INTRODUCTION:

The use of chemicals, such as fertilizers and pesticides for the enhancement of productivity of crops has become the order of the day. After the initiation of green revolution the crop productivity has increased manifold to meet the demand of ever growing population the world over. But the use of these chemicals have caused great damage to the environment in the form of pollution to air, water and soil and also led to various diseases to human beings, animals and plants. Over the last few decades there is change in the attitude of the farmers to revert back to the old system of farming using organic manures, panchagavya, vermicompost and other natural modes of fertilizers and pesticides, particularly in India.

This is a welcome change for the sustainable development of agriculture and at the same time saving the nature from pollution. The present study is one such attempt to prove the efficacy of these age old methods of crop development without harming the nature. Green revolution had led to intensified agriculture to meet the ever increasing demand for food and fibre. The extensive use of chemical fertilizers in different combinations increased the crop yield many folds as compared to the earlier local farming practices. Although green revolution has met the needs of food, it has its own inherent side effects like ecological degradation, high rate of pollution of air water and soil ultimately leading to overall health problems to the population, organisms and plants.¹ The indiscriminate use of chemical pesticides in modern agriculture resulted in the development of several problems such as development of pesticide resistant insects, resurgence of target and non-target pests, eutrophication of soil and water, destruction of beneficial organisms like honey bees, pollinators, parasitoids, predators, soil fertility depletion, increase in soil acidity, run-off of fertile soil, soil erosion and other socioeconomic problems etc. The use of pesticides has led to the contamination of food, feed and fodder. This awareness has resulted in the development of Integrated Pest Management (IPM). Organic farming has developed very rapidly in recent years.^2,3Indian agriculture has better chance to convert itself towards organic mode of farming because in India the use of chemical fertilizers and pesticides is still lower compared to the global standards. Cattle, particularly the cow is considered to be an indispensible animal for any Indian agriculturist.

Environmental degradation is a major threat confronting the world, and the rampant use of chemical fertilizers contributes largely to the deterioration of the environment, loss of soil fertility, less agricultural productivity and soil degradation. On one hand tropical soils are deficient in all necessary plant nutrients and on the other hand large quantities of such nutrients contained in domestic wastes and agricultural by products are wasted. It is estimated that in cities and rural areas of India nearly 700 million organic wastes are generated annually which is either burned or land filled. In natures laboratory there are a number of organisms that have the ability to convert organic waste into valuable resources containing plant nutrients and organic matter which are essential for maintaining soil productivity. Microorganism and earthworms are important biological organisms helping nature to maintain nutrient flow from one system to another and also minimize environmental degradation. The earthworm population is about 8-10 times higher in uncultivated area. This clearly indicates that earthworm population decreases with soil degradation and thus can be used as a sensitive indicator of soil degradation. In this report a simple biotechnological process disposal of waste as well as the most needed plant nutrients for sustainable productivity is described, which could provide a ‘win-win’ solution to tackle the problem of safe disposal of waste as well as the most needed plant nutrients for sustainable productivity.⁴

Organic vegetable cultivation gets a special attention ground nut (Arachis hypogaea L.) is an important oilseed, grown approximately due to its bio-efficacy, sustainability and eco-friendly 24 million ha throughout the world. It is a valuable application of vermicompost as an organic source for cash crops by millions of small farmers because of ideal for soil and crop management. Its kernels are rich leading countries where vermicompost is widely applied source of edible oil (43.55%) and protein (25.28%). Use of organic amendments two third of world production is crushed for oil and (such as vermicompost) has been found effective for remaining one third is consumed as food. Groundnut improving soil aggregation, structure, and fertility, cakes obtained after oil extraction is a high protein animal increasing soil microbial diversity, populations and feed. China, India, United States, Nigeria, Indonesia, enzymes, improving moisture-holding capacity of soils, Burma and Senegal are the major producers of groundnut. Increasing cation-exchange capacity (CEC) and finally Asia with 63.4 % area produces 71.7 % of world also crop yields.⁵

