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Physiological Responses of Eggplant (Solanum Melongena L.) in Salt Stress
Physiological Responses of Eggplant (Solanum Melongena L.) in Salt Stress
) IN SALT
STRESS
By:
Muhammad Ibnu Nazari B1K014005
Nadyatul Hanifah
B1K014006
Gabriella Ashari T.
B1K014008
Clotter
: II
Group
:2
Assistant : Azhar Faturohman Abidin
2015
I. INTRODUCTION
A. Background
Eggplant (Solanum melongena L.) is one commodity that is common and popular
vegetables grown in the tropical and subtropical regions, especially in Asia and
Africa and has a pretty good tolerance to biotic and abiotic. Eggplant is a species
with morphological and physiological variability of characters wide, so it is
necessary to get the sources of higher resistance of wild species (Sunantara, 2000).
Basically, every plant has a certain range tolerance of their environmental
factors. That principle is known as Shelford's Tolerance Law, which states that
"Every organism has an ecological minimum and maximum, which is the lower limit
and the upper limit of the tolerance range of the organism to environmental factors
condition". Every living thing has a range of optimum or optimum range to
environmental factors for its growth. The conditions above or below the limits of the
tolerance range will makes the organism experience physiological stress. At this
physiological stress conditions, the population will decline. If this stress condition
continues for a long time and has reached the limit of tolerance survival, the
organism will die (Lakitan, 1995).
According to Nilsen (1996) to understand the reactions of a particular
organism in a certain situation, individual external influences, so-called
environmental factors, are usually considered separately, if at all possible.
Environmental factors can be of abiotic and biotic nature. Biotic environmental
factors, resulted from interactions with other organisms, are, for example, infection
or mechanical damage by herbivory or trampling, as well as effects of symbiosis or
parasitism. Abiotic environmental factors include temperature, humidity, light
intensity, the supply ofwater and minerals, and CO2; these are the parameters and
resources that determine the growth of a plant.
Plants have different resistance to the presence of salts in the soil. High levels
of salinity will causing a decrease in the plant production. Moreover, salinity is also
suppress the growth processes in plants by inhibiting the effect of enlargement and
cell division, protein production and increase plant biomass. Plants that are stressed
salts generally do not show a response in the form of direct damage but subdued
growth and change slowly (Sipayung, 2007).
The concentration of dissolved salts are quite high in plants and saline soils will
cause salt stress in plants. The level of stress experienced by the plants is different in
various species with tolerance is not the same for different salt concentrations. Effect
of salt stress due to salinity did not show a response in the form of direct damage but
growth depressed by changes slowly. In the face of the influence of salinity, various
crops perform various forms of adaptation to the mechanisms of morphological and
physiological mechanisms. Adaptation to salinity is needed primarily to improve
water balance in order to maintain the water potential and turgor, and all the
biochemical processes for growth and normal activities (Sipayung, 2007). Adaptation
to salinity is complex which is the formation of glandular structures found on the
surface of the leaf or stem epidermis (Salisbury & Ross, 1995).
Vegetable cropsare very important they have high nutritional value which rich
source of minerals and vitamins, but most of the vegetable crops are known as
glycophyta plant, they have high sensitivity to salt stress. Thus, knowledge of the
mechanism of salt tolerance in vegetable crops is necessary so as to increase their
sustainable production on saline soils. Eggplant (Solanum melongena L.) so-called
terong in Indonesia is widely reported to be sensitive to salinity (Shaheen, 2012).
Beside their role we use eggplant in this lab because eggplant plant are are easily
obtained, easy maintenance, the growth media is not too difficult and easy to observe
growth. According to FAO (2012), eggplant is cultivated on an area of about 4
million acres (1.6 million ha) world-over. However, there is a dire need to enhance
its yield and introduced more varieties which could thrive well under stressful
habitats including saline stress. If the salt tolerance mechanism of eggplant plants is
explicitly elucidated, it maybe possible to minimize salt-induced injury in this crop.
In view of all this, we hypothesized that salt stress alters some key physiological
attributes such as water relations, gas exchange characteristics, etc., in eggplant
plants. So, the study was aimed to study the regulation of a variety of physiological
attributes in eggplant plants exposed to varying salinity regimes and find the salt
tolerance on eggplants (Shaheen, 2012).
