Assessment of selected organic manures as plant growth hormones and their impact on the growth attributes of Alium cepa Lin.

 

Perumal, 1* K, K. Praveena2, V. Stalin1 & B. Janarthanam1

 

1Shri AMM Murugappa Chetttiar Research Centre, Tharamani, Chennai 600 113, India,

email: energy1@vsnl.com

 

2 Srimad Andavan Arts & Science College, Department of Biotechnology, Bharathidasan University, Tiruchirappalli 620 024

 

 

 

Key words: Alium cepa, biodynamic compost, organic manure, plant growth regulators and vermicompost.

 

 

 

 

 

 

 

 

 

 

 

 

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*For correspondence (email: energy1@vsnl.com)

 

Abstract

 

In the current research work we have explored the presence of plant growth hormones in selected organic manures. Twelve types of organic manures such as vermicompost, NADEP compost, cow pat pit (CPP), biodynamic compost, cow horn manure (BD 500), biodynamic herbal preparations such as BD 502 to 507 were collected from Shri AMM Murugappa Chettiar Research Centre, Tharamani and Kurinji Organic Food Pvt. Ltd., Genguvarpatti Tamil nadu. Among the 12 organic manures analyzed for the enumeration of microorganisms, cow pat pit manure contained highest bacterial load (4.5x 106) and beneficial bacteria such as Rhizobium sp. (1.9x106), Azospirillium sp. (0.2x106) and Azotobacter sp . ( 0.8x106). All the 12 organic manures were analyzed for the presence) of plant growth hormones such as indole acetic acid (IAA), gibberillic acid (GA3), kinetin and abscissic acid (ABA)  found that there were appreciable differences in the content of plant growth hormones. The cow pat pit manure contained three plant growth hormones such as IAA (28.6 mg/ Kg of manure) and kinetin (7.6mg/ Kg of manure) and Gibberllic acid (23.6 mg/ Kg of manure) whereas the other manures did not contain Gibberlic acid. Biodynamic herbal preparations BD 504, BD 505 and BD 506 contained IAA, GA3 and abscissic acid (ABA). Trials under hydroponics growth condition amended with different organic manures revealed remarkable differences on root and sheath growth of onion (Alium cepa). The cow pat pit manure, cow horn manure (BD 500) and herbal preparation (BD 503) have influenced the onion root growth with a maximum number of roots (38) and rootlets (20). The roots of Alium cepa grown in various organic manures were collected and the individual cell size measured through cell squash method. The roots of onion grown in the cow pat pit manure, BD 500, and BD503 recorded with the maximum cell size of 41.25µ, 40.2µ, and 40.16µ respectively. The anatomical structure of onion root grown in CPP had 2 metaxylem whereas the others had only one metaxylem.

INTRODUCTION

Green revolution technologies involving greater use of synthetic agro chemicals such as fertilizers and pesticides with adoption of nutrient responsive high yielding varieties of crops have boosted the production output per hectare in most cases. However, this increase in production has slowed down and in many cases there are indications of decline in productivity and production. Moreover, the success of industrial agriculture and the green revolution in recent decades has often marked significant externalities affecting natural resources and human health as well as agriculture itself. Environmental and health problems associated with agriculture have been increasingly well documented. Increasing consciousness about conservation of environment as well as of health hazards caused by agro chemicals has brought a major shift in consumer preference towards food quality, particularly in the developed countries. Consumers are increasingly looking forward to organic foods that are considered safe and hazard free. Organic farming technology is often understood as a form of agriculture with use of only organic inputs for the supply of nutrients and managements of pest and diseases. The organic agriculture technology is a method of farming that aims to treat the farms as a living system which interacted with the environment to produce nourishing, vitalized food, thus benefiting mankind (Proctor, 2002).

 

Addition of organic manures like farmyard manure, crop residues, biogas slurry, crop wastes, oil cakes, earthworms and compost improved fertility of soil. The underlying principle of organic farming is the development of live humus out of dead organic materials so as to make biologically active humus as the basis of all living soil. Organic manures are important sources of plant nutrients. They maintain and restore active soil life and health. The chemical composition of organic residues which is a part of manures will influence the rate and extent of decomposition by the soil micro flora and thus potential for increasing soil organic matter.

