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ROLE OF ORGANIC FERTILIZERS ON CHLOROPHYLL CONTENT IN RICE (ORYZA SATIVA L.)

Morteza Siavoshi* and Shankar L. Laware

Lecturer, Department of Agricultural Science, Payame Noor University, I. R. of Iran.
Associate Professor, Department of Botany, Fergusson College, Pune, India.
(*Corresponding author, [email protected])

ABSTRACT
In order to study the role of organic fertilizers on chlorophyll content in rice, an experiment was carried out in 2008
and 2009, in randomized block design based on 4 replications. Cow manure, poultry manure, rice straw and husk
were used for formulation of organic fertilizers. The treatments of organic fertilizers CM, PM, CMR, PMR, and
CPMR were used alone at 4t/ha in five treatments. Half dose of CPMR was used with half dose of RDF (N-50, P-25,
K-25 kg/ha) at one level and RDF (N=100, P=50, K=50 kg/ha) alone at one level. The plants without treatments
considered as control. Chlorophyll content and Grain yield were significantly increased in all the treatments over
control. The maximum grain yield (4776.52 kg/ha) was noted in plants treated with CPMR 2t +½ RDF which had
more chlorophyll content. An increase in the grain yield at the abovementioned treatments was may be due to the
increase of chlorophyll content which provides the energy necessary for plant growth and reproduction.
KEYWORDS: Chlorophyll content, Grain yield, Rice, Organic fertilizers.

INTRODUCTION
In a study on nutrient absorption by crop plant and its relation to chlorophyll content by Follet et al. (1981) clearly
indicated that chlorophyll coloration is related to the amount of nutrients absorbed by the plant from the soil. Roy and
Singh (2006) also reported similar results in barley with vermi-compost applications. They also observed that
application of organic manure resulted in higher leaf chlorophyll content than inorganic fertilizer. Deshpande and
Devasenapathy (2010) observed that application of green manure along with FYM and poultry manure significantly
increased protein and carbohydrate content in rice.

There are some reports, which indicate that organic fertilizers when supplemented with chemical fertilizers give more
positive influence on crop growth and increase organic constituents in test plants. In this respect Jogloy et al. (2006)
reported that the application of animal manure along with chemical fertilizer gave the highest chlorophyll in
(Helianthus tuberosus L.). Xu et al. (2008) found that the organic manure application with chemical fertilizers
increased the yield more than chemical fertilizers alone because it increased nitrogen use efficiency of rice. It is
documented that higher chlorophyll content observed in Helianthus tuberosus due to organic manures + NPK can be
attributed to differences in nitrogen content of the organics manures alone and in combination with NPK and its uptake.

The greater chlorophyll values in leaves treated with organic manure + NPK are of importance because photosynthetic
activity and crop yield may increase with increased chlorophyll content of leaves (Ramesh et al., 2002).

MATERIALS AND METHODS

This investigation was carried out at Baykola Research Center, Neka, Mazandaran, Iran during years 2008 and 2009.
The experimental farm is geographically situated at 36°, 60'N latitude and 53°, 13'E longitude at an altitude of 4 m
above mean sea level.

Formulation of organic fertilizers

Cow manure, poultry manure, rice straw and husk were used for formulation of organic fertilizers. These materials
were mixed in required proportions and composted for about 40 days. After completion of composting, these were
sieved thorough 2 mm mesh and then analysed for their nutrients. The proportions of raw materials used are given in
Table-1.

