditions, while bumblebee visitation remained unaffected. In pollinator-excluded flowers, invertase-deficient plants harboured greater fungal diversity and compositionally distinct communities, indicating that nectar sugar profiles act as ecological filters shaping the nectar microbiome. This loss-of-function allele was found rarely and only at the heterozygote state in wild sunflowers and was fixed in 35% of cultivated lines indicating a positive selection during domestication. Our findings establish a causal link between a single gene and nectar chemistry with cascading ecological effects in a plant-pollinator system, illustrating how subtle genetic changes scale up to alter nectar traits, microbial assembly and pollinator foraging behaviour.