International Journal of Agriculture Innovations and Research

Volume 5, Issue 4, ISSN (Online) 2319-1473

Manuscript Processing Details (dd/mm/yyyy) :
Received: 28/01/2017 | Accepted on: 03/02/2017 | Published: 11/02/2017

Effect of Fermented Fish waste (Gunapaselam)

Application on the Soil Fertility with Special Reference
to Trace Elements and the Growth Characteristics of
Vigna radiata
B. Thendral Hepsibha1 and A. Geetha2*
1Department of Biochemistry, Ethiraj College for Women, Chennai – 600 008, Tamil Nadu, India.
2Department of Biochemistry, Bharathi Women’s College, Chennai – 600 108, Tamil Nadu, India.

*Corresponding author: Tel.: +91 9444902506

Abstract – The soil fertility largely depends on the nature [2] and to regulate the activities of enzymes involved in
and the quantity of trace elements. Application of chemical N2 fixation and assimilation. Integrity of the cell
fertilizers to increase the food production slowly robs the soil membrane, seed formation, elongation of roots and sugar
nutrients, harm the soil fauna and microbes and cause side metabolism are governed by Boron [3]. Copper performs
effects to consumers. Gunapaselam is prepared from fish vital role in respiration, photosynthesis, N2 and cell wall
waste by fermentation. The aim of the present study is to metabolism, carbohydrate distribution and seed production
assess the level of trace elements, microbial population and [4]. Legumes have very low boron requirements for root
the growth characteristics of Vigna radiata, green gram after
growth and root system architecture [5], stem, leaf, and
applying Gunapaselam. The soil was amended with either
chemical fertilizer (25kg N2: 50kg P2O5: 25kg K2O: 20kg vascular tissue elongation [6] and at the onset of flowering
S/ha) or Gunapaselam (1:100 diluted) on alternative days. and during seed development [7].
The soil was analyzed for trace elements Iron (Fe), Copper Beneficial plant-associated rhizobacteria also enhances
(Cu), Zinc (Zn), Manganese (Mn), Boron (B), Nickel (Ni) plant growth by providing trace elements (such as iron and
before and after applying Gunapaselam. Amendment of zinc), fixing nitrogen, and synthesizing substances
Gunapaselam to soil has improved the quantity of trace including siderophores and indole acetic acid [8] and
elements, macro nutrients and plant growth promoting providing protection against soil pathogens [9].
microbial status of soil. Spearman rank correlation test Manganese has profound influence on electron transport in
reveals positive correlation with the growth traits and
photosynthesis, photo destruction of chloroplast and
formation of number of nodules. The results suggested that
the addition of Gunapaselam may be considered as a novel chlorophyll, nitrogen fixation [7]. Hussain et al. [10] has
liquid biofertilizer as it promotes growth and increase reported that Nickel is critical for root and shoot growth,
nodulation probably by enriching the soil with trace elements photosynthesis, protein synthesis, enzyme activity and
and microbial community. maintenance of membrane integrity. Deficiency of zinc
results in reduced photosynthesis [11], short internodes
Key words – Fermented Fish Waste, Gunapaselam, formation, decrease in leaf size and delayed maturity [12].
Nodules, Plant Height, Root Length, Vigna radiata (Green Inadequate availability of the trace elements in the soil not
gram). only disturb the plants productivity and yield but also
affects the quality of food and feed entering the food chain
I. INTRODUCTION which causes global burden on animal [13] and human
health [14]. The concentration of these trace elements
Fertile soil is the greatest terrestrial stock that supplies should not exceed the critical limit as they are toxic.
all essential nutrients for the growth of healthy and Therefore awareness needs to be created and strategies
productive plants. Continuous harvesting of the crops have to be adopted to correct the trace element deficit
results in depletion of the soil fertility. Addition of through natural means.
chemical fertilizers, supply mainly the macronutrients but Legumes are the second largest group of feed crops
not all the essential elements for the proper growth and grown globally contributing 25% of crop production with
yield of the plants. Successful and sustained production of 61 million tons of grains. Among legumes, Vigna radiata
crops needs mineral nutrients like carbon (C), hydrogen (Green gram / Mung bean) is a short duration and cover
(H), oxygen (O), nitrogen (N2), phosphorus (P), potassium growing crop. It serves as an excellent intercrop that can
(K), calcium (Ca), magnesium (Mg) and Sulphur (S), as add organic matter, cycle the nutrients from depths to soil
well as zinc (Zn), iron (Fe), manganese (Mn), copper (Cu), surface and fix the atmospheric nitrogen by nitrogen-
boron (B), molybdenum (Mo), chlorine (Cl) and nickel fixing bacteria housed within nodules. The bacteria
(Ni). These micronutrients fulfill the requirements of lodging in the rhizosphere were efficient drivers of
plants for their growth and development. nutrient availability and sustainable crop production.
Trace elements are essential for physiological and Agronomists are currently focused on integrated plant
biochemical processes of plants besides the nutrient management system involving plant growth
macronutrients. Iron (Fe) has been found to promote promoting rhizobacteria (PGPRB) towards plant growth.
legume-rhizobia symbiosis [1] helps in nodule formation The yield and nodule forming ability of mung bean is
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 International Journal of Agriculture Innovations and Research
Volume 5, Issue 4, ISSN (Online) 2319-1473

