WO2011099020A1

WO2011099020A1 - Composition and method of preparation of fungal based product for controlling nematodes living in soil and damage to crops

Info

Publication number: WO2011099020A1
Application number: PCT/IN2010/000178
Authority: WO
Inventors: Chetan S. Patel
Original assignee: PATEL BABUBHAI C
Current assignee: PATEL BABUBHAI C
Priority date: 2010-02-09
Filing date: 2010-03-24
Publication date: 2011-08-18
Anticipated expiration: 2012-08-09

Abstract

This invention is based upon composition and method of preparing innovative combination of dormant spore of naturally occurring Paecilomyces lilacinus and Arthrobotrys sp. fungus with enzymes, fats and growth promoting molecules. Various nematode like root-knot nematode, sting nematode, spiral nematode, lesion nematode, stant nematode and most of the nematode causing injury to plant can be efficiently controlled. Upon each application, the dormant spore of Paecilomyces lilacinus and Arthrobotrys sp. gets active and during its growth, it adopts various mechanisms by which it controls nematode.

Description

Composition and Method of Preparation of Fungal Based Product for controlling Nematodes living in soil and damage to crops.

PREAMBLE OF INVENTION- This invention is particular described the nature of the invention and the manner in which it is to be performed.

FIELD OF INVENTION- The present invention relates generally to a composition and method of making fungal based product containing Paecilomyces lilacinus and Arthrobotrys sp. for controlling nematodes living in soil and preventing it from damaging the crops.

PRIOR ART-

In the existing system as given in United States Patent Application No. 4753799 - Granted Patent wherein the invention relates to Method and composition for an insecticide comprising a hydrated hydrogel capsule containing an insecticidal effective amount of at least one nematode capable of infecting an insect host, which capsule has sufficient hydration to maintain the viability and infectivity of said nematode, Also disclosed are methods for providing said insecticidal compositions.

In the existing system as given in United States Patent Application No. 6474259 - Granted Patent wherein disclosed are an apparatus and methods for in vivo mass production of insecticidal nematodes, which the efficiency and volume of nematode production. In the method, nematodes are cultured within a natural insect host. The apparatus comprises at least one harvesting area, a water dispensing system that promotes harvest of nematodes from the host organisms, and a water collection and concentration system for nematode collection and storage. The harvesting area comprises reusable stackable perforated trays, which allow passage of dispensed water while retaining the nematode hosts. The perforations are sized to retain the host organisms and facilitate the passage of harvested nematodes carried within the dispensed water.

In the existing system as given in United States Patent Application No. 20090298902 - Patent Application wherein the invention relates to a pesticide composition in the form of a water absorbent granular material and to the use of such compositions for combating arthropod pests, in particular soil-living arthropod pests, and nematodes, with particular preference given to soil-living insect pests. The pesticide composition according to the invention as a water absorbent granular material, which contains: i) from 0.001 to 10% by weight of at least one pesticide compound, in particular a pesticide compound that is active against said soil-living arthropod pest, ii) from 80 to 99.999% by weight of at least one granular super absorbent polymer, and iii) water, wherein the % by weight are based on the total weight of the composition, except for water and wherein the components i) and ii) make up at least 90% by weight of the composition except for water, and wherein the water absorbent granular material is obtainable by a process which comprises the treatment of superabsorbent polymer granules with a liquid aqueous composition containing the at least one pesticide compound..

In the existing system as given in United States Patent Application No. 20090048311 - Patent Application wherein the invention relates to compositions and processes for controlling nematodes are described herein, e.g., nematodes that infest plants or animals. The compounds include oxazoles, oxadiazoles and thiadiazoles.

In the existing system as given in United States Patent Application No. 20090018215 - Patent Application wherein the invention provides a fumigant for wood parasitic nematodes comprising methyl iodide dissolved in liquefied carbon dioxide, in place of methyl bromide the use of which is globally regulated as ozone depleting substance or a fumigant using methyl iodide dissolved in liquefied carbon dioxide in combination with one or more compounds selected from the group consisting of sulfuryl fluoride, methyl isothiocyanate, phosphine, ethylene oxide, carbonyl sulfide and propylene oxide, and a fumigation method for killing wood parasitic nematodes by using said fumigant.