Treatments
Organic fertilizers CM, PM, CMR, PMR, and CPMR were used alone at 4t/ha in five treatments. Half dose of CPMR
was used with half dose of RDF (N-50, P-25, K-25 kg/ha) at one level and RDF (N=100, P=50, K=50 kg/ha) alone at
one level. The plants without treatments considered as control. The recommended chemical fertilizer dose for rice is
N-100: P-50: K-50. This combination was termed as RDF and used in one treatment. Based on RDF amounts the NPK
values were calculated from the organic fertilizers analysis and 4 t/ha dose was decided based on NPK content of CMR,
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PMR and CPMR and similar volume (i.e. 4 t/ha) of CM and PM organic fertilizers was used for treatment. Nitrogen in
the form of urea was used three times during growth season (1 st dose at the time of transplanting, 2nd dose at tillering
time and 3rd dose at the time of flowering). Hand weeding was done after 3 weeks of transplanting. The pests and
diseases were controlled by application of insecticides and tricycasol was used for rice blast.

Table 1. Raw materials used in formulation of organic fertilizers.

Sr. Formulation Ingredients Total
No. (Abbreviated Name)
Cow Manure Poultry Rice Rice
(kg) Manure Straw husk
(kg) (kg) (kg)
1 CM 1900 - - 100 2000
2 CMR 1500 - 400 100 2000
3 PM - 1900 - 100 2000
4 PMR - 1500 400 100 2000
5 CPMR 500 900 500 100 2000
(1) CM: Cow manure+ rice husk, (2) CMR: Cow manure+ rice straw and husk,
(3) PM: poultry manure+ rice husk, (4) PMR: poultry manure+ rice straw and husk,
(5) CPMR: Cow manure+ poultry manure+ rice straw and husk

Table 2. Details of the field experiment.

1. Season May-August
2. Crop Oryza sativa L.
3. Variety Local Tarom
4. Plot size 5×2m
5. Crop duration 90 -95 days
6. Date of sowing 20. 04. 2008-9
7. Date of transplanting 24. 05. 2008-9
8. Date of harvesting 20-25.08.2008-9
9. Design RCBD

10. Number of 4
replications

Estimation of photosynthetic pigments

Chlorophyll were extracted and estimated by Arnon’s (1949) method and Coombs et al. (1985). One gram of the fresh
flag leaf tissue was cut into small pieces and placed into a specimen bottle containing10 ml of absolute ethanol and
stored in the dark for two weeks. 1 ml of the filtered extract was then diluted with 6 ml of absolute ethanol and the
absorbance of the chlorophyll solution measured using a spectrophotometer (Shimadzu- 1700) at 645 and 663 nm. The
chlorophyll a and chlorophyll b content in milligrams (mg) were estimated using the formula of Arnon (1949).

Grain yield (kg/ha): Samples from 1 M² area from each plot were used for Grain yield.

RESULTS
Chlorophyll a (mg g-1)
The results pertaining to effect of organic fertilizers, combination of chemical fertilizer with organic fertilizers and
recommended dose of NPK on flag leaf chlorophyll a content in rice is given in table 3. Among the treatments the half
dose of CPMR+ RDF, RDF and CPMR organic fertilizer show results at par with each other and better than CM, CMR,
PM and PMR in both the years and even in pooled means.

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Chlorophyll b (mg g-1)
The results pertaining to effect of organic fertilizers, combination of chemical fertilizer with organic fertilizers and
recommended dose of NPK on flag leaf chlorophyll b content in rice is given in table 3. Among the treatments the half
dose of CPMR+ RDF, RDF and CPMR organic fertilizer show results at par with each other and better than CM, CMR,
PM and PMR in both the years and even in pooled means.

Total Chlorophyll (mg g-1)

The results pertaining to effect of organic fertilizers, combination of chemical fertilizer with organic fertilizers and
recommended dose of NPK on flag leaf total chlorophyll content in rice is given in table 3. Among the treatments the
half dose of CPMR+ RDF, RDF and CPMR organic fertilizer show results at par with each other and better than CM,
CMR, PM and PMR in both the years and even in pooled means.