influenced by the availability of essential nutrients and with sterile distilled water. 0.1ml of each dilution was
interaction with rhizobium species. It was now reported spread on PVK containing insoluble tri calcium phosphate
that Indian soils are now facing deficiency of both macro and incubated at 30°c for 2 days. Colonies showing halo
and micro nutrients essential for crop production. Hence zones were conferred as phosphate solubilizer and
there is an urgent need to manage these deficiencies to enumerated.
ensure soil fertility and healthy production of crops. The 2.3. Site Description and Experimental Setup
escalating cost of fertilizers has forced us to recycle the Seeds of Mung bean (Vigna radiata- VBN GG2) were
organic waste and reutilize the nutrients. procured from Agricultural seed store, National Pulses
An organic fertilizer improves soil structure [15], slowly Research Centre, Tamil Nadu Agricultural University,
releases the nutrients, and increases beneficial microbial Vamban, Pudukkottai. Experiment was conducted in a
activity [16] and thus serves as a perfect alternative to field at Porur, Chennai, Tamil Nadu, India. The area of
chemical fertilizers [17]. In recent times, attention has each plot was 3 x 4 m2, keeping plant to plant distance of
been paid on safer organic foods with high quality. 10 cm and row to row distance of 30 cm with 3 m row
Environmental issues forces the reutilization of animal length. The treatment groups were as follows: Group 1-
manure, agricultural waste, animal waste etc., in to a better Water control; Group 2- Chemical fertilizer (basal
form of the nutrient as a good waste management practice treatment and before flowering); Group 3- 1:100 diluted
for sustainable agriculture. Composting and fermentation Gunapaselam (on alternate days). The treatments were
methods have regained focus as they are accepted arranged in randomized complete block design (RCBD)
ecologically. The obtained products are less expensive and with three replications. The source of chemical fertilizer
promote plant growth. They are non- polluting and includes urea as nitrogen fertilizer, single super phosphate
therefore safe to use in agriculture. as phosphate, muriate of potash as potassium and gypsum
More than 70% of fish waste is obtained from the fish as sulphur. Sowing was done at the end of January.
processing industry. Environmental problems caused by Irrigation was proceeded according to the propose design
fish waste can be minimized by transforming them in to throughout the growing season. Weeds were manually
useful products like fertilizers as they are rich in essential eradicated whenever observed in the field. The treatment
macro and micronutrients [18]. Fermentation of the fish groups received water, chemical and diluted Gunapaselam
waste is the right method with many advantages when as per the requirement for a period of 90 days.
carbon source, moisture and aeration were provided in 2.4. Assessment of Plant, Root Length and
right proportion. The effect of fermented fish waste as Nodulation Efficacy of Vigna Radiata
liquid fertilizer has been investigated on the yield of many Ten plants were randomly selected and removed from
vegetables and fruits [19, 20]. Vincent et al. [21] has each plot after 30 days (Flowering stage). The roots were
reported the fermentation of fish waste using jaggery and carefully washed to remove the soil particles and the
it was used under the trade name “Gunapaselam”. The formed nodules were counted manually. The plant height
growth promoting effect of Gunapaselam has been known and root length were recorded.
traditionally but the role played in improving the soil 2.5. Physicochemical Analysis of Experimental Soil
micronutrient status was not available. The aim of this Soil samples were taken randomly from the
study was to evaluate the beneficial role of Gunapaselam experimental site at a depth of 15 cm or 6 inches for
in improving the trace elements level in the soil and physiochemical analysis. The experimental soil used for
growth characteristics of Vigna radiata. the study was analysed at National Agro Foundation-
Laboratory Services Division, Anna University Taramani
II. MATERIALS AND METHODS Campus, Taramani before sowing and after the harvest of
Vigna radiata under different treatment groups. Soil pH,
2.1. Preparation of Gunapaselam electrical conductivity, moisture, organic matter, organic
Gunapaselam was prepared according to the procedure carbon and content of essential nutrients like nitrogen,
described by Vincent et al. [22]. Fermentation of fish potassium, phosphorus, calcium, magnesium, sulphur,
waste was carried out in a covered plastic container. The zinc, manganese, iron, copper, lead and boron
mixture contained 1.5 kg jaggery, 1 kg chopped fish offal exchangeable cations were determined [22].
and 5 lit water. The contents were swirled for 14 days 2.6. Microbial Analysis of Soil
followed by filtration. Then the filtrate obtained was The rhizosphere soil was collected by uprooting the
Gunapaselam which was used after 1:100 dilution. plants after 45 days of sowing and was serially diluted.
2.2 Analysis of Trace Elements and Phosphate The enumeration of total bacteria and rhizobium was done
Solubilisers in Gunpaselam by the spread plate technique. 0.1 ml of the diluted sample
The prepared liquid manure was analysed for essential was spread on nutrient agar and incubated for 48 hrs for
elements like sulphur, zinc, manganese, iron, copper, lead bacteria. 0.1 ml of the sample was spread on Congo red
and boron at National Agro Foundation- Laboratory yeast extract mannitol agar medium and incubated for 72
Services Division, Anna University Taramani Campus, hrs for rhizobium growth. Colonies showing pink colour
Taramani according to the procedure described by Tanden were counted and expressed as log (CFU/gm). Each plate
[22] and potential of Gunapaselam to solubilise insoluble was replicated 3 times.
phosphate was checked on Pikovskaya’s agar medium
(PVK) [23]. 1ml of the Gunapaselam was serially diluted
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 International Journal of Agriculture Innovations and Research
Volume 5, Issue 4, ISSN (Online) 2319-1473

