PREPARATION ABLE TO PRODUCE BIOPESTICIDE AND/OR REPELLENT FOR CONTROLLING PLANT PATHOGENS

Abstract

A preparation able to produce a biopesticide and/or repellent for controlling plant pathogens, comprising at least one nanoporous material and at least one active enzyme, the at least one active enzyme being configured to transform at least one precursor into a biopesticide and/or repellent product. The preparation able to produce a biopesticide and/or repellent for controlling plant pathogens is remarkable in that the at least one active enzyme is immobilized within the at least one nanoporous material.

Claims

1-15. (canceled)

16. A preparation able to produce at least one insecticides and nematicides for controlling plant pathogens, said preparation comprising: a) at least one nanoporous material, and b) at least one active enzyme selected from the group of glycosidase, the at least one active enzyme selected from the group of glycosidase being configured to transform at least one precursor into at least one of insecticides and nematicides product, wherein the at least one active enzyme selected from the group of glycosidase is immobilized within the at least one nanoporous material.

17. The preparation according to claim 16, wherein the at least one nanoporous material is embedded within one hydrophilic gel,

18. The preparation according to claim 17, wherein the hydrophilic gel is made of at least one of agarose, agar-agar, alginate, pectin, starch and gelatine.

19. The preparation according to claim 16, wherein the at least one active enzyme from the group of glycosidase is thioglucosidase.

20. The preparation according to claim 16, wherein the at least one active enzyme, selected from the group of glycosidase, immobilized within the at least one nanoporous material, is one of covalently bounded to the at least one nanoporous material, physically adsorbed to the at least one nanoporous material, encapsulated within the at least one nanoporous material, and entrapped within the at least one nanoporous material.

21. The preparation according to claim 16, wherein the active enzyme, selected from the group of glycosidase, immobilized within the at least one nanoporous material is cross-linked with glutaraldehyde or with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide.

22. The preparation according to claim 16, wherein the at least one nanoporous material is a silica-based mesoporous material.

23. The preparation according to claim 16, wherein the preparation one of comprises or is adapted to receive, at least one of the at least one precursor, at least one mineral, and at least one cofactor.

24. The preparation according to claim 23, wherein at least one of the at least one mineral and the at least one cofactor is one of a phosphate buffer and an ascorbic acid.

25. A fibre adapted to control plant pathogens, wherein the fibre is coated with at least one preparation able to produce at least one of insecticides and nematicides for controlling plant pathogens, wherein the preparation comprising a) at least one nanoporous material, and b) at least one active enzyme selected from the group of glycosidase, the at least one active enzyme selected from the group of glycosidase being configured to transform at least one precursor into the at least one of insecticides and nematicides product, wherein the at least one active enzyme selected from the group of glycosidase is immobilized within the at least one nanoporous material.

26. The fibre according to claim 25, wherein the at least one preparation is further coated with a protective layer.

27. The fibre according to claim 26, wherein the protective layer is a hydrophobic layer.

28. A pest protective apparatus, said apparatus comprising at least one of at least one fibrous network and at least one fibrous cover, wherein the at least one of the at least one fibrous network and the at least one fibrous cover is made of fibres adapted to control plant pathogens, wherein the fibre is coated with at least one preparation able to produce at least one of insecticides and nematicides for controlling plant pathogens, wherein the preparation comprising a) at least one nanoporous material, and b) at least one active enzyme selected from the group of glycosidase, the at least one active enzyme selected from the group of glycosidase being configured to transform at least one precursor into the at least one of insecticides and nematicides product, wherein the at least one active enzyme selected from the group of glycosidase is immobilized within the at least one nanoporous material.

29. The pest protective apparatus according to claim 28, wherein the pest protective apparatus further comprises a means for distributing water to at least one of the fibrous network and the fibrous cover.

30. The pest protective apparatus according to claim 29, wherein the means comprises a cartridge configured for being in fluid connection with at least one of the at least one fibrous network and the at least one fibrous cover and for being fed with water.

