Nov 6, 2011

Biofertilizer Potential of Traditional and Panchagavya Amended


Biofertilizer Potential of Traditional and Panchagavya Amended with Seaweed Extract
Sangeetha. V* and Thevanathan, R**

*Post Graduate and Research Department of Botany, Presidency College,
Chennai-5, Tamil Nadu, India. sangeethadotv@gmail.com 
**Centre for Advanced Study in Botany, University of Madras (Guindy campus), 
Chennai-25, Tamil Nadu, India. thevanathan@gmail.com

ABSTRACT
The potential of utilizing panchagavya as biofertilizer was tested on the pulses Vigna radiata, Vigna mungo, Arachis hypogea, Cyamopsis tetragonoloba, Lablab purpureus, Cicer arietinum and the cereal Oryza sativa var. ponni by growing in soil amended with dried traditional and seaweed based panchagavya. Experimental seedling recorded higher rates of linear growth of both shoots and roots as compared to controls. These seedlings produced 264 to 390% more lateral roots than the control and maximum lateral root production was always observed in seedlings grown in soil amended with seaweed based panchagavya at low concentrations (1:100; panchagavya: soil). A similar observation was made on the number of leaves produced, leaf area and the number of root nodules formed in the pulses by rhizobia. A marked decrease in Chlorophyll a/b ratio, C/N ratio in the plants grown in seaweed based panchagavya indicating high chlorophyll b levels and a better nitrogen use efficiency in these plants respectively. [The Journal of American Science. 2010;6(2):39-45 (ISSN 1545-1003)]

Key words: Panchagavya, Seaweed based, biofertilizer potential, pulses, cereal

 INTRODUCTION
The current global scenario firmly emphasizes the need to adopt eco-friendly agricultural practices for sustainable agriculture. Chemical agriculture has made an adverse impact on the health-care of not only soil but also the beneficial soil microbial communities and the plants cultivated in these soils. This eventually has lead to a high demand for organic produce by the present-day health conscious society and sporadic attempts are being made by farmers all over the world to detoxify the land by switching over to organic farming dispensing with chemical fertilizers, pesticides, fungicides and herbicides. In India, organic farming was a well developed and systematized agricultural practice during the past and this ‘ancient wisdom’ obtained through Indian knowledge systems such as ‘Vedas’ specify the use of ‘panchagavya’ in agriculture for the health of soil, plants and humans. In Sanskrit, panchagavya means the blend of five products obtained from cow, namely cow dung, cow urine, cow milk, curd and ghee (Sugha, 2005). The Vriskshayurveda systematizes the use of panchagavya. Few farmers in the southern parts of India have used modified formulations of panchagavya and found them to enhance the biological efficiency of  the crop plants and the quality of fruits and vegetables (Natarjan, 2002). In the past three decades, crude extracts from seaweeds have been show to exhibit many bioactivities that include biostimulant, fertilizer and antimicrobial properties. Different forms  of seaweed preparations such as LSF (Liquid Seaweed Fertilizer), SLF (Seaweed Liquid Fertilizer), LF (Liquid Fertilizer) and a manure prepared by using either whole or finely chopped seaweeds have been experimented and all of them have been reported to produce beneficial effects on cereals, pulses and flowering plants (Radley, 1961; Stephenson, 1974; 1981; Smith and van Staden, 1983; 1984; Tay et al., 1985; Temple
and Bomelle, 1989; Sekar et al., 1995). In this paper, we present the results of an investigation made to evaluate the fertilizer potential of traditional and modified form of panchagavya amended with liquid
seaweed preparations using some pulses and paddy as experimental plants. 