2.7. Statistical Analysis They play multi-factorial role in improving the soil
All data are reported as mean ± standard error of mean. fertility and plant growth. Phosphate solubilisers improve
Statistical analysis was done using SPSS 12.0 for windows the soil fertility and plant growth by facilitating
package. The statistical significance of differences mobilization of phosphorus, trace elements , enhancing
between groups was assessed by one-way analysis of nitrogen fixation and producing phytohormones [29].
variance (ANOVA) followed by Tukey’s multiple Vincent et al. [21] has reported the ammonifying and
comparison test. All statistical analysis was done using nitrifying effects of Gunapaselam. Moreover Gunapaselam
computerized Graph Pad Prism version 5.0, Software in its liquid state can be easily absorbed by plants and can
package (Graph Pad Software Inc., San Diego, CA, also be applied as foliar spray. Thus Gunapaselam, a
U.S.A.). Spearman rank’s correlation was conducted biological preparation with essential trace elements and
between the trace elements and the number of nodules. inoculants of plant growth promoting microbes may serve
P<0.05 was considered as statistically significant. as a cost effective and environmental friendly biofertilizer.
The physiochemical parameters of the soil like pH, EC,
III. RESULTS AND DISCUSSION Organic matter; carbon and CEC, macronutrients and
micronutrient levels are found improved after amending
India is considered to be the center of origin and major the soil with Gunapaselam (Table 3.2.). The pH of
producer of green gram. Due to the poor management of Gunapaselam treated group was 7.56±0.47 when
crops without proper soil enrichment the average compared with chemical (7.89±0.74) and water treated
productivity is poor [24]. Indian soil are poor in nutrient (8.32±0.57). Compared with water control the organic
status as indicated by the deficit of Zn, Fe, Mn, Cu, B, Mo matter and organic carbon were significantly increased in
and S at the rate of 48, 12, 5, 4, 33,13 and 4% [14] It needs Gunapaselam treatment. Chemical and Gunapaselam
fortification in case of inorganic fertilizers but cost and treatment have significantly increased the macronutrients
safety are the limiting factors [13]. Hidden hunger of trace N, P, and K when compared with Group I. The content of
elements can be corrected by agronomic rehabilitation Zn, Mn and Fe have shown a significant improvement in
practices like organic amendments, biofertilizers, and Group III when compared to
traditional crop rotation models including legumes, cereals other groups. The levels of B and Cu been increased
and vegetables to restore the soil fertility. Fish wastes are significantly (P<0.05) by irrigation with Gunapaselam.
the important resources with high nutritive value. Lead remains unaffected in all the treatment groups. Ni
Fermentation utilizing the microbes break down the level was significantly high (P<0.01) in Group III and
complex materials in to simple forms and makes the remains unaltered in Group II and Group I.
nutrients available to the soil.
The proximate trace elements present in Gunapaselam Table 3.1. Trace element Composition of Gunapaselam
(Fermented Fish waste) are shown in Table 3.1. The level (Fermented Fish Waste)
of micronutrients in Gunapaselam is Zn (3.21±0.32), Mn Parameter Unit Results
(1.34±0.10), Fe (15.69±1.36), Cu (0.55±0.05), Pb Iron mg/Kg 15.69±1.36
(0.69±0.06), Ni (Below detectable limit) and B Zinc mg/Kg 3.21±0.32
(1.76±0.14) (mg/Kg). The amount of sulphur and sodium Manganese mg/Kg 1.34±0.10
were 0.04±0.00 and 0.07±0.00 (%) respectively. The Copper mg/Kg 0.55±0.05
presence of the above mentioned microelements in Boron mg/Kg 1.76±0.14
Gunapaselam are the essential nutrients recovered from Sulphur % 0.04±0.00
the fish waste by fermentation process. Ghaly et al. [25] Lead mg/Kg (0.69±0.06
has reported that fish fillets are excellent sources of Ca, Sodium % 0.07±0.00
Mg, P, K, Na, Fe, Zn, Mn and Cu. Hence this method Nickel mg/Kg BDL
would be the best way to recover the plant growth BDL- Below detectable limit; Data represent Mean ±
promoting substances from fish waste. Nearly 25 trace Standard Error Mean
elements are found to be the critical components in
governing the growth and yield of plants. Moses et al. [26] The decrease in soil pH of the Gunapaselam treated plot
also observed similar results with different poultry was due to the decomposition of organic matter in the fish
manures. The content of trace elements found in waste in to humus which is further digested by soil
Gunapaselam is always within the permissible limits of a microbes to produce organic acids. Slightly acidic soil pH
fertilizer and they also satisfy the requirement of the plants favors the growth of green gram. This property increases
for their healthy growth [27]. the cation exchange capacity of the soil which is
The phosphate solubilising potential of Gunapaselam is responsible for the mobilization of calcium, potassium for
shown in Fig. 3.1 and the count was log 7.32 CFU/ml. plant growth. The variation in the level of trace elements
These phosphate solubilisers when added to soil will in various treatments may be attributed due to the variation
mobilize the insoluble phosphates in the soil and hence in microbial population caused by changes in the
improve the uptake by the plants and crop growth [28]. physiochemical properties of the soil like pH, moisture
Among the beneficial growth promoting microbes, and organic content, different in the concentration of
phosphate solubilisers are promising candidates in nutrients exuded by mung bean and by the nutrients
agriculture as they promote crop production and yield. provided by Gunapaselam. Intensive use of inorganic
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Volume 5, Issue 4, ISSN (Online) 2319-1473

