Title of Invention

A SYNERGISTIC COMPOSITION USEFUL IN PROMOTING PLANT GROWTH AND A PROCESS FOR THE PREPARATION THEREOF

Abstract The present invention relates to a synergistic composition useful as plant and soil health enhancer, comprising urine, neem and garlic, individually or in all possible combinations, with the treatment showing it has the ability to stimulate accumulation of nutrients in the plant biomass, proliferation of plant growth promoting, phosphate solubilizing, abiotic stress tolerant and antagonists towards plant pathogenic fungi, control phytopathogenic fungi in the rhizosphere of plants, and enhances the total phenolic contents of the plants. The said composition comprises bovine urine, crushed neem leaves and crushed garlic bulbs along with one or more carriers.
Full Text A SYNERGISTIC FERMENTED PLANT GROWTH PROMOTING, BIOCONTROL
COMPOSITION
Field of the present invention
The present invention relates to a synergistic composition useful as plant and soil
health enhancer using cow urine and application thereof for promoting plant growth
and controlling plant pathogenic fungi, said composition comprising urine, neem and
garlic, individually or in all possible combinations, with the treatment showing
stimulation of the accumulation of nutrients in the plant biomass, proliferation of
plant growth promoting, phosphate solubilizing, abiotic stress tolerant and antagonists
towards plant pathogenic fungi in the rhizosphere of plants, and enhances the total
phenolic contents of the plants; further, a method of producing said composition
thereof.
Background and prior art of the present invention
Farming is the oldest wealth-creating business known to man. Current scientific
strategies to maintain and improve yields in support of high-input agriculture place
great emphasis on 'fail-safe' techniques for each component of the production
sequence with little consideration of the integration of these components in a holistic,
systems approach. Research for sustainable agricultural practices requires a far greater
emphasis on such an approach than now is fashionable, despite all the rhetoric given
politically to sustainability.
The populations of the world's poorest countries have been growing rapidly,
increasing the demand for food. At the same time environmental degradation - both
natural and man made - has reduced the ability of farmers to grow food in many
areas. A lot has been written about the significant contribution due to "Green
Revolution" and correctly so, especially considering our failure to control
unsustainable population growth. Thanks due to the high yielding varieties we are still
self-sufficient in rice and wheat, but for how long? Hardly any one argues that
modern agriculture is sustainable. Besides, high input agriculture is increasingly
recognized as an environment degrading and not profitable. We now recognize that
technical progress may have social and environmental costs we cannot pay. People
are now seriously concerned with the protection of the environment and even more
about safeguarding their health. As now people realize that by consuming the standard
agriculture based food products they are constantly taking in small quantities of
poison of various kinds and much of this comes from the chemical pesticides that are
used to produce food crops.
Modern farming requires large inputs of chemical fertilizer and stimulants to increase
yields from hybrids. However for poor rural marginal farmers the use of chemical
fertilizers and pesticides have made agriculture very expensive and to maintain yields
in deteriorating soils increasing doses of modern chemical inputs have had to be used.
The time has now come to consider alternative means of sustaining our agriculture
and to protect the farmer from low prices, high indebtedness and to ensure that
production incentives remain. For small farmers, organic farming is most suitable as
considerable vertical integration is possible and appreciable cost savings could be
achieved through the recycling of waste and other materials that are available within
the system.
A considerable amount of literature is available on the practice of organic farming.
Where organic farming is practiced, the farmer will use natural processes to enhance
productivity, maintain the nutritive status of the soil to be less dependent on external
resources and to keep his costs down. This will strengthen his social and financial
position in the society. Organic farming uses natural materials which are the by
products of the farm and are environmentally safe, it enhances the nutritive qualities
of the soil and it nurtures the organisms in the soils, which are generally destroyed by
the use of chemical manures and pesticides, and significantly reduces cost. Therefore
at this juncture further work on the development of agriculture biotechnology
products based on cow offers immense potential as viable alternative for sustainable
agriculture.
We have observed serendipitously that when cow urine is applied to seedlings of
plants, it enhances overall growth of the plant and protects plants from plant
pathogenic fungi. Experiments were undertaken to investigate the significance of
these observations. According to Hindu mythology as well as the Indian traditional
agricultural practices Vrikshayurveda of Surapala, an ancient Sanskrit text on the
science of plant life describes the use of milk in changing the flower color and
enhancing fruit taste [N. Sadhale (1996) Surapala's Vrikshayurveda (translated by N.
Sadhale) Secunderabad, India: Asian Agri-History Foundation]. Panchgavya, a
mixture of five cow products namely, dung, urine, milk, curd and ghee (clarified
butter) is used in human medicine, to improve soil health and to protect plants from
diseases [S. N. Singh (1971) Krishi-Parashar (translated by S. N. Singh). Varanasi,
India: Jai Bharat Press]. Systematic collection and use of urine for fertilization
purposes only dates back approximately one century. More than 90% of the total N
content in urine is NH4
+-N. Also the K is predominantly present in inorganic form.
This means that urine is comparable to commercial inorganic N and K fertilizer. The
average K content in cattle urine is 0.7%, and when urine is applied to old grass sods
strong effects are often seen - effects that have often been confused with an N effect -
although it is primarily a K response.
Plants have remained central to every civilization as the primary source of life, due to
their numerous applications in daily life. Plants are composed of chemical substances
of which some are not directly beneficial for the growth and development of the
organism. These secondary compounds have usually been regarded as a part of the
plants' defense against plant-feeding insects and other herbivores [G. A. Rosenthal
and D. H. Janzen (eds.) 1979 Herbivores: their interaction with secondary plant
metabolites. Academic Press, New York]. The pesticidal properties of many plants
have been known for a long time and natural pesticides based on plant extracts such
as rotenone, nicotine and pyrethrum have been commonly used in pest control.
Jacobson has reviewed literature on pesticides from more than 3000 plant species [M.
Jacobson and D. G. Crosby (eds.) 1971. Naturally occurring insecticides. Dekker Inc.
New York].
Neem (Azadirachta indicd) is so far the most promising example of plants currently
used for pest control. Neem has remained for numerous purposes in Indian society
and is known since ancient times in Sanskrit as "Arishta" meaning health bestower. A
summary of how neem products are used as bio-pesticides, the mode of action, effect
on pests and natural enemies has been prepared by Schmutterer [H. Schmutterer
(1990) Annual Review of Entomology 35: 271-297]. Many agrochemical,
therapeutic, and medicinal uses of neem are known [U.P. Singh and D. P. Singh
(2002) Journal of Herbal Pharamcotherapy, 2: 13-28]. Disease caused by various
microorganisms such as fungi, bacteria, and viruses not only damage the plant as a
whole but also severely affect quality of the crop. A number of physiological and
biochemical alterations in the plants have been reported due to infection of the fungi,
bacteria, and viruses [H. Schmutterer. Neem products for integrated pest management.
In The Neem Tree: Source of unique natural products for integrated pest management,
medicine and other purposes (Schmutterer, H., Ed.). VCH Verlagsgesellschaft,
Weinheim, Germany. 1997; pp. 367-477]. Furthermore, control measures adopted to
combat plant diseases have serious public concern because of the indiscriminate use
of the synthetic pesticides. This has resulted in the intensive search to find the
alternative methods for disease control and use of plant products in controlling plant
pathogens could be a viable alternative [U. P. Singh and B. Prithiviraj (1997)
Physiological and Molecular Plant Pathology 51: 181-194]. Aqueous extracts of
various parts of neem, e.g., leaf, bark, seed, pulp and inflorescence have been used
successfully in vitro to inhibit the growth of various plant pathogenic fungi [U. P.
Singh, R .B. Singh, and H. B. Singh (1980) Mycologia 72: 1077-1093; U. P. Singh
and H. B. Singh (1981) Australian Journal of Plant Pathology 10: 66-67].
Like neem, the antibacterial and antifungal properties of garlic (Allium sativum L)
against human and plant pathogens are also well known [U. P. Singh, B. Prithviraj, B.
K. Sarma, M. Singh and A. B. Ray (2001) Indian Journal of Experimental Biology 39:
310-322]. Oil of garlic in natural and synthetic forms has been reported to suppress
the activity of many air and soil-borne fungi [N. B. K. Murthy and S. V. Amonkar
(1973) Indian Journal of Experimental Biology 12: 208-209]. Several other workers
have observed the antimicrobial activity of extracts of garlic [M. R. Tansey and J. A.
Appleton (1975) Mycologia 67: 409-413; H.B. Singh and U.P. Singh (1981)
Australian Journal of Plant Pathology 10: 66-67]. Singh et al. [U. P. Singh, K. K.
Pathak, M. N. Khare and R. B. Singh (1979) Mycologia 71: 556-564] have shown that
even garlic-leaf extract significantly reduced the growth of Sclerotinia sclerotiorum
and Fusarium oxysporum f. sp. ciceri. Padwick causing wilts in gram (Cicer
arietinum L.). Inhibition of growth and sclerotium formation in Rhizoctonia solani by
garlic oil has also been reported [H. B. Singh and U. P. Singh (1980) Mycologia 72:
1022-1025].
Improving soil fertility is one of the most common tactics to increase agricultural and
forest production. Soil organisms, especially bacteria have a key role in determining
the rate of organic matter decomposition and thereby nutrient mineralization. These
processes determine the rate of nutrient supply to primary producers, largely
determining the rate of biomass production and other fundamental ecosystem
processes like interactions among different functional groups of organisms that
constitute ecosystems [J. D. Bever, K. M. Westover and J. Antonovics (1997) Journal
of Ecology 85: 561-573]. Therefore, elucidation of the mechanisms that determine
species composition in plant communities is important. Rhizobacteria, once
considered passive bystanders of the root environment, are now known to affect plant
health, development, and environmental adaptation, both beneficially and
detrimentally, and the importance of these bacteria in agriculture is expected to grow
[D. J. O'Sullivan and F. O'Gara (1992) Microbiology Review 56: 662-676.; R. J. Cook
(2000) Annual Review of Phytopathology 38: 95-116]. A variety of mechanisms have
been identified as being responsible for such plant growth promoting activity. For
example, certain microorganisms indirectly promote plant growth by inhibiting the
growth of deleterious microorganisms; or directly enhance plant growth by producing
growth hormones; and/or by assisting in the uptake of nutrients by the crops, e.g.,
phosphorus (?) [C. S. Nautiyal et al., FEMS Microbiology Letters, Volume 182, pp.
291-296 (2000)].
Plant disease suppression mechanisms involved includes antibiotic and siderophoremediated
suppression, and successful root colonization. A clear relationship has been
established between the suppression of soil-borne diseases by bacteria and their
densities in the rhizosphere [C. T. Bull, D. M. Weller and L. S. Thomashow (1991)
Phytopathology 81: 954-959; B. J. Lugtenberg, L. Dekkers, L. and G. V. Bloemberg
(2001) Annual Review of Phytopathology 39: 461-490]. We have observed that the
effectiveness of plant growth-promoting rhizobacteria (PGPR) strains in controlling
soil-borne plant pathogens is generally related to their efficiency of root colonization
[C. S. Nautiyal (1997) Current Microbiology 33: 1-6; C. S. Nautiyal (1997) Current
Microbiology 35: 52-58; C. S. Nautiyal (1997) FEMS Microbiology Ecology 23: 145-
158; C. S. Nautiyal (2002) U. S. patent 6495362].
The ecology of rhizosphere competent bacteria is not yet well enough understood to
predict the behavior and efficacy of PGPRs in phytosphere (leaf, stem, rhizosphere,
and endorhizosphere) colonization and of the existence of crop specificity [C. S.
Nautiyal (2000) In Biocontrol potential and its exploitation in sustainable agriculture.
Edited by R.K. Upadhyay, K.G. Mukerji, and B.P. Chamola. Kluwer
Academic/Plenum Publishers, New York. pp. 9-23]. Therefore, these findings suggest
that rhizosphere microbial population is an important indicator of plant and soil health
[C. S. Nautiyal, J. K Johri and H. B. Singh. Canadian Journal of Microbiology 48:
588-601 (2002)].
Microbial population studies can provide valuable information concerning the impact
of introduced seed or soil treatment on indigenous microbial populations. Ideally,
microbial population studies should be linked to broader aspects of ecosystem
functioning, such as effects on plant growth, plant health, and nutrient cycling [M. N.
Schroth and J. G. Hancock (1981) Annual Review of Phytopathology 35: 453-476; C.
S. Nautiyal, J. K Johri and H. B. Singh. Canadian Journal of Microbiology 48: 588-
601 (2002)]. Phenols have known to occur in all plants investigated so far. Some of
them occur constitutively while others are formed in response to pathogen ingress and
associated as part of an active defense response in the host [R. L. Nicholson and R.
Hammerschmidt (1992) Annual Review of Phytopathology 30: 369-389]. There are
also reports on a sudden increase in phenolic concentrations following inoculation
with non-pathogenic organisms to the plants.
Direct reduction in fungal growth due to changes in phenolics in the tomato in
response to inoculation with Verticillium albo-atrum is available in literature [M. A.
Bernards and B, E. Ellis (1989) Journal of Plant Physiology 135: 21-26]. Seed
bacterization with rhi/obacteria results in greater accumulation of phenolic
compounds or mediated induced systemic resistance (ISR) in hosts offer a practical
way of immunizing plants against pathogen ingress [G. Wei, J. W. Klopper and S.
Tazun (1991) Phytopathology 81: 1508-1512] Recently we have reported
rhizobacteria elicited alterations in phenolics of chickpea infected by Sclerotium
rolfsii [B. K. Sarma, D. P. Singh, S. Merita, H. B. Singh and U. P. Singh (2002)
Journal of Phytopathology 150: 277-282].
Recently, effectiveness of cow's milk against zucchini squash (Cucurbita pepo)
powdery mildew (Sphaerotheca fulgined) has been demonstrated in greenhouse
conditions [W. Bettiol (1999) Crop Protection 18:489-492]. Cow milk may have more
than one mode of action in controlling zucchini squash powdery mildew. Fresh milk
may have a direct effect against S. fulginea due to its germicidal properties [A. J.
Salle (1954) Fundamental principles of bacteriology. New York: McGraw-Hill]. Milk
contains several salts and amino acids. These substances have been shown to be
effective in controlling powdery mildew and other diseases [A. J. Salle (1954)
Fundamental principles of bacteriology. New York: McGraw-Hill]. Several authors
have shown that sodium bicarbonate, oxalate, dibasic or tribasic potassium phosphate,
and other salts and amino acids have been efficient in the induction of systematic
resistance [A. J. Salle (1954) Fundamental principles of bacteriology. New York:
McGraw-Hill: van Andel (1966) Annual Review of Phytopathology 4;349-368; M.
Reuveni, V. Agapov, R. Reuveni (1995) Plant Pathology 44:31-39].
India is one of the few countries in world, which has contributed richly to the
International livestock gene pool and improvement of animal population in world.
Cattle and buffalo contribute nearly 15% of the gross national income. The country
possesses 23% of world bovine population. Sahiwal is one of the most popular breeds
of cow of the subcontinent. It has been exported to Sri Lanka, Kenya and many
countries in Latin America and West Indies where a new breed called Jamaica Hope
has been evolved out of Sahiwal and Jersey crossbreeds [P.N. Bhat, Handbook of
Animal Husbandry, Directorate of Publication and Information on Agriculture. Krishi
Anusandhan Bhawan, Pusa, New Delhi, India (1997)]. Thus the traditional
information about the use of cow urine an important bio matter which can be used
methodically to get better result in controlling plant pathogenic fungi and promoting
plant growth should not be ignored.
While work on use of urine for promoting plant growth and controlling plant
pathogenic fungi has been conducted in past there has been no clear indication
heretofore that any detailed study has been conducted to demonstrate that urine from
cow might act as stimulator of the accumulation of nutrients in the plant biomass,
proliferation of plant growth promoting, phosphate solubilizing, abiotic stress tolerant
and antagonists towards plant pathogenic fungi in the rhizosphere of plants, and