MATERIAL AND METHODS
Traditional panchagavya Traditional panchagavya was prepared following the procedures outlined by Pandurang Vaman Kane, 1941. It contained fresh cow dung - 0.5 kg; cow urine - 1.0 L; cow milk - 7.0 L; curd - 1.0 L; ghee - 1.0 L and water - 1.0 L. These ingredients were taken in a 25.0 L concrete pot, mixed well and allowed to stand in shade for 21 days with intermittent stirring. After 21 days, the preparation was allowed to dry for 180 days in shade. Dried panchagavya was mixed with sterilized garden soil at a ratio of 1: 100 (panchagavya : soil) and used.   Seaweed based Panchagavya Seaweed based panchagavya is a modified preparation (Natarajan, 2002) containing the aqueous extract of the alga,  Sargassum wightii. The preparation contained Cow dung - 5.0 Kg; cow urine - 3.0 L; cow milk - 2.0 L; cow curd - 2.0 L; cow ghee - 1.0 Kg; sugarcane juice  - 3.0 L; tender coconutwater - 3.0 L; banana - 12 nos; yeast powder -100 g;jaggery - 100 g; water - 2.0 L. The above composition gives approximately 20.0 L of panchagavya. Cow dung and cow ghee were mixed together in a 25.0 L concrete pot and kept for 3 days with intermittent stirring to exhaust methane gas. On the fourth day all the other ingredients were added to the cow dung - ghee mixture along with spores of Lactobacillus sporogenes (one SPOROLAC tablet having 60 million spores / tablet) and mixed thoroughly. The mouth of the container was covered with a thin cloth and kept in the open in shade. This mixture was stirred twice everyday and after 18 days, 5.0 g of the algal extract residue was added to the preparation and used in experiments. Algal extract residue was prepared by extracting 100.0 g of shade dried  Sargassum wightii with 5.0 L of boiling water for 30 minutes. The extract was allowed to cool, filtered through a layer of muslin cloth and dried in vacuo and the dry residue was used.  Assay of panchagavya preparations on the germination and development of seedlings of pulses and rice The effect of panchagavya preparation on germination and development of rice  Oryza sativa var. ponni and the pulses,  Vigna radiata, Vigna mungo, Arachis hypogaea, Cyamopsis tetragonoloba, Lablab purpureus, Cicer arietinum seedlings were studied. The growth medium was a mixture of dry panchagavya residue and soil at a ratio of 1: 100 (traditional), 1: 50 (seaweed based) and 1: 100 (seaweed based) v/v. Seeds were surface sterilized with 1.0% mercuric chloride, washed several times in running water, soaked overnight in sterile water and allowed to germinate in dark. Germinating seeds were implanted in soil preparations kept in pots of the size 5.2” tall and 3.5” radius. Seedlings raised in sterilized garden soil were used as control. Ten replicates were used for all experimental plants. The seedlings of pulses were inoculated with  Rhizobium R4 (approximately 1x10 9  cells/mL of sterile nutrient solution) twice, i.e. on the 5 th day and then on the 11th day in both control and test pots (except for Oryza sativa  var. ponni). All the developing seedlings except  Oryza sativa were watered on alternate days with Wilson’s N-free nutrient solution. For  Oryza sativa, normal water was used. Twenty one days old seedlings were carefully removed and their linear growth, the number of leaflets produced, leaf area, number of lateral roots formed, number of root nodules, and other biochemical parameters as
detailed below were determined. Rhizobium (R4) was used in the inoculation of plants were grown on YMA (Yeast Mannitol Agar) medium as specified by J.M. Vincent with slight modifications (Thevanathan, 1980). Purity of the organism was checked frequently using 0.05% Congo Red YMA medium. Wilson’s nitrogen-free nutrient solution (Wilson and Reisenauer, 1963) was used in the preparation of the inoculum as well as for watering the developing seedlings. An effective 72 hours old strain of Rhizobium R4 strain was used for inoculating the seedlings. The seedlings were inoculated twice, first on the 5th day after germination followed by a second inoculation on the 11th day.  Ten milliliters of a thick suspension of the inoculum (approximately 1x10 9
 cells/mL of sterile 
nutrient solution) was pipetted out around the base of
each seedling.  During the first few days after
inoculation, care was taken in watering the plants so
as to avoid washing the inoculum out of the soil.
Wilson’s nitrogen-free nutrient solution was used to
water the developing seedlings on alternate days.
Estimation of photosynthetic pigments
Pigments from leaves of the developing
seedlings were extracted with 80% acetone and the
amounts of chlorophylla
 and chlorophyllb were
determined as described by Arnon (1949) and
Yoshida et al.(1976). 
Chlorophylla = 12.21 A663 – 2.81 A646 mg/g tissue 
Chlorophyllb = 20.13 A646 – 5.03 A663 mg/g tissue
Where, A663 and A646 represent the optical
density (OD) values at the respective wavelengths.  
Estimation of nitrogen and carbon
At different stages of development, whole
plants were carefully removed from sand, cleaned
and dried. Shoot, root and nodules formed in pulses
were dried separately in an oven at 90
o
C, until the
weights remained constant.  Then the dry weights of Marsland Press                                         The Journal of American Science, 2010;6(2):61-67
these parts were determined. The dried plant material
was ground in a glass mortar with glass pestle and the
nitrogen content was determined by modified microKjeldahl method (Nesslerisation) (Umbreit  et al.,
1972). Carbon content was calculated from total dry
matter yield of the seedlings (Kvet et al., 1971; Terry
and Mortimer, 1972; Turgeon and Webb, 1975;
Causton and Venus, 1981). 