31. A method for controlling plant pathogens with a preparation able to produce at least one of insecticides and nematicides for controlling plant pathogens, wherein said method comprises the following steps: activating at least one precursor by addition of water to the preparation able to produce at least one of insecticides and nematicides for controlling plant pathogens, wherein the preparation comprises: a) at least one nanoporous material, and b) at least one active enzyme selected from the group of glycosidase, the at least one active enzyme selected from the group of glycosidase being configured to transform at least one precursor into the at least one of insecticides and nematicides product, wherein the at least one active enzyme selected from the group of glycosidase is immobilized within the at least one nanoporous material; and applying the at least one of the insecticides and the nematicides product to plants in order to prevent pest attacks.

32. The method for controlling plant pathogens according to claim 31, wherein the at least one precursor is comprised into the preparation able to produce the at least one of the insecticides and the nematicides for controlling plant pathogens.

33. The method for controlling plant pathogens according to claim 31, wherein the preparation also comprises at least one of at least one mineral and at least one cofactor.

34. The method for controlling plant pathogens according to claim 31, wherein the at least one precursor is incorporated into the cartridge of a pest protective apparatus, wherein the pest protective apparatus comprises at least one of at least one fibrous network and at least one fibrous cover, wherein the at least one of the at least one fibrous network and the at least one fibrous cover is made of fibres adapted to control plant pathogens, wherein the fibre is coated with at least one preparation able to produce the at least one of insecticides and nematicides for controlling plant pathogens, wherein the preparation comprising a) at least one nanoporous material, and b) at least one active enzyme selected from the group of glycosidase, the at least one active enzyme selected from the group of glycosidase being configured to transform at least one precursor into the at least one of insecticides and nematicides product, wherein the at least one active enzyme selected from the group of glycosidase is immobilized within the at least one nanoporous material.

35. The method for controlling plant pathogens according to claim 34, wherein the at least one precursor is incorporated into the cartridge of the pest protective apparatus with at least one of at least one mineral and at least one cofactor.

Description

DRAWINGS

[0049] FIG. 1 illustrates a preparation able to produce a biopesticide and/or repellent for controlling plant pathogens in accordance with various embodiments of the invention.

[0050] FIG. 2 illustrates fibres adapted to control plant pathogens in accordance with various embodiments of the invention.

[0051] FIG. 3 illustrates a pest protective apparatus in accordance with various embodiments of the invention.

DETAILED DESCRIPTION

[0052] The present invention relates to a pest management biocontrol system made of active biocatalysts able to produce natural repellents and/or biopesticides by means of controlled enzymatic reactions.

[0053] The present invention offers to the user an easy-to-handle and safe way to control plant pest by producing a constant flux of biopesticide and/or repellent products, being insecticides, fungicides, bactericides and/or nematicides.

[0054] FIG. 1 depicts a preparation 100, able to produce a biopesticide and/or repellent for controlling plant pathogens, according to the present invention.

[0055] The biopesticide and/or repellent compounds produced are those involved in the natural reaction of the plants when facing a pathogen attack. They have a low half-life in the environment and do not affect the quality of the plants which are treated.

[0056] The present invention relates to an innovative product that entraps enzymes 4 into a nanoporous material 2, in particular a silica sphere, in various instances into a mesoporous silica matrix, more particularly into a silica-based mesoporous material, the whole being glued or embedded into a hydrophilic gel 6, to degrade natural molecules or extracts into products 10 having biopesticide and/or repellent activity.

[0057] The mesoporous materials 2 of the present invention are ordered silica-based mesoporous particles with a narrow pore size distribution, a well-defined pore geometry and a well-defined pore connectivity.

[0058] The pore size distribution and the global geometry of the mesoporous materials 2 are defined according to the properties of the biopesticide-producing enzymes and/or repellent-producing enzymes that are immobilized into the mesoporous structure.

[0059] The pore size of the silica-based mesoporous material is comprised between 5 nm and 30 nm, more particularly 5 and 15 nm for thioglucosidase.

[0060] The immobilization procedure should allow to optimize the catalytic activities of the enzymes in comparison with the free enzymes and to enhance their reusability in order to produce a constant and sufficient flux of biopesticide and/or repellent product allowing a sufficient biopesticide and/or repellent activity against the targeted pathogens.

[0061] The silica-based mesoporous material will be synthesized in acidic conditions using a silica precursor that could be among other tetraethyl orthosilicate (TEOS) and/or tetramethyl orthosilicate (TMOS) and/or methyltrimethoxysilane (MTMOS) and/or (3-aminopropyl)triemthoxysilane (APTMOS), in combination with tri-block copolymer mixtures such as PluronicF127 and/or PluronicF123 as structure-directing agents. Numerous methods are known by the skilled person in the art to achieve this synthesis.