An estimated 70% of Indian arable land is rainfed. This increases the usefulness of introducing low cost organic farming techniques as a viable alternative to high cost conventional chemical farming. One of the main questions raised with regard to organic farming practices includes the ability of organic methods to meet the nutrient requirements of the crops while increasing yield at low-cost.⁶

Traditional composting of organic matter wastes has been known for many years but new methods of thermophilic composting have become much more popular in organic waste treatment recently since they eliminate some of the detrimental effects of organic wastes in the soil. Composting has been recognized as a low cost and environmentally sound process for treatment of many organic wastes.⁷

Furthermore, the rapid decomposition and raised temperatures during composting produce a relatively homogeneous, odor-free, pathogen-free and easy-to-handle product. It has been reported that compost-treated soils had lower pH and increased levels of organic matter, primary nutrients, and soluble salts.^8,9,10

However, composting and vermicomposting are quite distinct processes, particularly concerning the optimum temperatures for each process and the types of microbial communities that predominate during active processing (i.e. thermophilic bacteria in composting, mesophilic bacteria and fungi in vermicomposting). The wastes processed by the two systems are also quite different. Edwards and Aranon (1988) reported that vermicomposts have a much finer structure than composts and contain nutrients in forms that are readily available for plant uptake.¹¹There have also been reports of production of plant growth regulators in the vermicomposts.^12,13

Vermicompositing appears to be the most promising as high value bio fertilizer which not only increases the plant growth and productivity by nutrient supply but also is cost effective and pollution free. Use of vermicompost promotes soil aggregation and stabilizes soil structure. This improves the air- water relationship of soil, thus increasing the water retention capacity and encourages extensive development of root system of plants. The mineralization of nutrients is observed to be enhanced, therefore results into boosting up of crop productivity. The vermicomposts have a higher base exchange capacity and more exchangeable calcium, magnesium, potassium than the soil in which worms live. Pot experiments were conducted by Mathivannan et al, 2012 on the effect of vermicompost on growth and yield of groundnut (Arachis hypogaea L.).¹⁴ The highest root length, shoot length, total leaf area, number of root nodules, fresh weight, dry weight, chlorophyll, Carotenoids were recorded in ground nut crop grown in the application of 200g of vermicompost at various stages of its growth (25, 50, 75 and 100 DAS). The highest number of pods (65 per plant), number of seeds (116 per pods), hundred seed weight (288.5 g) were recorded in the crop grown with 200 grams of vermicompost application.¹⁴

With the progressive increase in the size of the world’s population and the adoption of intensive animal husbandry production, large volumes of organic wastes produced all over world creating a serious disposable problem and environment pollution.¹⁵ On the other hand the use of synthetic fertilizers causes a great impact on the environment and the cost of these fertilizers is increasing over the years. Now it is a well-established fact that the organic fertilizers provide a enough requirements for a proper growth of the crop and enhance the uptake of nutrients increase the assimilation capacity and will stimulate the hormonal activity as well. Microorganisms and earthworms are important biological organisms helping the nature to maintain the nutrient flow from one system to another and also minimize the environmental degradation.

Traditional composition of organic waste matter has been known for many years but new methods of thermophilic composting have been much more popular. Composting has been recognized as a low cost and environmentally sound process for treatment of many organic wastes. Furthermore, the rapid decomposition have raised temperatures during composting procedure a relatively homogenous, odour-free, pathogen free and easy to handle product. Vermicomposting is a simple biotechnological process of composting, in which certain species of Earth worms are used to enhance the process of waste conversion and produce a better end product. It is nutritionally rich natural organic fertilizer which releases nutrients relatively slowly in the soil and improve the quality of the plants along with physical and biological properties of soil.¹⁶The vermicompost had much larger population of bacteria (5.7×10⁷) fungi (22.7×10⁴) and actinomycetes (17.7×10⁶). Compared with those in conventional composts earth worms acts as mechanical blenders and by fragmenting the organic matter they modify its physical and chemical status by gradually reducing the ratio of C:N and increasing the surface area exposed to microorganisms. Thus making it much more favorable for microbial activity of further decomposition.¹⁷

