Abstract

It has long been known that certain ionic substances can be extremely detrimental to the basal
metabolism of plant species, when they reach a high enough concentration. This phenomenon
has severe consequences in areas of the third world, with vast tracts of land unsuitable for
crop growth due to salinization from poor farming practice and drought. However, this
detrimental effect varies greatly between ionic substance and plant species and the
differential toxicity of the ions was not known. With this in mind, solutions containing
different ions were made up and added to 5 samples of the species Arabidopsis thaliana, and
5 samples of species Thellungiella halophila, and the effect was observed in terms of
reduction of leaf matter mass over a period of time, then the difference in effect of each salt
on each species was compared.

Introduction
Of all abiotic stress factors, soil salinity is one of the most detrimental to plant growth. It severely
affects crop production in arid lands and is quickly becoming a great threat to sustainable agriculture.
One main source of soil salinity arises from the practice of irrigation without first having proper
drainage facilities. This leads to water evaporating off the surface of the soil and depositing salt
behind which then accumulates and results in the loss of valuable agricultural areas. The effect
salinity has upon the growth of a plant varies depending on the type of ions, the concentrations and
the species of plant involved.

Salinity Stress in Plants

Sodium chloride (NaCl), the most abundant salt found naturally and the most studied, causes damage
to plants in two different ways, by osmotic stress and by ion toxicity. Osmotic stress is the disruption
of the ordinary difference in osmotic pressure between the plant and its environment. Maintaining an
osmotic pressure that is higher relative to the soil solution is crucial for plants in order to take up
water and minerals through their roots. A high NaCl concentration in the soil increases the osmotic
pressure of the soil solution therefore hindering and sometimes even blocking the uptake of essential
molecules like water or potassium ions. Ion toxicity arises when high levels of sodium and chlorine
ions directly enter the plant, where they disrupt cell membranes and metabolic activity. Some
enzymes are critically affected by the presence of high sodium ion concentrations and high Na/K
ratios in the cytosol. Sequential secondary damage then occurs in the plants as a consequence of these
impacts. This is characterised by the immediate production of abscisic acid due to osmotic stress
leading to closure of stomata which results in the abating of photosynthesis. Prolonged exposure to
salt causes growth inhibition, accelerated development, senescence and then eventually cell death.
Interestingly, research shows some of the stress can be lessened by the addition of calcium ions.
Calcium works by conserving the transport of potassium and increases the effectiveness of ion pumps,
allowing cells to better maintain low cytosolic Na concentrations and low Na/K ratios.

Arabidopsis and Thellungiella

Arabidopsis, like most plants, is a glycophyte. This means it does not tolerate salt well and can only
withstand NaCl concentrations of between 100-200 mM. However Thellungiella, a halophytic relative
of Arabidopsis, is able to grow in salt concentrations of up to 500 mM. Halophytes have gained
tolerance through a number of specific adaptations which allow them to cope with high salt
environments. In halophytes, salt stress is accompanied by a large increase in cytosolic
,
activating sodium pumps and increasing discrimination between potassium and sodium. Na that
accumulates in the cell is actively pumped into and stored in the vacuole to protect cytosolic enzymes
from damage. Lastly, large amounts of benign compatible osmolytes are synthesised and allowed to
build-up in the cytosol to offset the increased osmotic pressure caused by extracellular and vacuolar
ions. These are but a few examples of the methods plants use to deal with sodium chloride stress.
Little is known about the relative toxicity of other salts on top of sodium chloride. In this report we
will examine the effects of

References

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2881266/
http://www.faculty.ucr.edu/~jkzhu/articles/2007/ELS%20Zhu.pdf
http://thellungiella.org/
http://tjeas.com/wp-content/uploads/2012/09/7-10ok.pdf
http://www.plantphysiol.org/content/147/3/1168.full

Methodology
25 Arabidopsis thaliana plants and 25 Thellungiella halophila plants were grown to maturity, and
then starved of water for a week. The plants were split into 5 experiment groups containing 5
individuals of each species. 5 measurements of 5.84g of NaCl were dissolved in 5 separate flasks
containing 500ml of distilled water, creating a 200mM solution of NaCl. 3.68g of CaCl2, 1.86g of KCl,
5.08g of MgCl2 and 3.56g of Na2SO4 were each dissolved in 50ml of distilled water then each
solution was added to a different flask of 200mM NaCl. Each of the resulting solutions was then
added to a different batch of the 2 plant species.

EXPERIMENT NO.

SALT SOLUTION ADDED

1 (control)
2
3
4
5

200mM NaCl in 500ml distilled H2O

200mM NaCl+ 50mM CaCl in 500ml distilled H2O
200mM NaCl+ 50mM KCl in 500ml distilled H2O
200mM NaCl+ 50mM MgCl2 in 500ml distilled H2O
200mM NaCl+ 50mM Na2SO4 in 500ml distilled H2O

The experiment groups were then safely stored in a growth room for 1 week and starved of water
once again. After this the experiments were taken out of the growth room, photographed to observe
the morphological differences, then the fresh weight of each plant was measured. In order to do
this a weighing boat was place on a balance and the weight was set to 0 by pressing tare. The
entire plant was excised from the soil using a volunteers hands, and the root matter was clipped off
using scissors, leaving only the leaf matter. The leaf matter of each was individually placed in the
weighing boat; the weight was measured and noted. The leaf matter was then placed on aluminium
foil, and the weighing boat was placed back on the scales and set to 0. This process was repeated
for each specimen, with each sample (group of five plants) placed in the same tin foil wrap which
was carefully folded. Each wrap was the weighed on the weighing boat and the combined weight of
the foil and plants was noted down. The ten samples were then labelled with the group and sample
name, then placed in an oven to dry for one week.

Figure 1- The 10 sample groups, shortly after removal from the growth room

After the samples had been in the oven for one week, they were taken out and the dry weight was
measured. This was again done using a weighing boat and set of scales. The weighing boat was
placed on the scales, set to 0and then each sample was measured and the weight noted. The dry
weight of each sample was then measured. The standard deviation and standard error were then
calculated, and then noted down for each sample. The average water content of each individual was
then calculated by subtracting the dry weight of each sample from the wet weight. The percentage
of water content of the average individual was then calculated using the following formula:

After this, the results were collated, and placed in tables and graphs in order to be analysed more
efficiently.

Results
The results were collated into the following graphs and figures for facilitation of analysis.

Plant Weight (g) Mean

0.9
0.8
0.7
0.6
0.5
Mass (g)
0.4

Plant Weight (g) Mean

0.3
0.2
0.1
0
A

Control

T
KCl

MgCl2

CaCl2

Na2SO4

Figure 2- Comparison of average fresh weight of each sample. The x-axis

shows the solutions and the y-axis shows the mass.

As seen in figure 2, there was a dramatic difference in mean fresh weight between plant
species. For example, in the control experiment there is an average difference of 0.35g
between the two species. A notable difference between the species is seen between the
,
and
experiments. It can be seen here that the average fresh weight of
Arabidopsis is comparatively reduced in the
experiment whereas the average fresh
weight of Thellungiallis increases. Also, between the control experiment and the
experiment, the fresh weight of the Arabidopsis sample is seen to be comparatively higher in
the latter. The lowest average fresh weight of Arabidopsis can be observed in the
experiment, and the lowest average fresh weight of Thellungiallis can be seen in the
samples.