University of Bahrain College of Science Department of Biology 2nd Semester 2011/2012

BIOLS 320 Project Report

Effect of Enviornmental Stress on Plants

Name:
Asma Tirmizi Bisma Bashir Fatima Abdul Waheed Rukaya Abdul Ghafoor

IDs
20092655 20094704 20092655 20094703

Effect of Enviornmental stress on Plants BIOLS 320

Table of Contents: 1. Summary. 2. Introduction. 3. Methods & Materials. 4. Results 5. Discussion & Conclusion. 6. Recommendations 7. References.

2|Page

Effect of Enviornmental stress on Plants BIOLS 320

Summary:

Introduction:

Methods & Materials:

Water, soil and plants samples from Tubli and Sitra with 3 Replicates were collected & were used for the following analysis. 1. Total Soluble Salts (TSS): Water & soil samples from the two stations had 3 replicates. The TSS for each of the 3 replicates from the station was determined by using a refractometer. And the readings were recorded. 2. pH: Three replicates of the water & soil samples from the two stations were present. The pH of the soil & water samples were measured by a pH meter and the pH were recorded. 3. Water potential measurements: Eight beakers were labeled with the different sucrose solutions i.e.0.1M, 0.2M, 0.3M, 0.4M, 0.5M, 0.6M, 0.7M and distilled water for the two stations (Sitra & Tubli). Then a set of two leaves from each station were washed with distilled water. Then the leaves were dried using tissue papers and weighed and the weight was noted. After this the dried leaves were added to different labeled beakers. And the beakers were covered with parafilm. The leaves were left in beakers for 90 min. After 90 min, the leaves from each of the beakers were dried again and reweighed and the weight was recorded.
Tubli Sitra Figure 2: pH meter Figure 1: Hand Refractometer

3|Page

Figure 3: leaves soaked in sucrose solution

Effect of Enviornmental stress on Plants BIOLS 320

4. Chloride ion accumulation: Chloride Ion content for the following samples was measured: Leaf Sap Water from the leaf surface Sediment Pond water

Sample preparation: Water from leaves surfaces: The leaves were washed with distilled water. 0.2ml of the water used to wash the leaf surface was then diluted with 49.8 ml of distilled water. Leaf sap: The leaf sap was obtained by grinding the leaves and then adding 1 ml of distilled water. Then 0.2 ml was pipetted from the grounded laves, and was diluted with 49.8ml of distilled water. Sediment: 10 g of soil sediment was weighed and dissolved in 40 ml of distilled
Figure 4: leaf sap preparation

water, and was left to settle down for few minutes. After the sediment was settled, 0.2 ml of water from the solution was pipetted and then diluted with 49.8 ml of distilled water.
Tubli Sitra

Figure 5: sediment soaked in water

Pond water: 0.2 ml of pond water was pipetted and diluted with 49.8 ml of distilled water. All these diluted sample solutions were titrated with 0.02N AgNO3. Five drops
Figure 6: titration

of the indicator K2CrO4 was added to the samples before each titration. Titration was done until the solution became reddish-brown in color, and the volume of AgNO3 used was recorded. 5. Chlorophyll Content Index (CCI): Chlorophyll Content Index was measured using a Chlorophyll Content Meter. 6. Chlorophyll Fluorescence (CF): To measure the Chlorophyll Fluorescence of the leaves obtained from two different areas. The leaves were 1 st clipped to provide darkness to the leaves. After 30min. the window of the clips were opened and the fluorescence was measured using a Fluorometer.
4|Page

Effect of Enviornmental stress on Plants BIOLS 320

Tubli Sitra

Figure 7: Dark adaptation for leaves

7. Moisture Content: The leaves were weighed freshly and then after keeping them in the oven for 24h, their dry weight was taken (lab handout, 2012). DATA ANALYSIS: Simple linear regression to estimate water potential. Using ANOVA single factor to determine any significant.

Results:
Generally observing the plants, following observations were made:
Table 1: General Observations

Tubli Height # of leaves Leaf thickness Sediment texture # of branches Taller More Thin Soft More

Sitra Shorter Less Thick Hard Less

Plants in Tubli were much taller than those in Sitra. Their number of leaves and branches were also more. While the leaves of Sitra were thick, they were much thinner in Tubli area. The sediment texture varied in both areas, it was hard in Sitra whereas it was soft in Tubli. 1. Total Soluble Salts (TSS): Total Soluble Salts measured using a refractometer was recorded and its mean was calculated, the following bar graph was made:
5|Page

Effect of Enviornmental stress on Plants BIOLS 320

60 50 TSS () 40 30 20 10 0 soil water Figure8: Salinity of soil & water samples of Tubli & Sitra P > 0.05 i.e. there is no statistical difference. tubli sitra

TSS analysis revealed that the salinity in Sitra was higher than that in Tubli. Statistical analysis showed that there is no statistical difference between the TSS of soil whereas water samples did show statistical difference. 2. pH: pH of the water and soil samples were recorded and following graph was made from those values:
9 P > 0.05 i.e. there is no statistical difference.

