Lodine-Potassium Lodide (I-KI) Dye Solution

This product can be used to display starch in plant tissues or to identify pollen fertility. The principle is that iodized starch is formed when iodized potassium iodide solution interacts with starch, which shows a special blue reaction. Iodized potassium iodide solution is a classical method to identify starch and an important reagent for plant histochemical identification. This product contains two components: iodine-potassium iodide staining solution and plant solid green staining solution. After staining, the starch grains were blue or blue-black, and the cell walls were green. If used to identify pollen fertility, fertile pollen grains are round, large and plump, with a uniform dark blue color.

Store and transport at room temperature; the iodine-potassium iodide staining solution should be stored sealed away from light and valid for 12 months.

1. Plant tissue section observation of starch granules (prepared anhydrous ethanol, xylene, neutral gum, etc.)

(1) Paraffin sections dewaxed to water: histochemical strokes encircle tissues,

(2) Iodine potassium iodide staining: circle the tissues with histochemical strokes, and add iodine potassium iodide solution to cover the tissues for 5-10 minutes. Quickly wash twice for 2 seconds each time.

(3) (Optional) Fast Green fixation staining: the slices were dehydrated quickly with 2 cylinders of absolute ethanol for 10 seconds each time. Immerse in the plant fast green dye for 10 s. The slices were dehydrated rapidly by 3 cylinders of absolute ethanol for 10 seconds each time.

(4) Transparent and mounting: xylene is transparent for 5 min, and the section is sealed with neutral gum. It is very easy to fade, and it is recommended to take a mirror inspection as soon as possible and take photos to save the results.

2. Identification of pollen fertility (taking rice pollen as an example)

(1) Anther collection: when the rice is heading, the spikelets flowering at the early stage of the rice spike are taken and fixed in 75% ethanol, and stored at 4 ℃. Generally, it is required to take the upper, middle and lower parts of the rice spike, and the anther length is more than 2/3 of the length of the glume as the mature anther.

(2) Staining microscopy: use pointed tweezers to take the anthers and put them on the slide, add 1-2 drops of iodine potassium iodide staining solution to cover the anthers, use tweezers to crush the anthers, so that the pollen grains can be fully released. Then cover the cover glass, gently press the cover glass with tweezers, and let it stand for 2-3 minutes. Observe under a 10 fold microscope.

(3) Generally, fertile pollen grains are round, large and plump, dark blue, and evenly colored; There are four types of sterile pollen, including pollerless type, typical abortion, round abortion and dye abortion. Pollen free type refers to no pollen or only residual pollen wall observed under microscope; The typical abortion means that the pollen is not stained and its shape is irregular, such as triple tube, polygon, etc; Round abortion refers to the round appearance of pollen grains without stained starch grains; Pollen abortion refers to that most pollens are normal in morphology, but their coloration is shallow or uneven, and some pollens are deeply stained, but their grain shape is obviously different from that of normal fertile pollens.

1. After using the reagent, please tighten the cap in time to prevent solvent volatilization.

2. Each set of staining solution can be used to stain approximately 400 sections. Replace the stain with a new one when the tissue or cell coloring is significantly lighter or abnormal.

3. Wear a lab coat and disposable gloves during operation.

For Research Use Only!