Pollen Development and Stainability in Vicia faba L. and Lupinus angustifolius L.

Abstract

Commercially, leguminous crops (Fabaceae) are the second most important group of cultivated plants, just after grasses (Poaceae). This study focuses on the analysis of pollen development and stainability in two species belonging to the Fabaceae family: Vicia faba L. and Lupinus angustifolius L. Morphological analysis of the anthers at various stages of flower development allowed us to trace the processes of microsporogenesis and microgametogenesis. Nine different cell staining protocols with diverse mechanisms of action, including acetocarmine, Alexander’s dye, aniline blue in lactophenol, Calcein AM, FDA, MTT, TTC, Lugol’s iodine, and aceto-orcein, were tested for their suitability in assessing the viability of microspores as well as pollen grains in both species. Among the applied dyes, four allowed for the discrimination between viable and nonviable microspores in V. faba, and six dyes allowed for this in L. angustifolius. For mature pollen grains, all dyes enabled differentiation between viable and nonviable cells in both species. The highest viability indications for V. faba microspores were obtained with acetocarmine (94.6%), while for mature pollen, aniline blue in lactophenol, MTT, and aceto-orcein yielded the highest viability indications (90.8–96.3%). In L. angustifolius, the highest percentages of viable microspores (64.9–66.5%) were obtained with the acetocarmine, aniline blue in lactophenol, and TTC dyes. For mature pollen, the highest viability indications (83.4%–92.9%) were obtained with acetocarmine, aniline blue in lactophenol, Lugol’s iodine, and aceto-orcein. The viability of V. faba pollen grains in an in vitro germination test showed that the highest pollen germination (61.3%) was observed on the BK medium (rich in minerals with 10% sucrose). In L. angustifolius, the highest pollen germination was observed on the media containing boric acid and 5% sucrose (70.5%) and on the medium containing 10% sucrose only (74.2%).