B. Objectives
1. To understand that plant growth is influenced by internal and external factor.
: Plantae (Plants)
Sub kingdom
Super Division
Division
Class
Sub Class
: Asteridae
Order
: solanales
Family
Genus
: Solanum
Species
Land with high salt levels will inhibit some of the activities that are essential for
respiration and photosynthesis (Harjadi & Yahya, 1988).
Salinity can affect inhibit plant growth in two ways: a. By damaging the cells
growing so disrupted crop growth. By limiting the amount of supply of essential
metabolic outcomes for cell growth through the formation of tyloses. Salinity
suppress the growth processes in plants by inhibiting the effect of enlargement and
cell division, protein production and increase plant biomass. Plants that are stressed
salts generally do not show a response in the form of direct damage but subdued
growth and change slowly (Mengel & Kirkby, 1987).
Salts that cause plant stress among others are NaCl, NaSO 4, CaCl2, MgSO4,
MgCl2 dissolved in water. In the soil solution, these salts affects the pH and electrical
conductivity. Ssaline soils have a pH <8.5 with electrical conductivity> 4 mmhos /
cm. In most species, the influence of other types of salt are generally not typical for
plant growth, but more dependent on the total concentration of salt. Salinity is not
determined by the salt Na Cl alone but by different types of salts impact and stress on
the plant. In this context stressed plants salt when excessive salt concentrations high
enough to lower the water potential of 0.05 to 0.1 MPa. Salt stress is different from
the stress of ions that are not so pressing water potential. Tolerance to salinity is
diverse with a wide spectrum among species of plants ranging from sensitive to fairly
tolerant. Proposed five-level effects of soil salinity on plants, ranging from nonsaline to very high levels of salinity (Shaheen et al., 2012).
Plant eggplant (S. melongena) is one species of plants that can not withstand
high salinity (Glycophyta). Tolerance to salinity is the ability to maintain growth and
metabolism in an environment rich in NaCl (Munns et al., 1995). Salt affects plant
growth mainly through: (a) poisoning caused by the absorption of building blocks of
salt in excess, such as sodium, (b) a decrease in the absorption of water, known as
water stress and (c) a decrease in the absorption of the essential elements for plants,
especially potassium. Early signs of salinity damage are (a) the color of the leaves
that become darker than normal colors are green-bluish, (b) the size of the leaves are
smaller and, (c) the stem with leaf stalk distance is shorter. If the problem gets worse,
the leaves will, (a) to yellow (chlorotic) and, (b) edge to dry dead leaves exposed to
"burning" (burning, becomes brownish) (Dwidjoseputro, 1994).
Analysis of variance (ANOVA) is a collection of statistical models used to
analyze the differences among group means and their associated procedures (such as
A. Material
The tools that used in this practicum are pounder (mortar and pestle),
ruler, scissors, polybag, spectrophotometer, filter paper, measuring glass,
beaker glass, Erlenmeyer glass,, cuvette, analytical balance, pipet, magnetic
stirer, oven, microscope, and reaction tube.
The materials that used in this practicum are egg plats (S. melongena),
NaCl solution with concentration 0%, 25%, 50%, 75%, 100%, Aceton, HVS,
label paper aquades and soil.
B. Methods
The methods that used in this practicum are;
3.1 Plant height
gram) (B).
Make the leaf pattern of 2nd leaf sample on the square paper. The
square paper cut down follow the leaf pattern, next the leaf paper
Leaf area =
Details:
A = Square paper area (cm2)
A
xC
B
Separated the medium or soil from the plant espsecially from the
roots. Cleaned the soil that still attacht in roots with water flow until
clean.
Cut the plant part, roots, stem, and leaves.
Measured the wet weight of each part, and noted the obtained data.
After plant performed wet weight the each part then ovened until dry
by warpped by paper.
Measured the dry weight of each part, and noted the obtained data.
Used paper filter to filter the nathan, and placed the supernathan in
reaction tube.
