Patent Application
20160058001
The invention relates to a method for improving the utilization of the production potential of transgenic plants and for controlling pests such as insects and/or nematodes.
In recent years, there has been a marked increase in the proportion of transgenic plants in agriculture.
Transgenic plants are employed mainly to utilize the production potential of respective plant varieties in the most favourable manner, at the lowest possible input of production means. The aim of the genetic modification of the plants is in particular the generation of resistance in the plants to certain pests or harmful organisms or else herbicides and also to abiotic stress (for example drought, heat or elevated salt levels). It is also possible to modify a plant genetically to increase certain quality or product features, such as, for example, the content of selected vitamins or oils, or to improve certain fibre properties.
Herbicide resistance or tolerance can be achieved, for example, by incorporating genes into the useful plant for expressing enzymes to detoxify certain herbicides, so that a relatively unimpeded growth of these plants is possible even in the presence of these herbicides for controlling broad-leaved weeds and weed grasses. Examples which may be mentioned are cotton varieties or maize varieties which tolerate the herbicidally active compound glyphosate (Roundup®), (Roundup Ready®, Monsanto) or the herbicides glufosinate or oxynil.
There has also been the development of useful plants comprising two or more genetic modifications (“stacked transgenic plants” or multiply transgenic crops). Thus, for example, Monsanto has developed multiply transgenic maize varieties which are resistant to the European corn borer (Ostrinia nubilalis) and the Western corn rootworm (Diabrotica virgifera). Also known are maize and cotton crops which are both resistant to the Western corn rootworm and the cotton bollworm and tolerant to the herbicide Roundup®.
It has now been found that the utilization of the production potential of transgenic useful plants can be improved even more by treating the plants with one or more compounds of the formula (I) defined below. Here, the term “treatment” includes all measures resulting in a contact between these active compounds and at least one plant part. “Plant parts” are to be understood as meaning all above-ground and below-ground parts and organs of plants, such as shoot, leaf, flower and root, by way of example leaves, needles, stalks, stems, flowers, fruit bodies, fruits and seed, and also roots, tubers and rhizomes. The plant parts also include harvested material and also vegetative and generative propagation material, for example cuttings, tubers, rhizomes, slips and seed.
One aspect refers to a method for improving the utilization of the production potential of a transgenic plant and/or for controlling/combating/treating pests, characterized in that the plant is treated with an effective amount of at least one compound of the formula (I)
One preferred embodiment refers to the method described above, characterized in that the compound of the formula (I) is formula (I-1):
One preferred embodiment refers to the method described above, characterized in that the compound of the formula (I) is selected from the group consisting of compound (I-2), (I-3), (I-4) or (I-5):
One preferred embodiment refers to the method described above, characterized in that the compound of the formula (I) is compound (I-5).
Further preferred embodiments refer to the method described above, characterized in that the plant has at least one genetically modified structure or a tolerance according to Table A or Table B or Table C.
Further preferred embodiments refer to the method described above, characterized in that the transgenic plant contains at least one cry-gene or a cry-gene fragment coding for a Bt toxin.
One preferred embodiment refers to the method described above, characterized in that the transgenic plant is a vegetable plant, maize plant, soya bean plant, cotton plant, tobacco plant, rice plant, sugar beet plant, oilseed rape plant or potato plant.
One preferred embodiment refers to the method described above, characterized in that the use form of the compound of the formula (I) is present in a mixture with at least one mixing partner.
One preferred embodiment refers to the method described above, characterized in that the Bt toxin of a Bt-plant is encoded by a bt-gene or fragment thereof comprising event MON87701.
Another aspect refers to a synergistic composition comprising a Bt toxin and a compound of formula (I) as described above.
One preferred embodiment refers to said synergistic composition, characterized in that the Bt toxin is encoded by a cry gene or a cry-gene fragment selected from the group consisting of cry1, cry2, cry3, cry5 and cry9.
One preferred embodiment refers to said synergistic composition, characterized in that the Bt toxin is encoded by a cry gene or a cry-gene fragment selected from the group consisting of especially preferred are cry1Ab, cry1Ac, cry3A, cry3B and cry9C.
One preferred embodiment refers to said synergistic composition, characterized in that the Bt toxin is encoded by a cry gene or a cry-gene fragment selected from the subgroup cry1A, preferably cry1Aa, cry1Ab, cry1Ac or a hybrid thereof (e.g., a hybrid of cry1Ac and cry1Ab).
One preferred embodiment refers to said synergistic composition, characterized in that the Bt toxin is encoded by a bt-gene or fragment thereof comprising event MON87701.
A Bt plant, preferably a Bt-soybean plant comprising event MON87701 or a Bt-soybean plant comprising event MON87701 and MON89788, characterized in that at least 0.00001 g of a compound of formula (I) is attached to it.
The preferred embodiments may be combined as long as such a combination would not contravene existing natural laws.
