UNIVERSITY use of acidified organic amendment and diammonium phosphate



M.Sc. (Hons.) Soil Science)

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TITLE: Enhancing
P uptake in maize(zia mays L.)
through integrated use of acidified organic amendment and diammonium phosphate


Name of Student


Haider Raza

Registration No.



Name of Supervisor


Dr. Muhammad Naveed







Low organic matter, fixation of phosphorous with CaCO3
and high pH are major causes of low availability of Phosphorous (P) to plants
in Pakistani soils. For higher production excessive fertilizer are applied to
soil which are mostly lost by fixation into the soil which make it unavailable
for plants. P uptake can be increased by organic amendment and combination of
(DAP). For this purpose, an experiment will be conducted at Research Area of
Soil and Environment Sciences, University of Agriculture Faisalabad. In this
field trial, P uptake in
maize yield through integrated use of acidified organic amendment and
diammonium phosphate (DAP) will be investigated. We will use randomized
complete block design (RCBD) with 8 treatments.

1-     CONTROL

2-     DAP

3-     PRODUCT

4-     PRODUCT

5-     PRODUCT
(SOLID)               +DAP(

6-     PRODUCT

7-     PRODUCT
(SOLID)               +½

8-     PRODUCT

three replications. Agronomic parameters such as total biomass, 1000 grain
weight and phosphorous contents, comb fresh weight, dry weight, fresh weight,
no of grains per treatment, plant height. Soil samples will be collected from
0-15 and 15-30cm depths with the help of auger and physical properties of soil
such as bulk density, porosity, hydraulic conductivity, and soil moisture
contents. Data will be statistically analyzed by ANOVA and treatments will be
compared by using LSD test.











(Synopsis for M.Sc. (Hons.) Soil Science)





TITLE: Enhancing P uptake in
maize(zia mays L.) through integrated
use of acidified organic amendment and diammonium phosphate (DAP).



a) Date of Admission



b) Date of Initiation



c) Probable Duration


6 months




a)    Name of


b)  Registration


c)  Name of



Haider Raza




Dr. Muhammad Naveed







Dr. Muhammad Naveed



Dr. Hafiz Naeem Asghar




Dr. Irfan Afzal










Phosphorous is an essential macro nutrient for plants
(Goldstein et al., 1988) applied to crops after nitrogen as fertilizer.
Its application significantly affect grain yield (Ali et al.,2002) as
well as dry matter yield and plant characters like height, number of leaves and
leaf area (Ayub et al., 2002). It has its important role in living
things due to part of RNA and energy as ATP (Havlin et




al., 2004). It is important for plant
growth, consumption of sugar and starch, involved inphotosynthesis, nucleus
formation and cell division, fat and albumen formation and carbohydrates
metabolism (Ayub et al., 2002). Hence, P deficiency may affect plant’s
physiological processes and metabolic pathways negatively which ultimately
decreases plant growth (Din et al., 2011).


Phosphorous limit crop
production on more than 30% of world agricultural lands (Vance et al., 2003),
its supply to plants depend upon soil (Grant et al., 2005). In many
agricultural soilsits availability is very low despite having large reservoir
of phosphorous (often hundred times more than required by the plants) (Al-Abbas
and Barber, 1964). Phosphorous present in soil solution is less than 1% of the
total P taken up by plants and approximately 99% of P taken up by plants being
bound to soil constituents before uptake (Morel, 2002). Main reason for low
available P to plants is the formation of insoluble complexes; in acidic
conditions with Al and Fe while in alkaline conditions with Ca and Mg.


About 80-90% of arid and
semi-arid region soils are deficit in plant available P (Memon et al.,
1992; NFDC, 2001). Pakistani soils are alkaline and calcareous (Memon et al.,
1992) andP is present as Ca complexes with different solubility (Rahmatullah et
al., 1994). Such soils are naturally poor in available P (Barber, 1995). In
these soils there are problems with phosphorous availability of chemical
fertilizer by fixation (Sayin et al., 1990) and only 20% of the applied
P is taken up by plants (Ju et al., 2007).