A. Result
Table 1. ANOVA Height of Plant 1
ANOVA
Sources of
Variation
Db
Perlakuan
Galat
Total
JK
KT
4 2,5784
0,6446
20
15,68
0,784
24 18,2584 0,760767
Fhitung
n
0,822194 s
FTable
0.05
0.01
2,87
4,43
Df
Perlakuan
Galat
Total
JK
KT
4 2,2424
0,5606
20
7,892
0,3946
24 10,1344 0,422267
Fhitung
n
1,420679 s
FTable
0.05
0.01
2,87
4,43
Df
4
20
Total
24
JK
KT
6,6856
1,6714
25,3
1,265
31,985
6 1,332733
Fhitung
1,321265 ns
0.05
2,87
FTable
0.01
Df
JK
KT
4 67,6576 16,9144
20 20,0216 1,00108
24 160,7824 6,699267
Fhitung
16,89615 **
FtabLE
0.05
0.01
2,87
4,43
4,43
Table 5. Data of Tukeys HSD (honest significant difference) test plant length IV
Tukey's HSD (honest
significant difference) test
0
Mean
3,36
0
3,36
0 NS
25
3,5
25
3,5
0,14 NS
50
75
100
2,88
1,52
1,42
0,48 NS
1,84 **
1,94 **
0,62
1,98
2,08
50
2,88
N
S
N
S
**
**
75
1,52
0 NS
1,36 *
1,46 **
100
1,42
0 NS
0,1 NS
0 NS
Df
Perlakuan
Galat
4
20
Total
24
JK
KT
183,613
6
58,5736
253,446
4
Fhitung
45,9034
2,92868
10,5602
7
FTable
0.05
0.01
*
15,67375 *
2,87
4,43
Table 7. Data of Tukeys HSD (honest significant difference) test plant length V
Tukey's HSD (honest
significant difference) test
0
25
Mean
4,9
4,22
0
4,9
0 NS
25
4,22
0,68 NS
50
75
100
4,12
1,24
1,44
0,78 NS
3,66 **
3,46 **
0,1
2,98
2,78
50
4,12
N
S
N
S
**
**
0 NS
2,88 **
2,68 **
75
1,24
0 NS
0,2 NS
100
1,44
0 NS
0 mM
25 mM
50 mM
75 mM
100 mM
15
10
5
0
1
Df
perlakuan
galat
20
total
24
JK
KT
4650,87
7
10126,8
3
1162,71
9
506,341
4
615,737
7
14777,7
F hitung
2,296315
n
s
0,05
FTable
0,01
2,87
4,43
Df
perlakuan
galat
20
total
24
JK
KT
449,067
5
27407,5
4
27856,6
1
112,266
9
1370,37
7
1160,69
2
FTable
0,05
0,01
F hitung
0,081924
n
s
2,87
4,43
150
100
50
0
1
Df
JK
4
20
2,082
2,108
24
4,182
FTable
0.05
0.01
4,93833 **
2,87
4,43
KT
Fhitung
0,5205
0,1054
0,1742
5
Table 11. Data of Tukeys HSD (honest significant difference) of Dry & Wet
Weight
Tukey's HSD (honest
significant difference) test
0
25
4,18
Mean
2
2,082
0 4,182
0 NS
2,08
25 2,082
2 **
0
2,10
50 2,108
8 ** 0,026
3,62
75 0,556
6 ** 1,526
10
3,22
0 0,956
6 ** 1,126
50
75
100
2,108
0,556
0,956
ns
ns
0 ns
**
1,552 **
0 ns
**
1,152 **
0 ns
0 ns
25
50
75
100
Concentration
FTable
Df
JK
KT
Fhitung
0.05
4
20
24
2,285
2,639
3,399
0,57125
0,13195
0,141625
4,329291
4 *
0.01
2,87
4,43
0,517 0,1628 NS
0,06 NS
0,607
2 0,0726 NS 0,1502 NS
50
0,527
8
0 NS
0,010
8 NS
0,079
4 NS
75
0,517
100
0,607
2
0 NS
0,0902 NS
0 NS
2
1.5
1
0.5
0
0
25
50
75
CONCENTRATION
100
B. Discussions
Different salt regimes (0, 25, 50, 75, and 100 mM NaCl)of the root growing
medium significantly suppressed the growth of eggplant plants in terms of shoot and
root fresh and dry weights as well as shoot lengths. Although growth reduction in
most plants is a common effect of salinity stress, however, the actual physiological
mechanisms involved in growth reduction are still not well determined (Munns &
Tester 2008; Shaheen, 2012).