pH

7.5

Tubli Sitra

6 Water Soil Figure 9: pH of water & soil in Tubli & Sitra

pH analysis showed that there is no difference in the pH in soil and water of Tubli and Sitra. Statistical analysis also proved this as it showed no statistical difference. 3. Water potential measurement: The percent change in weight of the potato tuber cylinders was calculated using the formula: Percent change (%)

6|Page

Effect of Enviornmental stress on Plants BIOLS 320

With these values, graph in figure 10 was plotted. According to this graph, 0% change in weight was obtained at 0.97M solute concentration for Tubli and 2.32M in Sitra.
35 30 % change in weight (g) 25 20 sitra 15 10 5 0 0.000 -5 0.500 1.000 1.500 2.000 2.500 tubli

Concentration of solutes in plants (M)

Figure 10: % change in weight of leaves incubated in different sucrose solutions at 22C

Solute potential of leaves can be calculated as, s = -RTCs, Where R=0.0083143L MPa mol-1 K-1 C=0.21M
-1 -1

T= (273+22C) =295K s=1

Water Potential (MPa) 0 -1 -2 -3 -4 -5 -6 sitra tubli

For Tubli: s = - (0.0083143 L MPa mol K 295K 0.97M 1) = -2.37MPa For Sitra: s = - (0.0083143 L MPa mol-1 K-1 295K 2.32M 1) = -5.69MPa Calculated value of w is given below: w = s + p + g (here, p=0MPa and g=0MPa) w = s = -2.37MPa (for Tubli) & -5.69MPa (for Sitra) Hence, the w for Sitra is lower than that of Tubli. 4. Chloride ion Accumulation: To calculate mg/l of Cl- following formula was used: Mg/l = dilution factor

P < 0.05 i.e. there is statistical difference.

Figure 11: Difference in w in Tubli & Sitra

7|Page

Effect of Enviornmental stress on Plants BIOLS 320

Where, A = volume of AgNO3 consumed for sample B= volume of AgNO3 consumed for distilled water.
25 20 15 tubli 10 5 0 plant sap plant surface Area water soil sitra

N= Normality of AgNO3 = 0.2N MW of = 35.450gmol-1

Chloride content (mg/l)

P < 0.05 i.e. there is statistical difference.

Figure 12: Measurements of chloride content in Tubli and Sitra

Chloride accumulation rate was calculated by the formula: Accumulation rate = Calculated accumulation rate for Cl- was then plotted in bar graph shown in figure 13. Similar results were obtained, i.e. Sitra showed to accumulate more Cl- than Tubli.
0.18 0.16 0.14 0.12 0.1 0.08 0.06 0.04 0.02 0

Chloride accumulation ratio

P < 0.05 i.e. there is statistical difference.

tubli

sitra

Figure 13: Chloride Accumulation Ratio

5. Chlorophyll Content Index (CCI): Mean values for CCI were plotted in a bar graph and they also showed no statistical difference.

8|Page

Effect of Enviornmental stress on Plants BIOLS 320

50 45 40 35 30 25 20 15 10 5 0 Sitra

Chloride Content Index (CCI)

P > 0.05 i.e. there is no statistical difference.

Tubli

Figure 14: CCI of leaves in Tubli & Sitra

6. Chlorophyll Fluorescence (CF): Measuring the chlorophyll Fluorescence in the leaves of Tubli and Sitra, after providing them darkness by the help of clip, also showed no statistical difference. It means the PSII is working efficiently in both the areas.
0.84 Chlorophyll Fluorescence (Fv/Fm) 0.82 0.8 0.78 0.76 0.74 0.72 0.7 Tubli Sitra Figure 8: CF of leaves in Tubli & Sitra P > 0.05 i.e. there is no statistical difference.

7. Moisture Content: To calculate the moisture content, the following formula was used: %W = 100

Where: %W = Percentage of moisture in the sample, A = Weight of wet sample (grams),

9|Page

B = Weight of dry sample (grams)

Effect of Enviornmental stress on Plants BIOLS 320

55 50 45 40 35 30 25 20 15 10 5 0 Tubli

P > 0.05 i.e. there is no statistical difference.

Moisture Content (%)

Sitra

Figure 9: Moisture Content of leaves in Tubli & Sitra

From the graph, it is clear that there exist no difference between the moisture content of plants in Tubli and Sitra.

Discussion & Conclusion:

Recommendations:
On-site measurements of CCI & CF. Photosynthetic rate. ABA accumulation. Measuring the rate of evapo-transpiration (stomatal conductance). Soil & Xylem water potential measurements. TEM.

References:
1.

10 | P a g e

Effect of Enviornmental stress on Plants BIOLS 320

Summary of results: Statistical analysis showed that TSS, Solute concentration and Chloride accumulation was higher in Sitra than in Tubli. So, this could be the reason behind growth difference in the 2 areas, i.e. salinity. High solute concentration is characterized by low solute potential and eventually lower water potential. However, pH, CCI, CF and the moisture content was almost similar in the plants growing in the 2 nearby areas. So plants in both the areas were photosynthesizing equally.

11 | P a g e