Objective
This practical experiment was carried out to determine the factors that damage the cells that contain bactecyanin pigment. The idea about this experiment emerged during the previous laboratory experiment using the beetroot seeds. The seeds stained the hands when they were stressed.
Introduction
In the cells of beetroot plant, there is a pigment that gives it its red color. The pigment can drain from the cell like an ink. This situation can help in determining which factors can influence the integrity of the cells. Beetroot pigments are not stable when the temperature is high. A number of parameters such as the oxygen concentration, the pH, and the length of heating also affect the beetroot cells.
Materials
The materials used were; scalpel, test tubes, water, refrigerator, acetone or alcohol, forceps, beaker, spectrophotometer, graduated cylinder and hot plate.
Methods
The assumption of this practical experiment was that the increase in temperature and addition of organic solvent would lead to the leakage of the pigment in the beetroot cell.
The beet was cut into three cubes of equal sizes using a scalpel. After washing the cut pieces, they were placed in a test tube filled with water. The tubes were then placed at different temperatures. After some time, the beetroots were removed, and measurements taken using a spectrophotometer at 460nm wavelength.
In the second part, the beetroot was cut into two cubes of the same size using a scalpel. After washing the beets, they were placed into separate test tubes. Each tube contained acetone of different concentration. The observation was then done on the remaining liquid after removing the beetroot pieces. The color was measured on a scale of 1 to ten at 460nm wavelength spectrophotometer.
Results
The color of the pigment leaked in from the cells. In the first test tube of the first part of the experiment, treatment temperature was 40o C, and the absorbance was observed to be 0.04nm and color intensity as 1. The second one was at 5o C, and the absorbance was observed to be 0.03nm and color intensity as 0. In the third test tube, the temperature was -5o C and absorbances were 0.64nm and color intensity as 8.
In the second part of this experiment the results from the first test tube that contained 1% acetone was 3 for light intensity and 0.05 for absorbance. The second test tube contained 50% acetone, and the results were 7 for light intensity and 0.47 for absorbance.
Discussions and Conclusions
The beetroot cell could withstand a small amount of stress, but extreme changes in temperature and organic solvent concentration damaged the cells. The damage led to leakage of the red pigment.
Increase in temperature is known to weaken the cell membrane structure. Low temperature also decreases the fluidity of the membrane. Therefore, there is a certain temperature where the cell membrane functions normally.
When the temperature is high, many betalain molecules are lost from the vacuole as compared to when the temperature is low. At high temperature, the betalain molecules are converted into kinetic energy, which leads to release of the red pigment through the cell membrane into the water. The pigment molecules are saturated in the vacuole of the cell, their instability due to temperature, or any stress may lead to their leakage through the cell membrane.
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