[0062] The mesoporous structure can be produced hydrothermally and/or by sol-gel synthesis and can be functionalized.

[0063] The enzymes 4 that can be immobilized into the mesoporous material are those involved in the natural response of the plants when facing a pathogen attack. The biopesticide and/or repellent compounds produced during the enzymatic reaction can be a bactericide, a fungicide, an insecticide and/or a nematicide.

[0064] The enzyme from the glycosidase, the oxidoreductases, the transferases, the hydrolases, the lyases, the isomerases and the ligases can be immobilized into the mesoporous material.

[0065] Exemplary transferases are those belonging to the glycosidases class, i.e. the enzymes hydrolysing O-glycosyl and S-glycosyl compounds.

[0066] Exemplary lyases are, for example, carbon-sulfur lyases, in particular alliin lyase (also known as alliinase).

[0067] Enzymes from the PF10604 family, i.e. the lachrymatory-factor synthase, can also be used in the present invention.

[0068] An exemplary glycosidase is thioglucosidase. This is the enzyme of choice which has been tested for transforming a precursor into a biopesticide and/or repellent product.

[0069] The immobilization procedure can be performed through the following techniques known in the art: cross-linking methods, covalent binding, physical adsorption, encapsulation and entrapment.

[0070] The immobilization includes the penetration of the enzymes inside the mesoporous material, through the nanopores and an eventual step of cross-linkage between the enzymes using gluturaldehyde or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide. Numerous methods are known by the skilled person in the art to achieve the immobilization step.

[0071] The mesoporous biocatalyst will then be embedded in a hydrophilic gel 6 made of agarose, agar-agar, alginate, pectin, starch, gelatine taken alone or in mixes, in various instances alginate.

[0072] The use of alginate allows a gelation without heating. Only CaCl.sub.2 must be added for the gelation step.

[0073] The hydrophilic gel 6 allows the diffusion of the products of the enzymatic reaction, the diffusion of the substrates 8 of the enzymes 4 and the confinement of all chemicals required to maintain the enzymes 4 in a working status.

[0074] The hydrophilic gel 6 can contain all minerals and cofactors indispensable for the enzymatic activities. This can be a phosphate butter (pH=6.1) at a concentration comprised between 10 mM and 500 mM, in various instances at a concentration of 100 mM and/or ascorbic acid at a concentration comprised between 50 M and 1500 M, in various instances at a concentration of 500 M.

[0075] In case the minerals and cofactors are not added before the gelation step, they can be added by diffusion within the gel by soaking into an aqueous buffer comprising these compounds.

[0076] The pH of the preparation 100 able to produce biopesticide and/or repellent for controlling plant pathogens is in various instances comprised between 4 and 11. In particular, the pH of the hydrophilic gel allows an optimal activity of the immobilized enzymes.

[0077] The hydrophilic gel with the silica-based mesoporous material embedded within the hydrophilic gel, the silica-based mesoporous comprising immobilized thioglucosidase, forms a biopesticide and/or repellent preparation 100 adapted to control plant pathogen.

[0078] Upon addition of the substrate 8 for the enzyme 4 and upon addition of water, and, upon addition of minerals and cofactors for activating the enzyme 4 if they are not formerly included within the hydrophilic gel 6, the enzyme will proceed to the synthesis of a product 10 which will present biopesticide and/or repellent properties. This product 10 will be released and will act to eliminate (in case of biopesticide activity) and/or to keep away (in case of repellent activity) the pest and/or the plant pathogens from the living plants, such as bacteria, fungi, insects, bugs, and/or nematodes.

[0079] The scheme on FIG. 1 represents in particular a precursor 8 composed of two chemical moieties, one being the biopesticide and/or the repellent product 10, and the other being a compound 22.

[0080] In various instances, the product 10 of the enzymatic reaction is a volatile organic compound.

[0081] The biopesticide and/or repellent preparation 100 can further be coated onto fibres 200 or natural fibres. This is schematically depicted on FIG. 2.

[0082] Such fibres might be for example hemp, flax, nettle, cotton, jute, ramie, sisal, and/or any other.

