Rapid determination of seed germination rate—BTB method

Rapid determination of seed germination rate—BTB method

   Seed germination rate refers to the percentage of seeds that germinated in the specified number of days under the most suitable conditions. It is the main basis for determining seed quality and practical value, and it is directly related to the amount of seed used when sowing. However, the conventional method (direct germination) requires a long time to determine the germination rate, especially for emergency needs, there is not enough time to determine the germination rate, and it is impossible to know when it encounters dormant seeds. The rapid determination method can obtain the result in a short time.
Bromothymol blue method (BTB method)
Principle: All living cells must have respiration, absorb O2 in the air and release CO2. CO2 soluble
Increase, bromothymol blue (BTB) can be used to determine the change in acidity. The discoloration range of BTB is pH 6.0-7.6, acid is yellow, alkaline is blue, and the middle passes through green (the discoloration point is pH 7.1). The color difference is significant and easy to observe.
Instruments and drugs: incubator, balance, petri dish, beaker, tweezers, funnel, filter paper, agar
0.1% BTB solution: Weigh 0.1g of BTB and dissolve it in boiled tap water (the water used to prepare the indicator should be slightly alkaline to make the solution blue or blue-green, distilled water is slightly acidic and not suitable), and then use filter paper Filter out the residue. If the filtrate is yellow, add a few drops of dilute ammonia to make it blue or blue-green. This liquid can be stored in a brown bottle for a long time.petri dish,petri dish 90mm sterile,disposable cell culture 90x15mm petri dish,plastic petri dish,disposable petri dish
1% BTB agar gel: Take 100ml of 0.1% BTB solution and place it in a beaker, cut the agar into small pieces and add it, heat it over a small fire and stir constantly. After the agar is completely dissolved, pour it into several dry petri dishes while it is hot to form a uniform thin layer, and cool it for later use.petri dish,petri dish 90mm sterile,disposable cell culture 90x15mm petri dish,plastic petri dish,disposable petri dish
Steps:
1. Seed soaking
Soak the tested seeds in 30-35℃ warm water (barley, wheat, indica for 6-8 hours, corn for about 5 hours, and japonica for 2 hours) to enhance the respiration strength of the embryo and make the color development fast.
2. Color rendering
Take 200 swollen seeds and bury them neatly in the prepared agar gel petri dish. Place the seeds flat with a distance of at least 1 cm. Then place the petri dish at 30-35°C for 2-4 hours, and observe under a blue background. If there is a darker yellow halo near the seed embryo, it is a live seed, otherwise it is a dead seed.petri dish,petri dish 90mm sterile,disposable cell culture 90x15mm petri dish,plastic petri dish,disposable petri dish
The seeds killed by boiling water were treated in the same way, and the observation was made for comparison.

3. Count the number of live seeds with a yellow halo near the embryo to calculate the bud rate.petri dish,petri dish 90mm sterile,disposable cell culture 90x15mm petri dish,plastic petri dish,disposable petri dish

Rapid determination of seed germination rate---BTB method


Post time: Nov-17-2021

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