 

Application of these soil amendments would change the rhizosphere environments by affecting porosity, aeration temperature, water holding capacity and soil microflora. These manures contain ingredients necessary for the crops such as nitrogen, phosphorus, potash, sulphur, calcium and magnesium. Soil fertility and nutrient supplying capacity of a soil can be maintained on a long-term basis only by replenishing, through addition of external inputs (manures), nutrients removed by cropping and those lost through physical, chemical and biological processes. In addition to replenishing plant nutrients with organic inputs the application of growth hormones such as Indole acetic acid (IAA), Gibberlic acid (GA3) Cytokinin and Abscisic acid (ABA) are incorporated by the farmers during various stages of crop growth. These growth hormones are commercially available at high cost and are sporadic in nature. Therefore many farmers find it difficult to source and purchase the synthetic growth hormones. In rural areas, farmers generally use cow dung in their field and also for covering the top of plant cuttings or grafting during plantation/cropping. According to Proctor application of cow pat pit (CPP) manures as tree paste in tea plantations stimulated numerous young buds and branches. The application of vermivash on sugarcane field, paddy, and vegetables increased better plant growth (Ismail, 2000). Preliminary study conducted by Perumal (2003) on organic manure indicated the presence of IAA (61 g per kg ) and GA (45 g per kg ) in cow pat pit manure obtained from Kurinji organic Foods Pvt.Ltd. Hitherto, limited scientific rational is available on how these cow dung, vermivash and cow pat pit stimulated plant growth. Therefore the present study aims on enumeration of fungi and bacteria from various organic manures, extraction, estimation of growth hormones from organic manures and application on onion (Alium cepa) root and shoot growth under hydroponics growth condition.


METHODS

Sample Collection

 

The organic manure such as Vermicompost, NADEP, Panchakavya, Biodynamic (BD) compost and cow pat pit (CPP) were obtained from Shri AMM Murugappa Chattier Research Centre, Tharamani. The biodynamic herbal preparation such as cow horn manure (BD500) and biodynamic herbal preparations (BD502-507) were obtained from Kurinji Organic Foods Pvt. Ltd, Genguvarpatti, Tamil Nadu. All these manures were brought to the laboratory and stored in a wooden box as per the procedure described by Proctor, 2002.

 

Physicochemical analysis

 The manures physicochemical properties such as pH, EC, N, P, K and organic carbon were estimated by following standard method as described by Muthuvel and Udayasoorian (1999). Lowry et al., (1951) method, determined protein content of organic manure.

 

Microbial analysis of manure

Using standard dilution plate count techniques (Waksman, 1952) the microbial analysis of organic manure was carried out. The media such as Nutrient agar (total bacteria), Azospirillium isolation agar medium for Azospirillum, Burk’s medium for Azotobacter, Yeast extract mannitol agar medium (YEMA) for Rhizobium and Rose Bengal agar medium (total fungi) were prepared and used for enumeration of total bacteria and fungi respectively. All the 12 samples were serially diluted using sterile distilled water and dilutions were prepared from 10-2 to 10-7. One ml of 10-5 to 10-6 dilutions were taken as inoculum and after inoculation the plates were incubated at 37° C for 3 days and observed periodically for enumeration of bacteria and fungi respectively. All the experimented analysis was carried out in triplicates and the average values were recorded.

 

Extraction and estimation of Plant growth hormones from manure

Extraction and estimation of growth hormones like Indole acetic acid (IAA), Gibberlic acid (GA3), kinetin and Abscisic acid in twelve organic manures were performed according to the method of Unyayar et al., 1996. The extraction and estimation procedure are shown in

 figure 1. Spectrophotometer techniques were used to determine the amount of growth hormones. Two grams of organic manure was individually weighed and crushed into fine paste by using a clean mortar and pestle by adding 60 ml of combined extract. The extract contains Methanol: Chloroform: Ammonium (2N) solution in the ratio 12:5:3 v/v/v. The combined extract is filtered and the filtrate is centrifuged at 6,000 rpm for 15 minutes at 27ºC. Combined extract filtrate was treated with 25 ml of distilled water. The chloroform phase was discarded. The water- methanol phase was evaporated. The water phase was adjusted to the extract pH of 2.5 or 7 or 11 with 1N HCl or 1N NaOH respectively and 15 ml of ethyl acetate was added at each of the three steps as mentioned above. The sample is then, hydrolyzed at 70º C for 1 hour.

 

Quantification of PGR

Hydrolyzed sample was dissolved in 2ml of methanol and optical density (O.D) was determined by using UV-visible spectrophotometer at different wavelengths. The amount of total growth hormones in the manure was determined from the standard graph prepared with different concentration (10µg to 100µg) of Indole acetic acid (IAA), Kinetin, Abscisic acid (ABA) and Gibberlic acid (GA3) obtained from Himedia India. The growth hormones from organic manure were compared with growth hormone extracted from wheat grass which contained IAA- 0.5 g/kg, Kinetin-3 g/kg, ABA- 2.62 g/kg, GA3- 4.35 g/kg).

 Flow Chart

                            Plant growth hormones                               Wavelength

                                            IAA                                                250,222nm

                                            GA3                                                    254nm

                                            ABA                                                 263 nm

                                            Kinetin                                           269nm

 

 

Effect of manure on onion growth under Hydroponics conditions

Selection of onion (Alium cepa)

Onion bulb (Alium cepa) was obtained from local market Adyar in Chennai. The individual onion bulb was brought to the laboratory and dried in sunlight for 1 day. The clean and healthy bulbs were selected weighed and used for the following study.