enhances the total phenolic contents of the plants, per se. Nevertheless, a cow urinemediated
promotion of plant growth and controlling plant pathogenic fungi, if one
were discovered, could find immediate application, e.g., in soils affected by
phytopathogens, poor nutrient availability in a desired improvement in crop
development. We have found by direct comparison on a variety of plant types that the
unique combination of selected plants with cow urine is effective in the enhancement
of plant growth and soil health.
The present invention relates to usage of urine from cow which acts as plant and soil
health enhancer and application thereof for promoting plant growth and controlling
plant pathogenic fungi, said composition comprising urine, neem and garlic,
individually or in all possible combinations, the treatment showing stimulation of the
accumulation of nutrients in the plant biomass, proliferation of plant growth
promoting, phosphate solubilizing, abiotic stress tolerant and antagonists towards
plant pathogenic fungi in the rhizosphere of plants, and enhances the total phenolic
contents of the plants; and a method for producing the composition.
Objects of the present invention
The main object of the present invention is to develop a synergistic fermented
composition useful in promoting plant growth, soil health and bio-controlling.
Another object of the present invention is to develop a process of preparing
synergistic fermented composition comprising cow urine, crushed neem leaves and/or
crushed garlic bulbs, optionally along with carrier(s), useful in promoting plant
growth.
Yet another object of the present invention is to develop a method of promoting plant
growth using bovine urine and/or crushed neem leaves and/or crushed garlic bulbs,
optionally along with carrier(s).
Summary of the present invention
The present invention relates to a synergistic composition useful as plant and soil
health enhancer, comprising urine, neem and garlic, individually or in all possible
combinations, with the treatment showing it has the ability to stimulate accumulation
of nutrients in the plant biomass, proliferation of plant growth promoting, phosphate
solubilizing, abiotic stress tolerant and antagonists towards plant pathogenic fungi,
control phytopathogenic fungi in the rhizosphere of plants, and enhances the total
phenolic contents of the plants.
Detailed description of the present invention
Accordingly, the present invention relates to a synergistic fermented composition
useful in promoting plant growth, soil health and bio-controlling, said composition
comprising bovine urine; crushed neem leaves of concentration ranging between 10 to
750 grams/liter of bovine urine and/or crushed garlic bulbs of concentration ranging
between 1 to 500 grams/liter of bovine urine, optionally along with carrier(s).
In still another embodiment of the present invention, wherein the concentration of
neem is preferably 250 grams/liter of bovine urine.
In still another embodiment of the present invention, wherein the concentration of
garlic is preferably 100 grams/liter of bovine urine.
In still another embodiment of the present invention, wherein the bovine urine is fresh
bovine urine.
In still another embodiment of the present invention, wherein the bovine urine is cow
urine.
In still another embodiment of the present invention, wherein the carrier is selected
from a group comprising vermicompost, soil, peat, rice husk,- vermiculite,
carboxymethyl cellulose, perlite, polyvinyl-pyrrolidone, talc, and fermented pres mud.
In still another embodiment of the present invention, wherein the carrier is preferably
vermicompost or fermented pres mud.
In still another embodiment of the present invention, wherein the concentration of
carrier is ranging between 10 to 1000 gm/ liter of bovine urine.
In still another embodiment of the present invention, wherein a process of preparing
synergistic fermented composition comprising cow urine, crushed neem leaves of
concentration ranging between 10 to 750 grams/liter of bovine urine, and/or crushed
garlic bulbs of concentration ranging between 1 to 500 grams/liter of bovine urine,
optionally along with carrier(s), useful in promoting plant growth, said process
comprising steps of:
• collecting fresh urine from healthy bovine,
• adding crushed garlic bulbs and neem leaves to the collected urine,
• fermenting resultant mixture of step (b) to obtain the synergistic composition,
and
• optionally, adding carrier to the synergistic composition.
In still another embodiment of the present invention, wherein the concentration of
neem is preferably 250 grams/liter of bovine urine.
In still another embodiment of the present invention, wherein the concentration of
garlic is preferably 100 grams/liter of bovine urine.
In still another embodiment of the present invention, wherein the bovine is cow.
In still another embodiment of the present invention, wherein fermenting the resultant
mixture for about 30 days.
In still another embodiment of the present invention, wherein the carrier is selected
from a group comprising vermicompost, soil, peat, rice husk, vermicuiite,
carboxymethyl cellulose, perlite, polyvinyl-pyrrolidone, talc, and fermented pres mud.
In still another embodiment of the present invention, wherein the carrier is preferably
vermicompost or fermented pres mud.
In still another embodiment of the present invention, wherein the concentration of
carrier is ranging between 10 to 1000 gm/ liter of bovine urine.
In still another embodiment of the present invention, wherein a method of promoting
plant growth using bovine urine and/or crushed neem leaves of concentration ranging
between 10 to 750 grams/liter, and/or crushed garlic bulbs of concentration ranging
between 1 to 500 grams/liter, optionally along with carrier(s), said meth'od consisting
step of exposing plant part(s) to bovine urine and/or neem and/or garlic.
In still another embodiment of the present invention, wherein the concentration of
neem is preferably 250 grams/liter.
In still another embodiment of the present invention, wherein the concentration of
garlic is preferably 100 grams/liter.
In still another embodiment of the present invention, wherein the bovine is cow.
In still another embodiment of the present invention, wherein the garlic and/or neem
is crushed in urine or water.
In still another embodiment of the present invention, wherein the carrier is selected
from a group comprising vermicompost, soil, peat, rice husk, vermiculite,
carboxymethyl cellulose, perlite, polyvinyl-pyrrolidone, talc, and fermented pres mud.
In still another embodiment of the present invention, wherein the carrier is preferably
vermicompost or fermented pres mud.
In still another embodiment of the present invention, wherein the concentration of
carrier is ranging between 10 to 1000 gm/ liter of bovine urine.
In still another embodiment of the present invention, wherein the method controls
plant pathogenic bacteria.
In still another embodiment of the present invention, wherein the method promotes
accumulation of nutrients in plant biomass.
In still another embodiment of the present invention, wherein the method promotes
accumulation of nitrogen in plant biomass.
In still another embodiment of the present invention, wherein the method promotes
accumulation of phosphorus in plant biomass.
In still another embodiment of the present invention, wherein the method promotes
phosphate solubilization.
In still another embodiment of the present invention, wherein the method promotes
abiotic stress tolerance.
In still another embodiment of the present invention, wherein the method promotes
antagonists towards plant pathogenic fungi.
In still another embodiment of the present invention, wherein the method promotes
antagonists towards plant pathogenic fungi in rhizosphere of plants.
In still another embodiment of the present invention, wherein the fungi are selected
from a group comprising Fusarium sp., Alternaria sp., Phytophthora palmivora,
Sclerotinia sclerotiorum, Sclerotium rolfsii, Colletotrichum sp., Penicillium sp.,
Aspergillus niger, Rhizoctonia solani, Pythium aphanidermatum, Curvularia lunata,
and Phoma sorghi.
In still another embodiment of the present invention, wherein the method enhances
total phenolic content of the plant.
In still another embodiment of the present invention, wherein the method protects
plants from soil borne plant pathogens forming sclerotia/chlamydospores.
In still another embodiment of the present invention, wherein promoting plants
growth by soil drenching.
In still another embodiment of the present invention, wherein promoting plants
growth by aerial/foliar spray.
In still another embodiment of the present invention, wherein promoting plants
growth by seed soaking.
In still another embodiment of the present invention, wherein promoting plants
growth by furrow treatment.
In still another embodiment of the present invention, wherein the method stimulates
proliferation of plant growth promoting microorganisms in the rhizosphere of plants.
In still another embodiment of the present invention, wherein the method stimulates
proliferation of phosphorus solubilizing microorganisms in the rhizosphere of plants.
In still another embodiment of the present invention, wherein the method stimulates
proliferation of abiotic stress tolerant microorganisms in rhizosphere of plants.
In still another embodiment of the present invention, wherein the neem and/or garlic
and/or urine are in boiled state.
In still another embodiment of the present invention, wherein the plants are selected
from a group comprising chickpea, maize, wheat, and pea.
In still another embodiment of the present invention, wherein the neem and/or garlic
and/or urine in earthen and copper vessel promote plant growth.
In still another embodiment of the present invention, wherein the method using
copper and/or earthen vessel promotes plant growth increases plant dry weight by
about 110%.
In still another embodiment of the present invention, wherein the neem and/or garlic
and/or urine is diluted in the ratio ranging between 1:5 to 1: 1000.
In still another embodiment of the present invention, wherein the neem and/or garlic
and/or urine is diluted preferably in the ratio of about 1:10.
In still another embodiment of the present invention, wherein the combination of
neem, garlic, and urine is most effective in promoting plant growth.
In still another embodiment of the present invention, wherein the synergistic
combination of neem, garlic, and urine show about 85% increase in wheat growth.
In still another embodiment of the present invention, wherein the method promotes
plant growth by inhibiting sclerotia and chlamydospores of pathogenic fungi in about
2 to 4 hours.
In still another embodiment of the present invention, wherein the method promotes
plant growth by protecting plant from soil-borne plant-pathogens.
In still another embodiment of the present invention, wherein the method promotes
plant growth as combination of neem, garlic, and urine is showing 100% biocontrol
activity against collar rot.
In still another embodiment of the present invention, wherein the method promotes
plant growth by controlling leaf spot disease.
In still another embodiment of the present invention, wherein the method promotes
plant growth by increasing dry weight of the plant by about 50%.
In still another embodiment of the present invention, wherein the method promotes
plant growth by increasing nitrogen accumulation by about 50%.
In still another embodiment of the present invention, wherein the method promotes
plant growth by increasing phosphorus accumulation by about 35%.
In still another embodiment of the present invention, wherein the method promotes
plant growth by reducing pathogenic bacterial population by about 1 log unit.
In still another embodiment of the present invention, wherein the method promotes
plant growth by reducing pathogenic fungal population by about 0.7 log unit.
In still another embodiment of the present invention, wherein the method promotes
plant growth by reducing actinomycetes population by about 1 log unit.
In still another embodiment of the present invention, wherein the method promotes
plant growth by increasing antagonism by about 150% towards fungi.
In still another embodiment of the present invention, wherein the method promotes
plant growth by increasing abiotic stress tolerance by about 100%.
In still another embodiment of the present invention, wherein the method promotes
plant growth by increasing phosphate solubilization by about 120%.
In still another embodiment of the present invention, wherein the method shows
increase in gram-positive bacteria by about 40%.
In still another embodiment of the present invention, wherein the method shows
decrease in gram-negative bacteria by about 20%.
In still another embodiment of the present invention, wherein the method shows
increase in gram-positive bacteria.
In still another embodiment of the present invention, wherein the carrier increases
plant growth by 30 to 50%.
In still another embodiment of the present invention, wherein the carrier increases
antagonism towards plant-pathogenic fungi in the range of 30 to 45%.
In still another embodiment of the present invention, wherein the method promotes
plant growth by increasing phenolic content in the range of 120 to 130%.
In still another embodiment of the present invention, wherein accordingly, the present
invention relates to a synergistic composition useful as plant and soil health enhancer,
comprising urine, neem and garlic, individually or in all possible combinations, with
the treatment showing it has the ability to stimulate accumulation of N and P in the
plant biomass, proliferation of plant growth promoting, phosphate solubilizing, abiotic
stress tolerant and antagonists towards plant pathogenic fungi, control
phytopathogenic fungi in the rhizosphere of plants and enhances the total phenolic
contents of the plants.
In an embodiment of the present invention, a synergistic composition useful as plant
and soil health enhancer, comprising urine and plants, individually or in all possible
combinations.
In still another embodiment of the present invention, wherein urine is collected from a
bovine group of animal comprising cow.
In still another embodiment of the present invention, wherein plants are selected from
a group comprising neem and garlic.
In still another embodiment of the present invention, wherein amount of neem and
garlic is about 250 gm and 100 gm/liter urine.
In still another embodiment of the present invention, wherein total amount of neem is
250 gm/liter urine and preferably 10-750 gm/liter urine.
In still another embodiment of the present invention, wherein total amount of garlic is
100 gm/liter urine and preferably 1-500 gm/liter urine.
In still another embodiment of the present invention, wherein said synergistic
composition enhances plant growth.
In still another embodiment of the present invention, wherein plant for growth
promotery activity is selected from a group comprising chickpea, maize, wheat and
pea.
In still another embodiment of the present invention, wherein said synergistic
composition protects plants from pathogens which has the potential as biopesticide for
the biocontrol of wide host range of economically important pathogenic fungi.
In still another embodiment of the present invention, wherein pathogenic fungi for
biological control activity is selected from a group comprising Fusarium sp.,
Alternaria sp., Phytophthora palmivora, Sclerotinia sclerotiorum, Sclerothim rolfsii,
Colletotrichum sp., Penicillium sp., Aspergillm niger, Rhizoctonia solani, Pythium
aphanidermatum, Curvularia lunata and Phoma sorghi.
In still another embodiment of the present invention, wherein said synergistic
composition protect plants from soil borne plant pathogens forming
sclerotia/chlamydospores.
In still another embodiment of the present invention, wherein said synergistic
composition protects plants by seed soaking treatment, from seed borne plant
pathogens.
In still another embodiment of the present invention, wherein said synergistic
composition protect plants by soil drenching treatment, from plant pathogens.
In still another embodiment of the present invention, wherein said synergistic
composition protect plants as an aerial spray from plant pathogens.
In still another embodiment of the present invention, wherein said synergistic
composition is used with carriers to disperse the subject treatment, wherein preferred
carriers are selected from a group comprising vermicompost and fermented press
mud.
In still another embodiment of the present invention, wherein said synergistic
composition enhances accumulation of nutrients in the plant biomass.
In still another embodiment of the present invention, wherein said synergistic
composition stimulate proliferation of plant growth promoting microorganisms in the
rhizosphere of plants.
In still another embodiment of the present invention, wherein said synergistic
composition stimulate proliferation of phosphate solubilizing microorganisms in the
rhizosphere of plants.
In still another embodiment of the present invention, wherein said synergistic
composition stimulate proliferation of abiotic stress tolerant microorganisms in the
rhizosphere of plants.
In still another embodiment of the present invention, wherein said synergistic
composition stimulate proliferation of antagonists towards plant pathogenic fungi in
the rhizosphere of plants.
In still another embodiment of the present invention, wherein said synergistic
composition enhances the total phenolic contents of the plants.
