PESTICIDE COMPOSITIONS CONTAINING MUSTARD BRAN

Abstract: The invention provides pesticide precursor compositions comprising bran from a mustard of the genus Brassica and methods for controlling soil pests by application of the pesticide precursor compositions to the soil. The invention further provides pesticides comprising aqueous suspensions or extracts of mustard bran.

Description: PESTICIDE COMPOSITIONS CONTAINING MUSTARD BRAN Field of the Invention The present invention relates to compositions and methods for controlling pests. More particularly, the invention relates to pesticide precursor compositions comprising mustard bran and methods for using these compositions. Background of the Invention Soil pests affect most cultivated crops and cause extensive damage, for example, to fruit crops, vegetables, cereals and ornamental plants, with resulting economic loss. Efforts have been made to control soil pests by introducing pesticidal formulations onto or into the soil, before planting, at the time of planting or after planting. For example, the synthetic compound methyl isothiocyanate has been used as a soil fumigant against a variety of pests, including insects, nematodes, mites and various microorganisms. In recent years, however, it has been realised that many of the pesticides currently in use are toxic and environmentally undesirable and there is a need to find practical alternatives which are more environmentally friendly. Many Brassica species contain glucosinolates which can be converted by the endogenous plant enzyme myrosinase to a variety of compounds. For example, the glucosinolate sinigrin, which is found in mustards, can be converted by myrosinase to allyl isothiocyanate (AITC) . As early as 1945, mustard oil and AITC were examined as a means of control of nematodes (Ellenby (1945), Annals of Applied Biol., v. 32, pp. 67-70; (1951), Annals of Applied Biol., v. 38, pp. 859-875). Mustard oil did not, however, prove to be a practical pesticide for agricultural application due to its handling reguire ents, since it is an irritant liquid producing noxious fumes, and due to its iπmiscibility with soil moisture. There remains a need for a practical and cost- effective treatment for soil pests which is at the same time environmentally- and user-friendly. Summary of the Invention In the commercial preparation of mustard condiments, whole mustard seed is milled to strip off the outer seed coating or husk from the kernel or seed tissue. The kernel is ground into mustard flour which is sold as dry mustard condiment powder or is processed into other mustard condiments. The milled husk or bran is a waste product previously thought to have no commercial value and therefore discarded. The present inventors have found, surprisingly, that mustard bran from some mustard species is a significant source of sinigrin and AITC and can be used as an inex¬ pensive, effective and safely handled pesticide or soil fumigant, to control pests such as insects, nematodes, mites, fungi and bacteria. Mustard bran can be applied to the soil in a manner similar to the application of a granular fertiliser. The mustard bran may be mixed with a suitable agricultural carrier to optimise its handling properties. On contact with water in the soil, the sinigrin of the mustard bran is converted by myrosinase from the bran to the active pesticide, AITC. Mustard bran may also be used to prepare pesticidal aqueous extracts or aqueous suspensions which can be applied to pests or to the soil to achieve control. In accordance with one embodiment of the present invention, there is provided a pesticide precursor compo¬ sition comprising bran from a mustard of the genus Brassica. In accordance with a further embodiment of the pres¬ ent invention, a method is provided for controlling a pest in soil which comprises applying to the soil an effective amount of a pesticide precursor composition comprising a bran from a mustard of the genus Brassica, wherein the bran contains an effective amount of sinigrin capable of conversion to allyl isothiocyaate on contact with water in the soil. In accordance with a further embodiment of the invention, there is provided a pesticide comprising a pesticidally effective amount of an aqueous extract of bran from a mustard of the genus Brassica. In accordance with a further embodiment of the invention, there is provided a pesticide comprising a pesticidally effective amount of an aqueous suspension of bran from a mustard of the genus Brassica. In accordance with a further embodiment of the invention, a method is provided for controlling a pest which comprises applying to the pest an effective amount of an aqueous extract of bran from a mustard of the genus Brassica. In accordance with a further embodiment