 

Preparation of Hydroponics

One gram of manure was added in to a glass container containing 125 ml of distilled water, stirred well, and completely dissolved. Individual manures were added and hydroponics set was used for onion growth studies. Onion bulbs were placed at the top of the container and confirm that the root initiation part of the onion bulb touched the aqueous layer. All the setup were kept at 30º C and maintained at the laboratory up to 30 days. At every 5 day interval the sheath length, root length and numbers of roots were recorded.

 

Cytological studies

Cross section of onion root

Twenty day old onion roots were collected and the roots were washed with distilled water. A cross section of root was sliced using a sharp blade, where the root tip was previously placed in between the thermacol. The cross section was stained using safranin stain for few minutes and washed with distilled water. The cross section was mounted in a clean glass slide and placed a glass cover slip over the specimen. Examined the slide under the microscope (40 x objectives). The cross sections of individual onion root were observed under white illumination and photomicrographs were taken using Minnesota camera attached with microscope.

 

Root tip squash preparation

Onion root tips were placed in the fixative (Acetic acid and alcohol). Two or three times the root was washed and all the trace of fixative removed thoroughly with distilled water. The root tips were hydrolyzed in 1N HCl at 60º C for 7 minutes. Two or three times the root is washed to remove all the traces of HCl with distilled water. The root tips were trimmed by using a blade and stained in 2% acetocarmine. The root tip in a clean glass slide was mounted and a glass cover slip placed over the specimen. The meristem was squashed with 2% acetocarmine on the slide by applying gentle pressure on the cover slip and examined under the microscope and measures the cell size by using ocular/and stage micrometer.

 

RESULTS AND DISCUSSION

According to Lixiju et al. (1998) the content of IAA and cytokinins in soil treated with different organic manure were analyzed in an apple pot trial and the observation revealed that organic manure increased the soil content of growth regulators and stimulated plant growth. Besides higher plants, fungi synthesize IAA, GA3, ABA and Zeatin. Many researchers established the presence of IAA in Dipodoscopsis uninucleata, GA3 is produced by Gibberella fujikuroi, ABA is produced by Cercospora rosicola, Cytokinin is produced by Fusarium moniliforms and F. colmorum. It has been found that IAA, GA3, ABA and zeatin can be obtained from P.chrysoporium and synthesized IAA, GA3, ABA and zeatin as primary and secondary metabolites. Recovery of IAA, GA3, ABA and zeatin were respectively 55.5 + 10%, 74.6% + 8%, 51.6 ±10% and 56.63± 6% (Unyayar et al., 1996). In the present investigation on all the 12 manures studied had revealed presence of growth hormones and had influence on onion root and sheath growth tested under laboratory conditions.

 

Cow horn manure (BD 500)

BD 500 is called cow horn manure. It is made from fresh lactating cow dung packed into cow horns, buried over the winter for fermentation in the earth. Chemically it was found that an increase of nitrate nitrogen from 0.06% to 1.7 %. i.e. about 28.31 times of original content (Pfeiffer, 1958). BD 500 is unique formulation activates regulates the soil by increasing its humus and bacterial content. BD 500 also encourages earthworm activity and allows for better nourishment of the plants (Purple Hills 2003). The important effect of BD 500 is stimulation of root growth, particularly fine hair roots. In our study the BD 500 had a pH of 7.2 and EC 0.17 %. It was found that it has the phosphorus content of 1.10 %, Potassium of 2.50 % and organic carbon of 24.50 % (Perumal and Vatsala 2002) was recorded (Table 1). BD 500 analyzed for the presence of growth hormone such as Indole acetic acid (IAA) 21.6 mg /Kg and Kinetin 2.9 mg /Kg was recorded (Table 2). The microbial analysis  shows that  4.0x106 bacteria in nutrient agar,0.2X106 Azoto bacter in Burk’s medium, 0.2x106Azospirillum, 0.8x106 fungi in Rose Bengal agar, 3.0x106 Rhizobium in YEMA medium was recorded in (Table 3).The BD 500 had a protein content of 2.48 gm / Kg of manures (Figure 2). The effect of BD500 on root /sheath growth was studied under laboratory condition. Onion treated with BD500 had 41 roots with an average length of rootlets 8.2 cm (Table 4). There were six sheaths, with an average length of 16.4 cm was recorded on day 30. The microscopic observation of onion root treated with CPP showed that cell size was 40.2μ in length and 20.3 μ in breadth (Table 5). The cross section of onion roots revealed the presence of one metaxylem, protoxylem groups in pentarch, phloem and parenchymatous cortex.  (Figure 1 b, c).