The experiments were conducted using urine from Gir, Kankrej, Nagpur, Tharparkar,
and Haryana cows. The difference in the plant biomass was within the range of 2%.
By far, Sahiwal is the best breed of the subcontinent. Therefore, urine from Sahiwal
cow was used for further work.
In still another embodiment of the present invention, applicants have discovered a
novel method of using cow urine to select those plants that are useful as plant and soil
health enhancer and showing the ability of the accumulation of nutrients in the plant
biomass, proliferation of plant growth promoting, phosphate solubilizing, abiotic
stress tolerant and antagonists towards plant pathogenic fungi in the rhizosphere of
plants, and enhances the total phenolic contents of the plants.
The above-stated invention is further elaborated in the form of examples and should
not be construed to limit to limit the scope of the invention
1. Collecting fresh urine from healthy indigenous (Sahiwal) cows in morning into
clean plastic container. Pouring the 1 liter urine into 2 liter capacity either copper
or earthen pot buried in soil up to its neck for its fermentation up to 30 days. After
30 days, using the fermented product either directly or boiling it to concentrate to
0.25 liter, before its application. In another combination fermenting cow urine
along with neem (250 gm leaves/liter) and/or garlic (100 gm crushed bulbs/liter).
In yet another combination fermenting neem and garlic with water. Thus
preparing 12 treatments using urine, neem and garlic individually in copper or
earthen pot, each, to a final total of 24 treatments.
2. Screening of 24 treatments prepared in step 1 in the greenhouse having potential
for promoting plant growth as follows: growing chickpea plants in the presence of
individual treatment as prepared in step 1 in the greenhouse in a concentration of
about 1:100 dilution in non-sterile soil; growing control chickpea plants as above
but without addition of the treatment; and selecting as plant growth promoter
those treatments which cause the treated plants to exhibit greater dry weight.
3. Further evaluation of the treatment selected in step 2 as plant growth promotion
ability of chickpea, maize, wheat and pea in greenhouse using different dilution in
the range of 0 to 1:1000 and selecting those treatments which cause the treated
plants to exhibit greater dry weight.
4. Screening the treatments urine + neem and urine + neem + garlic fermented in
earthen vessel, obtained by the procedure outlined in Example 1 were screened for
the potential to inhibit growth of Fusarium oxysporum f. sp. ciceri, Fusarium
oxysporum f. sp. gladioli, Fusarium moniliforme, Alternaria solani, Alternaria
alternata, Phytophthora palmivora, Phytophthora nicotianae, Sclerotinia
sderotiorum, Sclerotium rolfsii, Colletotrichum falcatum, Colletotrichum capsici,
Penicillium sp., Aspergillus niger, Rhizoctonia solani, Pythium aphanidermatum,
Curvulana lunata and Phoma sorghi, under in vitro conditions as follows:
transferring an agar plug inoculum of the fungi to be tested (5-mm square) to the
center of the plate individually from a source plate of the fungi on nutrient agar
plates (NA), boring 4 holes of 5 mm diameter on each NA plate; adding in each
hole 100 uL of the treatments were individually in 3 holes while adding sterile
water in the 4th hole as negative control; incubating plates for 5 to 8 days and
selecting the treatments having the biocontrol activity which inhibited fungal
growth.
5. Evaluating effect of various concentrations of neem and garlic in various
combinations and amounts (gm/liter) as indicated on the product containing
fermented in earthen vessel under in vitro conditions for ability to suppress
Phytophthora palmivora and promote growth of wheat as follows: selecting
amount and combination of neem and garlic demonstrating better ability to inhibit
P. palmivora and promote plant growth of wheat, with respect to dry weight.
6. Evaluating the product containing urine + neem + garlic fermented in earthen
vessel, obtained by the procedure outlined in Example 1 for its ability to inhibit
growth of the sclerotia of Sclerotium rolfsii, Rhizoctonia solani and Sclerotinia
sderotiorum and chamydospores of Fusarium sp. under in vitro conditions as
follows: dipping three-week old similar sized sclerotia/chamydospores were
collected from NA plates after surface sterilized in various dilutions of the
treatments individually, for 2 hrs; monitoring the viability of the
sclerotia/chamydospores by removing and inoculating 3 batches of 15
sclerotia/chamydospores on NA after incubation at 25°C for 7 days; recording the
viability of sclerotia/chamydospores in terms of number of germinating
sclerotia/chlamydospores by visual examination of mycelial growth after 7 days.
7. Screening of the 24 treatments, obtained by the procedure outlined in Example 1
for their ability to inhibit growth of Sclerotium rolfsii by seed soaking and soil
drenching treatment to control collar rot of chickpea seedlings in greenhouse as
follows: soaking seeds by dipping in the 10% dilution of the treatments
individually, for 2 hrs; for soil drenching treatment adjusting the dry pot soil
moisture to 20% with the product diluted 1:100 individually.
8. Evaluating effect of various concentrations of the product containing urine + neem
+ garlic fermented in earthen vessel by foliar spray treatment for its ability to
control leaf spot disease of field grown betelvine caused by Alternaria alternata
as follows: preparing one-meter wide beds with a width of 35 cm and 10 cm high
ridges, planting cuttings from single node cuttings on a field plot naturally
infected with Alternaria alternata; first spraying of the product at the time of
appearance of disease symptoms followed by 5 more sprays at 15 days' interval;
noting disease intensity 15 days after 5th spray of the product.
9. Evaluating effect of addition of boiled urine + neem + garlic fermented in earthen
vessel on the rhizosphere microorganisms and accumulation of N and P of
chickpea plants as follows: growing chickpea plants for 4 weeks in the presence
and absence of 1:10 diluted boiled urine + neem + garlic fermented in earthen
vessel; determining the dry weight, microbial population, and N and P content of
the plants.
10. Elucidating effect of addition of boiled urine + neem + garlic fermented in earthen
vessel on rhizosphere microorganisms of chickpea plants was evaluated by
determining the percentage of bacteria among the resident population,
demonstrating plant growth promotion, phosphate solubilization, abiotic stress
tolerance and antagonism towards plant pathogenic fungi as follows: growing
chickpea plants for 4 weeks in the presence and absence of 1:10 diluted boiled
urine + neem + garlic fermented in earthen vessel; determining changes in the
percentage of bacteria among the resident population demonstrating plant growth
promotion, phosphate solubilization, abiotic stress tolerance and antagonism
towards plant pathogenic fungi.
1 1 . Elucidating effect of addition of boiled urine + neem + garlic fermented in earthen
vessel on the total phenolic contents of chickpea plants as follows: growing
chickpea plants for 4 weeks in the presence and absence of 1:10 diluted boiled
urine + neem + garlic fermented in earthen vessel; determining changes in the
total phenolic compounds in the leaf and root of treated chickpea was observed,
compared with un-treated control.
The synergistic compositions comprising urine, neem and garlic, individually or in all
possible combinations, selected by the above process have the ability of promoting
plant growth and controlling plant pathogenic fungi, stimulate accumulation of N and
P in the plant biomass, proliferation of plant growth promoting, phosphate
solubilizing, abiotic stress tolerant and antagonists towards plant pathogenic fungi in
the rhizosphere of plants, and enhances the total phenolic contents of the plants.
In accordance with this discovery, it is an object of the invention to provide
synergistic compositions comprising urine, neem and garlic, individually or in all
possible combinations, selected by the above process have the ability of promoting
plant growth and controlling plant pathogenic fungi, stimulate accumulation of N and
P in the plant biomass, proliferation of plant growth promoting, phosphate
solubilizing, abiotic stress tolerant and antagonists towards plant pathogenic fungi in
the rhizosphere of plants, and enhances the total phenolic contents of the plants.
It is also an object of the invention is to provide a means for screening synergistic
compositions comprising urine to select those plants that have the ability of promoting
plant growth and controlling plant pathogenic fungi, stimulate accumulation of N and
P in the plant biomass, proliferation of plant growth promoting, phosphate
solubilizing, abiotic stress tolerant and antagonists towards plant pathogenic fungi in
the rhizosphere of plants, and enhances the total phenolic contents of the plants.
A further object of the invention is to provide a means for screening synergistic
compositions comprising urine to select those plants that have the ability of promoting
plant growth and controlling plant pathogenic fungi, stimulate accumulation of N and
P in the plant biomass, proliferation of plant growth promoting, phosphate
solubilizing, abiotic stress tolerant and antagonists towards plant pathogenic fungi in
the rhizosphere of plants, and enhances the total phenolic contents of the plants for
commercial production useful as plant and soil health enhancer for plants, seeds, and
soil.
Yet another object of the invention is to provide a means for screening synergistic
compositions comprising urine to select those plants that have the ability to protect
plants from soil borne plant pathogens forming sclerotia/chamydospores.
Still another object of the invention is to provide a means for screening synergistic
compositions comprising urine to select those plants that have the ability to protect
plants by seed soaking and soil drenching treatment, from plant pathogens.
Another object of the invention is to provide a means for screening synergistic
compositions comprising urine to select those plants that have the potential to protect
plants as an aerial spray, from plant pathogens.
Other objectives and advantages of the invention will become apparent from the
ensuing description. It should be understood, however, that the detailed description
and the specific examples, while indicating preferred embodiments of the invention,
are given by way of illustration only, since various changes and modifications within
the spirit and scope of the invention will become apparent to those skilled in the art
from this detailed description.
It has been discovered that usage of urine from cow acts as plant and soil health
enhancer. Application of cow urine results in promoting plant growth and controlling
plant pathogenic fungi, said composition comprising urine, neem leaves and garlic,
individually or in all possible combinations, stimulation of proliferation of plant
growth promoting, phosphate solubilizing, abiotic stress tolerant and antagonists
towards plant pathogenic fungi in the rhizosphere of plants and enhances the total
phenolic contents of the plants.
Therefore one aspect of the present invention relates to method for screening useful
synergistic compositions comprising urine to select those plants that have the
potential for promoting plant growth and controlling plant pathogenic fungi, with the
treatment showing stimulation of the accumulation of N and P in the plant biomass,
proliferation of plant growth promoting, phosphate solubilizing, abiotic stress tolerant
and antagonists towards plant pathogenic fungi in the rhizosphere of plants, and
enhances the total phenolic contents of the plants.
First fresh urine from healthy indigenous (Sahiwal) cows was collected in morning
into clean plastic container. The 1 liter urine was poured into 2 liter capacity copper
vessel buried in soil up to its neck for its fermentation up to 30 days, along with neem
(250 gm leaves/liter) and/or garlic (100 gm crushed bulbs/liter). After 30 days, using
the fermented product either directly or boiling it to concentrate to 0.25 liter, before
its application. In yet another combination cow urine was fermented along with neem
leaves and/or crushed garlic bulbs for 30 days in 2 liter capacity earthen vessel buried
in soil up to its neck. After 30 days, using the fermented product either directly or
boiling it to concentrate to 0.25 liter, before its application. In yet another
combination fermenting neem leaves and/or crushed garlic bulbs with water. After 30
days, using the fermented product either directly or boiling it to concentrate to 0.25
liter, before its application. Thus 12 treatments were prepared using urine, neem
leaves and/or crushed garlic bulbs individually in copper or earthen pot, each, to a
final total of 24 treatments.
Twenty-four treatments prepared in the previous step are screened to select those,
which at a particular concentration promote plant growth under greenhouse conditions
as described earlier [C. S. Nautiyal, J. K Johri and H. B. Singh. Canadian Journal of
Microbiology, Volume 48, pp. 588-601 (2002)]. In this test seeds of chickpea are
grown in non-sterilized soil and treated individually, at a concentration of 1:100
dilution. Barthen pots (of 25 cm diameter) have been found to be of a convenient size
to grow chickpea and other plants for the greenhouse test. Each pot was filled with
non-sterilized soil. Although sterile soil or any other plant growth supporting material
for example like vermiculite may also be used instead of non-sterile soil, it is
preferred that non-sterile soil from the field where these treatments are intended to be
applied is used in greenhouse test. Tap water was added to each hole before planting
seeds to adjust the dry soil to 20% moisture with the 24 treatments individually
diluted to 1:100. Tap water for control and diluted treatment was added before
planting seeds to adjust the dry soil to 15 to 30% moisture. Preferred soil moisture is
20%. Six seeds were added per pot. Data was recorded after 21 days of plant growth
with respect to plant height and weight and as plant growth promoter those treatments
were selected which cause the treated plants to exhibit greater dry weight.
Next the treatment selected in the previous step is evaluated for its plant growth
promotion ability of chickpea, maize, wheat and pea in greenhouse using different
dilution in the range of 0 to 1:1000 and selecting those treatments, which at a
particular concentration promote plant growth under greenhouse conditions as
described earlier [C. S. Nautiyal, Current Microbiology, Volume 34, pp. 12-17
(1997)]. In this test seeds of chickpea, maize, wheat and pea are grown in nonsterilized
soil. Trays (35x35 cm.) with 16 (4x4) places per tray (each space was of 7
cm. width, 10 cm. depth and 1 cm. apart from each other) have been found to be of a
convenient size to grow chickpea, maize, wheat and pea and other plants for the
greenhouse test. Each place was filled up to 8 cm. with non-sterilized soil.
Although sterile soil or any other plant growth supporting material for example like
vermiculite may also be used instead of non-sterile soil, it is preferred that non-sterile
soil from the Held where these treatments are intended to be applied is used in
greenhouse test. Data was noted after 30 days of plant growth with respect to plant
height and weight and as plant growth promoter those treatments were selected which
cause the treated plants to exhibit greater dry weight.
Next, the treatments urine + neem and urine + neem + garlic fermented in earthen
vessel showing good plant growth promotion are subjected to a screening for their
ability to suppress phytopathogenic fungi suppressing growth of Fusarium oxysporum
f. sp. ciceri, Fusarium oxysporum f. sp. gladioli, Fusarium moniliforme, Alternaria
solani, Alternaria alternata, Phytophthora palmivora, Phytophthora nicotianae,
Sclerotima scleroiiorum, Sclerotium rolfsii, Colletotrichum falcatum, Colletotrichum
capsici, Penicillium sp., Aspergillus niger, Rhizoctonia solani, Pythium
aphanidermatum, Curvularia hmata and Phoma sorghi under in vitro conditions as
described earlier [C. S. Nautiyal (1997) Current Microbiology 35: 52-58; C. S.
Nautiyal (1997) FEMS Microbiology Ecology 23: 145-158].
An agar plug inoculum of the fungi to be tested (5-mm square) was then transferred to
the center of the plate individually from a source plate of the fungi. Four holes (5 mm
diameter) were bored on each NA plate. In each hole 100 uL of the treatments were
individually added and in the 4th hole sterile water was added as a negative control.
After incubation for 5 to 8 days inhibition zones were readily observed in the case of
treatments having the biocontrol activity as the fungal growth around the hole was
inhibited. While in case of treatments not having biocontrol activity and the hole
sterile water was added as a negative control, fungal growth around the hole was not
inhibited and the fungi grew around the edge of the hole.
Concentrations of neem (50, 100, 250 and 500 gm/liter) and garlic (10, 50, 100 and
250 gm/liter) in urine were further improved upon by using several combinations and
amounts in earthen vessel under in vitro conditions for optimization of its ability to
suppress pathogenic fungi and promote plant growth. Amount of neem -used is in the
range of 50 to 500 gm/liter and for garlic 10 to 250 gm/liter. Preferred amount for
neem and garlic is 250 and lOOgm/liter, respectively.
Next the treatment containing urine + neem + garlic fermented in earthen vessel are