Plants growing in low available
P obtain it from adsorbed, sparingly soluble and organic complexes that exist
in soils by biochemical mechanisms (Raghothama and Karthikeyan, 2005).
Carboxylates such as citrate and malate are the major root exudates release
under P deficiency for mobilizing its uptake (Neumann and Römheld, 1999;
Dechassa and Schenk, 2004). The quantity of carboxylates released dependent on
the plant species (Ohwaki and Hirata, 1992; Dechassa and Schenk, 2004).
Genotypes of the same crop species are also differ in their ability to release
organic anions and in their ability to mobilize P from sparingly soluble
compounds (Dong et al., 2004; Corrales et al., 2007). Released
anion binds phosphate from P complexes by ligand exchange in soil and makes it
available for plants (Raghothama and Karthikeyan, 2005).







Root exudation mainly depends
upon nutrient deficit conditions of soils. Plants species release
phytosiderophores due to deficiency of P, Fe and Zn (Haynes, 1990; Jones and Darrah,
1994). Under P stress, C3 plant releases
more citric acid and C4 plants releases
more tartaric acid and malic acid (Vranova et al., 2013). Components of
root exudates also vary under aerobic and anaerobic condition also (Jackson and
Ilamurugu, 2014).


Considering these facts, a
hydroponics experiment will be conducted in order to investigate rice growth
and organic acids production like Citrate, Malate and Oxalate under normal and
phosphorous stressed environment.




Screening of Phosphorous efficient rice genotypes.


To determine exudates amount secreted in phosphorous deficit




Jackson and Ilamurugu (2014) conducted an experiment to
study the chemical composition of rice root exudates under anaerobic conditions.
Analysis was performed with HPLC and GC. They reported that rice root exudates
contain amino acids, sugars and organic acids at various crop growth stages. At
early stages rice roots exude higher amount of organic acids (0.95M g-1
root of acetic acid), sugars (Glucose-1.77 M g-1
root) and amino acids (Alanine-0.34 M g-1
root) while root exudation decreased in later stages (acetic acid-0.21 M g-1
root, glucose-1.10 M g-1


Xin-Bin et al. (2012.)
conducted an experiment using two rape cultivars (genotype LG and genotype HG)
that differed in P-uptake due to root morphology and organic acid release under
sparingly soluble phosphate (Fe-P and Al-P). He reported that root exudates
have ability to activate insoluble P under P stress, parameters like root
length, root tips and surface area of genotype HG were significant than
genotype LG.


Zhang et al. (2011)
conducted an experiment to determine P uptake efficiency of two rapeseed
genotypes under two sparingly soluble P sources. P efficient genotypes under
low P conditions cause acidification of Rhizosphere due to H+
ion efflux, H+-ATPase
activity and




exudation of carboxylates. He reported that genotype 102 is
P efficient while genotype 105 is P inefficient.


Carvalhais et al. (2011)
conducted an experiment on maize plants grown under N, P, K, Fe deficiency and
reported that Plant grown under Fe-deficiency release a higher amount of
ribitol, glucose, citrate and glutamate. Amino butyric acid and carbohydrates
were released under P-deficiency, k deficiency releases less sugar while under
N-deficiency lower amount of amino acids are produced in exudates. He concluded
that release of root exudates is mechanism to subsist under deficiency of
nutrients in plants.


Long et al. (2008)
stated that organic acids increase availability of carbonate fixed P because
organic acids compete P for binding sites. A dialysis tubing analysis was
performed to quantify dissolved organic acids in vegetative (sea grass) and
non-vegetative (bulk pore-water concentrations) medium. He reported that
phosphate concentration is positively related to organic acids prouction,
organic acids increased productivity of sea grass under P-limited environment.


Pearse et al. (2007)
conducted a comparative experiment between six crops (Wheat, Sarsoon,
Chickpea, Pea, white lupin, Blue Lupin and Lupinus cosentinii) to
testify their ability to utilize slowly soluble P(AlPO4,
FePO4 and Ca5OH(PO4)3).
These crops were grown in sand and P was applied @ 40 mg P kg?1.
Plant species varied in their ability to utilize P due to release of


Singh and Pandey (2003)
conducted a study to check both qualitative and quantitative differences in
root exudation of different maize and green gram genotypes, availability of P
increased by exudation of organic acids while green gram released more organic
acids than maize. Genetic variability was found in exudation of organic acids
of maize and green gram genotypes. He reported that there is a positive
correlation between P uptake rate and total root exudation.


