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Abstract
Phosphorus has been acquired by the plants in the soluble form which is found in the soil in the
minimum quantity. The available phosphorus is found in the soil, which reacts with the other
minerals available in the soil like calcium, iron and aluminium causing the precipitation of the
compounds leaves the phosphorus in the insoluble form. The role of phosphorus-solubilizingbacteria are found to be very important in the conversion and decomposition in order to produce
a soluble form of phosphorus to the plants so that it can be utilized for plants growth and
development. Moreover, the observable and gradual decrease in the pH indicates the activity of
PSB for phosphorus immobilization by showing the traces of inorganic acids. The current study
aimed to isolate the phosphate solubilizing bacteria by analyzing the four soil samples, i.e.
banana, ground, garden and graveyard soil and analyzing the rhizosphere under the PKV medium
and tricalcium phosphate. The halozone colonies indicated the presence of Pseudomonas
fluorescens and the chemical reaction indicated the production of organic acids and the
experiment indicate the immobilization of phosphorus.
Contents
List of Tables.................................................................................................................. 2
CHAPTER 1: INTRODUCTION......................................................................................... 2
CHAPTER 2: LITERATURE REVIEW.................................................................................2
2.1 Isolation of Strains................................................................................................... 2
2.2 Formation of Phosphate-Solubilizing-Bacteria PSB........................................................2
2.3 Phosphorus Solubilizing Mechanism.............................................................................2
CHAPTER 3: METHODOLOGY........................................................................................ 2
3.1 Sample.................................................................................................................. 2
3.2 Preparation of medium.............................................................................................. 2
3.3 Isolation of PSM...................................................................................................... 2
3.4 Morphological Characterization...................................................................................2
3.5 Gram Staining......................................................................................................... 2
CHAPTER 4: RESULTS AND DISCUSSION.........................................................................2
4.1 Isolation of PSB...................................................................................................... 2
CHAPTER 5: CONCLUSION............................................................................................ 2
References..................................................................................................................... 2
List of Tables
Sr. No
1
Title
Table 1 Prepartion of Medium
Page No.
24
CHAPTER 1: INTRODUCTION
Plants containing adequate amounts of phosphorus entail appropriate growth and development.
Phosphorus plays a vital role by being a component of plant structure which is responsible to
regulate the protein synthesis mechanism in plants. The mechanism of cell division, distribution
and transformation of energy in terms of glucose takes place under the sufficient presence of
phosphorus in plants. Moreover, the process of tissue development is also buffered paving way
for the proper growth in plants. On the contrary, the deficiency of phosphorus can cause revert
the growth mechanism in plants while changing the normal color of leaves to dark-green color or
brown and make the plant vulnerable (Schachtman, et al., 1998).
The detailed work of Holford (1997) infers that the soil may contain high quantity of
phosphorus, but plants, doesn't necessarily absorb the required phosphorus quantity due to its
insoluble form present in soil. Therefore, the potential solubility of phosphorus takes place under
the manifestation of the enzyme known as phosphatase that has been originated in number of
microorganisms found in soil. Phosphorus carries a negative charge which makes it to react with
the minerals found in the soil mainly known as aluminium, calcium, iron and magnesium and
these cations precipitate and form soluble phosphorus that is absorbed by the plants. In the
presence of phosphate solubilizing bacteria, the conversion of phosphorus is enhanced from the
insoluble phosphorus and the process takes place when these bacteria and fungi release organic
acids and enzymes consisting of phosphorus.
According to Chen, et al. (2006), when the adequate amount of phosphorus is available in the
soil then its presence ensure the proper absorption of phosphorus to the plant which causes its
proper development in terms of providing vulnerable to weathering, growth towards maturity
5
Al(OH)2H2PO4(insoluble) +2H+
The study also exhilarates that the lack of phosphorus in plants under the circumstances
in which soils are not found to be rich in minerals affects the overall yield of the crops by
postponing the maturity stage of the plants, producing low quality of crops and reduction in the
vulnerability of plants to encounter diseases.