 
RESULTS
 The potential of utilizing panchagavya as
biofertilizer was tested on the pulses  Vigna radiata,
Vigna mungo, Arachis hypogea, Cyamopsis
tetragonoloba, Lablab purpureus, Cicer arietinum
and the cereal Oryza sativa var. ponni by growing in
soil amended with dried traditional and seaweed
based panchagavya. Traditional panchagavya was
used at a proportion of 1: 100 (panchagavya: soil)
while seaweed based panchagavya was tried at 1: 50
and 1: 100 dilutions with soil. After 21 days, the
seedlings were harvested and studied for their growth
and development.
Effect on the linear growth of experimental
seedlings:
Effect on shoot and root growth
 Soil amended with seaweed based
panchagavya increased the linear growth of both
shoot and root systems in all the pulses and rice as
compared to respective controls (Figures 1 & 2).
Enhancement in the growth of root and shoot systems
in the experimental plants was more pronounced in
seedlings grown in soil amended with seaweed based
panchagavya rather than with traditional
panchagavya. At a ratio of 1: 100 (panchagavya:
soil), shoots of the rice seedlings grown in seaweed
based panchagavya exhibited nearly 100 % more
growth than that of the control plants. In pulses, the
percent increase in the linear growth of shoots over
control plants in  Vigna radiata,  Vigna mungo and
Cicer arietinum was relatively low in the range of
only 16 – 20% as compared to other experimental
plants, in which it was 64 – 98%.  Even in seedlings
grown in traditional panchagavya, the shoots of the
treated plants exhibited 3 – 50% more growth than
the controls. Linear growth of root in the
experimental plants too exhibited a similar response
to treatment with panchagavya (Figure 2). In seaweed
based panchagavya treatment, a two fold increase in
the linear growth of root could be observed in Vigna
radiata, Vigna mungo and Oryza sativa as compared
to controls. Roots of other experimental plants
responded to the same treatment with an increase in
growth ranging from 37 to 89%. Increasing the
concentration of panchagavya (1: 50 dilution)
decreased the effect on linear growth of both shoots
and roots of the experimental plants. Nevertheless,
the effect of seaweed based panchagavya was more
than that observed for treatment with traditional
panchagavya.
Effect on lateral root growth
 As observed for the linear growth of shoots
and roots, the number of lateral roots formed also
was more in the seedlings raised in soils amended
with both traditional and seaweed based panchagavya
(Figure 3). However, use of the latter produced more
lateral roots than the former in all the cases. Effect on
lateral root formation was more pronounced in
Arachis hypogea as compared to other experimental
plants. Seedlings grown in soil amended with
panchagavya produced 264 to 390% more lateral
roots than the control and maximum lateral root
production was always observed in seedlings grown
in soil amended with seaweed based panchagavya
(1:100; panchagavya: soil). Treatment with seaweed
based panchagavya was higher than that observed for
both control and traditional panchagavya.
Effect on leaf development and growth
 Plants grown in soil amended with
panchagavya produced more leaflets ( in pulses) or
leaves (in Oryza sativa) than their respective controls
(Figure 4) and the effect was more pronounced in
seedlings grown in soil amended with low levels of
seaweed based panchagavya (1: 100, panchagavya :
soil ). In  Cyamopsis tetragonoloba,  Lablab
purpureus and  Oryza sativa, the number of leaflets
formed in seedlings grown in seaweed based
panchagavya at 1: 100 dilution was observed to be
twice of that recorded for their respective controls.
Arachis hypogea, Cicer arietinum and  Oryza sativa
also exhibited positive response to the treatments
with both traditional and seaweed based panchagavya
while Vigna radiata showed poor response. Seedlings
grown in soil amended with traditional panchagavya
produced less number of leaves per plant than those
grown in seaweed based panchagavya. In all the
plants, increasing the levels of seaweed based
panchagavya in soil resulted in decreased production
of leaves (Figure 4). Apart from the large number of
leaf or leaflet production, the leaf area or the lamina
of the leaves in plants grown in soil amended with
panchagavya was always larger than those of the
control plants (Figure 5). 