Table-3: Effect of different levels organic fertilizer, recommended dose of chemical fertilizer and
combination of organic and chemical fertilizers on rice (O. sativa) leaf chlorophyll a, chlorophyll b and total
chlorophyll contents (mg g-1) and grain yield (kg/ha).
Chlorophyll a Chlorophyll b (mg g-1) Total Chlorophyll Grain yield (kg/ha)
-1
Treatments (mg g ) (mg g-1)
(Per hectare) Pooled PIOC Pooled PIOC Pooled PIOC Pooled PIOC
Control 1.07 0.00 0.707 0.00 1.777 0.00 3943.90 0.00

RDF (NPK) 1.46 36.45 0.898 27.02 2.358 32.70 4582.32 16.19
100:50:50 kg
CM 4t 1.26 17.76 0.779 10.18 2.039 14.74 4044.92 2.56

CMR 4t 1.30 21.50 0.807 14.14 2.107 18.57 4135.21 4.85

PM 4t 1.36 27.10 0.868 22.77 2.228 25.38 4377.07 10.98

PMR 4t 1.39 29.91 0.884 25.04 2.274 27.97 4594.59 16.50

CPMR 4t 1.45 35.05 0.894 26.45 2.339 31.63 4699.31 19.15

CPMR 2t + 1.50 40.19 0.931 31.68 2.431 36.80 4776.52 21.11

½ RDF
CD (0.05) 0.037 - 0.046 - 0.042 - 96.36 _

CD (0.01) 0.066 - 0.082 - 0.075 - 170.13 _

PIOC: Percent Increase over Control

Grain yield (kg/ha)

The organic fertilizers treatments had significant effect on the grain yield. The results in table 3 clearly indicate that
application of organic fertilizers increased the grain yield significantly in all the treatments over control. Among the
treatments the half dose of CPMR+ half dose of RDF CPMR, PMR and RDF show results at par with each other and
significantly better than CM, CMR and PM alone in both the years and even in pooled means.

DISCUSSION
The fertilizers added to soil provides macronutrients and micronutrients, which are assimilated by plants and utilized
for various metabolic activities to synthesize chlorophyll, required for their normal growth and developments. Various
micronutrients are needed for catalytic activities of enzymes essential for respiration, photosynthesis, flowering, fruit
setting and seed filing as well as fight against abiotic and biotic stresses. In present investigation, it was planned to
study the effect of organic fertilizers alone and in combination with chemical fertilizer on leaf and grain biochemical
attributes in rice. The organic fertilizers based on cow manure (CM, CMR) and poultry manure (PM, PMR)
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supplemented with rice husk and straw and a composite organic fertilizer (CPMR) were given at the dose of 4 t/ha. A
half dose of organic fertilizer CPMR and half dose of RDF, RDF alone were also used for fertilization in rice and
investigated for chlorophylls and grain yield. The results pertaining to flag leaf biochemical parameters like
photosynthetic pigments are given in tables 3.

Photosynthetic pigments content increased in all the plants treated with organic fertilizers and chemical fertilizers as
compared to the control. The plants treated with CPMR + RDF showed maximum content as compared to control and
other treatments. The present investigation indicated that rice plants respond better with respect to chlorophyll content
with combination treatment of CPMR+NPK compared to NPK and organic fertilizers alone.

Photosynthesis is a process, which provides the energy necessary for plant growth and reproduction. Among pigments
chlorophylls, the green pigments with magnesium at core of heterocyclic protoporphyrin ring, represents the principal
pigment responsible for light absorption and photosynthesis (Nelson and Cox, 2004). Photosynthesis however, is a
complex process that is sensitive to macronutrients and micronutrients availability (Marschner, 1995).

Chlorophyll content increased in all treatments compared to the control. It showed maximum content in combination
treatment as compared to control and other treatments. The present investigation indicated that rice plants responded
better with respect to Chlorophyll content with combination treatment of organic fertilizer CPMR + RDF (NPK)
compared to NPK and organic fertilizers alone. Amujoyegbe et al. (2007) studied the effects of inorganic fertilizers
(NPK), poultry manure alone, and in combination on the chlorophyll component of maize and sorghum; they noted that
combination treatment exhibited higher chlorophyll a and b in maize and poultry manure alone enhanced those in
sorghum. According to them increase in total chlorophyll pigments due to addition of chemical fertilizers to poultry
manure increased both chlorophyll a and b and thereby increased the total chlorophyll.

