Acid Indigo Staining for Seed Viability Detection Experiment

Summary

Knowledge of the principles and methods of determining seed viability by the staining method

Operation method

Acid Indigo Staining for Seed Viability Detection Experiment

Principle

The protoplasmic membrane of the embryonic cells of living seeds is semi-permeable and has the ability to selectively absorb external substances, so that dyes cannot enter the living cells and the embryonic parts cannot be stained. The loss of vitality of the seed, the embryonic cell protoplasmic membrane has lost the ability to selective absorption, dyes can freely enter the cell and will be the dead embryo staining (such as acid indigo, red ink, etc.), so the embryonic part of the seed according to whether it is stained to determine the vitality of the seed.

Materials and Instruments

Barley Soybean
Acid indigo aqueous solution
Petri dishes, blades, tweezers, beakers.

Move

I. Materials, equipment and reagents

1. Materials: New seeds of barley, soybean and other crops and old seeds stored for more than three years.

2. Equipment: Petri dishes, blades, tweezers, beakers.

3. Reagents: 0.1% aqueous solution of acid indigo (0.1 g dissolved in 100 ml of water).

II. Experimental steps

1. Seed treatment

First, barley and soybean seeds were soaked in water for 1~2 h, and after fully absorbing and expanding, a part of them was taken out and boiled in boiling water for 3~5 min as dead seeds. Select 100 seeds each of water-soaked new, stale and dead seeds, in the case of soybeans, carefully peel off the seed coat with tweezers, in the case of wheat use a razor blade to cut into two halves longitudinally along the midline of the embryo, and use one half of them for the assay (need to have the embryo).

2. Staining

Spread the treated seeds evenly in a petri dish and add 0.1% acidic indigo solution to submerge the seeds.

3. Rinse the seeds

After 15-20 minutes of dyeing, pour out the solution and rinse the seeds repeatedly with tap water until the dyed color no longer washes out.

III. Observation

Compare and observe the coloring of the embryo of fresh, old and dead seeds after rinsing. Where viable seeds have no coloring of the embryo at all, or have insignificant lightly colored spots above the cotyledons and radicle, if the entire radicle is fully colored or there are heavily colored spots on the radicle and cotyledons, they are seeds that have lost all or most of their viability. The percentage of viable seeds was counted as an estimate of the germination rate and compared with the actual germination rate (determined beforehand by the ordinary germination test method).

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