 

BD 502

BD 502 is made from yarrow blossoms (Achiella milefolium) contained in a stag’s bladder and hung over the summer. The effect BD 502 on plant is, it attracts trace element from the cosmos. Compost prepared using yarrow will provide potash and sulphur (Perumal, 2002). The analysis of the available minerals shows a decrease of potassium from 1.05% to 0.13%, an increase of calcium from 0.05% to 0.375% and phosphate remains stable at about 0.06% and organic carbon 22.6%. The major increase is observed in nitrate nitrogen from 0.07% to 2.5% (Pfeiffer, 1958). In our study the BD503 had a pH of 5.3 and EC 0.26% was recorded (Table-1). BD502 analyzed for the presence of growth hormones such as Indole acetic acid 9.7 mg /Kg and kinetin 6.9 mg /Kg was recorded (Table-2). The microbial analysis  shows that 1.2x106 bacteria in nutrient agar, 0.3x106 in Azotobaceter medium, 0.1x106 in Azospirillum medium, 0.3x105 fungi in Rose Bengal agar and 1.0x106 in Rhizobium, YEMA medium was recorded (Table-3). The BD 502 had a protein content of 0.84 gm / Kg of manures (Figure 2).

 

 

 
The effect of BD 502 on root /sheath growth was studied under laboratory condition. Onion treated with BD 502 had 23 roots with an average length of 6.5 cm (Table 4). There were three sheaths, with an average length of rootlets 7.5 cm was recorded on day 30. The microscopic observation showed that cell size was 22.6μ in length and 13.75 μ in breadth (Table 5). The cross section of onion root revealed the presence of parenchmatous cortex, one metaxylem and phloem.

 

BD 503

The preparation is made from Matricaria recutita, chamomile blossoms, contained in the bovine small intestine fermented over the winter bedded in earth. Effect of BD 503, on plant is it regulates the decomposition of plant protein and in preservation of plant nitrogen and provides calcium (Perumal, 2002). It was found that it has increase level of calcium, from 0.05% to 0.47%, nitrogen from 0.04% to 3.1%, phosphate from 0.08% to 0.75%, Potassium 1.05%, OC 27.38% (Pfeiffer, 1958) was recorded (Table 1). In our study the BD 503 had a pH of 5.7 and EC 0.29% was recorded (Table 2). BD 503 analyzed for the presence of growth hormone such as Indole acetic acid (IAA) 7.8 mg /Kg, Kinetin 3.9 mg /Kg and absence of Abscisic acid (ABA) and Gibberlic acid (GA3) was recorded (Table 2). The microbial analysis shows that 3.9x106 bacteria in nutrient agar, 1.2x106 Azotobacter in Burk’s medium, 0.8x106 fungi in Rose Bengal medium and 2.0x106 Rhizobium in YEMA medium (Table-3). The BD 503 had a protein content of 0.74 gm / Kg of manure (Figure 2). The effect of BD503 on root /shoot (sheath) growth was studied under laboratory condition. Onion treated with BD503 had 22 roots with an average length of rootlets 6.8 cm (Table 4). There were six sheaths, with an average length of 16.4 cm was recorded on day 30. The microscopic observation showed that cell size was 40.16μ in length and 21.4 μ in breadth (Table 5). The cross section of onion root revealed the presence one metaxylem, phloem and parenchymatous cortex (Figure1 f, g).

 

BD 504

BD 504 is made from stinging nettle (Urtica dioreca) leaves and stem buried about 2 feet deep in humus earth, separated from the soil by a thin layer of peat mosses. The effect on plant is that, it mediates iron, which is important for plant chlorophyll and helps strengthen the plants. It also provide certain quantity of magnesium and sulphur, and because of the formic acid presence, it very good for humus formation. (Perumal,, 2002). It has been found that, BD 504 has boron of 0.1%, copper of 0.01%, manganese of 0.001 to 0.01%, calcium between 1 to 10 %, magnesium 5%, Nitrogen 0.5%, potassium 1.05%, phosphorus 0.05% and nitrate content is ½ that of the green leaves (Pfeiffer, 1958). In our study, BD 504 had a pH of 8.1 and electrical conductivity 0.23 % and organic carbon 28.32 % was recorded (Table-1). BD 504 analyzed for the presence of growth hormone such as IAA-8.1 mg /Kg, Kinetin-5.6 mg /Kg, ABA-18.5 mg /Kg and absence of Gibberlic acid GA3 was recorded (Table-2). The microbial analysis shows that 3.5x106 bacteria in nutrient agar, 0.5x106 Azotobacter in Burk’s medium, 0.2x106 fungi in Rose Bengal medium, 2.9x106 Rhizobium in YEMA medium (Table-3). The BD 504 had a protein content of 0.94 gm / Kg of manures (Figure 2). The effect of BD 504 on root /shoot (sheath) growth was studied under laboratory conditions. Onion treated with BD 504 had 27 roots with an average length of rootlets 6. 2 cm (Table 4). There were four sheaths, with an average length of 6.3 cm was recorded on day 30. The microscopic observation showed that cell size was 39.2μ in length and 15.5 μ in breadth. The cross section of onion root revealed the presence of one metaxylem, phloem and the parenchmatous cortex.

