Ultrastructural Evidence Elucidates the Mode of Action of Sulfur in Preventing Pollen Tube Development in Stigma of Citrus cv. Nadorcott and Other Horticultural Species

Abstract

Citrus species have a “wet”-type stigma with abundant exudate, and their style contains numerous canals. For successful seed formation, the process includes pollen grain germination on the stigma, pollen tube development and growth through the stigma and style and, ultimately, successful ovule fertilization. However, preventing the fertilization process can be useful for many agronomic and plant-breeding purposes, such as seedless fruits or for developing new varieties. Several studies have recently shown the inhibition effect of sulfur on pollen tube development inside the Nadorcott mandarin stigma and its effective application to obtain seedless mandarins. However, when applied to the stigma, how can sulfur inhibit pollen tube growth? Moreover, does sulfur have the same effect on other species? The main objective of the present study is to clarify the mode of action of sulfur on the ultrastructure of the Nadorcott mandarin stigma and style. To fulfill this goal, untreated flowers and flowers treated with sulfur were pollinated 24 h later. The treated and untreated stigmas were analyzed and compared with several microscopy techniques. The main results showed that sulfur specifically caused an alteration to the outer layer of stigma papillary cells. This marked alteration resulted in papillary cells losing their functionality due to the deterioration and degradation of their cellular structure. Basal papillae, the stigmatic tissue and stylar canals also underwent major alteration. Sulfur also modified the quantity and uniformity distribution of the stigmatic exudate. All these alterations collectively prevented pollen tube development inside the stigma. These effects have been observed in several Citrus species and varieties, and in some other horticultural species, which suggests a generic (non species-specific) action.