 The growth of leaves in all the experimental
plants was high when grown in seaweed based
panchagavya. In  Lablab purpureus, the lamina size
was nearly 27% larger with seaweed based
panchagavya as compared to those grown in soil
amended with traditional panchagavya (Figure 5). As
compared to control, these seedlings produced leaves
http://www.americanscience.org    41 americansciencej@gmail.comBiofertilizer Potential of Panchagavya   Sangeetha and Thevanathan
http://www.americanscience.org
which had 93% more surface area than that of their
respective controls. Percent  increase over control in
the leaf area of the seedlings of Vigna radiata, Vigna
mungo, Arachis hypogea, Cyamopsis tetragonoloba
and  Cicer arietinum grown in soil amended with
seaweed based panchagavya at a ratio of 1: 100 was
27%, 35%, 46%, 140% and 37% respectively. In
Oryza sativa, the percent increase in leaf area in
response to the same treatment (1: 100 panchagavya:
soil) was 96%.
    42 americansciencej@gmail.com
Effect on photosynthetic pigments  
 Panchagavya amended soil had a profound
effect on the quantities of chlorophylls too in leaves
of the experimental plants (Figure 6). A low
chlorophyll a/b ratio as compared to the control was
recorded in the leaves of all the plants grown in
panchagavya soil preparations. Seaweed based
panchagavya was more effective than the traditional
panchagavya irrespective of the dilutions tried. The
ratio decreased with a decrease in the levels of
seaweed based panchagavya in soil. In other words,
low levels of the panchagavya in soil effectively
decreased chlorophyll a/b ratio without decreasing
the levels of the individual pigments. The treatment
increased the quantities of chlorophyll b in the leaves
of the experimental plants resulting in a low
chlorophyll a/b ratio.
Effect on nodule formation
 Since all the experimental plants except rice
were legumes, the effect of panchagavya on root
nodule formation was also studied in these legumes. 
 Root nodule formation was also enhanced in
the presence of panchagavya.  Again, the effect was
marked in the seedlings grown in soil amended with
low levels of seaweed based panchagavya
(panchagavya: soil; 1: 100). The effect was
maximum in  Arachis hypogea and minimum in
Cyamopsis tetragonoloba. Even traditional
panchagavya treatment could cause an increase from
18% to 62% in the formation of root nodules (Figure
7).
Effect on C/N ratio
The seedlings raised in soil amended with
panchagavya registered a low C/N ratio as compared
to their respective controls (Figure 8). Percent
reduction in the C/N ratio of seedlings grown in
panchagavya amended soils was in the range of 1 –
22%. The C/N ratio of  Arachis hypogea grown in
seaweed based panchagavya preparation was 22%
less than that of its control.  Oryza sativa recorded a
value that was 16% less than that of the control
seedlings (Figure 8). Lowest values for C/N ratio in
the experimental plants were recorded in seedlings
grown in soil amended with low levels of seaweed
based panchagavya (1: 100; panchagavya: soil). Even
at a concentration of 1:  50 (Panchagavya: soil), the
seaweed based preparation was able to reduce the
C/N ratio by about 3 to 17% than the controls. Marsland Press                                         The Journal of American Science, 2010;6(2):61-67
http://www.americanscience.org    43 americansciencej@gmail.com
DISCUSSION
The biofertilizer potential of panchagavya
prepared in the traditional way and a modified
preparation amended with seaweed extract have been
evaluated for their fertilizer potential using the pulses
Vigna radiata,  Vigna mungo,  Arachis hypogea,
Cyamopsis tetragonoloba,  Lablab purpureus, Cicer
arietinum  and the cereal  Oryza sativa var.  ponni  as
the experimental plants. Some farmers in the
southern parts of India use a modified panchagavya
that contains many other plant products to boost
fermentation and to support the growth of beneficial
microorganisms.  In the past three or four decades,
the potential of seaweeds and their liquid extracts in
agriculture as a biofertilizer and a source of growth
promoters have been indicated by many (Bentley,
1960; Bhosle  et al., 1975; Williams  et al., 1981;
Jeanin  et al., 1991; Immanuel and Subramaniam,
1999; Thevanathan  et al., 2005). These reports
formed the basis for a new formulation of
panchagavya with the inclusion of an aqueous extract
of the brown alga,  Sargassum wightii and assess for
its biofertilizer potential in the present investigation. 
Panchagavya is normally advocated as foliar
nutrition (Caraka-Samhita, 1981; Susruta Samhita,
1985; Chauhan, 2002 b; 2004 b; 2005; Fulzele et al.,
2001; Joshi, 2002; Garg and Chauhan, 2003 a;
Saxena et al., 2004) and has not been tried as manure.
Nevertheless dried and powdered seaweeds have
been shown to be a good source of manure when
mixed with soil in small quantities for the cultivation
of vegetable crops and tea. In the present
investigation dried traditional panchagavya and
seaweed based panchagavya were tested as manure.