Shoot dry weight decreased as a result of stagnant stress caused by the
condition of the roots that were damaged as a result of the decrease in the ability of
plants to absorb water and mineral nutrients in the soil. Furthermore, the decreasing
mass of plant is caused by reduced water availability due to a decreased ability of
roots as an absorbent organs. Salisbury (1995) states that the water stress caused
nutrients transport in plants disrupted resulting in a biochemical process which is
shown by a low dry weight of plants.
According to Harjadi (1988), tress can be defined a condition or situation
unfavorable environment for plants, will have a negative impact on plant growth,
reproduction and survival of the plant itself. The plant stresses could be induced by
several factors. These factors could be limited availability water, salinity,
temperature, non-optimal soil pH or herbicides. There is a conspicuous difference in
the sensitivity of plants to salinity. Species that grow in saline environments would
be met with a high osmotic pressure in the soil. Physiology of salt tolerance in plants
will increase because of the salt deposits in the top layer of soil irrigation (Jenks &
Hasegawa, 2005).
Salinity is a measure of the content of salts in soil or water. Salts are highly
soluble in surface and groundwater and can be transported with water movement.
Large salt deposits are a natural feature of vast areas of the Australian landscape,
stored deep in soils or as surface salt deposits and salt lakes. This natural distribution
of salt in the landscape is referred to as 'primary salinity'. In normal circumstances,
the deep roots of native plants absorb most water entering the soil before it reaches
the salt contained in groundwater below the plant root zone. However, widespread
vegetation clearance, poor land use, irrigation and industrial practices have made it
easier for salt to be transported to the soil surface or to waterways. The additional salt
from these altered land use and management practices is referred to as 'secondary
salinity (Campbell, 2011).
Salinity will affect the physiological and morphological processes of water
balance in the plant body. These effects may includes reduction in size and number of
leaves and a decrease in the number of stomata per leaf unit, which eventually will
reduce the production of plant. Generally the high soil salinity level has a double
effect on the plants, which are reduces the water potential on the tissues due to
increased osmotic potential, and gives the toxic effects directly due to the Na and Cl
ions accumulated highly in plant tissues (Yuniati, 2004). This project perform NaCl
salt stress in Eggplant (S. melongena) with different regimes 0 mM, 25 mM, 50 mM,
75 mM, 100mM. The lab parameters measurement includes plant height, leaf surface
area, chlorophyll content, wet weight and dry weight. This observation was
conducted over 6 weeks and watering every three days once with NaCl based on
varying concentrations.
Environmental stress can be either external factors or internal factors. External
factors include environmental conditions that do not support the growth and
development of plant parts such as nutrient deficiency and excess, deficiency and
excess water, the temperature is too low or too high. The internal factors are the
individual genes (Purwadi, 2011).
According to Lakitan, (1996), in general, environmental stress in plants are
grouped into two:
1
Biotic stress
a Intra species competition and inter species competition
Competition, whether intra or interspecies, are leading plants into
stress condition. The plant that lost on the competition to take nutrient
b
metabolic pathway.
Abiotic stresses
a Temperature (high and low)
Plants vary greatly in their abilities to cope with freezing or hot
temperatures. Plants experience unfavorable temperature in many
different ways and adaptation or acclimation to unfavorable
temperature occurs over different time scales and levels ofplant
organization (Jenks & Hasegawa, 2005).
high
water
potential
under
water-limiting
conditions
and
Salinity
Salt stress occurs with the presence of salinity or the concentration of
dissolved salts in the plant redundant. This generally occurs in plants
that grow on saline soils. Salt stress increases with increasing salt
concentrations up to a certain level of concentration which may result
in death of the plant. Salts that cause plant stress are NaCl, NaSO 4,
CaCl2, MgSO4, MgCl2 dissolved in water Stress due to excess Na + can
affect several physiological processes ranging from germination to
growth of plants (Sipayung, 2006).
The kind of stress that we observed on this mini project is salt stress on
eggplant (S. melongena). This plant performs several responses due to saline
condition. According to Mulyani (2006), generally, the plant response to salinity can
be seen in two forms of adaptation:
1
Morphological responses
Morphological and anatomical adaptations forms that can be derived is unique
and can be found on the type halophyta that evolved through natural selection in the
area of coastal forest and salt marshes. Salinity cause structural changes that improve
the water balance in the water potential of plants so that the plants can maintain
turgor and the whole biochemical process for normal growth and activity. Structural
changes include a smaller leaf size, smaller stomata per unit leaf area, increased
succulent, thickening of the cuticle and the waxy coating on the surface of the leaves,
and roots lignification earlier (Mulyani, 2006).