In the existing system as given in United States Patent Application No. 5162376 - Granted Patent wherein the invention relates to methods of combating nematodes or of preventing crop plants from being attacked by nematodes, wherein an effective amount of an alkylbenzene of the general formula I ##STR1## where R.sup.l, R.sup.2 are each C.sub.l -C.sub.4 -alkyl; R.sup.3 is C.sub.l -C.sub.6 -alkyl, C.sub.3 -C.sub.6 -cycloalkyl, C.sub.2 -C.sub.4 -alkenyl or C.sub.2 -C.sub.4 -alkynyl; R.sup.4 is hydrogen, C.sub. l - C.sub.6 -alkyl or C.sub.2 -C.sub.4 -alkenyl; and R.sup.5 is C.sub. l -C.sub.6 -alkyl, C.sub.3 -C.sub.6 -cycloalkyl, C.sub.2 -C.sub.4 -alkenyl or C.sub.2 -C.sub.4 -alkynyl is allowed to act on the pests or their habitat, and the use of alkylbenzenes of the formula I for combating nematodes..

In the existing system as given Indian Patent Application No. 1519/ OLNP/2004 - Granted Patent wherein the invention relates to an excellent Meloidogyne-resistance gene and a method of using this gene. Namely, a novel Meloidogyne-resistance gene having qualitative resistance which shows no high-temperature sensitivity and is widely applicable to various species and strains of Meloidogyne; and Meloidogyne-resistant recombinant plants carrying the gene transferred thereinto

In the existing system as given in WIPO Patent Application No. PCT/US2001/002848 - Granted Patent wherein the invention relates to the methods and compositions for the control of nematodes are disclosed. Specifically, the subject substituted oxadiazole anthelmintic compounds have been found to advantageously control nematodes at concentrations which are non-phytotoxic. The anthelmintic compounds can be used in conjunction with other nematicidal agents such as free fatty acids, fatty acid salts, avermectins, ivermecin, and milbemycin. In another embodiment, the subject invention further provides methods for killing the eggs of nematodes. Thus, the subject invention further relates to the surprising discovery that certain compounds have ovicidal activity against nematode eggs.

In the existing system as given in WIPO Patent Application No. PCT US 1998/016952- Granted Patent wherein the invention concerns pest-control compositions and advantageous methods of using these compositions. More specifically, this invention relates to the use of bacteria to control plant-parasitic nematodes.

OBJECT OF THE INVENTION Traditionally, the known bacterial or fungal product for control of plant parasitic nematodes has different method of its preparation and composition. The present invention is based upon composition and method of making fungal based product containing Paecilomyces lilacinus and Arthrobotrys sp. for controlling all the nematodes living in soil and preventing it from damaging the crops.

STATEMENT OF INVENTION- The inventor has invented a composition and method of making a fungal based product to control plant-parasitic nematodes

DETAILED DESCRIPTION OF INVENTION-

Background of Invention: This invention relates to a method for utilizing fungal based product containing Paecilomyces lilacinus and Arthrobotrys sp. to control all the nematodes living in soil and preventing it from damaging the crops.

Nematodes are worms that survive in the ground and water, causing diseases. They attack human, animals and vegetate. They destroy roots of beetroot and cabbage - flower, e.g., causing a global damage for 100 million dollars by year on agriculture.

Damage to plants caused by nematodes is a prevalent and serious economic problem. Nematodes cause wide-spread and serious damage in many plant species.

The fungal based product containing Paecilomyces lilacinus and Arthrobotrys sp. controls plant-parasitic nematodes and prevents it from damaging crops.

