Seed assay by triphenyltetrazolium chloride (TTC) method

Summary

Viable seed embryos undergo continuous redox reactions during respiration, and the hydrogen shed reduces the coenzymes (NAD or NADP). When TTC penetrates into the living cells of the seed embryo and acts as a hydrogen acceptor to be reduced by hydrogen on the reducing coenzymes (NADH+H+ or NADPH+H+), it changes from colorless triphenyltetrazolium chloride (TTC) to reddish triphenylmethyl filth (TTF).

Operation method

Seed assay by triphenyltetrazolium chloride (TTC) method

Principle

The respiration of viable seed embryos undergoes a continuous redox reaction in which the hydrogen shed reduces the coenzymes (NAD or NADP). When TTC penetrates into the living cells of the seed embryo and acts as a hydrogen acceptor to be reduced by hydrogen on the reducing coenzymes (NADH+H+ or NADPH+H+), it changes from colorless triphenyltetrazolium chloride (TTC) to reddish triphenylmethyl filth (TTF).

Materials and Instruments

Wheat, corn, peanut, rice seeds
TTC solution
Thermostat Petri dish Tweezers Blades

Move

I. Materials and equipment

Wheat, maize, peanut or rice seeds

Thermostat, Petri dish, blade, tweezers

0.1% TTC solution: weigh 0.1 g of TTC and add a small amount of 95% ethanol to dissolve and then add distilled water to 100 ml.

Experimental steps

1. Seed dipping: same as BTB method.

2. Color development: take 10 sucked seeds, with a razor blade along the center of the seed embryo longitudinal cut into two halves, one half of each seed, placed in a petri dish, add the appropriate amount of 0.1% TTC solution, submerged seeds, in the 45 ℃ incubator for 20 minutes, take out and observe, where the embryo is stained red is the living seed. The other half is kept for the red ink method.

Can be taken with boiling water to cook dead seeds for the same treatment, control observation.

3. Count the seeds with colored embryos and calculate the percentage of viable seeds.


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Categories: Protocols