fertilizers has brought deleterious effects like worsening of recorded significantly longest root length and plant height
soil physical and chemical properties, creating imbalance and more number of nodules per plant. Application of
in soil nutrients, reducing crop yield [30]. The chemical treatment registered the next best results when
improvement in soil properties shown by Gunapaselam compared to water treatment. Spearman rank correlation
application will restore back the soil fertility otherwise lost test reveals the nodules formed in Gunapaselam treatment
due to chemical fertilizers. was in positive correlation with the level of trace elements
The rhizobacterial and rhizobium populations of soil Fe, Zn, Cu, and Mn essential for nodulation (Table 3.5).
under various treatments are presented in Table 3.3. The The number of nodules produced in chemical fertilizer
total rhizobacterial load in the soil of Group I and Group II treatment was lower than Gunapaselam treated plants.
plots were significantly lower than Group III (8.30±0.21 The root system play dynamic role in water and nutrient
log(CFU/gm)). The lowest rhizobium count was recorded absorption. The presence of humic substances in organic
in Group I (6.39±0.05) than in Group II and Group III. matter of the soil and availability of nutrients especially
Microbial populations have major influence on plant ammonium and nitrate modify the root morphology by
growth and productivity [31]. These rhizobacteria inducing the proliferation of root cells leading to root
contributes to seed germination and influences α- amylase elongation, branching of axial roots and elongation of
activity that mobilizes the storage reserve and thereby lateral roots [34, 35] . The rhizobacteria also contributes
provides energy for development and growth of roots and in increasing the root length, number of branches and
shoots. surface area of roots [36]. Our results prove that
Gunapaselam treatment has lead to long and extensive root
system which can explore large volume of the soil to
acquire water and more nutrients. It might have favored
the accumulation of rhizobacteria which adopts the
potential mechanism of sorption, precipitation and
occultion to mobilize the trace elements to enhance the
root growth and proliferation of lateral roots. Similar
results are reported with the root growth of nectarine [37],
raddish [38] and peach [39] by using cow manure and fish
emulsion. The increase in plant height of Vigna radiata
induced by Gunapaselam was due to the attainment of
threshold turgor for stem cell elongation under good
Fig. 3.1. Insoluble Phosphate solubilising potential of moisture condition and the plant will harvest better
Gunapaselam on Pikovskaya’s agar sunlight for more photosynthates. The increase in plant
height was contributed by boron which promotes division
Continuous application of chemical fertilizers may lead and differentiation of cells controlling cellular and
to depletion of beneficial microbial community in the soil. metabolic activities [40], copper functions as chlorophyll
Microbial population is controlled by pH, salinity, activator 41]. Sufficient presence of Zinc leads to
temperature and organic matter [32] The increase in the development of long internodes, therefore increase in plant
rhizobacterial population in Gunapaselam treated soil height and leaf size [12] and nitrogen increases the number
confirms the favorable atmosphere for multiplication of and length of the internodes which results in progressive
