The estimation of pollen viability in rice

Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor

“…We observed that incubation time was important when detecting viability, the MTT assay having an optimum incubation time of 120 min. These results are in accordance with previous work: the time for the MTT reaction can vary from 15 min for pollen (Khatun and Flowers, 1995) to 24-48 hours for spores of phytopathogenic fungi (Sutherland and Cohen, 1983). These authors also consider that MTT concentration and incubation temperature and time are important factors in the assay.…”

“…In agreement with these authors, we also observed that it was difficult to discriminate between living and dead cells. In contrast, the MTT assay based on the cell metabolic activity changes the colour of living cells by the reduction of the salt (MTT), forming an insoluble compound of red or purple colour (formazan) through the action of the mitochondrial dehydrogenase (Tisserat and Manthey, 1996), whereas the black colour or no change in colour is a sign of cell death (Khatun and Flowers, 1995).…”

“…Tetrazolium salts are reduced in viable cells to a red coloured water-insoluble formazan by the action of dehydrogenases (Tisserat and Manthey, 1996;Verleysen et al 2004). Black coloured or noncoloured cells are considered dead (Khatun and Flowers, 1995).…”

“…For example, cell suspensions of Glycine max infected by Fusarium solani were stained with 0.1% phenosafranin, the living cells acquiring a red colour (Li et al 1999). Khatun and Flowers (1995) estimated the viability of pollen of Oryza sativa with fluorescein diacetate, aniline blue, TTC and MTT (0.1-1%); the best results were obtained with MTT, with viable cells staining light red to red and yellow, non-stained or black pollen were considered non-viable.…”

Evaluation of Pollen Viability of Nakdongbyeo, Two Transgenic Rice Lines, Its Hybrids with Weedy Rice, and Subsequent Selfed Progenies: F2and F3

“…To make 0.9% final concentration of 3-{4,5dimethylthiazolyl-2}-2, 5-diphenyl monotetrazolium bromide (MTT), 0.1125 gram of MTT was dissolved in 5% sucrose solution to make total volume of 12.5 ml of MTT in 5% sucrose solution [9,15].…”