of the invention, a method is provided for controlling a pest which comprises applying to the pest an effective amount of an aqueous suspension of bran from a mustard of the genus Brassica. Detailed Description of the Invention The terms listed below have the following meanings: "Mustard" means a species or a plant of the genus Brassica. "Mustard condiment" means a food product or condiment prepared from a plant or from the seeds of a plant of the genus Brassica. "Bran" or "mustard bran" means the husk or outer seed coating from the seeds of a plant of the genus Brassica removed from the kernel by milling or other suitable process. Mustards with a high seed kernel concentration of sinigrin are valued for human consumption as seasonings and are used for the preparation of mustard condiments and other food products. The mustard seed is first milled to remove the outer husk, typically by an abrasive process, and the kernel is ground into flour for condiment production. Until the work of the present inventors, the residual bran was considered a waste product and was discarded. It has now been shown that mustard bran can be a useful and effective pesticide precursor, providing sinigrin and myrosinase. Under moist conditions, the sinigrin is converted by the myrosinase to the pesticidal compound, AITC. This is exemplified in Example 2 in which aqueous extracts of mustard bran were compared with extracts of mustard flour, which is known to contain high levels of sinigrin, with respect to nematicidal activity. Bran was found to have excellent nematicidal activity. In con- trast, extracts of whole mustard seed or cracked mustard seed showed very low nematicidal activity. In accordance with one embodiment of the invention, a method is provided for controlling soil pests by apply¬ ing a pesticide precursor composition comprising mustard bran to the soil. Mustard bran may be applied to the soil in a similar manner to the application of materials such as granular fertilisers and using similar commercially available agricultural equipment. For example, the bran may be scattered on the soil surface or incorporated into the soil with a disk or rototiller. Application may be prior to planting, at the time of planting or after planting, as desired. In accordance with a further embodiment of the invention, mustard bran may be mixed with an agriculturally acceptable carrier to improve its handling properties before application to the soil. Those skilled in the art will be familiar with carrier materials which are suitable for formulation of pesticidal compositions for application to soil. Such carriers include finely ground clay, starch-based or plastic polymers, surfactants or organic carriers such as ground corn cobs. Cracked mustard seed may also be used as a carrier. Lower grade mustard seed of insufficient quality for processing into flour for condiment preparation may be employed for this purpose. In accordance with a further embodiment, mustard bran may be formulated with a mixture of carriers, and optionally may be formulated in a variety of forms; for example, it may be formulated into layered granules, to obtain a desired rate of release of pesticide into the soil. Such methods of formulation are known to those skilled in the art. Mustard bran is an environmentally friendly pesti¬ cide, being a natural and biodegradable product. It is also much more easily and safely handled than pesticides such as synthetic methyl isothiocyanate, as the dry bran is non-toxic and the effective pesticide, the irritant and toxic compound AITC, is formed from the sinigrin contained in the bran only after application of the composition to the soil and its contact with water in the soil. The dry bran may be formulated, handled and applied without the precautions which would be required for deal¬ ing with AITC. In accordance with a further embodiment of the invention, a pesticide comprising an aqueous extract or an aqueous suspension of mustard bran may be prepared and applied to pests or to the soil to achieve pest control. Application may be carried out, for example, by spraying the suspension or extract on the pests or on to the soil. As will be understood by those skilled in the art, mustard bran effective as a pesticide precursor may be obtained from any Brassica species or variety having a husk sinigrin content sufficient to give a desired level of pest control on conversion to AITC. As exemplified herein, a mustard having a sinigrin content of about 2.5% by weight provides an effective level of pesticide activ- ity. Sinigrin may be measured by a standard technique, such as those described by Minchinton, I. et al., (1982) , J. Chromatog. , v. 247, pp. 141-148; Sang, J.P. et al., (1984), Can. J. Plant.Sci., v. 64, pp. 77-93; Betz, J.M. and