MATERIALS AND METHODS:

Plant Description:

Ground Nut (Arachis hypogaea L.) is an important oil seed crop in India of commonly known as poor man’s nut (Fig 1). Ground nut kernels contains 42% to 50% oil, 26% protein, 18% carbohydrate are also rich in source of riboflavin, thiamine, nicotinic acid and vitamin E.¹⁸ Ground nuts for edible purposes require considerable processing and sorting to ensure high quality. So the present study was to determine the effect of different ratio of vermicompost on the growth and nutrient content of Ground nut (Arachis hypogaea L) under laboratory conditions.

Fig 1 Arachis hypogaea

EXPERIMENTAL DESIGN:

A pot culture experiment was conducted in the laboratory using the vermicompost and vermicompost enriched with biofertilizers red soil was collected from the agricultural lands sieved and weighed in the black plastic bags with 20cm in diameter and 20cm in depth. The experimental was conducted in a complete randomized block design with 10 treatments and 1 control as follows, BAG 1: 10% vermicompost+90% red soil (SAMPLE 1), BAG 2: 20% vermicompost+ 80% red soil (SAMPLE 2), BAG3: 30% vermicompost +70% red soil (SAMPLE 3), BAG4: 40% vermicompost+60% red soil (SAMPLE 4), BAG5: 50% vermicompost+50% red soil (SAMPLE 5), BAG6: 60% vermicompost+40% red soil (SAMPLE 6), BAG7: 70% vermicompost+30% red soil (SAMPLE 7), BAG8: 80% vermicompost+20%red soil (SAMPLE 8), BAG9: 90% vermicompost+10% red soil (SAMPLE 9), BAG10: 100% vermicompost (SAMPLE 10), BAG11: 100% red soil (control).

Ground nut seeds (Arachis hypogaea. L.) were soaked in water for one day and three seeds were sown in each plastic bag. The experimental plants were kept under observation with the proper level of irrigation. The chemical analysis of control and vermicompost plants was tabulated.

PLANT ANALYSIS:

The ground nut growth parameters such as shoot length (SL), root length (RL), total length (TL), fresh weight of shoot (Fwt S), fresh weight of root (Fwt R), dry weight of shoot (Dwt S), dry weight of root (Dwt R), Total dry weight (Dwt TP), leaf area index (LAI), chlorophyll a, b and total biochemical parameters such as protein (%) and amino acid (%) were analyzed.

LEAF AREA INDEX (LAI):

Leaf area index (LAI) is the ratio of leaf area per plant to the land area occupied by the plant and calculated by using the formula as,

LAI =

FRESH WEIGHT:

The seedlings were separated into root &shoot. They were rolled in a blotting paper to remove water after that each seedling’s fresh weight were taken using single pan balance.

DRY WEIGHT:

The seedlings were kept in hot air oven at 80^°C for 24 hours. Then their dry weight was weighed and recorded with the help of an electronic single pan balance.

GERMINATION PERCENTAGE:

The germination percentage was calculated by using the formula,

GI =×100

VIGOUR INDEX:

The vigor index of seedlings was calculated by using the formula proposed by Abdul-Baki and Anderson (1973).¹⁹

VI = germination percentage × length of the seedlings

BIOCHEMICAL STUDIES:

The 15^th day seedlings were separated into leaves, shoot and root. They were used for biochemical analysis like chlorophyll-a chlorophyll-b, total chlorophyll, carotenoids, amino acids, proteins some enzyme like catalase, peroxidase by the following methods.

