Determination of pollen tube growth in plants
Familiarize yourself with methods of determining pollen tube growth and the major factors affecting growth.
Principle
Mature pollen is highly viable and can germinate and grow under suitable culture conditions. Pollen germination and growth are related to plant species, pollen maturity, climate and culture conditions. By changing the culture conditions and measuring the length of pollen tubes in an orthogonal experiment, we can find the optimal culture conditions to promote the growth of pollen tubes.
Operation method
Determination of pollen tube growth in plants
Principle
Mature pollen is highly viable and can germinate and grow under suitable culture conditions. Pollen germination and growth are related to plant species, pollen maturity, climate and culture conditions. By changing the culture conditions and measuring the length of pollen tubes in an orthogonal experiment, we can find the optimal culture conditions to promote the growth of pollen tubes.
Materials and Instruments
Materials: Mature flowers that have just opened or will open. Move The basic procedure for the determination of plant pollen tube growth can be divided into the following steps: 1. medium preparation 2. Experimental steps (1) Take a mature flower that has just opened or is about to open. (2) Prepare the culture chamber by placing a glass ring 15 mm in diameter and 5 mm in height on a clean, dry slide (Fig. 32-1). o (3) Place 1 drop of medium solution in the center of a clean, dry coverslip and scatter a few pollen grains on the medium. o (4) After 45-60 min of incubation in a 25 °C incubator, the growth was terminated with 1 mol・L-1NaOH solution and placed under a low magnification microscope for observation. (4)After 45~60 min of incubation at 25°C, the growth was terminated with 1 mol-L-1NaOH solution, placed under a low-power microscope, and the length of pollen tubes was calculated by micrometer. The length of pollen tubes was calculated by micrometer. 50 pollen tube lengths were observed in each treatment, and then the average value was calculated.
Reagents: culture medium (when preparing the medium, the concentration of agar remains the same and the concentration of boric acid and sucrose is changed to see what combination is more suitable for pollen germination and growth. If the medium is not used on the same day, it needs to be autoclaved).
Equipment: thermostat, microscope, eyepiece micrometer, objective micrometer, pollen culture chambers, shackles, slides, coverslips.
Caveat
Pollen should be spread evenly over the medium so as not to interfere with later observations.
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