Egle et al. (2003)
conducted an experiment with and without P fertilization to check qualitative
and quantitative exudation of organic acids from six cultivars of three lupin
species, two cultivers of each species were grown on sand for 21 days, root
exudates were collected in 0.05 mM CaCl2.
Under P-deficiency eight organic acids (citric, 2-oxoglutaric, malic, succinic,




lactic, formic, acetic and fumaric acids) from three lupin
spp. L. angustifolius had highest carboxylate efflux. Under P-deficiency
protoid roots developed and citrate exudation rate on average 67% for L.
albus, 37% for L. angustifolius and 72% for L. luteus were


Lambers et al. (2002)
investigated carboxylates exuded from of Banksia grandisL. Plants were grown
on sand with phosphate @ 25 ?g P g?1
using different P sources like K-phosphate, glycerol phosphate, Fe-phosphate or
Al-phosphate. Citrate, malate and trans-aconitate were major carboxylates and
were released 60%, 25% and 14% respectively when P added as Al-phosphate. While
Fe-phosphate carboxylates released relatively in less amount i.e. citrate
(31%), malate (14%) and trans-aconitase (12%). Monocarboxylates were also
released by these plants like Lactate (30%) and acetate (12%).


Aulakh et al. (2001)
conducted an experiment to check the exudation of ten rice cultivars at
different stages from seedling to maturity. Exudation rate was generally lowest
at seedling stage increased till flowering and decreased at maturity. Among
organic acids, malic acid was released in utmost amount.


Neumann and Römheld (1999)
conducted a comparative experiment in solution culture to check P induced
metabolic changes (exudation of carboxylic acids and protons) in wheat (cv.
Haro), tomato (cv. Moneymaker), chickpea and white lupin (cv. Amiga). Proton
exudation from tomato, chickpea and white lupin was increased under
P-deficiency while release of carboxylic acids with proton increased in roots
of chickpea and white lupin but not in wheat and tomato. Citric acid was one of
important carboxylic acid investigated in all species in P-stressed environment
and this was due to increased activity of PEP carboxylase required for citrate
synthesis. The results suggest that P deficiency induced exudation of carboxylic
acids depends on the ability to accumulate carboxylic acids in the root tissue.
In some plant species such as white lupin, there is specific transport
mechanism (anion channel) of root exudation in high amount of citric acid.




A field experiment will be conducted at field area Institute
of Soil and Environment Sciences, University of Agriculture Faisalabad.
Experiment will consist of eight treatments of phosphate fertilizer (DAP) and
acidified amendment with dose before sowing of maize (zea mays L.)

1-     CONTROL

2-     DAP

3-     PRODUCT

4-     PRODUCT

5-     PRODUCT
(SOLID)               +DAP(

6-     PRODUCT

7-     PRODUCT
(SOLID)               +½

8-     PRODUCT

At first water same dose will
be applied in solid form and at the time of second water fertigate will be




height (cm)


Total biomass (g)


1000 grain weight (g)


Comb fresh weight (g)


Dry weight (g)


Fresh weight (g)


No of grains per treatment


Phosphorous contents


VII.Statistical analysis


The data obtained will be
statistically analyzed using computer based software; STATISTIX 8.1®;
an analytical statistics program. After ANOVA table formation, Tukey’s HSD
(honest significant difference) test will be applied to check treatment
significance (Steel et al., 1997).




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M.A. Nadeem, M.S. Sharar and N. Mahmood. 2002. Response of maize (Zea mays
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Wirén. 2011. Root exudation of sugars, amino acids, and organic acids by maize
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X.Y. Jin, Q.P. Wang, Y.M. Lin, L. Gan and C.X. Tang. 2007. Confirmation and
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and M.K. Schenk. 2004. Exudation of organic anions by roots of cabbage carrot
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Dong, D., X.
Peng and X. Yan. 2004. Organic acid exudation induced by phosphorus deficiency
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Xin-Bin, Z.,
H. Jian-Guo, Z. Yong-xiang and S. Wei-Ming. 2012. Genotypic variation of rape
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