 

BD 505

BD 505 is prepared from oak bark (Quercus incana), buried together with skull of ruminants. The effect of BD 505 on plants is that, the oak bark provides a connection between the living and dead. This is very effective against plant disease (Perumal, 2002). It has been found that, BD 505 has high calcium content, Nitrogen more than 10 %, potassium content from 0.05 to 0.013%, phosphate from 0.01 to 0.03%, alum from 0.1 to above 1%, magnesium from 0.1 to 1 %, nitrate of 0.073% (Pfeiffer, 1958). In our study BD 505 had a pH of 7.9 and EC 0.12% and organic carbon 16.85 % was recorded (Table 1). BD 504 analysed for the presence of growth hormones such as Indole acetic acid IAA 8.9 mg /Kg, Kinetin-0.03 mg /Kg Abscisic acid ABA 18.6 mg /Kg and absence of Gibberlic acid (GA3) was recorded (Table 2). The microbial analysis shows that, BD 505 has 0.8x106 bacteria in nutrient agar, 1.3x106 Azotobacter in Burk’s medium 0.2x106 Azospirillum, 0.3x106 fungi in Rose Bengal agar and 0.7x106 Rhizobium in YEMA medium  (Table 3). The BD 505 had a protein content of 2.0 gm / Kg of manure (Figure 2). The effect of BD 505 on root growth was studied under laboratory condition. Onion treated with BD 505 had 3 roots with an average length of rootlets 6.5 cm was recorded on day 30 (Table 4). The microscopic observation showed that cell size was 21.05μ in length and 12.5 μ in breadth (Table 5). The cross section of onion root revealed the presence of one metaxylem, protoxlem groups in hexarch, phloem and parenchymatous cortex.

 

BD 506

BD 506 is prepared from dandelion (Taraniam officinalis) contained within the bovine mesentery. Due to the sensitive nature of the flower, Steiner suggests that when added it provides silica for the plant development (Perumal, 2002). It has been found that, BD 506 has an increased content of calcium 10%, magnesium (10%), nitrate nitrogen from (0.336 -3.38%), phosphate from (0.46 to 0.72%), alum from (0.1 to 1%), boron (1%), barium (0.1%), sodium (1%), potassium from (1.25 to 0.27 %) etc (Pfeiffer 1958). In our study, BD 506 had a pH of (6.1) and EC of (29%) and organic carbon 11.42% was recorded (Table-1). BD 506 analyzed for the presence of growth hormone such as Indole acetic acid (IAA) 3.1 mg /Kg , Kinetin 5.7 mg /Kg, Abscecic acid (ABA18.6 mg /Kg and absence of Gibberlic acid (Table-2). The microbial analysis  shows that , 3.7x106 bacteria in nutrient agar, 0.3x106 Azotobacter in Burk’s medium, 0.6x106 fungi in Rose Bengal agar and 2.0x106 Rhizobium in YEMA medium were recorded (Table-3). The BD 506 had a protein content of 0.32 gm / Kg of manure (Figure 2) the effect of BD500 on root growth was studied under laboratory conditions.  Onion treated with BD 506 had 23 roots with an average length of rootlets 2.5 cm was recorded on day 30 (Table 4) The microscopic observation showed that cell size was 27 u in length and 12.3 u in breath (Table 5).

 

BD 507

This preparation is the pressed out extract from valerian blossoms (Valeriana officinalis). Valerian stimulates the phosphorus process. It may thus provide fast protection for flowering fruit trees, and also promotes the development of the flowers and fruit. When it is sprayed on to compost heaps it provides protective enveloping, warmth and is also beneficial to earthworms. It has found that BD 507 has nitrate nitrogen of 0.145%, magnesium 0.1 to 10 % etc. (Pfeiffer, 1958). In our study BD 507 had a pH of 6.8, EC 0.01%, potassium 1.23% and organic carbon 26.90% (Table-1). BD 507 analysed for the presence of growth hormones such as IAA 6.3 mg /Kg Kinetin 5.8 mg /Kg and absence of ABA and GA3 was recorded (Table-2). The microbial analysis shows that 3.0x106 bacteria in nutrient agar, 0.3x106 Azotobater in Burk’s medium, 0.1x106 Azospirillum, 0.3x106 fungi in Rose Bengal agar and 1.0x106 Rhizobium in YEMA medium (Table-3). The BD 507 had a protein content of 0.7 gm / Kg of manure (Figure 2). The effect of BD 506 on root growth was studied under laboratory condition. Onion treated with BD 507 had 26 roots with an average length of rootlets 7.1 cm was recorded on day 30 (Table 4). The microscopic observation showed that cell size was 27μ in length and 12.3 μ in breadth (Table 5). The cross section of onion roots revealed the presence of one metaxylum, protoxlem groups in hexarch, phloem and parenchmatous cortex.