subjected for its ability to inhibit growth of the sclerotia of Sclerotia rolfsii,
Rhizoctonia .solani and Sclerotinia sclerotiorum and chamydospores of Fusarium sp.
under in vitro conditions by dipping three-week old similar sized
sclerotia/charnydospores collected from NA plates after surface sterilized in various
dilutions of the treatments individually, for up to 6 hrs; monitoring the viability of the
sclerotia/chamydospores by removing and inoculating 3 batches of 15
sclerotia/chamydospores on NA after incubation at 25°C for 7 days; recording the
viability of sclerotia/chamydospores was recorded in terms of number of germinating
sclerotia/chamydospores by visual examination of mycelial growth after 7 days.
The 24 treatments were further tested for their ability to inhibit growth of Sclerotium
rolfsii by seed soaking and soil drenching treatment to control collar rot of chickpea
seedlings in greenhouse by dipping in 10% dilutions of the treatments individually, as
indicated for 2 hrs; for soil drenching treatment adjusting the dry pot soil moisture to
20% with the product diluted 1:100 individually, as stated.
Various concentrations of the treatment containing urine + neem + garlic fermented in
earthen vessel by foliar spray treatment for its ability to control leaf spot disease of
field grown plant.
Effect of addition of boiled urine + neem + garlic fermented in earthen vessel on the
plant health, rhizosphere microorganisms and accumulation of N and P of chickpea
plants was evaluated. Plant health and the population of rhizosphere microflora was
estimated by a previously described procedure [C. S. Nautiyal, J. K Johri and H. B.
Singh. Canadian Journal of Microbiology 48: 588-601 (2002)] Total N was
determined by the Kjeldahl method [J. M. Bremner and C. S. Mulvancy (1982)
Nitrogen-total, p. 595-624. In A. L. Page (ed.), Methods of soil analysis, part 2, 2nd
ed.. Agronomy, no. 9, American Society of Agronomy, Madison, Wis., U.S.A.]. Plant
tissues were digested in a mixture of 15 ml of HC1O4 and 5 ml of HNOs and
phosphorus was determined colorimetrically by the vanado-molybdate method [H. L.
S. Tandon, M. P. Cescas and E. H. Tyner (1968) Soil Science Society of America
Proceedings 32: 48-51].
Next boiled urine + neem + garlic fermented in earthen vessel is added-to the soil to
elucidate its effect on rhizosphere microorganisms by determining the percentage of
bacteria among the resident population, demonstrating plant growth promotion,
phosphate solubilization, abiotic stress tolerance and antagonism towards plant
pathogenic fungi. Chickpea plants were grown for 4 weeks in the presence and
absence of 1:10 diluted boiled urine + neem + garlic fermented in earthen vessel.
To isolate rhizosphere bacteria chickpea roots were thoroughly washed with tap water
for two minutes to remove all loosely adhering soil particles, followed by washing
with sterile 0.85% (w/v) saline Milli Q water (MQW). The roots were then macerated
in 0.85% saline MQW with a mortar and pestle. Serial dilutions of the homogenate
were then plated on NA agar plates as described earlier [C. S. Nautiyal (1997) Current
Microbiology 33: 1-6]. One thousand bacterial representatives of the predominant
morphologically distinct colonies present on the plates were selected from NA plates
and purified by sub culturing an individual strain on NA plates to obtain a pure
culture for further screening.
Each isolate was stored in an aqueous solution of 30% glycerol at -25°C. One
thousand bacterial strains were used to determine the % of gram-positive and gramnegative
bacteria present in the heterogenous population of the bacteria. Out of 1000
bacterial strains 100 bacterial representatives of the predominant morphologically
distinct colonies were randomly used to determine the changes in the % of bacteria
among the resident population demonstrating plant growth promotion, phosphate
solubilization, abiotic stress tolerance and antagonism towards plant pathogenic fungi.
Individual strains of the 100 bacteria were subjected to screening for selecting
bacteria having potential for plant growth promotion. Bacterial inoculum for maize
(Zea mays) seeds was prepared by scraping 48 h grown culture from plates with 10 ml
of 0.85% saline water. Maize seeds were surface sterilized by gently shaking (80
R.P.M. on a reciprocal shaker at 28°C) with 70% ethanol (5 min.), 20% bleach
Chlorox (10 min.), followed by three rinses in sterile water. After surface sterilization
seeds were soaked in the bacterial suspension (IxlO8 CFU/ml) for 4h at 28°C on a
reciprocal shaker at 100 R.P.M. Control seeds (non bacterized) were soaked in 0.85%
saline water washed from uninoculated plates. Bacterization levels of seeds were
determined by agitating 4 seeds from each treatment and plated after serial dilution on
NA plate. Mean cfu/seed were determined by averaging the cfu/gm values of three
populations in three replicates per treatment after 48 h incubation of the plates at
28°C. Tap water was added to each hole before planting seeds to adjust-the soil to 15
to 30% moisture.
Preferred soil moisture is 20%. Trays (35x35 cm.) with 16 (4x4) places per tray (each
space was of 7 cm. width, 10 cm. depth and 1 cm. apart from each other) were used,
to grow maize. Bach place was filled up to 8 cm. with non-sterilized soil. Tap water
(25 ml.) was added to each hole before planting seeds to adjust the soil to 20%
moisture. Four bacterized seed was added per hole. The experiment in greenhouse
was carried out in four different sets of 16 maize seedlings each, for non-bacterized
(control) and bacterized seeds. In each set, data of 21-days-old seedlings was noted
with respect to dry weight of plants. For the bacterial strains to be plant growth
promoter, the bacterized seedlings must have averaged at least 10% higher dry weight
than comparable non-bacteri/ed plants [C. S. Nautiyal, J. K Johri and H. B. Singh.
Canadian Journal of Microbiology 48: 588-601 (2002)].
Quantitative estimation of phosphate solubilization capability of the 100 bacterial
strains was carried out in triplicate by using National Botanical Research Institute's
Phosphate solubilizing medium (NBRIP) in broth. The method has been described by
Mehta and Nautiyal [S. Mehta and C. S. Nautiyal (2001) Current Microbiology 43:51-
56],
Next individual strains of the 100 bacteria were subjected to screening for abiotic
stress tolerance. The stress tolerance of the strains towards salt (NaCl), pH, and
temperature was tested by growing them individually on nutrient broth (NB) under
various stress conditions, e.g., like 6% salt (NaCl), pH (3 and 11), and at temperature
(5 and 55°C) stress were grown overnight (14-16 hrs). For 6% salt (NaCl), pH (3 and
11) stress cultures were grown overnight at 30°C. Serial dilutions of each sample were
spotted (25 ul) onto NA plates, and incubated at 30°C in triplicate as described earlier
[C. S. Nautiyal, J. K Johri and H. B. Singh. Canadian Journal of Microbiology 48:
588-601 (2002)]. Growth of bacterial strains having the abiotic stress tolerance ability
was clearly visible with in 2-3 days of incubation of the plates at 30°C.
Individual strains of the 100 bacteria were subjected to screening for bacteria having
potential for suppressing phytopathogenic fungi namely Fusarmm oxysporiim f. sp.
ciceri, Colletolnchum falcatum, Sclerotium rolfsii, Alternaria solani, Penicillium sp.,
Pythium aphanidermatum, Phytophthora palmivora, Curvularia lunata, Sclerotinia
sclerotiorum, Aspergillus niger and Phoma sorghi under in vitro conditions as
described earlier [C. S. Nautiyal (1997) Current Microbiology 35: 52-58]. Bacterial
colonies on NA plates were streaked around the edge of a 90-mm diameter petri plate
and the plates were incubated at 28°C for 1 to 2 days. An agar plug inoculum of the
fungi to be tested (5-mm square) was then transferred to the center of the plate
individually from a source plate of the fungi. After incubation for 5 to 8 days
i n h i b i t i o n /ones were readily observed in the case of bacterial strains having the
biocontrol activity.
Boiled urine + neem + garlic fermented in earthen vessel is added to the carriers to
disperse the subject treatment such as peat, vermicompost and fermented press mud to
15 to 45% moisture. Preferred moisture is 35% before its addition to soil as seed or
furrow treatments. For furrow-treatment, formulation was directly applied all over the
open furrow containing seeds and covered after treatment C. S. Nautiyal, J. K Johri
and H. B. Singh. Canadian Journal of Microbiology 48: 588-601 (2002)]. Chickpea
plants were grown for 4 weeks to elucidate its effect on rhizosphere microorganisms
as described above by determining the percentage of bacteria among the resident
population, demonstrating plant growth promotion, phosphate solubilization, and
antagonism towards plant pathogenic fungi.
Next effect of addition of boiled urine + neem + garlic fermented in earthen vessel
was evaluated on the total phenolic contents of chickpea plants. In this test, seeds of
chickpea are grown in non-sterilized soil and treated individually, at a concentration
of 1:10 dilution. Earthen pots (of 25 cm diameter) have been found to be of a
convenient size to grow chickpea and other plants for the greenhouse test. Each pot
was filled with non-sterilized soil. Tap water was added to each hole before planting
seeds to adjust the dry soil to 20% moisture with the 24 treatments individually
diluted to 1:10, Tap water for control and diluted treatment was added to each hole
before planting seeds to adjust the dry soil to 20%. Four seeds were added per hole.
Data was noted after 30 days of plant growth with respect to total phenolic
compounds in the leaf and root of treated chickpea, compared with un-treated control
and total phenolics were estimated spectrophotometrically using the Prussian Blue
method as modified by Graham [H. G. Graham (1992) Journal of Agriculture Food
Chemistry Volume 40: 801-805].
In addition to the other properties noted above, the unexpected and surprising
attributes of these synergistic compositions include the following characteristics. All
the treatments have been prepared from the urine of Sahiwal cow. The treatments
promote plant growth and inhibit the growth of many pathogenic fungi of plants.
These treatments promote plant growth and reduce the plant disease in soil both under
greenhouse and field conditions. The treatments of the present invention are capable
of stimulation of the accumulation of nutrients in the plant biomass, proliferation of
plant growth promoting, phosphate solubilizing, abiotic stress tolerant and antagonists
towards plant pathogenic fungi in the rhizosphere of plants. Moreover the treatment
enhances the total phenolic contents of the plants.
It is within the compass of the invention to composition useful as plant and soil health
enhancer using cow urine along with neem and garlic and application thereof for
promoting plant growth and controlling plant pathogenic fungi, said composition
comprising any type of urine or plant having the ability to stimulate the accumulation
of nutrients in the plant biomass, proliferation of plant growth promoting, phosphate
solubilizing, abiotic stress tolerant and antagonists towards plant pathogenic fungi in
the rhizosphere of plants, and enhances the total phenolic contents of the plants.
Another aspect of the invention is directed to a method of controlling plant diseases
and promoting plant growth of plants in soil both under greenhouse and field
conditions which have the potential as biopesticide for the biocontrol of wide host
range of economically important pathogenic fungi.
The treatment can be applied directly to the seeds or plants, can be present in the soil
before planting or can be distributed, e.g., by spreading, spraying or the like, over the
crop or soil top or in soil furrow where the crop has been planted.
Sclerotia/chlamydospores of pathogenic fungi can be treated by treatment by soaking,
or other method known in the art for applying treatment to sclerotia and
chlamydospores.
Seeds can be treated by soaking with a composition containing the subject treatment
by spraying with the liquid, or other method known in the art for applying treatment
to seeds by well-known methods.
According to a further aspect the invention, seed-borne diseases can be prevented by
soaking the seeds with treatment by spraying with the liquid, or other method known
in the art for applying treatment to seeds by well-known methods.
The carriers that may be used to disperse the subject treatments would include all
those commonly used for inoculating crops and would include carriers such as peat,
vermicompost and fermented press mud.
According to this embodiment of the invention the optimum carrier may vary
depending on the treatment used. Any of the above compositions, liquids, powders,
oils or solid bases such as peat, soil, rice husk, vermiculite, caboxymethyl cellulose,
peat, vermicompost, perlite, polyvinyl-pyrrolidone, and talc and the like may have
nutrients included therein or and any other carrier agents. However, as demonstrated
by the examples below, vermicompost and fermented press mud are preferred.
Further aspect of this invention relates to a process whereby the synergistic
composition thus produced of the present invention may be used in any manner
known in the art for example, including pretreatment of soil or seeds or pregerminated
plant roots alone or in combination with other chemicals which is harmless to the
growth and survival of bacteria for example plant growth promoting compounds,
pesticides, fertilizers, peat, vermicompost, and fermented press mud.
The invention is further elaborated with the help of examples. However, the examples
should not be construed to limit the scope of the invention.
EXAMPLE 1
Collection and fermentation of cow urine11
Fresh urine from healthy indigenous (Sahiwal) cows was collected from Gajaria farm,
Department of Animal Husbandry, Government of Uttar Pradesh, Lucknow in
morning into clean plastic container. One liter urine was poured into 2 liter capacity
either copper or earthen pot buried in soil up to its neck. Pot was covered on top and
urine was fermented at temperature of 25-35°C, for 30 days. Level of the urine in the
pot was maintained by adding water as and when necessary. After 30 days, fermented
product was either used directly or boiled and concentrated to 0.25 liter, before its
application. In another combinations cow urine was fermented along with neem (250
gm leaves/liter) and/or garlic (100 gm crushed bulbs/liter). In yet another
combinations neem leaves and/or crushed garlic bulbs were fermented with water.
Like wise 12 treatments using urine, neem leaves and/or crushed garlic bulbs were
individually prepared in copper or earthen pot, each, to a final total of 24 treatments.
Initial work done at Go-Vigyan Anusandhan Kendra, Deolapar, Nagpur
EXAMPLE 2
Effect of 24 treatments on plant growth promotion ability of chickpea in
greenhouse
The experiment to examine the effect of 24 treatments on plant growth promotion
ability of chickpea in greenhouse was carried out in four different sets of 30 chickpea
seedlings each, for treated and non-treated seeds (control). Earthen pots (of 25 cm
diameter) in the replicate of 4 were used, to grow chickpea. Each pot was filled up
with non-sterilized soil. Tap water was added to each hole before planting seeds to
adjust the dry soil to 20% moisture with the 24 treatments individually diluted to
1:100. Six treated seeds were added per hole. In each set, data was noted after 21 days
of plant growth with respect to plant dry weight. The results are tabulated in the
following Table I . Treated plants demonstrated better plant dry weight, compared
with un-treated control. Among the treated plants, boiled urine+neem fermented in
earthen pot showed best results in terms of better seedling dry weight by showing
106% increase, compared with un-treated control.
TABLE 1
(Table Removed)
"initial work done at Go-Vigyan Anusandhan Kendra, Deolapar, Nagpur
It was discovered that treated plants demonstrated better plant dry weight, compared
with un-treated control. Among the treated plants, boiled urine+neem fermented in
earthen pot showed best results in terms of better seedling dry weight, compared with
un-treated control.
EXAMPLE 3
Effect of different dilution of boiled urine + neem fermented in earthen vessel on
plant growth promotion ability of chickpea, maize, wheat and pea in greenhouse
The experiment to examine the effect of boiled urine + neem fermented in earthen
vessel as described in Example 1, on plant growth promotion ability of chickpea
(Cicer iirietimun), maize (Zea mays), wheat (Trilicum vulgare) and pea (Pisum
sativwn) in greenhouse was carried out. Trays (35x35 cm.) with 16 (4x4) places per
iray (each space was of 7 cm. width, 10 cm. depth and 1 cm. apart from each other)
were used, to grow chickpea, maize, wheat and pea. Each place was filled up to 8 cm.
with non-sterilized soil. Tap water was added to each hole before planting seeds to
adjust the dry soil to 20% moisture with the product diluted as stated. Four seeds were
added per hole. The experiment in greenhouse was carried out in four different sets of
16 chickpea, maize, wheat and pea seedlings each, for non-treated (control) and
treated seeds. In each set, data of 30-days-old seedlings was noted with respect to dry
weight of plants. Results demonstrated that 1:10 dilution of the product urine + neem
fermented in earthen vessel treated plants demonstrated best plant dry weight,
compared with un-treated control, followed by 1:100 dilution (Table 2).
TABLE 2
(Table Removed)