Researchers like Berova and Karanatsidis (2009) and Fernéndez-Luqueño et al. (2010) studied the effect of organic
fertilizers on the amount of chlorophyll pigments and rate of photosynthesis. These researchers concluded that the
application of organic fertilizers can not only enhances the synthesis and amount of chlorophylls but also increases the
rate of photosynthesis. Similarly, potash application enhances the uptake of N and hence there is an improvement in
growth characters. Phosphorus helps in early establishment of crop and formation of fibrous and strong root system.
The strong root system absorbs more nutrients including micronutrients from the soil and finally contributes towards
rapid plant growth (Hari et al., 2006). Micronutrients are also reported to have vital role in photosynthesis. Magnesium
is principal element required for chlorophyll synthesis, as it is the part of chlorophyll structure. Iron complexes with
proteins to form important enzymes in the plant and is associated with chloroplasts, where it has some roles in the
synthesizing chlorophyll, hence iron is necessary for the maintenance and synthesis of chlorophyll and RNA
metabolism in the chloroplasts. Calcium and copper must be present for iron to function properly. Boron has a vital role
in nitrogen assimilation. Fertilizers with micronutrients can increase chlorophyll because the availability of Mg and
uptake of Mg by plant (Suntoro, 2002). Magnesium is involved in numerous physiological and biochemical processes
activating more enzymes than any other mineral nutrient, thus, making a significant contribution to plant growth and
development (Epstein and Bloom, 2004). Zinc enhances the cation-exchange capacity of the roots, which in turn
enhances the absorption of essential nutrients, especially of nitrogen that is responsible for higher protein content. The
increase in nitrogen uptake might be due to increase in sulphur content resulting in the stimulation of protein synthesis
and due to its synergistic effect on the utilization of nitrogen (Singh et al., 2010).

This suggests that nutrients released by combination treatment were available to synthesize more chlorophyll and
develop the site of photosynthesis. A promotion effect of organic and inorganic fertilizers on chlorophyll contents
might be attributed to the fact that N is a constituent of chlorophyll molecule. Moreover, nitrogen is the main
constituent of all amino acids in proteins and lipids that act as a structural compound of the chloroplast (Arisha and
Bradisi, 1999).

Regarding to the key role of nutrients, such as nitrogen (protein, enzymes and chlorophyll synthesis), potassium (N
uptake), phosphorous (strong root system), magnesium and iron (chlorophyll synthesis), zinc, manganese (cation-
exchange and enzyme activities), calcium and copper (for functioning of iron) and boron (N assimilation); these
nutrients might have absorbed by plants through fertilizers, which worked in coordination and contributed in
chlorophyll structure and synthesis. And this could be the reason for more chlorophyll in plants treated with half dose
of CPMR and RDF, where RDF might have supplied NPK during early growth and CPMR might have supplied

Volume-2 Issue-3 (2013) ISSN: 2319–4731 (Print); 2319–5037 (Online) © 2013 DAMA International. All rights reserved. 16
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micronutrients essential for synthesis and increasing leaf chlorophyll pigments. Increased photosynthetic pigment in
2 t/ha CPMR+½RDF, NPK and CPMR organic fertilizers treatment may be attributed to improved soil physical and
chemical properties and optimum nutrients availability in fertilizers.

CONCLUSION
From aforementioned data, we conclude that CPMR supplemented with RDF fertilizer can fulfil the requirement of
essential macro and micronutrients at proper growth stages. In present investigation the half doses of CPMR and RDF
showed significant enhancement for chlorophyll a, chlorophyll b and total chlorophyll which is followed by RDF and
CPMR. Hence more productivity in rice better grain yield was observed in these treatments.

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