 

BD compost

The biodynamic (BD) compost was made from locally available biomass. Blending with alternative layers of carbonaceous and nitrogenous materials built the heap. 1 gram of each biodynamic herbal preparation (BD 502-507) was inserted in to various places at the heap. Moisture content is maintained and repeated turning was done at 60 days interval for up to 5 months. The quantity obtained from 5 times of biodynamic compost is 320 kg. They were utilized for the farm model and kitchen gardening activities in the lab premises. It was also used for nursery preparation of carrot and other seedlings (Perumal and Vatsala, 2002). In our study BD compost had a pH of 7.3, EC 0.03 %, nitrogen 0.5%, phosphorus 0.03%, potassium 0.75% and organic carbon 27.45% was recorded. (Table 1). BD compost analyzed for the presence of growth hormones such as IAA 6.1 mg /Kg, Kinetin 0.3 mg /Kg and absence of ABA and GA3 was recorded (Table 2). The microbial analysis shows that 2.7x106   bacteria in nutrient agar, 2.4x106 Azotobacter in Burk’s medium, 0.1x106 Azospirillum,         0. 6x106 fungi in Rose Bengal agar, 3.0x106 Rhizobium in YEMA medium (Table 3). The BD compost had a protein content of 0.58 gm / Kg of manure (Figure 2). The effect of BD compost on root growth was studied under laboratory condition. Onion treated with BD compost had one root with an average length of rootlets 6 cm was recorded on day 30 (Table 4). The microscopic observation showed that cell size was 23.5μ in length and 14.6 μ in breadth (Table 5). The cross section of onion roots revealed the presence of one metaxylum, protoxlem groups in hexarch, phloem and parenchmatous cortex.

 

Cow Pat Pit (CPP)

Manure from a cow pat pit is a useful vehicle or medium for spreading the influence of herbal preparation such as BD 502-507 in the compost. The lactating cow dung, crushed eggshells and rock phosphate are mixed thoroughly and added with herbal preparation in a defined pit. CPP provides nutrient and stimulate plant growth by enhancing microbial population and protecting against fungal disease (Perumal, 2002). CPP manures as tree paste stimulated numerous young buds in tea plantation (Peter Proctor, 2003).It has been found that, the CPP has nitrogen content of 0.092%, phosphorus of 6.84%, potassium of 4.68 and organic carbon of 16.4% (Perumal, 2002). In our study CPP had a pH of 8.6 and EC 0.11 % (Table 1). CPP analyzed for the presence of growth hormones such as IAA 28.6 mg /Kg, Kinetin 7.6 mg /Kg , GA3 23.6 mg /Kg and absence of ABA was recorded (Table 2). The microbial analysis shows that 4.8x106bacteria in nutrient agar, 0.8x106 Azotobacter in Burk’s medium, 0.2x106 Azospirillum, 1.1x106 fungi in Rose Bengal agar and 1.9x106 Rhizobium in YEMA medium was recorded (Table-3). The CPP had a protein content of 4.96 gm / Kg of manure (Figure 2). The effect of CPP on root/sheath growth was studied under laboratory condition. Onion treated with CPP had 30 roots with an average length of rootlets 7.0 cm (Table 4).  There were 23 sheaths with an average length of 18.5 cm was recorded on day 30 (Table 4).  The microscopic observation showed that cell size was 41.25μ in length and 20.5 μ in breadth (Table 5). The cross section of onion roots revealed the presence of two metaxylum, protoxlem groups in hexarch phloem and parenchmatous cortex (Figure. 2 d, e).

 

Vermicompost

Vermicompost play an important role in the organic farming. It is one of the important methods of compost preparation. Vermicomposting utilizes earthworms for the purpose of producing value added manure. Moreover earthworms ingest litter, dung and other organic matter and grind it in to fine particles, there by increasing the surface area and promoting faster decomposition. The material passes through the body of the earthworm to produce vermicast. Soil with vermicasts has roughly 100 times more bacteria than soil without worms. Moreover plant growth promoting substances have been reported to be present in vermicast (Ismail, 2002). It has been found that vermicompost has nitrogen content of 2.12% phosphorus of 2.01%, potassium of 2.27% and organic carbon of 27.38% (Perumal, 2002).In our study, vermicompost had a pH of 6.6 and EC 0.04% was recorded (Table 1) .Vermicompost analyzed for the presence of growth hormones such as IAA8.2 mg /Kg, kinetin 5.7 mg /Kg and absence of ABA and GA3 was recorded (Table 2). The microbial analysis shows that 2.8x106 in nutrient agar, 2.6x106 Azotobacter in Burk’s medium, 0.9x106 Azospirillum, 0. 3x106 fungi in Rose Bengal agar and 1.6x106 Rhizobium in YEMA medium was recorded (Table 3). The Vermicompost had a protein content of 0.92 gm / Kg of manure (Figure 2). The effect of vermicompost on root/sheath growth was studied under laboratory condition. Onion treated with vermicompost had 28 roots with an average length of 7.6 cm was recorded on day 30 (Table 4). The microscopic observation showed that cell size was 23μ in length and 11.5 μ in breadth (Table 5). The cross section of onion roots revealed the presence of one metaxylem, protoxlem groups in hexarch, phloem and parenchmatous cortex.