It was discovered that 1:10 dilution of the product urine + neem fermented in earthen
vessel treated plants demonstrated best plant dry weight, compared with un-treated
control, followed by 1:100 dilution.
EXAMPLE 4
Effect of product containing urine + neem and urine + neem + garlic fermented
in earthen vessel under in vitro conditions for ability to suppress wide range of
plant pathogenic fungi
The treatments urine + neem and urine + neem + garlic fermented in earthen vessel,
obtained by the procedure outlined in Example 1 were screened for their ability to
inhibit growth of Fusarium oxysporum f. sp. ciceri, Fusarium oxysporum f. sp.
gladioli, Fusarium moniliforme, Alternaria solani, Alternaria alternata, Phytophthora
palmivora, Phytophthora nicotianae, Sclerotinia sclerotiorum, Sclerotium rolfsii,
Collelotrichum falcatum, Colletotrichum capsici, Penicillium sp., Aspergillus niger,
Rhizoctonia solani, Pythium aphanidermatum, Curvularia lunata and Phoma sorghi
under in vitro conditions as follows: An agar plug inoculum of the fungi to be tested
(5-mm square) was transferred to the center of the plate.
Four holes (5 mm diameter) were bored on each NA plate. In each hole 100 uL of the
treatments were individually added and in the 4th hole sterile water was added as a
negative control. In each hole 100 uL of the products from 24 treatments were
individually added. After incubation for 7 days inhibition zones were readily observed
in the case of treatments having the biocontrol activity as the fungal growth around
the hole was inhibited. While in case of treatments not having biocontrol activity,
fungal growth around the hole was not inhibited and the fungi grew around the edge
of the hole (Table 3). It was elucidated that the product containing urine + neem +
garlic fermented in earthen vessel demonstrated better ability to inhibit wide range
pathogenic fungi, under in vitro conditions, compared with the urine+neem product
fermented in earthen vessel, indicating its potential as biopesticide for the biocontrol
of wide range of economically important pathogenic fungi.
TABLE 3
(Table Removed)
It was discovered that the product containing urine + neem + garlic fermented in
earthen vessel demonstrated better ability to inhibit wide range pathogenic fungi,
under in vitro conditions, compared with urine+neem fermented in earthen vessel.
Therefore the use of product has the potential as biopesticide for the biocontrol of
wide host range of economically important pathogenic fungi.
EXAMPLE 5
Effect of various concentration of neem and garlic on the product containing
fermented in earthen vessel under in vitro conditions for ability to suppress
Phytophthora palmivora and promote growth of wheat11
Effect of various concentrations of neem and garlic in various combinations and
amounts (gm/liter) as indicated in Table 4 on the product containing fermented in
earthen vessel under in vitro conditions for ability to suppress Phytophthora
palmivora was evaluated as follows: An agar plug inoculum of the P. palmivora (5-
mm square) was transferred to the center of the plate. Four holes (5 mm diameter)
were bored on each NA plate. In each hole 100 uL of the treatments were individually
added and in the 4th hole sterile water was added as a negative control. While in case
of treatments not having biocontrol activity and the hole sterile water was added as a
negative control, fungal growth around the hole was not inhibited and the fungi grew
around the edge of the hole. In each hole 100 uL of the products from 24 treatments
were individually added. After incubation for 5 days inhibition zones were readily
observed in the case of treatments having the biocontrol activity as the fungal growth
around the hole was inhibited. While in case of treatments not having biocontrol
activity. /'. palmivora growth around the hole was not inhibited and the fungi grew
around the edge of the hole (Table 4).
The experiment in greenhouse was carried out in four different sets of 16 wheat
seedlings each, for non-treated (control) and treated seeds as described in Example 3
using 1:100 dilution of the treatments, individually. It was elucidated that the product
containing urine + neem (500 gm) + garlic (100 gm) and urine + neem (500 gm) +
garlic (250 gm) fermented in earthen vessel demonstrated better ability to inhibit P.
palmivora, under in vitro conditions, compared with urine + neem (250 gm) + garlic
(100 gm). There was not much difference among the treatments [Urine + neem (250
gm)+ garlic (100 gm) and Urine + neem (500 gm)+ garlic (100 gm)] with respect to
dry weight of wheat. Growth of wheat was 37% more in the presence of Urine + neem
(250 gm)+ garlic (250 gm), compared with 80% more in Urine + neem (250 gm)+
garlic (100 gm) and 83% more in Urine + neem (500 gm)+ garlic (100 gm)
treatments, compared with untreated wheat plants.
TABLE 4
(Table Removed)