ESTIMATION OF CHLOROPHYLL:

leaf material (0.5g) was ground with pestle & mortar using 10ml of 80% acetone. The homogenate was centrifuged at 3000 rpm for 10 minutes. The supernatant was collected and then the pellet was re-extracted with 5ml of 80% acetone. All the supernatant were collected & utilized for chlorophyll determination. The absorbance was measured at 645nm & 663nm using a spectrophotometer. The chlorophyll content was determined using the formula,

CHLOROPHYLL a=

CHLOROPHYLL b=

ESTIMATION OF CAROTENOID

The procedure for the measurement of carotenoid was same as that of chlorophyll estimation. The extract was measured at 480nm in spectrophotometer. The amount of carotenoid present in the extract was calculated by using the formula, carotenoid (mg.g-1)=A480-0.114XA663-0.638XA645

ESTIMATION OF TOTAL FREE AMINO ACIDS:

The plant material (0.5 g) was ground with 10ml of 80% ethanol with the help of pestle and mortar. the extract was centrifuged at 8000rpm for 10minutes.the supernatant was collected and make upto 10ml with 80% ethanol. 1ml of extract was taken and add a drop of methyl red, 1ml of 0.1N NaOH and 1ml of ninhydrin reagent, it is heated in a water bath for 30minutes.then the solution was made upto 20ml with distilled water. The absorbance was measured at 570nm in a spectrophotometer.

ESTIMATION OF PROTEIN:

The plant material (0.5 g) was ground with 10ml of 20% TCA. The homogenate was centrifuged for 10 minutes at 8000rpm. The supernatant was collected and the pellet was re-extracted with 5ml of 0.1N NaOH. One ml of the supernatant was used for protein estimation using Lowry method. The absorbance was calculated at 660nm in a spectrophotometer.

ENZYME ASSAY:

PEROXIDASE TEST:

1g of fresh plant tissue extracted in 3ml of 0.1M phosphate buffer pH 7 by grinding with a pre-cooled mortar and pestle. Centrifuge the homogenate at 18000rpm at 5^°C for 15 minutes. The supernatant containing enzyme source was used.

Pipette out 3ml of 0.1M phosphate buffer solution (pH 7.0),0.05ml of 20mM guaiacol solution,0.1ml enzyme extract and diluted 30% of 0.03ml hydrogen peroxide solution in a cuvette and mixed well. the cuvette was placed in spectrophotometer until the absorbance increases from by 0.05 to 0.1. the time interval was calculated as t.

Enzyme activity (units/litre) =

CATALASE ACTIVITY:

The plant tissue was blended with the M\150 phosphate buffer and centrifuged at 18000rpm for 15 minutes at 1-4°C. The supernatant was collected and treated for catalase activity. An aliquot containing 3ml of hydrogen peroxide and 0.01-0.04ml of sample was taken and shaken well in a test tube and the absorbance was measured using spectrophotometer at 240nm. The absorbance should decrease from 0.45 to 0.40. The time interval for the decrease of the absorbance was calculated.

RESULTS AND DISCUSSION:

Table 1 shows the effect of Vermicompost on Leaf Area Index. Table 2 and 3 indicate the effect of Vermicompost on Fresh Plant Weight and dry weight, respectively. Table 4 indicates the effect of Vermicompost on Germination Percentage and Vigour index. Table 5: Indicates the effect of vermicompost on shoot and root length of Arachis hypogaea. Table 6: Indicates the chlorophyll contents in Aarachis hypogaea. Table 7: Indicates the levels of carotenoid content of Arachis hypogaea L. Table 8 and 9 indicates the effect of Amino acid and Protein contents respectively. Table 10 indicates Peroxidases and Catalase enzyme activities in Arachis hypogaea L. Figure1. Plant of Arachis hypogaea. Figure. 2. indicates the effect of Vermicompost on the Protein levels of experimental plants.

Table 1: Changes in leaf area index of arachis hypogaea l.

Number of Sample	Leaf Area Index (LAI)
SAMPLE 1	2
SAMPLE 2	2.33
SAMPLE 3	3
SAMPLE 4	3
SAMPLE 5	4.6
SAMPLE 6	5.67
SAMPLE 7	6.3
SAMPLE 8	7.67
SAMPLE 9	7.33
SAMPLE 10	8.67

The leaf area index (LAI) shows linear variations that it increases with the increase of the vermicompost composition.