microbes when compared with chemical and water increase in plant height. Similar results were reported by
treatment. They are the chief driving forces in recycling Asha [42] and Brajeshwar [43] on treatment with
the soil nutrients and make them available for plant Kunapajala, a fermented liquid from animal wastes. A
growth. healthy root growth is the basis for maintaining a high
The presence of appreciable amount of symbiotic nutrient status, shoot growth and yield. The well
nitrogen fixing rhizobium species in Gunapaselam developed root system in Gunapaselam treated plants
treatment establish complex interplay with roots of Vigna might have resulted in increased absorption of water and
radiata resulting in the formation of more number of nutrients and hence transportation of the same to the shoot
nodules and hence fix atmospheric nitrogen when system. In the present study, the changes in the nodule
compared with other treatments. In the present study numbers under various treatments might be due to the
addition of Gunapaselam with phosphate solubilizers influence of microbial community and concentration of
augment plant growth not only by solubilising insoluble essential trace elements. The plant growth promoting
inorganic phosphate, also by promoting nitrogen fixation, rhizobes facilitate the available forms of all essential
increasing the availability of trace elements and producing elements in surplus amounts to influence legume root
plant hormones [33]. Thus a favourable environment for infection and nodulation [44]. Sufficient supply of B
the beneficial rhizobacteria, phosphate solubilizers and ensures formation and upholding of functional sym-
rhizobium species provided by Gunapaselam treatment biosomes [45]. Our results shows that, lesser concentration
will be a natural biological approach for plant nutrient of iron content in soil in water treated group limits the root
management. nodule production in Vigna radiata, as Fe is essential for
Amendment of Gunapaselam has positively influenced symbiotic partnership of root nodule bacteria and
the plant length, root length and number of nodules multiplication [46], nodule development and their
formed (Table. 3.4). Gunapaselam treated plants has
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functions [47]. Fe is also required for establishing the Gunapaselam has promoted the plant growth, root length
synthesis of key enzymes involved in N2-fixation and nodulation when compared with chemical fertilizer
(nitrogenase, leghemoglobin) and in nitrogen assimilation and untreated water control. The results suggested that
[48]. Deficiency of iron has resulted in lesser nodules, preparation of Gunapaselam by treatment of fish waste
bacteriods, symbiosomes and reduced plant growth as will reduce the environmental burden of solid wastes and
evidenced by Balestrasse et al. [49]. The increase in the also it would be an inexpensive alternative to chemical
soil nitrogen content after the harvest of Vigna radiata was fertilizers to promote the growth of legumes.
due to increased formation of nodules facilitated by
surplus of Fe and Ni essential for nodulation aided ACKNOWLEDGMENT
transport of ureides to upperparts of the plants, Mn for
nitrate reduction and concomitant nitrogen fixation by This work was supported by the University Grants
microbes. This improvement induced by Gunapaselam in Commission, New Delhi, India under minor project
the vegetative character and symbiotic relationship might scheme.
have been due to the improvement in soil properties with
enhanced nutrients.