It was discovered that the product containing urine + neem (500 gm) + garlic (100
gm) and urine + neem (500 gm) + garlic (250 gm) fermented in earthen vessel
demonstrated better ability to inhibit P. palmivora. under in vitro conditions,
compared with urine + neem (250 gm) + garlic (100 gm). However, in view of the
cost of the material involved and dry weight of wheat seedlings, product consisting of
urine + neem (250 gm) + garlic (100 gm) was used for further work.
EXAMPLE 6
Effect of various concentrations of the product containing urine + neem + garlic
fermented in earthen vessel for ability to inhibit growth of the sclerotia of
Sclerotium rolfsii, Rhizoctonia solani and Sclerotinia sclerotlorum and
chamydospores of Fusarium sp. under in vitro conditions
The product containing urine + neem + garlic fermented in earthen vessel, obtained by
the procedure outlined in Example 1 was screened lor its ability to inhibit growth of
the sclerotia of Sclerotium rolfsii, Rhizoctonia solani and Sclerotinia sclerotiorum and
chamydospores of Fusarium sp. under in vitro conditions as follows: Three-weeks old
similar sized sclerotia/chamydospores collected from NA plates were surface
sterilized and dipped in the treatment individually, as indicated for up to 6 hrs. In the
control set the sclerotia/chamydospores were suspended in sterile distilled water. The
viability of the sclerotia/chamydospores was monitored by removing and inoculating
3 batches of 15 sclerotia/chamydospores on NA after incubation at 28°C for 7 days.
Viability of sclerotia/chamydospores was recorded in terms of number of germinating
sclerotia/chamydospores by visual examination of mycelial growth after 7 days. After
incubation for 7 days mycelial growth was readily visible in the case of treatments
does not having biocontrol activity, as fungal growth around the scierotia was not
inhibited and mycelia grew around the sclerotia on the plate (Table 5). On the
contrary after incubation for 7 days mycelial growth was not visible in the case of
treatments having biocontrol activity, as fungal growth around the sclerotia was
inhibited and mycelia did not grew around the sclerotia on the plate (Table 5). Product
containing urine + neem + garlic fermented in earthen vessel demonstrated that
sclerotia of Sclerotium rolfsii, Rhizoctonia solani and Sclerotinia sclerotiorum and
chamydospores of Fusarium sp. was inhibited in 4, 2, 4 and 2 hrs respectively, under
in vitro conditions.
TABLE 5
(Table Removed)