The present invention is an innovative combination of dormant spore of naturally occurring Paecilomyces lilacinus and Arthrobotrys sp. Fungus with enzymes, fats and growth promoting molecules. Various nematode like root-knot nematode, sting nematode, spiral nematode, lesion nematode, stant nematode and most of the nematode causing injury to plant can be efficiently controlled. Upon each application, the dormant spore of Paecilomyces lilacinus and Arthrobotrys sp. gets active and during its growth, it adopts various mechanisms like mechanical trap, endozoice growth, parasitizing eggs masses etc. by which it controls nematode.

Material and Method used in preparing the above said innovative combination is as under Step-1 Nucleus culture (Maintain culture)

Pure culture of fungus (Paecilomyces lilacinus, Artherobotrys spp.) is inoculated aseptically on plate having 20 ml PAC-00 agar Media; such two to three plates are generally inoculated. Such inoculated plate is maintained in BOD incubator for at 26 ± 1 °C for 6 to 7 days. (Composition for one liter of PAC-00 Media is 300 gm potato, 20 gm dextrose, 20 gm agar agar and 1000 ml distilled water)

Step-2 Starter culture (In flask)

Culture grown on plate is inoculated aseptically in a flask (1000 ml capacity) having 500 ml PAC-00 broth media via cork borer (having 5 mm diameter cutting capacity), and allow to grow for at 26 ± 1 °C for at least 3 to 4 days. For superior growth, flask is put on shaker with agitation of media at 150 RPM. Such two flasks are maintained. (Composition for one liter of PAC-00 broth Media is 300 gm potato, 20 gm dextrose and 1000 ml distilled water)

Step-3 Seed Fermentor (In small fermentor)

One (1) lit culture grown in step-3 is inoculated aseptically in small fermenter having 10 litter PAC-00 media (Composition for one liter of PAC-00 broth Media is 300 gm potato, 20 gm dextrose and 1000 ml distilled water), and allow for further growth at 26 ± 1 °C for at least 3 to 4 days with agitation by motor at 100 RPM

Step-4 Add to solid substrate (poly bags)

20 ml culture form fermenter is inoculated aseptically in previously sterilized poly propelin polybag (12 x 10 inch) having 180 gm broken wheat gain. After inoculation of culture, mouth opening of poly bag seal with sponge and allow to develop. Such 500 bag get inoculated from 10 lit broth recovered from fermentor Step-5 Allow to develop

After inoculation each and every bag transfer to growth room and allows developing at 26 ± 1 °C for 9-10 days. During growth (four days after growth) all bag are crush smoothly by hand to break the clump of mycelium develops on grains. After 10 days of growth all content of bags are removed in trays and transfer to cold room where they maintain at 5 °C till the grinding of grain or submerging of grain having fungal growth in Ground nut oil.

Step-6 Grind/ Submerging

After sufficient cooling and drying (in cold room at 5 °C) for 10 to 15 days all the grain with the growth of microbes are grind in grinder or they were brought to submerge technique whereby all the grain were directly submerge in definite quantity of Ground nut oil and then oil-grain mixture is agitate by using commercial in tank agitator.

Step-7 Sieving

Grinding material sieve from 75 micron net with the help of vibro shifter and powder was collected by using cyclone separator, especially design for collecting of spore of microbes. And in case of submerge technique oil-grain mixture sieve from 300 micron sieve and collected material which have fungal spore and mycelium in oil brought to homogenizer.

Step-8 Mixing (Liquid carrier, emulsifier)

10 % fungal spore powder is than mixing in mineral oil (ground nut oil) and tween- 80 emulsifier mixture (90 % ground nut oil + 10 % tween-80) or in case of submerge technique, homogenize mixture having spore of fungus where again homogenized by adding 10 to 15 % tween-80 which act as a emulsifier.

Step-9 Curing

Mix formulation is allow to cure for two to three days in drums and than send for storage or for packing

Step-10 Packging

Form above prepared mixture 50 ml, 100 ml pouch is prepared by using pouch packing machine

Step-11 - Distribution to farmer, retailer, distributor etc. The final material prepared has to be used in following form: Dose in one Acre: Liquid Form: 50 ml to 100 ml

Claims

CLAIMS I claim,

1. A composition and method of preparing innovative combination of dormant spore of naturally occurring Paecilomyces lilacinus and Arthrobotrys sp. fungus with enzymes, fats and growth promoting molecules to control plant-parasitic nematodes, the method comprising steps of nucleus culture, starter culture, seed fermentor, addition to solid substrate, allowing to develop, grinding, sieving, mixing, curing and packing the final output.