 

NADEP compost

NADEP preparation is made from agricultural waste, cow dung dried soil, water etc. Decomposition processes follows through aerobic method and it requires about 90 to 120 days for obtaining the finished products. The main advantage is it improves the physical, chemical and biological condition of the soil. It also supplies the major and micronutrients to the plant growth. It was found that the NADEP manure has nitrogen content of 1.38%, phosphorus of 0.92%, potassium 2.50% and organic carbon of 30.35% (Perumal, 2002). In our study the NADEP had a pH of 6.7 and EC 0.05% was recorded (Table-1). NADEP analyzed for the presence of growth hormones such as IAA 21.4 mg /Kg , kinetin 5.4 mg /Kg and absence of ABA and GA3 was recorded (Table-2). The microbial analysis shows that 3.2x106 bacteria in nutrient agar, 0.5x106 Azotobacter in Burk’s medium, 0.3x106 Azospirillum, 0.4x106 fungi in Rose Bengal medium and 1.9x106 Rhizobium in YEMA medium was recorded (Table-3). The NADEP had a protein content of 4.4 gm / Kg of manure (Figure 2). The effect of NADEP on root/sheath growth was studied under laboratory condition. Onion treated with NADEP had 37 roots with an average length of 10.5 cm recorded on day 30 (Table 4). The microscopic observation showed that cell size was 27.15μ in length and 11.35 μ in breadth (Table 5). The cross section of onion roots revealed the presence of one metaxylem, protoxlem groups in hexarch, phloem and parenchmatous cortex.

 

Panchakavya

Panchakavya is prepared by using five products from cow dung, urine, milk, curd and ghee. It contains bacteria, fungi, protein, carbohydrates, fats and some growth promoting factors. In our study panchakavya had a pH of 3.7% and EC 0.40% nitrogen 1.28 %, phosphorus 0.72%, potassium 2.23 % and organic carbon 17.45% was recorded. (Table 1). Panchakavya analyzed for the presence of growth hormones such as IAA10.4 mg /Kg, kinetin 3.9 mg /Kg and absence of ABA was recorded (Table 2). The microbial analysis shows  that 3.8x106 bacteria in nutrient agar, 0.8x106 Azotobacter in Burk’s medium, 0.2x106 Azospirillum, 0.6x106fungi in rose bengal agar and 2.0x106 Rhizobium in YEMA medium was recorded (Table 3). The Panchakavya had a protein content of 4.8 gm / Kg of manure (Figure 2). The effect of panchakavya on root/sheath growth was studied under laboratory condition. Onion treated had 3 sheaths with an average length of rootlets of 3.5 cm recorded on day 30 (Table 4). The microscopic observation showed that cell size was 24.32 μ in length and 13.3 μ in breadth (Table 5). The cross section of onion roots revealed the presence of one metaxylum protoxlem groups in hexarch, phloem and parenchmatous cortex.

 

As the organic manures both directly and indirectly by benefit the soil and plants, their conservation and efficient use in agriculture assumed importance as essential components of nature friendly agriculture. Because of the need for increasing agricultural output in the developing countries it is not advisable to dispense with the use of chemical fertilizer abruptly, this strategy in the course of time can cause a significant reduction in the consumption of chemical fertilizer in agriculture without depressing the production potential of soil resource. It has been proved all over India, under ICAR multi-location trials, that the use of organic farming is essential not only for better utilization of applied fertilizer, but also to make the soil protective and agricultural sustainable. In this investigation the extraction and estimation of growth hormone like IAA, Kinetin, and ABA were recorded in 12 organic manures for the first time in India. These organic manures tested in hydroponics revealed better effect on the root- sheath growth of onion. Therefore, these research findings throw light on the use of organic manure as source of growth hormones. The farmers can directly prepare and use these growth hormone formulations like CPP and Cow horn manure, which are cost effective and locally available at right time for better crop growth.

 

ACKNOWLEDGEMENTS

We are grateful to Prof. P. N. Raju, The Acting Director, Shri AMM Murugappa Chettiar Research Centre, Tharamani, Chennai for providing laboratory facilities. We thank Mr.Peter Proctor for the comments on the manuscript. We also thank Mr. Jeyakaran, Kurinji organic Foods Pvt.Ltd. for providing various biodynamic herbal preparations used in this study.