IV. CONCLUSIONS
Gunapaselam, the fermented fish waste was found to be
a biological preparation epitomizing as a sufficient source
of micronutrients and microorganisms. The tangible
beneficial role of Gunapaselam was evidenced by
providing a proper rhizosphere with beneficial microbes
for the growth of Vigna radiata. The treatment with

Table 3.2. Soil fertility before the start of the study and after the growth of Mung bean
Parameter Unit Before applying Group I Group II Group III
pH - 8.85±0.63 8.32±0.57 aNS 7.89±0.74 aNS, bNS 7.56±0.47 aNS, bNS
Electrical conductivity mS/cm 0.24±0.01 0.38±0.03 a* 0.52±0.02 a***, b* 0.59±0.04 a***, b**
Texture Sandy Clay Loam Sandy Clay Loam Sandy Clay Loam Sandy Clay Loam
Organic matter % 1.21±0.05 1.27±0.11 aNS 1.41±0.11 aNS, bNS 2.04±0.15 a**, b**
aNS aNS, bNS
Organic Carbon % 0.32±0.03 0.35±0.00 0.41±0.03 0.53±0.03 a**, b**
aNS a**, b*
Available Nitrogen Kg/ha 204±16.11 240±22.80 357±21.66 394±34.34 a**, b**
aNS a*,b*
Available Phosphorus Kg/ha 9±0.81 10±0.59 15±1.35 20±1.38 a***, b***
aNS a**, b*
Available Potassium Kg/ha 98±7.64 150±14.7 219±14.5 250±19.75 a***, b**
aNS aNS, bNS
Available Zinc ppm 2.15±0.19 2.24±0.19 2.81±0.2 5.38±0.36 a***, b***
aNS aNS, bNS
Available Manganese ppm 13.61±1.21 14.05±1.47 14.05±1.47 29.99±1.73 a***, b***
aNS aNS, bNS
Available Iron ppm 13.10±1.14 14.24±1.03 15.26±1.12 22.37±1.8 a**, b*
aNS aNS, bNS
Available Copper ppm 1.93±0.19 1.99±0.17 2.08±0.14 2.87±0.19 a*, b*
aNS aNS, bNS
Available Boron ppm 0.8±0.07 1.0±0.08 1.0±0.10 1.37±0.05 a**, b*
aNS aNS, bNS
Available Lead ppm 1.47±0.11 1.34±0.12 1.31±0.07 1.67±0.15 aNS, bNS
aNS aNS, bNS
Available Nickel ppm 0.39±0.03 0.37±0.02 0.36±0.02 0.68±0.06 a**, b**
aNS aNS, bNS
CEC Meq/10 22.94±2.24 23.49±1.68 24.60±2.23 25.54±2.42 a*, b*
0g
(Group I- Water; Group II- Chemical Fertilizer (25kg N2: 50kg P2O5: 25kg K2O: 20kg S/ha); Group III- unapaselam (1:
100 dilution)

