Standardization of the fluorochromatic reaction test to assess pollen viability

Abstract

The fluorochromatic reaction that viable pollen grains exhibit when exposed to fluorescein diacetate (FDA) is an easy, quick and accurate tool for assessing pollen viability in many plants. Despite its widespread use, the method as initially proposed by Heslop-Harrison and Heslop-Harrison lacks specificity in some respects that are essential for comparing simultaneously the viability of different pollen sources. We have determined the time needed for the fluorochromatic reaction to take place and fade, and the lifetime and most effective concentration of the FDA working solution. Our results show that consistent records of pollen viability in samples of olive (bicellular pollen grains) and cherimoya (tricellular pollen grains) can be obtained within minutes, but that measurement must be completed in the first hour after pollen exposure to FDA. The lifespan of the FDA working solution is 1 h, whereas concentrations between 1.5x10(-5) M and 1.8x10(-3) M are equally valid. Pollen grains of both species responded similarly, although some differences were observed in the persistence of fluorochromasia.

Effect of seasonality and time after anthesis on the viability and longevity ofCannabis sativapollen

“…In this study, among all three staining techniques, the FCR test was considered reliable to test viability. In earlier studies also, FCR has also been regarded as the best cytochemical test to measure the viability and ability to germinate of pollen (Heslop-Harrison & Heslop-Harrison 1970;Shivanna & Heslop-Harrison 1981;HeslopHarrison et al 1984;Shivanna et al 1991;Dafni 1992;Kearns & Inouye 1993;Pacini et al 1997;Dafni & Firmage 2000;Pinillos & Cuevas 2008;Aguilera & Valenzuela 2013). However, the Alexander and TTC tests were discarded due to faulty results, as these staining tests did not allow discrimination in regard to the estimation of pollen grain viability.…”

Adaptor Protein Complex 2–Mediated Endocytosis Is Crucial for Male Reproductive Organ Development in Arabidopsis

“…The amount of pollen produced in the anthers appeared to be greatly reduced in ap2m plants. To look for defects in pollen quality, the viability of pollen grains was tested using propidium iodide and fluorescein diacetate (FDA), which stain dead and live pollen, respectively (Pinillos and Cuevas, 2008) (Figure 4A). Dead pollen grains amounted to 42% in ap2m-1 plants, compared with only 15% in the wild type ( Figure 4B).…”

Characterization of the procera Tomato Mutant Shows Novel Functions of the SlDELLA Protein in the Control of Flower Morphology, Cell Division and Expansion, and the Auxin-Signaling Pathway during Fruit-Set and Development

“…First, by looking at the number of pollen grains stained in red with carmine acetate solution. Second, by incubating extracted pollen in 2.4 3 10 25 M fluorescein diacetate solution for 10 min and measuring fluorescence intensity (l ex = 494 nm, l em = 521 nm) in a Perkin-Elmer LS50B spectrofluorimeter (Pinillos and Cuevas, 2008). Pollen germination tests were performed on glass slides coated with germination medium [0.292 M Suc, 1.27 mM Ca(NO 3 ) 2 , 1.62 mM H 3 BO 3 , 1 mM KH 2 PO 4 , and 0.5% agarose].…”