Preliminary Investigation of Foliar Application of Boron on Pollen Viability and Development in the Cultivation of Red Clover in the Field

Red clover (Trifolium pratense L.) is a crop used as a forage that possesses an exceptional nutritional profile and digestibility. Unfortunately, this crop has low seed yield. Within the framework of the “Legume Generation” EC-funded project, our team aimed to investigate the role of foliar boron application on pollen viability and pollen tube development, and to assess its overall effect on red clover cultivation. Plants of six commercial diploid red clover cultivars, Nika 11, Sofia 52, AberClaret, Milvus, Global, and S123, were field-grown and boron-treated by spraying with the commercial product “Lebasol”, 11% active water-soluble boron. To reach our purpose, the transcript levels of genes related to flower, pollen, and pollen tube development and boron transport were measured by qRT-PCR; pollen grain viability and count were assessed microscopically. For this research, eight genes were selected: Auxin Response factor (TprARF17); TprAPETALA3; Walls are thin (TprWAT1 and TprWAT2); NIPs genes (Nodulin Intrinsic Protein) TprNIP4;2, TprNIP7;1, TprNIP5;1, and TprNIP6;1. Additionally, total nitrogen content in leaves detached from field-grown boron-treated and untreated plants was assessed and compared with the expression levels of two TprNIP5;1 and TprNIP6;1 transporters. The fresh and dry biomass weight from the first and second cuts was evaluated, as well as the seed collected from the red clover plants. Seed germination percentage and vigor of seedlings were examined in vitro for both boron-treated and untreated groups of two specific cultivars. Collected data confirm that foliar application of boron affects pollen viability and plant development of red clover in the cultivation conditions of South East Europe.