Fox, W.D., (1994), in "Food Phytochemicals for Cancer Prevention", ACS Symposium Series 546, American Chemical Society, pp. 181-193. Mustards having a higher or lower husk sinigrin content may also be employed in the pesticide precursor compositions of the invention. It is within the expert- ise of those skilled in the art to determine whether the bran from any variety of mustard is effective to give a desired level of pest control. Suitable Brassica varieties may be found, for example, in the following species: Brassica juncea ; Brassica campestris ; and Brassica nigra . Brassica juncea , cv Forge and Brassica juncea , cv Cutlass are preferred varieties of B . juncea . Brassica species or varieties may also be bred or genetically engineered to increase husk content of sinigrin. Brassicas such as Brassica napus have already been shown to be amenable to successful genetic manipula¬ tion to improve various properties. Such techniques may be used to convert species or varieties with naturally low husk sinigrin content to a useful level or may be used to further enhance husk sinigrin in high-sinigrin varieties. Transgenic or hybrid Brassicas so produced may also be used in the compositions of the invention. As AITC is believed to act against pests by a simi¬ lar mechanism to that of methyl isothiocyanate, the pes¬ ticide precursor compositions and pesticides of the invention can be expected to combat a similar range of pests, including plant-parasitic nematodes, insects, mites, fungi and bacteria. The following examples are described for the purpose of illustration and are not intended to limit the scope of the invention. Example 1 Samples of mustard bran and mustard flour from vari¬ ous mustard types were obtained from a commercial mustard milling company. Sinigrin content was determined by extracting a portion of bran or flour in phosphate buffer, pH 7.0, at 100°C for 20 min. The aqueous extract was purified by chromatography on a Clδ-solid phase extraction cartridge and the purified sinigrin was determined by high perform¬ ance liquid chromatography. The available AITC was calculated as 23.9% of the sinigrin concentration, on the basis of the relative molecular weights and the demonstrated quantitative con- version of sinigrin to AITC by myrosinase (data not shown) . The results are shown in Table 1. TABLE 1 Mustard Sinigrin Cone, AITC Cone. Variety (mg/g) (mg/g) Bran Flour Bran Flour Oriental" 25.2 40.2 6.0 9.6 Yellow" nd 1.9 nd 0.5 Mixed c 9.3 24.7 2.2 5.9 Brassica juncea , cv. Forge (Oriental mustard) Brassica napus (Yellow mustard or rapeseed) Mixture of Oriental and Yellow mustard nd = not detected Example 2 Nematicidal efficacy of various mustard-derived materials was determined using aqueous extracts of the materials applied to the lesion nematode Pratylenchus penetrans . Materials extracted were from Brassica juncea cv. Forge, as follows: 1. Intact seed. 2. Cracked seed. 3. Mustard bran. 4. Mustard flour. 5. Formulated bran: 1 g mustard bran was mixed well with 1.5 g canola oil. After 30 minutes, 2.5 g powdered clay was added and the mixture was mixed well in a mor¬ tar. 6. Formulated bran: bran was formulated as in 5, except that Tween-20 was used instead of canola oil. Weighed portions of each sample were extracted with 50 ml water at room temperature, either for 30 mins or 60 mins, and filtered. A 1 ml portion of each filtered aqueous extract was mixed with 2 ml of a suspension of Pratylenchus penetrans (100 or 200 nematodes per 2 ml) and the nematodes were assessed for mobility and mortal- ity at various intervals. Mobility was assessed by microscopic observation. Mortality was determined after 68-72 hours of exposure to the extracts by looking for lack of movement on visual inspection and no reaction to mechanical stimulation. The results are shown in Tables 2 (30 minute extrac¬ tion) and 3 (60 minute extraction) . TABLE 2 Sample Weight of Mobility %Mobility- % Mortality Number sample 2 hrs. 4 hrs. 68-72 hrs. extracted treatment 1 231 all active 100.0 10.0 2 231 most inactive 4.4 14. 1 3 301 ditto, 1.3 45.8 sluggish 4 189 slight 5.1 54.9 movement 5 1506 slight 1.4 51.6 movement 6 1506 slight 1 1.4 46.0 movement TABLE 3 Sample Weight of Nematode % Mobility % Mortality Number sample Mobility- 4 hrs. 68-72 hrs. extracted 2 hrs. ' treatment treatment 1 231 all active 100.0 2.7 2 231 slight 3.6 4.0 movement 3 301 slight 12.5 84.5 movement 4 189 slight 15.8 72.8 movement 5 1506 slightly 5.7 69.3 more than above 6 1506 slightly 13.4 69 . 1 more than above Although only preferred embodiments of the present invention have been described, the present invention is not limited to the features of these embodiments, but includes all variations and modifications within the scope of the claims.