Effect of vermicompost on fresh weight:

Table 2: Changes in fresh weight of Arachis hypogaea l.

Number of Sample	Fresh Weight of Shoot (Fwt S)	Fresh weight of Root (Fwt R)	Fresh Weight of The Total Plant (Fwt TP)
Sample 1	0.58	0.19	0.59
Sample 2	0.6	0.3	0.90
Sample 3	0.78	0.39	1.18
Sample 4	1.52	0.76	2.28
Sample 5	1.88	0.94	2.82
Sample 6	2	0.99	2.99
Sample 7	2	1	3.0
Sample 8	2.1	0.9	3.0
Sample 9	2.08	1.04	3.13
Sample 10	3	0.96	3.96

The fresh weight of the shoot and root indicate that Vermicompost has a positive effect on the growth on this plant.

Effect of vermicompost in dry weight

Table 3: changes of dry weight in arachis hypogaea l.

Number of Sample	Dry Weight of Shoot (Dwt of S)	Dry Weight of Root (Dwt R)	Dry Weigh of Total Plant (Dwt TP)
Sample 1	0.007	0.003	0.01
Sample 2	0.009	0.003	0.12
Sample 3	0.677	0.003	0.68
Sample 4	0.49	0.34	0.83
Sample 5	0.71	0.05	0.76
Sample 6	0.3	0.69	0.99
Sample 7	1.17	0.73	1.90
Sample 8	1.11	0.85	1.96
Sample 9	0.44	0.69	1.13
Sample 10	0.56	0.90	1.46

The dry weight of the shoot and root shows linear increment due to the effect of Vermicompostin this plant.

EFFECT OF VERMICOMPOST ON GERMINATION PERCENTAGE AND VIGOUR INDEX

Table 4: Changes in the Germination percentage and Vigour Index of Arachis hypogaea L.

Number of Sample	Germination Percentage (G %)	Vigour Index (VI)
Sample 1	33.3	233.1
Sample 2	33.3	419.58
Sample 3	66.67	1040.52
Sample 4	83.3	1332.8
Sample 5	83.3	1790.95
Sample 6	83.3	1849.26
Sample 7	100	2370
Sample 8	100	2700
Sample 9	100	3000
Sample 10	100	3250

The vigour index increases with increase of the vermicompost composition as the germination yield shows steady increase with the same ratio.

Figure 2: Indicate the proportional growth of plants with increase in Vermicompost concentrations

Effect of vermicompost on the growth parameters

Table 5: changes of shoot and root length of Arachis hypogaea l.

Number of Sample	Shoot Length (SL)	Root Length (RL)	Total Length (TL)
SAMPLE 1	5	2	7
SAMPLE 2	7.7	4.9	12.6
SAMPLE 3	9.7	5.9	15.6
SAMPLE 4	10	6.0	16
SAMPLE 5	15.5	6.0	21.5
SAMPLE 6	16	6.2	22.2
SAMPLE 7	17	6.7	23.7
SAMPLE 8	21	7.0	27
SAMPLE 9	22.9	7.1	30
SAMPLE 10	24	8.5	32.5

The shoot and root length shows a gradual increase due to the treatment of vermicompost concentrations.

Effect of vermicompost on chlorophyll content

Table 6: Changes in chlorophyll contents of Arachis hypogaea l.

Number of Sample	chlorophyll a	Chlorophyll b	Total Chlorophyll
SAMPLE 1	0.0520	0.0295	0.0649
SAMPLE 2	0.0961	0.0165	0.1126
SAMPLE 3	0.0347	0.0080	0.0433
SAMPLE 4	0.0117	0.0208	0.0325
SAMPLE 5	0.0125	0.0223	0.0348
SAMPLE 6	0.0694	0.0133	0.0827
SAMPLE 7	0.0453	0.0091	0.0544
SAMPLE 8	0.0675	0.0117	0.0792
SAMPLE 9	0.0101	0.0170	0.1711
SAMPLE 10	0.0211	0.0147	0.9358

The chlorophyll contents did not vary due to the effect of vermicompost.

