Seed Viability Bromothymolan (BTB) Method Identification Experiment

Summary

Where viable seeds can constantly respire, absorbing O2 from the air while releasing CO2, CO2 dissolves in water to form H2CO3, which is unstable and dissociates into H+ and HCO3-.

Operation method

Rapid identification experiments of seed viability

Principle

Where viable seeds can constantly respiration, absorption of O2 in the air, while releasing CO2, CO2 dissolved in water to form H2CO3, H2CO3 unstable dissociation into H + and HCO3-. Due to the continuous dissociation of H2CO3, the acidity of the surrounding medium increases gradually. The change of acidity is determined with BTB.The molecular structure formula of BTB is shown in the figure: its color change range is between pH6.0-7.6, it is yellow in acidic medium, blue in alkaline medium, and passes through the green in the middle (the color change point is pH7.1).

Materials and Instruments

Corn Wheat Seed
BTB solution BTB agar gel
Thermostat Petri dish Tweezers Filter paper Drug balance Beaker Funnel Agar BTB Electric stove Measuring cylinder

Move

I. Materials and equipment

Corn or wheat seeds

Thermostat, Petri dish, forceps, filter paper, drug balance, beaker, funnel, agar, BTB, electric stove, 50 ml measuring cylinder

0.1% BTB solution: weigh 0.1 g of BTB, dissolve in boiled tap water, then filter off the residue with filter paper. If the filtrate is yellow, add a few drops of dilute ammonia to make it blue or blue-green, and store this solution in a brown bottle for a long time.

1.5% BTB agar gel: take 0.1% BTB solution 40 ml in a beaker, and weigh 0.5 grams of agar, cut it into pieces and add it to the cup, with a small fire (electric stove) heating and stirring. When the agar is completely dissolved, it can be slightly cooled and poured into a 9cm Petri dish while it is still hot, so that it will form a uniform thin layer, and then cooled completely and prepared for use.

Experimental Steps

1. Seed soaking: Soak the seeds to be tested in warm water at 30℃-35℃ for about 5 hours to enhance the respiratory strength of the seed embryo.

2. Color development: Take 10 sucked seeds, neatly buried in the prepared agar gel, pay attention to the embryo buried in the gel. The petri dish will be placed in the 35 ℃ temperature box in an hour visible results, more than two hours the results are more obvious. Observe the yellow halo around the seed embryo is live seed, otherwise it is dead seed.

3. Calculation: count the number of seeds with yellow halo around the embryo one by one, and calculate the percentage of viable seeds.

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