The relationship between in vivo pollen germination and vital staining was tested for 7 genotypes of deciduous azalea (Rhododendron sp.) differing in fertility. Fluorescein diacetate, 3-amino-9-ethylcarbazole (AEC), and 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyl tetrazolium bromide (MTT) all showed significant correlations between staining and germination. AEC produced the highest significant correlation (r = 0.96). AEC and MTT were the most suitable vital stains for azalea pollen.
A procedure is described for determining pollen viability in tomatoes (Lycopersicon esculentum Mill.) by growing pollen in a growth medium containing 0.29 M sucrose, 1.27 mm Ca(NO3)2, 0.16 mm H3 BO3, and 1 mm KNO3 (pH 5.2) to which 0.001% fluorescein diacetate (FDA) was added. Pollen viability can be evaluated within 30 min by determining percent fluorescing pollen in a sample. The procedure further allows the determination of percent germination in vitro and pollen tube growth within 1.5 hours. Neither the germination medium nor FDA has any adverse effects on germination and pollen tube growth. Percent fluorescent pollen and percent total pollen germination were highly correlated, suggesting that fluorescence is a good measure of pollen viability. The combined fluorescence-germination procedure is simple and adapted to routine screening of many samples.
cytoplasm; the PG considered viable were those whose cytoplasm remained stained and intact; and b) fluorescein diacetate ( Heslop-Harrison and Heslop-Harrison, 1970 ), to detect the esterase activity and plasmalemma integrity of the vegetative cell. For each
, University of Novi Sad, voucher No. 2-2069. Pollen viability test. A rapid method with fluorescein diacetate (FDA) ( Heslop-Harison and Heslop-Harison, 1970 ) was used to determine pollen viability after 1–2 h. FDA (2 mg·L −1 ) dissolved in acetone was
vitro plantlets were soaked in liquid media with different pH levels for 6 h. Then roots were stained with fluorescein diacetate/propidium iodide solution. Green and yellow cells were evaluated as viable, whereas red cells were scored as damaged. Bar
. angustipetala . Pollen viability. Pollen viability was assessed using a fluorescein diacetate (FDA) staining procedure developed by Heslop-Harrison and Heslop-Harrison (1970) . Flowers were collected on the day of anthesis from five randomly chosen hybrids per
pots (total volume of 1.5 L). Results for media respiration were expressed in μmol CO 2 /cm 2 /s. Determination of fluorescein diacetate hydrolysis. An estimation of the growing medium general microbial activity was performed 5 and 10 weeks after
was estimated in the hybrids and parents by fluorescein diacetate staining. ‘Annabelle’ and ‘Blue Diamond’ had 76.4% and 48.4% stainable pollen, respectively. Stainable pollen ranged from 0% to 76.5% among 14 hybrids ( Table 3 ). In previous studies
standard for comparison of pollen division patterns. Table 1. Genome constitution and cytological characteristics of nine Doritaenopsis hybrids. Pollen staining with fluorescein diacetate (FDA). Pollen viability analyzed by staining
A seasonal study was conducted to assess the freezing injury of `Boskoop Giant' black currant (Ribes nigrum L.) samples from Oct. 1991 through Mar. 1992. Buds were subjected to either differential thermal analysis (DTA) or one of a series of temperatures (0 to -36C). Freeze injury was then assessed either visually or with TTC. Results indicated that black currant floral buds have multiple low-temperature exotherms (LTE). Freeze injury in intact buds could not be visually quantified because of the lack of visible browning, nor assayed with TTC reduction. Excised floral primordia incubated in TTC, however, developed colored formazan following exposure to nonfreezing and sublethal freezing temperatures, but remained colorless when exposed to lethal temperatures. The percentage of floral primordia that were colored and colorless were tabulated and a modified Spearman-Karber equation was used to calculate the temperature at which 50% of floral primordia were killed (T50 The T50 temperature was correlated with the temperature at which the lowest LTE was detected (R2 = 0.62). TTC reduction assay using excised floral bud primordia was a good indicator of viability in frozen blackcurrant buds. Chemical name used: 2,3,5-triphenyltetrazolium chloride (TTC).