It was discovered that the product containing urine + neem + garlic fermented in
earthen vessel inactivated sclerotia/chlamydospores' under in vitro conditions.
Therefore the use of product has the potential to protect plants from soil borne plant
pathogens forming sclerotia/chlamydospores.
EXAMPLE 7
Effect of various concentrations of 24 treatments by seed soaking and soil
drenching treatment to control collar rot of chickpea seedlings in green house
caused by Sclerotiutn rolfsii
The 24 treatments, obtained by the procedure outlined in Example 1 were screened for
their ability to inhibit growth of S. rolfsii by seed soaking and soil drenching
treatment to control collar rot of chickpea seedlings in green house as follows: For
seed soaking treatment seeds were treated by dipping in 10% dilutions of the
treatments individually, as indicated for 2 hrs. For soil drenching treatment the dry pot
soil was adjusted to 20% moisture with the product diluted 1:100 individually, as
stated. Products containing urine + neem + garlic fermented both in copper or earthen
vessel, and used as soil drenching method, demonstrated best biocontrol activity
against S. rolfsii under in vitro conditions, as 0% mortality was observed.
TABU- 6
(Table Removed)
It was discovered that the products containing urine + neem + garlic fermented both in
copper or earthen vessel, and used as soil drenching method, demonstrated best
biocontrol activity against S. rolfsii under in vitro conditions. Therefore the use of
product has the potential to protect plants by seed soaking and soil drenching
treatment, from plant pathogens.
EXAMPLE 8
Effect of various concentrations of the product containing urine + neem + garlic
fermented in earthen vessel by foliar spray treatment to control leaf spot disease
of field grown betelvine caused by Alternaria alternata
The product containing urine + neem + garlic fermented in earthen vessel, obtained by
the procedure outlined in Example 1 was screened for its ability to control leaf spot
disease of field grown betelvine caused by Alternaria alternata as follows: The field
experiment for management of leaf spot disease of betelvine with the help of various
concentrations of the product containing urine + neem + garlic fermented in earthen
vessel was conducted in the conservatory at the National Botanical Research Institute,
Lucknow, India in a randomized block design. The conservatory in which the
experiments were carried out was about 5 meter high and covered on all the sides with
wire nettings supported on angled iron pillars. Paddy straw was used on these nettings
to provide suitable conditions of shade and moisture needed for betelvine cultivation.
One-meter wide beds were prepared with a width of 35 cm and 10 cm high ridges.
The setts for sowing were selected from single node cuttings from two year old vines
growing in beds of 4.5 x 3 m2 area.
The cuttings were planted on either side of the ridges in April 2001, on a field plot
naturally infected with Alternaria alternata and covered with paddy straw and
sprinkled with water 4-5 times a day for nearly one month creating suitable moisture
conditions for the proper germination of vines. In each bed 15 vines of the plants were
maintained for each treatment. Disease symptoms on lower leaves were visible by
July, 2001. First spraying of the product was done at that time and was repeated by 5
more sprays at 15 days' interval till September, 2001. Disease intensity was noted 15
days after 5'h spray of the product. The products containing urine + neem + garlic
fermented in earthen vessel demonstrated ability to control leaf spot disease of field
grown betelvine caused by Alternaria alternata at 20% concentration.
TABLE 7
(Table Removed)

