POLLEN GRAIN: MATERIALS, ACETOLYSIS AND POLLEN PARAMETERS
The materials used in this research study were :beakers, water bath, glass rod, sieves, syringe, light microscope, slides, cover slides, sulphuric acid (H2 SO4), Ethanol, acetic anhydride, glycerol, distilled water, centrifuge machine, digital camera for photomicrograph, tissue paper, measuring cylinder and ocular micrometer.
The flower bud used from different species of sterculia with anthers were teased into beaker,20ml of ethanol was added to the beakers. A glass rod was used to squash the anther to release the pollen grains. The solutions were sieved to remove plant revolution per minutes. The solutions were centrifuged for 15 minutes at 2000 tissues. The solutions were decanted to recover the precipitates. The precipitates were washed twice with distilled water and 45% of acetic anhydride was added to remove the fatty oil, each wash was followed by centrifuging at 2000 revolution per minute for 10 minutes and decanted.
The recovered precipitates were acetolysed using Edtman’s 1971 acetolysis method of 9:1 acetic anhydrate and concentrated sulphuric acid (H2SO4). The solutions were boiled in a water bath for 5 minutes at 100oC and then centrifuged at 2000 revolution per minute for 5 minutes. The solution were decanted to recover the precipitates.
The recovered precipitation were washed with distilled water, centrifuged and decanted again. The recovered pollen grains were stored in vials with 10 drops of glycerol. Temporal slides of the precipitates were prepared and examined with light microscope using X10,X40,andX100. photomicrographs of the different pollen samples were conducted.
Different parameters were evaluated under the light microscope. Quantitative character which include the Polar View Diameter (PVD), the Equatorial View Length (EVL), the Polar View Length (PVL), the width of aperture (W.APO), were measured. Qualitative characterization of the pollen grains conducted were shaped at both equatorial and polar view, number of apertures and ornamental.