Transcript Variation in Growth and Ion Uptake of Maize due to

Presented by:
Ummu M. S. (080610091)
Shelly F. N. (080610317)
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• INTRODUCTION
– KEY WORDS
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PROBLEM STATEMENT
PURPOSE
MATERIAL AND METHOD
RESULT
DISCUSSION
CONCLUSION
INTRODUCTION
Among various environmental stresses soil salinity limits
plant growth and production in many part of the world,
particularly in arid and semi arid area (Shannon, 1984).
Under salinity, growth depression results from increased
level of ethylene (Nukui et al., 2001), decreased
photosynthetic capacity (Drew et al., 1990), water
deficit and ion imbalance (Wyn Jones, 1981).
Many attemps have been made to reduce the drastic
effect mostly focusing on chemical amelioration and
saline agriculture. Recently, a biological approach using
microorganism was attempt.
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Key words:
• PGPR
• GROWTH
• IONS UPTAKE
• MAIZE
• SALINITY
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PGPR
Plant Growth Promoting Rhizobacteria are free
living microorganism having beneficial effects on
plants by colonizing their roots (Nadeem et. al,
2006) and via direct or indirect mechanisms
(Nelson, 2004)
Kloepper and Schroth (1978) describe as soil
bacteria that colonize the roots of plants
following inoculation onto seed and that enhance
plant growth.
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1.1 Beneficial effect of PGPR
• Production of phytohormones (auxin, cytokinins, and
giberelins) (Garcia de Salamone et al., 2001)
• Enhancing release of the nutrients (Nautiyal et al., 2000;
Idriss et al., 2002)
• Preventing the deleterious effects of environmental stress
(Kloepper and Schroth, 1978).
• Produce antibiotics that protects roots from deasease,
example: 2,4 –DAPG (2,4 – diacetylphloroglucinol) (Aragno
et al.,2004)
• Also produce exopolysaccharides (EPSs) to bind cations
including sodium (Geddie and Sutherland, 1993), thus help
alleviating salt stress in plants grown under saline
environment (Ashraf et al., 2004).
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THE KINDS OF PGPR
• Bacillus vallismortis
for mediated systemic resistance against
Potato Virus
• Bacillus subtilis
• Pseudomonas flourescens
• Bacillus polymixa
• Serratia marcescens
Scanning electron micrograph of wheat root-colonizing
Pseudomonas fluorescens OE 28.3
the role of Pseudomonas fluorescens cell envelope proteins (in
particular the major outer membrane protein OprF) in adhesion
to roots. Biocontrol of phytopathogens appears to be a major
mechanism of plant growth promotion by these bacteria
Product of PGPR
1.2 Environmental stress
Nilsen and Orcutt in they book “Physiology of Plant
Under Stress” explain several kinds of stress:
• salinity
• Temperature
• Water (Flooding and drought)
• pH
• Light
• nutrient
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SOIL SALINITY
The dynamic nature of soil salinity in the rootzone
affects performance of plants. Profile distribution of
salts is affected by leaching fraction and changes
with changing water content from irrigation and
rootwater extraction. Soluble salts in soils are highly
mobile and tranported by water through mass flow
and dispersion. Irrigation water management is one
of the keys in maintaing salt balance in the rootzone
(Salinity by Andre Lauchli and Ulrich Luttge).
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GROWTH
• Definition
An increase in the size of an organism or part of an organism,
usually as a result of an increase in the number of cells.
Growth of an organism may stop at maturity, as in the case of
humans and other mammals, or it may continue throughout
life, as in many plants.
• Growth is one of the best indices for evaluating plant
responses to environmental or biotic stress (Nilsen and
Orcutt, 1996)
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CLASSIFICATION OF MAIZE
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Kingdom
: Plantae
Plants Subkingdom
: Tracheobionta
Vascular plants Superdivision : Spermatophyta
Seed plants Division : Magnoliophyta -Flowering plants
Class : Liliopsida -Monocotyledons
Subclass
Commelinidae
Order
Cyperales
Family
Poaceae - Grass family
Genus
Zea L. – corn
Species
Zea mays L. – corn
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MORE
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PROBLEM STATEMENT
1. How does the effect of the strains of PGPR on
growth and ion uptake of maize?