Claims: WE CLAIM : 1. A pesticide precursor composition comprising bran from a mustard of the genus Brassica. 2. The composition of claim 1 further comprising an acceptable carrier. 3. The composition of claim 2 wherein the bran is from a mustard of a species selected from the group consisting of (a) Brassica juncea ; (b) Brassica campestris ; and (c) Brassica nigra . 4. The composition of claim 2 wherein the bran is from a mustard of the species Brassica juncea . 5. The composition of claim 4 wherein the mustard is of the variety B . juncea cv. Forge or B . juncea cv . Cutlass. 6. The composition of claim 4 wherein the carrier comprises cracked mustard seed. 7. The composition of claim 4 wherein the carrier is selected from the group consisting of finely ground clay, at least one polymer, at least one surfactant and at least one organic carrier. 8. The composition of claim 4 wherein the composition is nematicidal. 9. The composition of claim 1 wherein the bran contains an effective amount of sinigrin capable of conversion to allyl isothiocyanate on contact with water. 10. A method for controlling a pest in soil which comprises applying to the soil an effective amount of a pesticide precursor composition comprising a bran from a mustard of the genus Brassica, wherein the bran contains an effective amount of sinigrin capable of conversion to allyl isothiocyanate on contact with water in the soil. 11. The method of claim 10 wherein the composition further comprises an acceptable carrier. 12. The method of claim 11 wherein the bran is from a mustard of a species selected from the group consisting of (a) Brassica juncea ; (b) Brassica campestris ; and (c) Brassica nigra . 13. The method of claim 11 wherein the bran is from a mustard of the species Brassica juncea . 14. The method of claim 13 wherein the mustard is of the variety B . juncea cv . Forge or B. juncea cv. Cutlass. 15. The method of claim 13 wherein the carrier comprises cracked mustard seed. 16. The method of claim 13 wherein the carrier is selected from the group consisting of finely ground clay, at least one polymer, at least one surfactant and at least one organic carrier. 17. The method of claim 13 wherein the pest is selected from the group consisting of plant-parasitic nematodes, mites, insects, fungi and bacteria. 18. The method of claim 13 wherein the pest comprises plant-parasitic nematodes. 19. A pesticide comprising a pesticidally effective amount of an aqueous extract of bran from a mustard of the genus Brassica. 20. A pesticide comprising a pesticidally effective amount of an aqueous suspension of bran from a mustard of the genus Brassica. 21. The pesticide of claim 19 wherein the bran is selected from a mustard of a species selected from the group consisting of (a) Brassica juncea ; (b) Brassica campestris ; and (c) Brassica nigra . 22. The pesticide of claim 19 wherein the bran is from a mustard of the species Brassica juncea . 23. The pesticide of claim 22 wherein the mustard is of the variety B . juncea cv. Forge or B . juncea cv. Cutlass. 24. The pesticide of claim 20 wherein the bran is selected from a mustard of a species selected from the group consisting of (a) Brassica juncea ; (b) Brassica campestris ; and (c) Brassica nigra . 25. The pesticide of claim 20 wherein the bran is from a mustard of the species Brassica juncea . 26. The pesticide of claim 25 wherein the mustard is of the variety B. juncea cv. Forge or B. juncea cv. Cutlass. 27. A method for controlling a pest which comprises applying to the pest an effective amount of an aqueous extract of bran from a mustard of the genus Brassica. 28. A method for controlling a pest which comprises applying to the pest an effective amount of an aqueous suspension of bran from a mustard of the genus Brassica.

Inventors: CHIBA MIKIO, POTTER JOHN W, CHIBA, MIKIO, POTTER, JOHN, W.
Applicants: CANADA MAJESTY IN RIGHT OF, HER MAJESTY IN RIGHT OF CANADA, REPRESENTED BY THE
Application year: 1996
Main classification: A01N65
IPC codes: A01N65