[0083] In various instances, the fibres are hemp or flax.

[0084] The substrate 8 of the enzyme 4 is able to diffuse within the fibre 200. This will allow the substrate 8 to reach the preparation 100 able to produce biopesticide and/or repellent for controlling plant pathogens in order to be processed by the enzyme 4.

[0085] In order to protect such fibres 200 from drying, a protective layer, in various instances a hydrophobic layer 12 will be coated over the fibres 200 containing a coating of the preparation 100 able to produce biopesticide and/or repellent for controlling plant pathogens.

[0086] Such hydrophobic layer 12 is also configured to be permeable to the biopesticide and/or repellent product 10.

[0087] The hydrophobic layer 12 can be (3-aminopropyl)triethoxysilane, (3-mercaptopropyl)triethoxysilane, succinic anhydride, alkylketene dimer, 3-isopropenyl---dimethylbenzyl isocyanate, m-phenylene bismaleimide, vinyl trialkoxysilane, 3-metacryloyloxy propyl trimetoxysilane and/or any other.

[0088] The moiety 22, resulting from the cleavage of the precursor 8, stays therefore or diffuses slowly within the preparation 100 able to produce biopesticide and/or repellent for controlling plant pathogens and does not therefore pollute the environment.

[0089] As indicated on FIG. 3, the fibres 200 with the biopesticide and/or repellent preparation 100 might also be incorporated within a pest protective apparatus 300, more particularly by coating onto at least one fibrous network and/or at least one fibrous cover 14 of the pest protective apparatus 300.

[0090] The pest protective apparatus 300 of the present invention can also comprise means for distributing water to the at least one fibrous network and/or the at least one fibrous cover 14, the means in various instances comprising a cartridge 16. The cartridge 16 is configured for being in fluid connection with the at least one fibrous network and/or the at least one fibrous cover 14 and for being fed with water. The cartridge 16 can comprise an inlet 18 and an outlet 20.

[0091] Depending whether the pest protective apparatus 300 comprises or does not comprise the cartridge 16, the apparatus is configured either to transform at least one precursor 8, which is incorporated to the preparation 100 able to produce biopesticide and/or repellent for controlling plant pathogens by diffusion through the fibre 200, into a biopesticide and/or a repellent product 10; or to incorporate the precursor 8 into the cartridge 16.

[0092] Depending whether the pest protective apparatus 300 comprises or does not comprise the cartridge 16, the at least one mineral and/or the at least one cofactor indispensable for the enzyme activity are incorporated either directly within the hydrophilic gel 6 of the biopesticide and/or repellent preparation 100; or within the cartridge 16.

[0093] In the case of the preparation 100 able to produce biopesticide and/or repellent for controlling plant pathogens, in the case of the fibres, as well as in the case of the pest protective apparatus of the present invention, the enzymatic reaction is triggered by the addition of water, which plays the role of the solvent of the reaction, bringing subsequently the substrate 8 into contact with the fibres 200 of the preparation 100 able to produce biopesticide and/or repellent for controlling plant pathogens, and more particularly, into contact with the active site of the enzyme 4.

[0094] The second utility of water is to bring to the enzyme 4 the at least one mineral and/or the at least one cofactor indispensable for its activity.

[0095] Water can come from the rain and/or from artificial means, such for example a drain pump, a hosepipe and/or an irrigation system, and can be channelled to contact the preparation 100 able to produce biopesticide and/or repellent for controlling plant pathogens, triggering subsequently the enzymatic reaction.

[0096] When the pest protective apparatus 300 comprises a cartridge 16, the flux of water reaches the cartridge 16 through the inlet 18 which is provided on the cartridge.

[0097] Once the enzymatic reaction is over, the product 10, which presents biopesticide and/or repellent properties, is released. Generally, the product is a volatile organic compound.

[0098] This product acts to eliminate (in case of biopesticide activity) and/or to keep away (in case of repellent activity) the pests and/or the plant pathogens from the living plants, such as bacteria, fungi, insects, bugs, and/or nematodes.

[0099] The biopesticide and/or repellent product 10 resulting from the activation of the precursor by addition of water to the preparation 100 taught in the present invention is applied to plants, in various instances vegetables and/or fruits, in order to prevent pest attacks.