It was discovered that the products containing urine + neem + garlic fermented in
earthen vessel demonstrated ability to control leaf spot disease of field grown
betelvine caused by Alternaria allernata at 20% concentration. Therefore the use of
product has the potential to protect plants as an aerial spray from plant pathogens.
EXAMPLE 9
Effect of addition of boiled urine + neem + garlic fermented in earthen vessel on
the rhizosphere microorganisms, growth and nutrient accumulation of chickpea
plants
Effect of addition of 1:10 diluted product containing urine + neem + garlic fermented
in earthen vessel, obtained by the procedure outlined in Example 1, on the rhizosphere
microorganisms of chickpea plants was evaluated 4 weeks after planting as described
in Example 2. Dry weight of the treated plants was higher than that of control plants.
Reduction in the counts of treated microorganisms in the chickpea rhizosphere was
observed, compared with un-treated control. Bacterial population reduced by 1 log
unit, followed by reduction of 0.6 and 0.3 log unit of fungal and actinomycetes. Table
8 shows stimulation of the accumulation of N and P in the biomass of chickpea plants.
TABLE 8
(Table Removed)

Results demonstrate influence of boiled urine + neem + garlic fermented in earthen
vessel on the rhizosphere microorganisms. Therefore the use of product has the
potential as an effective agent to change the composition of soil microorganisms and
stimulation of the accumulation of nutrients in the biomass of plants.
EXAMPLE 10
Effect of addition of boiled urine + neem + garlic fermented in earthen vessel on
the percentage of rhizosphere microorganisms of chickpea plants demonstrating
plant growth promotion, phosphate solubilization, abiotic stress tolerance and
antagonism towards plant pathogenic fungi
Effect of addition of 1:10 diluted product containing urine + neem + garlic fermented
in earthen vessel, obtained by the procedure outlined in Example 1, on rhizosphere
microorganisms of 4 weeks old chickpea plants was evaluated by determining the
percentage of bacteria among the resident population, demonstrating plant growth
promotion, phosphate solubilization, abiotic stress tolerance and antagonism towards
plant pathogenic fungi. Changes in the percentage of bacteria among the resident
population, demonstrating plant growth promotion, phosphate solubilization, abiotic
stress tolerance and antagonism towards plant pathogenic fungi of treated
microorganisms in the chickpea rhizosphere was observed, compared with un-treated
control.
Percentage of gram-positive bacteria increased and that of gram-negative bacteria
decreased in the treated chickpea rhizosphere, compared with un-treated control.
Percentage of plant growth promotery and phosphate solubilizers and abiotic stress
tolerant bacteria increased in the treated chickpea rhizosphere, compared with untreated
control. Except for Curvularia lunata and Aspergillus niger percentage of
plant pathogenic fungal antagonistic bacteria against Fiisarium oxyxponim f. sp.
ciceri, Colletotrichum falcatum, Sclerotium rolfsii, Alternaria solani, Penicillium sp.,
Pythium aphanidermatum, Phytophthora palmivora, Sclerotinia sclerotiomm and
Phoma sorghi increased in the treated chickpea rhizosphere. compared with untreated
control.
TABLE 9
(Table Removed)

It was discovered that addition of boiled urine + neem + garlic fermented in earthen
vessel (on rhizosphere microorganisms of chickpea plants, resulted in stimulation of
proliferation of plant growth promoting, phosphate solubilizing, abiotic stress tolerant
and antagonists towards plant pathogenic fungi in the rhizosphere of plants. Therefore
the use of product has the potential to promote plant growth and protect plants from
plant pathogens,
EXAMPLE 11
Effect of addition of boiled urine + neem + garlic fermented in earthen vessel on
the percentage of rhizosphere microorganisms of chickpea plants demonstrating
plant growth promotion, phosphate solubilization and antagonism towards plant
pathogenic fungi in the presence of peat, vermicompost, fermented press mud
used as carrier and applied as furrow treatment'
Effect of addition of the product containing urine + neem + garlic fermented in
earthen vessel, obtained by the procedure outlined in Example 1, on the carriers to
disperse the subject treatment such as peat, vermicompost and press mud to 35%
moisture and applied as furrow treatment. For furrow-treatment, formulation was
directly applied all over the open furrow containing seeds and covered after treatment.
Chickpea plants were grown for 4 weeks to elucidate its effect on rhizosphere
microorganisms as described above by determining the percentage of bacteria among
the resident population, demonstrating plant growth promotion, phosphate
solubilization, and antagonism towards plant pathogenic fungi. Changes in the
percentage of bacteria among the resident population, demonstrating plant growth
promotion, phosphate solubilization and antagonism towards plant pathogenic fungi
of treated microorganisms in the chickpea rhizosphere was observed, compared with
un-treated control. As indicated in the Table 10, except for peat % population of plant
growth promotery, phosphate solubilizers and fungal antagonist bacterial population
increased in the treated chickpea rhizosphere, compared with control (without carrier).
TABLE 10
(Table Removed)

done in collaboration with Go-Vigyan Anusandhan Kendra, Deolapar, Nagpur
It was discovered that addition of boiled urine + neem + garlic fermented in earthen
vessel on rhizosphere microorganisms of chickpea plants, except for peat, along with
vermicompost and press mud resulted in stimulation of proliferation of plant growth
promoting, phosphate solubilizing bacteria, antagonists towards plant pathogenic
fungi in the rhizosphere of plants. Therefore the use of product along with carriers has
the potential to promote plant growth and protect plants from plant pathogens.
EXAMPLE 12
Effect of addition of boiled urine + neem + garlic fermented in earthen vessel on
the phenolic contents of chickpea plants
Effect of addition of 1:10 diluted product containing urine + neem + garlic fermented
in earthen vessel, obtained by the procedure outlined in Example 1, on the total
phenolic contents of chickpea plants was evaluated 4 weeks after planting. Increase in
the tolal phenolic compounds in the leaf and root of treated chickpea was observed,
compared with un-treated control.
TABLE 1 1
(Table Removed)

Results demonstrate influence of boiled urine + neem + garlic fermented in earthen
vessel on the total phenolic contents of the chickpea plants. Therefore the use of
product has the potential as an effective agent to enhance the total phenolic contents
44
of the plants. Greater accumulation of phenolic compounds or mediated induced
systemic resistance (ISR) in hosts offer a practical way of immunizing plants against
pathogen ingress
It is understood that the foregoing detailed description is given merely by way of
illustration and that modification and variations may be made therein without
departing from the spirit and scope of the invention.




We claim:
1. A synergistic composition useful in promoting plant growth, soil health and bio-controlling, wherein the said composition comprising:
[a] bovine urine - one liter;
[b] crushed neem leaves in concentration ranging between 10 to 750 grams/liter of bovine urine;
[c] crushed garlic bulbs in concentration ranging between 1 to 500 grams/liter of bovine urine;
[d] one or more carriers in concentration ranging between 10 to 1000 grams/liter of bovine urine.

2. A composition as claimed in claim 1, wherein the concentration of the neem leaves is 250 grams/liter of bovine urine.
3. A composition as claimed in claim 1, wherein the concentration of the crushed garlic bulbs is 100 grams/liter of bovine urine.
4. A composition as claimed in claim 1, wherein the bovine urine is cow urine.
5. A composition as claimed in claim 1, wherein the carrier is selected from the group consisting of vermicompost, soil, peat, rice husk, vermiculite, carboxymethyl cellulose, perlite, polyvinylpyrrolidone, talc, and fermented press mud.
6. A composition as claimed in claim 1, wherein the carrier is vermicompost or fermented press mud.
7. A process for the preparation of synergistic composition as claimed in claim 1, wherein the steps comprising:

a) collecting fresh urine from healthy bovine to provide bovine urine,
b) adding crushed neem leaves and crushed garlic bulbs to provide a resultant mixture, wherein the concentration of the crushed neem leaves is between 10 to 750 grams/liter of said bovine urine and the concentration of the crushed garlic bulbs is between 1 to 500 grams/liter of said bovine urine,
c) fermenting the resultant mixture of step [b] for a period of 30 days to obtain a composition, followed by adding one or more carriers to obtain the desired synergistic composition.

8. A synergistic composition useful in promoting plant growth and a process for the preparation thereof substantially as herein described with reference to the foregoing examples.



Documents:

2925-DELNP-2004-Abstract-(02-12-2008).pdf

2925-delnp-2004-abstract.pdf

2925-DELNP-2004-Claims-(02-12-2008).pdf

2925-DELNP-2004-Claims-(23-12-2008).pdf

2925-delnp-2004-claims.pdf

2925-DELNP-2004-Correspondence-Others-(02-12-2008).pdf

2925-DELNP-2004-Correspondence-Others-(23-12-2008).pdf

2925-delnp-2004-correspondence-others.pdf

2925-delnp-2004-description (complete).pdf

2925-DELNP-2004-Form-1-(02-12-2008).pdf

2925-delnp-2004-form-1.pdf

2925-delnp-2004-form-18.pdf

2925-DELNP-2004-Form-2-(02-12-2008).pdf

2925-delnp-2004-form-2.pdf

2925-DELNP-2004-Form-3-(02-12-2008).pdf

2925-delnp-2004-form-3.pdf

2925-delnp-2004-form-5.pdf

2925-DELNP-2004-Petition-137-(02-12-2008).pdf


Patent Number 227512
Indian Patent Application Number 2925/DELNP/2004
PG Journal Number 05/2009
Publication Date 30-Jan-2009
Grant Date 12-Jan-2009
Date of Filing 29-Sep-2004
Name of Patentee COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH
Applicant Address RAFI MARG,NEW DELHI-110 001,INDIA.
Inventors:
# Inventor's Name Inventor's Address
1 CHANDRA SHEKHAR NAUTIYAL NATIONAL BOTANICAL RESEARCH INSTITUTE LUCKNOW,INDIA.
2 HARIKESH BAHADUR SINGH NATIONAL BOTANICAL RESEARCH INSTITUTE LUCKNOW,INDIA.
3 SANGEETA MEHTA NATIONAL BOTANICAL RESEARCH INSTITUTE LUCKNOW,INDIA.
4 SUNIL BALKRISHNA MANSINGHKA NATIONAL BOTANICAL RESEARCH INSTITUTE LUCKNOW,INDIA.
5 SURESH HARIBHAU DAWLE NATIONAL BOTANICAL RESEARCH INSTITUTE LUCKNOW,INDIA.
6 NARYAN EKNATH RAJHANS NATIONAL BOTANICAL RESEARCH INSTITUTE LUCKNOW,INDIA.
7 PALPU PUSHPANGADAN NATIONAL BOTANICAL RESEARCH INSTITUTE LUCKNOW,INDIA.
PCT International Classification Number A01N 63/00
PCT International Application Number PCT/IN04/00078
PCT International Filing date 2004-03-31
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 60/45 8375 2003-03-31 U.S.A.