Mean value between the groups were analysed using one way ANOVA followed by tukey’s multiple comparison test in
graphpad prism 5.0. a*, a**, a*** indicates p value < 0.05, 0.01 and 0.001, respectively vs before study and b*,b**,
b*** indicates p value < 0.05, 0.01 and 0.001, respectively after study vs group I

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Table 3.3. Soil rhizobacterial and rhizobium population influenced by Gunapaselam(GP) on Vigna radiata
Group / Treatment Rhizobacterial load Rhizobium load
log (CFU/gm) log (CFU/gm)
Group I- Water 6.81± 0.15 6.39± 0.05
Group II- Chemical Fertilizer
7.69±0.23*
(25kg N2: 50kg P2O5: 25kg K2O: 20kg S/ha) 7.32± 0.03***
Group III- Gunapaselam
8.30±0.21**
(1: 100 dilution) 8.09± 0.04***
Mean value between the groups were analysed using one way ANOVA followed by tukey’s multiple comparison test in
graph pad prism 5.0. *, **, *** indicates p value < 0.05, 0.01 and 0.001, respectively vs group I

Table. 3.4. Vegetative growth and symbiotic parameter influenced by Gunapaselam(GP) on Vigna radiata
Group / Treatment Plant Length (cm) Root Length No. of nodules
(30th day) (cm) (30th day)
(30th day)
Group I- Water 22.02± 0.75 13.7± 0.85 6±0.1
Group II- Chemical Fertilizer 17.3± 0.73** 14±0.3***
(25kg N2: 50kg P2O5: 25kg K2O: 20kg 25.70±1.12*
S/ha)
Group III- Gunapaselam 19.7± 0.48** 32±0.7***
27.25±0.75**
(1: 100 dilution)
Mean value between the groups were analysed using one way ANOVA followed by tukey’s multiple comparison test in
graphpad prism 5.0. *, **, *** indicates p value < 0.05, 0.01 and 0.001, respectively vs group I

Table 3.5. Spearman rank Correlation test

Parameters rs Value P value
Iron Vs Number of nodules
1.000 p < 0.0025
Zinc Vs Number of nodules
0.943 p < 0.01
Copper Vs Number of
nodules 0.943 p < 0.01
Manganese Vs Number of
nodules 1.000 p < 0.01
Based on the critical values of rank correlation (Spearman’s rho- rs), null hypothesis of no correlation was rejected and
concluded that the aforementioned parameters have correlations

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