Effect of vermicompost on carotenoid content

Table 7: Changes in the carotenoid content of Arachis hypogaea l.

Sample	Carotenoid
SAMPLE 1	0.4132
SAMPLE 2	-0.0165
SAMPLE 3	0.0067
SAMPLE 4	0.157
SAMPLE 5	0.0534
SAMPLE 6	0.1115
SAMPLE 7	0.2794
SAMPLE 8	0.1635
SAMPLE 9	0.0227
SAMPLE 10	0.2281

The vermicompost treatment did not show much increase in carotenoid contents.

Effect of vermicompost on amino acids

Table 8: changes in the amino acids content of Arachis hypogaea l.

Sample	Amino acids
SAMPLE 1	0.43
SAMPLE 2	0.42
SAMPLE 3	0.41
SAMPLE 4	0.42
SAMPLE 5	0.43
SAMPLE 6	0.44
SAMPLE 7	0.42
SAMPLE 8	0.44
SAMPLE 9	0.44
SAMPLE 10	0.44

The amino acid content also did not vary much as a linear variation with the vermicompost composition.

Effect of vermicompost on protein

Table 9: changes in the protein content of Arachis hypogaea l.

Sample	Protein
SAMPLE 1	0.03
SAMPLE 2	0.06
SAMPLE 3	0.06
SAMPLE 4	0.08
SAMPLE 5	0.10
SAMPLE 6	0.12
SAMPLE 7	0.11
SAMPLE 8	0.12
SAMPLE 9	0.15
SAMPLE 10	0.18

The amounts of protein have increased with the increase of vermicompost concentration.

Effect of vermicompost on peroxidases enzyme activity

Table 10: changes in peroxidases enzyme activity on Arachis hypogaea l.

Sample	Peroxidase Activity
SAMPLE 1	17.6
SAMPLE 2	8.8
SAMPLE 3	20
SAMPLE 4	13
SAMPLE 5	11.1
SAMPLE 6	10.8
SAMPLE 7	10
SAMPLE 8	9.8
SAMPLE 9	9.2
SAMPLE 10	3.5

Effect of vermicompost on catalases enzyme activity

Table 11: changes in catalases enzyme activity on Arachis hypogaea l.

Sample	Catalases Activity
SAMPLE 1	46
SAMPLE 2	26
SAMPLE 3	30
SAMPLE 4	27
SAMPLE 5	23
SAMPLE 6	19
SAMPLE 7	12
SAMPLE 8	9
SAMPLE 9	8
SAMPLE 10	2

Table 10 and Table 11 indicate that the activities of both in vivo antioxidant enzymes, namely peroxidase and Catalase decreased with the increase in the fertilizer concentration. This decrease could be due to the reduced ROS concentrations in the plants due to Vermicompost treatment.

From the results it is understood that most of the growth parameters like leaf area index (LAI) , fresh weight and dry weights of plants, germination percentage and Vigor index, root and shoot lengths, etc. indicated a regular increase as the concentration of vermicompost increased which show a promising role of vermicompost on growth of ground nut plant. There was no change in the chlorophyll, carotenoid and amino acid contents whereas there is a regular increase in protein contents. The activities of antioxidant enzymes peroxidase and catatlase decreased as the concentration of vermicompost increased. The decrease of activities of these enzymes indicate that vermicompost reduces the oxidative stress in plant by reduced production of Reactive Oxygen Species (ROS). It could be possible that as the plants grow further there could be corresponding increase in chlorophyll and carotenoid contents.

CONCLUSION:

Our experiment clearly indicate that the Vermicompost is an effective, cost saving, easily affordable manure which is biocompatible and which does not contribute to pollution, as we find in case of chemical fertilizers.

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Received on 25.02.2019 Modified on 16.03.2019

Research J. Pharm. and Tech. 2019; 12(6):2895-2901.

DOI: 10.5958/0974-360X.2019.00488.8