The meta-study conducted by Richardson and Simpson (2011) confers that the large numbers of
microorganisms have the tendency of solubilizing the inorganic form of phosphorus being the
part of the soil. These microbes are found to be isolated while prevailing around rhizosphere and
able to possess the tendency to convert inorganic phosphorus in organic form. This study
determines that the microorganisms existing in basic or alkaline soil when examined in such
manner that they are evaluated by using calcium phosphates which makes them to discharge
soluble phosphorus. The absorption of phosphorus also depends on the lack of soil capacity for
absorption and capability of the soil to provide phosphorus to the plants. The phosphorus fixation
process is stimulated at the higher pace when soil is composed of coarse texture having its
contents highly patterned with clay. The fixation process takes place when the hydrogen
phosphate is met with calcium ions and the reaction of both chemicals results in the reversible
formation of calcium phosphate and hydrogen ions and the reaction.
The study also advocates that the absorption of phosphorus in the process is relatively at
the fastest pace and considerably reduces during the passage of time. Furthermore, the
availability of phosphorus should be made when the crops or plants are at their growth cycle and
if the phosphorus is applied at the beginning when the seeds are sown then the huge proportion
of phosphorus will be fixed even before the plant requirement which will ultimately cause the
deficiency of phosphorus in plants.
Fankem, et al. (2006) infers that the unavailability of phosphorus has become a crucial problem
because the phosphorus by means of fertilizers provided to the crops become fixed after its
application making the phosphorus insoluble for the plants. The phosphorus available in the soil
reacts with the soil minerals mainly consisting of calcium, aluminium and iron, which ultimately
results in the fixation of phosphorus and converts it in the insoluble form. It has been emphasized
that the presence of microorganisms which helps in the solubilization of insoluble phosphorus
has taken the utmost importance in order to enhance the plant growth and overall yield. These
microorganisms solubilize the inorganic or insoluble form of phosphorus so that it can be
absorbed the plants as required. The experiment was conducted in this regard by analyzing the
seven samples from the rhizosphere of oil-palm-tree. The soil samples were found to be highly
acidic in nature and the presence of iron and aluminium was highly noted. The dilution of
samples took place and isolates were formed which were observed in order to find the PSM and
further tests to calculate the pH were conducted which determined the considerable drop in pH,
which indicated the presence of organic acids which resulted in the formulation of carboxylic
acid at the highest pace. The PSM were identified as the Pseudomonas and were found to be
responsible for the decomposition of insoluble phosphorus into the soluble phosphorus.
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21
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CHAPTER 3: METHODOLOGY
3.1 Sample
The soil samples were collected from neighboring places consisting of banana soil, graveyard
soil, garden soil and ground soil and the samples were gathered from the site of cultivating areas.
The samples for each soil were taken 6 times from different sites at the depth of 17 inches. These
samples were dried and then all 6 samples were mixed together by making one sample for each
type of soil.
Gram/ml
10
0.5
0.5
0.1
5
0.2
0.2
0.002
0.002
10
Table 1 Preparation of Medium
25
Figure. 1 Halozone in
Banana Soil
for
the
immobilization of phosphorus
indicating the presence of
bacteria that are responsible
Figure. 2 Halozone in
Garden Soil
for the solubilization of phosphorus i.e. PSB. The halozone in banana soil can be seen in Figure
1.
The result obtained from the halozone of the garden soil sample proved its proficiency of the
immobilization of phosphorus by indicating the existence of phosphorus-solubiling-bacteria and
the garden soil demonstrated the dilution of 10^-4. The halozone is shown in figure 2.
The third soil sample consisting of graveyard soil also showed the positive result by having the
ability to immobilize phosphorus, which in turn indicated the presence of PSB and the banana
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Figure. 3 Halazone in
Graveyard Soil
CHAPTER 5: CONCLUSION
The study conducted an experiment by analyzing the four soil samples of banana, graveyard,
garden and ground soil in order to isolate the microorganisms that isolate phosphorus. The results
of the present study indicated that the three soil samples, i.e. Garden, graveyard and banana soil
in which the isolates were formed in the samples from garden soil, graveyard soil and banana
soil. These samples indicated the formulation of halozone which represented the
microorganisms colony lying under the PVK medium. These isolates were studied under the
microscope, which confirmed the existence of bacteria which are responsible to convert
27
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