2. How does the effect of salt concentration on
growth and ion uptake of maize?
3. How does the interaction between strain of
PGPR and salt concentration on growth and
ion uptake of maize?
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PURPOSE
1. To know the effect of the strains of
PGPR on growth and ion uptake of
maize
2. To know the effect of salt
concentration on growth and ion
uptake of maize
3. To know the interaction between
strain of PGPR and salt concentration
on growth and ion uptake of maize
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MATERIAL AND METHODS
1
Prepared the bacteria
Rhizosphere soil were collected from saltaffected field of maize.
 Several bacterial strains were isolated by
dilution plate technique using DF minimal
medium (Dworkin and Foster, 1958).
 Further streaking on fresh plates purified the
collected rhizobacterial strains
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S5
S5
S15
S20
S15
SHAKER
S20
Inoculum @
60 mL broth
Incubated for 72 hours under shaking
(100 rpm) condition 28 ± 1°C
An optical density of 0.5 measured at a
wavelength of 535 nm was achieved by
dilution to maintain uniform cell density (107108 cfu/mL)
N
E
X
T
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• The selected strain were characterized by measuring
ACC-deaminase activity (Honma and Shimomura, 1978)
by monitoring the amount of α-ketobutyrate produced
when the enzyme ACC-deaminase cleaves ACC
(Aminocyclopropane-1-Carboylic Acid )
• In vitro auxin production was determined as IAA
equivalent in the presence and absence of Ltryptophan by using protocol describe by Khalid et al.
(2004)
• Chitinase activity and phosphorus solubilizing activity
was determined qualitatively as describe by Chernin et
al. (1998) and Mehta and Nautiyal (2001).
• Colonization ability of these strains was studied under
axenic condition as describe by Simon et al. (1996) HOME
2
soil
Physicochemical
characteristic was
analyzed
- sandy clay loam
-pH
-Ece
-Organic matter
-Total nitrogen
-Olsen phosphorus
-Exchangable potassium
- surface disinfected
- Inoculated with peat mixed with
10% sugar solution
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Maize
seed
Uninoculated seed was coated with
sterilized peat in the sterilized broth
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Inoculated seed
POT
Containing 12 kg
soil
S5
A B
S15
C D
S20
-Pot arranged in wire house under
ambient light and temperature
according to CRD
-Recommended doses NPK fertilizer
were applied in each pot as Urea, DAP,
and SOP respectively
-Phosporus and Potassium was
applied as basal dose while Nitrogen
was applied in two split
A B
C D
A B
C D
UNINOCULATED
A B
C D
A, B, C, D are four ECe level 0, 4, 8, and
12 dS/m respectively
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For N and P analysis:
Thawed
andwith
1) Dried leaf sample were
digested
crushed
sulphuric acid and H2O2
2) N was determined by Kjeldhal method
3) P content were determined by
spectrophotometer after mixing with
Barton reagents.
The sap was collected
in polypropylene by
gilson pipette
Centrifuge at 6500
rpm for 10 minutes
Collected after 75 days
The supernatant sap was
Chlorophyll pigment analysis:
andand
analyzed
collected
Na and K were
1) 0.5 g of leaf samples of each Data were collected
determined
using sherwood
410
statistically using
a completely
randomized
treatment were homogenized with
Flame1980)
Photometer.
design (Steel and Torrie,
80% acetone
Chloride content was determined
2) The homogenate was filtered
by chloride analyzer
through filter paper
3) Absorbance of the resulting
solution was read at 663, 645, and
480 nm for chlorophyll a, b, and
FLAME
carotenoids.
FLAME
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RESULT AND DISCUSSION

Variation in growth
Shoot fresh weight
Shoot dry weight
Root fresh weight
Root dry weight
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Variation ion uptake
Photosintetic pigments (Clorophyll a, b, and
carotenoid contain),
Ion composition (N, P, K and NaCl
concentration)
Variation in growth
• SFWS20,12
dS m-1 64%
over c.
• SDW S20,12
dS m-1 102%
over c.
