Optimized qualitative and quantitative methods for barley viability testing using triphenyl tetrazolium chloride staining

Abstract

Background and objectives Barley viability is an important agronomic trait and has a direct influence on subsequent beer production. Traditional triphenyl tetrazolium chloride (TTC) test evaluating seed viability still has some drawbacks, for example, excessive instability and time consumption. In the present study, we optimized both the qualitative and quantitative TTC seed viability test methods, and established more accurate and convenient assays to evaluate barley seed viability. Findings In the qualitative evaluation method, we established the vigor index according to the proportion of staining embryos, allowing quick evaluation of both germination and mortality rates in <12 hr. In the quantitative evaluation method, we optimized TTC incubation time, extraction temperature, and solvent volume to tissue ratios making results more reliable. Finally, we developed a high‐throughput method by combining freeze drying and tissue lysing instead of the time‐consuming method of grinding on liquid nitrogen. There was a positive significant statistical correlation between germination rate and formazan content by the optimized quantitative evaluation method (R2 = 0.907; p < 0.01). Conclusions Both the optimized qualitative and quantitative TTC seed viability test methods allow a quick and reliable seed vigor evaluation for different varieties or different aged barley seeds. Significance and novelty The optimized seed vigor evaluation method can be used to effectively screen out low vigor barley seeds. The qualitative evaluation method is appropriate to quickly evaluation for a small number of samples in <12 hr. The high‐throughput quantitative evaluation method is fit for a large number of sample evaluations as much as 80 samples at the same time.

A comparative study on seed physiology and germination requirements for 15 species of Eucalyptus

“…quadrangulosa although all three species had similar TZ viability scores. In contrast, Ma et al (2019) observed a positive correlation between TZ viabilities and vigour however they were studying different batches of a single species. The TZ seed vigour test is not an effective measure of vigour when comparing different species, especially where there are significant differences in seed size, as with the 15 species of eucalypts studied here.…”

Seed Treatment with Iron Chlorine E6 Enhances Germination and Seedling Growth of Rice

“…After 2 days, when the seeds were just budding, seeds were collected and homogenized in water to produce crude enzyme extract. The seed viability was assessed using quantitative 2,3,5-triphenyltetrazolium chloride (TTC) assay [21]. The content of triphenyl tetrazolium formazan (TTF) produced by dehydrogenase per gram of seed in 1 h was used as seed vigor index.…”

“…Seed vigor is an important comprehensive index to reflect the quality of seeds, and it is a characteristic of high germination rate [24]. The TTF content in seeds is positively correlated to germination rate [21], which was used as the seed vigor index. In the…”

“…Seed vigor is an important comprehensive index to reflect the quality of seeds, and it is a characteristic of high germination rate [24]. The TTF content in seeds is positively correlated to germination rate [21], which was used as the seed vigor index. In the standard curve, TTF content (0.05-1 mg) had a linear relationship with absorbance at 485 nm (y = 0.58326x + 0.00066, R 2 = 0.9996).…”

“…Funding: This research was funded by the Jiangsu Agricultural Science and Technology Innovation Fund, grant number CX (21)1011.…”