Dried panchagavya (both traditional and seaweed
based) when mixed with soil at a ratio of 1:100
(panchagavya: soil v/v) and used as a growth medium
promoted the linear growth of both the shoot and
roots of the seedlings of both the pulses and the
cereal, the paddy (Figures 1 and 2).The effect was
pronounced in soils amended with seaweed based
panchagavya even at 1:50 (v/v) dilution. The effect
on all experimental plants except  Lablab purpureusBiofertilizer Potential of Panchagavya   Sangeetha and Thevanathan
http://www.americanscience.org    44 americansciencej@gmail.com
was moderate in the sense that the linear growth did
not mimic etiolation. Similarly, soils amended with
panchagavya (both traditional and seaweed based)
promoted the production of lateral roots, leaves,
leaflets and the growth of lamina in all the
experimental plants (Figures 3, 4 and 5). As
compared to control, the  seedlings produced leaves
which had 93% more surface area than that of their
respective controls. Percent  increase over control in
the leaf area of the seedlings of Vigna radiata, Vigna
mungo, Arachis hypogea, Cyamopsis tetragonoloba
and  Cicer arietinum grown in soil amended with
seaweed based panchagavya at a ratio of 1: 100 (v/v)
was 27%, 35%, 46%, 140% and 37% respectively.
Increased production of lateral roots would provide
more surface area for absorption of water and
minerals by the experimental seedlings than their
controls. Similarly, large number of leaves or leaflets
with greater surface area could be construed as an
indication of enhanced photosynthetic efficiency in
plants grown in soil amended with panchagavya. This
is further confirmed by the marked decrease in the
ratio of chlorophyll a to chlorophyll b (Figure 6) in
leaves of the plants grown in soil amended with both
traditional and seaweed based panchagavya. The C/N
ratio also was very low in these plants as compared to
controls (Figure 8). A low C/N ratio is normally
indicative of a better carbon and nitrogen use
efficiency than plants with high C/N ratio. Since all
the experimental plants except rice were legumes, the
effect of panchagavya on root nodule formation by
rhizobia was also studied in these legumes.
Panchagavya promoted the formation of root nodules
by the inoculated rhizobia (Rhizobium, R4) in the
experimental plants (Figure 7).  The effect was
marked in the seedlings grown in soil amended with
low levels of seaweed based panchagavya
(panchagavya: soil; 1: 100). The effect was more
pronounced in  Arachis hypogea. Even the use of
traditional panchagavya as manure was able to
increase nodule formation  by nearly 18% to 62%
(Figure 7). Though panchagavya has been claimed to
have antibacterial activities (Subramaniam, 2005;
Sugha, 2005), use of panchagavya as manure in the
present investigation was found to promote both the
survival ability and nodulating efficiency of the
inoculated strain of  Rhizobium (Figure 7). High
levels of trace elements and adequate amounts of
potassium and nitrogen have been shown to be
present in seaweeds (Smith and van Staden, 1983,
1984; Tay  et al., 1985; Temple and Bomelle, 1989;
Sekar  et al., 1995). But for the low levels of
phosphate, seaweed meal has been shown in this lab
to be an alternative to farm yard manure in raising
cereals, pulses and nodal cuttings of tea
(Thevanathan et al., 2005). This could be the reason
for the enhanced effect of seaweed based
panchagavya over traditional panchagavya in
promoting the growth and development of
experimental pulses and cereals. 
CONCLUSION
The biofertilizer potential of traditional and
a modified panchagavya containing liquid seaweed
preparation of Sargassum wightii were investigated. 
Soil amended with panchagavya at a
concentration of 1: 100 (panchagavya: soil v/v)
increased the linear growth of both shoot and root
systems of the seedlings of the pulses Vigna radiata,
Vigna mungo, Arachis hypogea, Cyamopsis
tetragonoloba, Lablab purpureus, Cicer arietinum
and the cereal  Oryza sativa var.  ponni.  Increase in
linear growth of the shoots and roots was associated
with a concomitant increase in the number of lateral
roots produced, the number  of leaves or leaflets
produced, increase in leaf area, nodule formation by
Rhizobium and a decrease in the chlorophyll a/b and
C/N ratio. The effect was further enhanced when
seaweed based panchagavya was used as manure at
the same concentration.  
CORRESPONDING AUTHOR
Thevanathan, R.
Centre for Advanced Study in Botany, 
University of Madras (Guindy campus), 
Chennai-25, Tamil Nadu, India.
thevanathan@gmail.com.
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