• RFW S20,12
dS m-1 114%
over c.
• RDW S20,12
dS m-1 102%
over c.
Pot experiment of maize was observed that
inoculation with these rhizobacterial strains
significantly improved shoot/root fresh weight
and shoot/root dry weight at all salinity levels.
 In general, growth was reduced with increase in
salinity. However, inoculation was effective
even in the presence of higher salinity levels
(12 dS m-1).
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Variation ion uptake
• C aS20,0
and 8 dS m-1 13
and 76% over c.
• C b S20,12
dS m-1 102%
over c.
• Car S20 et all
EC level up to
58% over c.
• N S20,12 dS
m-1 55% over c.
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Results of pot study showed that on overall
basis the increase in salinity level decreased
the photosynthetic pigments (chlorophyll a, b
and carotenoid contents) of the maize
seedlings. However, inoculation with PGPR
containing ACCdeaminase activity significantly
increased the pigments under salinity stress
compared to control.
Variation ion uptake
• PS20,12 dS
m-1 56% over c.
• K S20,12 dS
m-1 119% over c.
• Na+S20,12
dS m-1 16% lower
than c.
• Cl- S20,12 dS
m-1 40% lower
than c.
Data in Table 2 and 3 described that N, P and
K concentration significantly increased due to
inoculation with PGPR.
 Na+ and Cl- concentration significantly
increased with the increasing salinity
particularly in control treatment.
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Under salinity stress, ethylene is produced at
higher concentration and it’s caused
inhibitory to plant growth.
 Removing or blocking the effect of stress
ethylene results can decrease of the stress
effect.
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Mechanism of PGPR
Lowering the endogenous inhibitory levels of
ethylene in roots because of their ACC
metabolizing ability and also be due to the
expolysaccharides (EPS) activity of bacteria.
The PGPR strains can produce bacterial
exopoloysaccahrides which bind cations including
Na+ (Geddie and Sutherland, 1993) and decrease
the content of Na available for plant uptake, thus
helping decrease + salt stress in plants (Ashraf et
al., 2004).
CONCLUSION
Gift of PGPR inoculum were statistically non
significant on growth and ion uptake of maize.
 Gift of salt concentration were increased
significantly on growth and ion uptake of maize.
 The interaction between strain of PGPR and
salt concentration were increased significantly
on growth and ion uptake of maize.

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THANK YOU FOR THE ATTENTION
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DILUTION PLATE METHOD
• DEFINITON OF DILUTION
– the mixing of a small accurately measured sample with a
large volume of sterile water or normal saline called
(diluents or dilution blank)
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It has been known that if we use a larger volume we obtain a more accurate dilution.
So for better results, we use 1:1000 dilution. And that is by adding 1ml of sample to
999 ml of diluent. But practically we cannot use 999 ml of diluent. So we do what is
called a serial dilution.
Serial Dilution: is a dilution made of a series of smaller dilution, and the total
dilution is the product of each dilution in the series.
DILUTION
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ACC deaminase
• Hasnain and Sabri (1996) reported that inoculation
with Pseudomonas sp. stimulated plant growth by
reduction of toxic ion up take and production of stressspecific proteins in plant under stress. Certain
microorganisms contain an enzyme 1aminocyclopropane-1-carboylic acid (ACC) deaminase
that hydrolyses ACC, the immediate precursor of
ethylene thereby lowering the plant ethylene level
(Glick et al., 1998) which eliminates the potential
inhibitory effect of high ethylene concentration (Glick
et al., 1999).
ACC deaminase
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DUNCAN’S NEW MULTIPLE RANGE
TEST
• In statistics, Duncan's new multiple range test (MRT) is
a multiple comparison procedure developed by David
B. Duncan in 1955. Duncan's MRT belongs to the
general class of multiple comparison procedures that
use the studentized rangestatistic qr to compare sets of
means.
• David B. Duncan developed this test as a modification
of the Student-Newman-Keuls method that would have
greater power. Duncan's MRT is especially protective
against false negative (Type II) error at the expense of
having a greater risk of making false positive (Type I)
errors. Duncan's test is commonly used in agronomy
and other agricultural research.