2. A composition and method of preparing fungal based product according to claim 1, by inoculating aseptically pure culture of fungus namely Paecilomyces lilacinus, Arthrobotrys sp. on two to three plates having 20 ml PAC-00 agar Media and maintaining in BOD incubator for at 26 ± 1 °C for 6 to 7 days, wherein composition for one liter of PAC-00 Media containing 300 gm potato, 20 gm dextrose, 20 gm agar agar and 1000 ml distilled water.

3. A composition and method of preparing fungal based product according to claim 1, by inoculating aseptically culture grown on plate as per method cited in claim 2 in a flask of 1000 ml capacity having 500 ml PAC-00 broth media via cork borer having 5 mm diameter cutting capacity, and allowing to grow for at 26 ± 1 °C for at least 3 to 4 days, wherein for superior growth, flask is put on shaker with agitation of media at 150 RPM (two flasks are maintained), wherein, composition for one liter of PAC-00 broth Media contains 300 gm potato, 20 gm dextrose and 1000 ml distilled water.

4'. A composition and method of preparing fungal based product according to claim 1, wherein, one (1) lit culture grown as per method cited in claim 3 is inoculated aseptically in small fermenter having 10 litter PAC-00 media, wherein composition for one liter of PAC-00 broth Media contains 300 gm potato, 20 gm dextrose and 1000 ml distilled water), and allow for further growth at 26 ± 1 °C for at least 3 to 4 days with agitation by motor at 100 RPM

5. A composition and method of preparing fungal based product according to claim 1, wherein, 20 ml culture form fermenter is inoculated aseptically in previously sterilized poly propelin polybag (12 x 10 inch) having 180 gm broken wheat gain and after inoculation of culture mouth opening of poly bag seal with sponge and allowing to develop resulting in getting 500 bag inoculated from 10 lit broth recovered from fermentor

6. A composition and method of preparing fungal based product according to claim 1 , wherein, after inoculation each and every bag is transferred to growth room and is allowed to develop at 26 ± 1 °C for 9-10 days, wherein during growth (four days after growth) all bag are crushed smoothly by hand to break the clump of mycelium develops on grains and after 10 days of growth all content of bags are removed in trays and transfer to cold room where they are maintained at 5 °C till the grinding of grin or submerging of grain having fungal growth in Ground nut oil.

7. A composition and method of preparing fungal based product according to claim 1, wherein, after sufficient cooling and drying in cold room at 5 °C for 10 to 15 days, all the grain with the growth of microbes are grinded in grinder or they were brought to submerge technique whereby all the grain were directly submerge in definite quantity of Ground nut oil and then oil-grain mixture is agitate by using commercial in tank agitator.

8. A composition and method of preparing fungal based product according to claim 1, wherein, grinding material sieve from 75 micron net with the help of vibro shifter and powder is collected by using cyclone separator, especially design for collecting of spore of microbes. And in case of submerge technique oil-grain mixture sieve from 300 micron sieve and collected material which have fungal spore and mycelium in oil brought to homogenizer.

9. A composition and method of preparing fungal based product according to claim 1 , wherein, 10 % fungal spore powder is than mixing in mineral oil (ground nut oil) and tween- 80 emulsifier mixture (90 % ground nut oil + 10 % tween-80). or in case of submerge technique homogenize mixture having spore of fungus where again homogenized by adding 10 to 15 % tween-80 which act as a emulsifier.

1 0. A coinposition and method of preparing fungal based product according to claim 1, wherein, mixed formulation is allowed to cure for two to three days in drums and than send for storage or for packing in 50 ml, 100 ml pouch prepared by using pouch packing machine and same when applied in dose of 50 ml in liquid form in soil is effective against controlling plant-parasitic nematodes.