2
Physiological responses
Osmoregulation
Tolerant plants can make adjustments by lowering osmotic potential
without loss of turgor. To obtain water from the surrounding soil, xylem water
potential in the liquid to be extremely lowered by the voltage. Some halophyta
able to maintain osmotic potential to remain even become more negative during
he growing season in line with the absorption of salt. On the other halophyta
has the ability to regulate accumulation of salt (Na + and Cl) on the conditions
of salinity, such as mangroves are capable of removing 100% salt (Prihastanti,
2011).
b
control the salt concentration in the cytoplasm through membrane transport and
compartmentation. The salt is stored in the vacuole, accumulated within
organelles or excreted to the outside plant. Spending salt on the surface of the
leaves will help maintain a constant salt concentration in plant tissue.
(Salisbury, 1995).
c Membrane integrity
Semipermeable membrane system that encloses the cells, organelles and
compartments are the most important structures to regulate ion levels inside the
cells. The outermost layer of the cell membrane or plasmalemma separates the
cytoplasm and metabolic components of a solution from saline soils which are
chemically not match. The semi-permeable membrane function blocking free
diffusion of salt into the plant cells, and provide an opportunity for ongoing
active absorption of elements essential nutrient. Plasmalemma who deal
directly with the soil is the membrane that was first to suffer from the influence
of salinity. Thus, the relative resilience of this membrane becomes another
important element in salt tolerance (Armawan, 1998).
Several parameters of responses of S. melongena are observed, such as height,
leaf area, wet and dry weight, and also the chlorophyll content. The plants which
exposed by high salinity condition, both its wet and dry weight are decreasing if
compared than in normal condition. The higher salinity the smaller leaf area and leaf
mass become (Neto et al., 2004)
Actually, plant growth is altered by salt stress through two distinct ways,
osmotic effect of the salt on the soil and the toxic effect of salt within the salt.
Largely, the osmotic effect is causing the rate at which growing leaves expand is
reduced, new leaves emerge more slowly, and lateral buds develop more slowly or
remain quiescent, so fewer branches or lateral shoots form. The reduction in shoot
growth occurs in two phases, a rapid response to the increase in external osmotic
pressure, and a slower response due to the accumulation of Na + in leaves (Munns &
Tester, 2008).
Various levels (0, 25, 50,75 and 100 mM) of salt (NaCl) significantly
suppressed the plant height. The maximum reduction in shoots apical length was
observed at week 4 (Table. 4) and 5 (Table. 6), while in week 1 (Table. 1) there is no
significant changes, it also the same in week 2 (Table. 2) and week 3 (Table. 3). In
week 4 significantly suppressed at 100 mM NaCl (Table. 5), and in week 5
signicantly suppressed at 75 mM NaCl (Table.6). Application of different levels of
salinity treatment to eggplants are not significantly reduce the leaf area of the
eggplant plants, just a slight reduction. Wet and dry weights of shoot and root of
eggplant plants also significantly reduced. A maximal reduction in shoot and root
biomass was observed at 75 mM NaCl (Table 11). A slight reduction, not significant
reduction in total chlorophyll contents of eggplant plants was observed under
different saline regimes.
Appraisal of water relations in plants grown under stress conditions including
saline stress is necessary to identify up to what extent the cellular water content is
maintained, because almost all metabolic activities within the cell are dependent on
the availability of sufficient amount of water inside. However, in the present study,
leaf water and osmotic potentials of eggplant plants were decreased significantly,
while leaf turgor potential was increased due to addition of varying levels of salt to
the growth medium. These results are partially relevant in accordance to Shaheen et
al. (2012), that states in eggplant which they recorded had a significant decrease in
osmotic and water potentials under saline stress. Recently, in another study with
radish, Noreen et al. (2009) found a considerable reduction in leaf osmotic potential
due to increase in NaCl concentration in the growth medium.
Photosynthetic pigments are the key components of light reactions of the
photostynthesis mechanism. It is widely reported that almost all different types of
stresses including salinity stress cause damage to thylakoid membrane, the site where
all different types of photosynthetic pigments are accumulated. In several studies,
there is a close association exists between photosynthetic pigments (mainly
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