 

 

 

Table 1. Physico chemical properties of organic manures

 

Treatment

pH

EC

(%)

Nitrogen (N) %

Phosphorus

(P) %

Potassium

(k) %

Organic Carbon

(OC) %

BD 500

7.2

0.17

1.62

1.10

2.50

24.50

BD 502

5.3

0.26

0.07

0.05

1.05

    22.6

BD 503

5.7

0.29

0.04

0.08

2.27

27.38

BD 504

8.1

0.23

     0.5

0.05

1.05

28.32

BD 505

7.9

0.12

0.04

0.01

0.05

16.85

BD 506

6.1

0.29

0.33

0.46

1.25

11.42

BD 507

6.8

0.01

0.14

0.01

1.23

26.90

BD Compost

7.3

0.03

     0.5

0.03

0.75

27.45

CPP

8.6

0.11

2.09

6.84

4.68

    16.4

Vermicompost

6.6

0.04

2.12

2.01

2.27

27.38

NADEP

3.7

0.05

1.38

0.92

2.50

30.35

Panchakavya

 

3.7

0.40

1.28

0.72

2.23

17.45

 

Note: EC-Electrical conductivity

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table 2. Quantification Plant growth Hormone from organic manure

 

Treatment

IAA

mg/ Kg of manures

Kinetin

 mg/Kg of manures

ABA

mg/ Kg of manures

GA3

mg/ Kg of manures

BD 500

21.6

2.9

-

-

BD 502

9.7

6.9

-

-

BD 503

7.8

3.9

-

-

BD 504

8.1

5.6

18.5

-

BD 505

8.9

0.03

18.6

-

BD 506

3.1

5.7

18.6

-

BD 507 (2ml)

6.3

5.8

-

-

BD Compost

6.1

0.3

-

-

CPP

28.6

7.6

-

23.6

Vermicompost

8.2

5.7

-

-

NADEP

21.4

5.4

-

-

Panchakavya

 ( 2ml)

10.4

3.9

-

-

Wheat grass

0.5

3

2.6

4.4

 

Note: - No PGR compound

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table 3.  Microbial analysis of organic manures

 

 

Treatment

Nutrient agar x106

Azotobacter

x106

Azospirillum

x106

Fungi

x106

Rhizobium

x106

BD 500

4.0

0.2

0.2

0.8

3.0

BD 502

1.2

0.3

0.1

0.3

1.0

BD 503

3.9

1.2

0.1

0.8

2.0

BD 504

3.5

0.5

-

0.2

2.9

BD 505

8

1.3

0.2

0.3

0.7

BD 506

3.7

0.8

-

0.6

2.0

BD 507

3.0

0.3

0.1

0.3

1.0

BD Compost

2.7

2.4

0.1

0.6

3.0

CPP

4.8

0.8

0.2

1.1

1.9

Vermicompost

2.8

2.6

0.9

0.2

1.6

NADEP

3.2

0.5

0.3

0.4

1.9

Panchakavya

3.8

0.8

0.2

0.5

2.0

 

Note: - No microbial count

 

 

 

 

 

 

 

 

 

 

 

 

Table 4. Effect of different organic manures on growth of (Alium cepa).

Treatment

Root(30 th day)

Sheath(30 th Day)

Number of roots

Length of roots (Average3 times)

Number of Sheath

Length of Sheath

BD 500

41

8.2

6

16.4

 

BD 502

23

6.5

3

7.5

 

BD 503

22

6.8

-

-

 

BD 504

27

6.2

4

6.25

 

BD 505

3

6.5

-

-

 

BD 506

23

2.5

-

-

 

BD 507

26

7.1

-

-

 

BD compost

1

6

-

-

 

CPP(MCRC)

30

7

23

18.5

 

Vermicompost

28

7.6

-

-

 

NADEP

37

10.5

-

-

Panchakavya

12

9

3

3.5

 

Note: - No growth

 

 

 

 

 

 

 

Table. 5. Effect of organic manures on onion root (cell size-root tip squash method)

 

 

Cell size ( Onion)

 

Treatment

Length in(μ)

Breadth (μ)

BD 500

40.2

20.3

BD 502

22.6

13.75

BD 503

40.16

21.4

BD 504

39.2

15.5

BD 505

21.05

12.5

BD 506

27

12.3

BD 507

37.7

14.5

BD Compost

23.5

14.6

CPP

41.5

20.5

Vermicompost

23

11.5

NADEP

27.15

11.35

Panchakavya

24.32

13.32

Control

10

6

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Figure. 2. Estimation of protein in different organic manures

 

 

 

 

 

 

 

              

 

      

 

      a. BD 500     e. BD 505            i. CPP

      b. BD 502     f. BD 506            j. Vermicompost

      c. BD 503      g. BD 507           k. NADEP

      d. BD 504      h. BD compost   l. Panchakavya

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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