Compounds of the formula (I)
wherein
A represents individually halogen, cyano, nitro, hydroxyl, amino, C1-C8 alkyl group, substituted C1-C8 alkyl group having at least one substituent elected from the group consisting of halogen, hydroxy, cyano, nitro, amino, halo C1-C3 alkyl group, C1-C3 alkoxy group, halo C1-C3 alkoxy group, C1-C3 alkylthio group, halo C1-C3 alkylthio group, C1-C3 alkylsulfinyl group, halo C1-C3 alkylsulfinyl group, C1-C3 alkylsulfonyl group, halo C1-C3 alkylsulfonyl group and C1-C3 alkylthio, C1-C3 alkyl group; further, an arbitrary saturated carbon atom in said optionally substituted C1-C8 alkyl group;
n represents 0, 1, 2, 3 or 4, preferably 0, 1 or 2;
R1 represents hydrogen, halogen, cyano C1-C8 alkyl or C1-C8 haloalkyl;
R2 represents hydrogen, halogen, cyano C1-C8 alkyl or C1-C8 haloalkyl;
R3 represents O or S;
R4 represents O or S;
Y represents individually hydrogen, halogen, cyano, nitro, C1-C6 alkyl group, halo C1-C6 alkyl group, C2-C6 alkenyl group, halo C2-C6 alkenyl group, C2-C6 alkynyl group, halo C2-C6 alkynyl group, C3-C6 cycloalkyl group, halo C3-C6 cycloalkyl group, C1-C6 alkoxy group, halo C1-C6 alkoxy group, C1-C6 alkylthio group, halo C1-C6 alkylthio group, C1-C6 alkylsulfinyl group, halo C1-C6 alkylsulfinyl group, C1-C6 alkylsulfonyl group, or halo C1-C6 alkylsulfonyl group;
m represents 0, 1, 2, 3, or 4;
X represents a C1-C8 alkyl group or a substituted C1-C8 alkyl group having at least one substituent selected from the group consisting of halogen, hydroxy, cyano, nitro, amino, halo C1-C3 alkyl group, C1-C3 alkoxy group, halo C1-C3 alkoxy group
and their insecticidal action are known from the prior art (see, e.g., EP 0 919 542, W0 2004/018410, W0 2010/012442 or WO 2012/034472).
From these documents, the person skilled in the art will be familiar with processes for preparing and methods for using compounds of the formula (I) and with the action of compounds of the formula (I).
Preferred sub-groups and compounds of formula (I) mentioned above are listed below.
In a preferred embodiment of the present invention, the compounds of the general formula (I) is represented by compounds of formula (I-1):
wherein
Hal represents F, Cl, I or Br; and
X′ represents C1-C6 alkyl or substituted C1-C6 alkyl having at least one substituent selected from the group consisting of halogen, hydroxy, cyano, nitro, amino, halo C1-C3 alkyl group, preferably a C1-C6 cyanoalkyl;
A′ represents C1-C3 alkyl, C1-C3 haloalkyl, halogen, preferably methyl, halomethyl, ethyl or haloethyl, more preferably methyl or ethyl;
n represents 0, 1, 2, 3 or 4, preferably 0, 1 or 2, more preferably 1.
In a more preferred embodiment of the present invention, a composition comprises at least one compound of the general formula (I) selected from the group consisting of compound (I-2), (I-3), (I-4) or (I-5):
Even more preferably, a compound of formula (I) is selected from the group consisting of compound (I-2) or compound (I-5).
In one preferred embodiment, the compound of formula (I) is compound (I-5).
According to the invention, “alkyl” represents straight-chain or branched aliphatic hydrocarbons having 1 to 8, preferably 1 to 6, more preferably 1 to 3, carbon atoms. Suitable alkyl groups are, for example, methyl, ethyl, n-propyl, i-propyl, n-, iso-, sec- or tert-butyl, pentyl or hexyl. The alkyl group may be unsubstituted or is substituted by at least one of the substituents mentioned here.
According to the invention, “halogen” or “Hal” represents fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine.
According to the invention, “haloalkyl” represents alkyl groups having up to 8 carbon atoms in which at least one hydrogen atom has been replaced by a halogen. Suitable haloalkyl groups are, for example, CH2F, CHF2, CF3, CF2Cl, CFCl2, CCl3, CF2Br, CF2CF3, CFHCF3, CH2CF3, CH2CH2F, CH2CHF2, CFClCF3, CCl2CF3, CF2CH3, CF2CH2F, CF2CHF2, CF2CF2Cl, CF2CF2Br, CFHCH3, CFHCHF2, CHFCF3, CHFCF2Cl, CHFCF2Br, CFClCF3, CCl2CF3, CF2CF2CF3, CH2CH2CH2F, CH2CHFCH3, CH2CF2CF3, CF2CH2CF3, CF2CF2CH3, CHFCF2CF3, CF2CHFCF3, CF2CF2CHF2, CF2CF2CH2F, CF2CF2CF2Cl, CF2CF2CF2Br, 1,2,2,2-tetrafluoro-1-(trifluoromethyl)ethyl, 2,2,2-trifluoro-1-(trifluoromethyl)ethyl, pentafluoroethyl, 1-(difluoromethyl)-1,2,2,2-tetrafluoroethyl, 2-bromo-1,2,2-trifluoro-1-(trifluoromethyl)ethyl, 1-(difluoromethyl)-2,2,2-trifluoroethyl. The haloalkyl group may be unsubstituted or is substituted by at least one of the substituents mentioned here.
“Production potential” as used herein refers to the yield of a transgenic plant under specific conditions. “Improving the utilization of the production potential of transgenic plants” thus refers to an increase of yield under unfavorable environmental conditions such as use of herbicides, drought stress, cold stress, stress induced by insects, nematodes, or fungus etc. compared to the yield of such plants under the same conditions without the use of the compounds of formula (I) as described herein.
The method can also be used for an increased control/an increased treatment of pests such as insects and/or nematodes. Thus, the combination of a transgenic plant such as a Bt-plant and a compound of formula (I) can show better treatment/control/combating of insects and/or nematodes compared to the expected effect.
According to the method proposed according to the invention, transgenic plants, in particular useful plants, are treated with compounds of the formula (I) to increase agricultural productivity and/or to control and/or to combat pests, especially nematodes and insects. Preferably, the invention refers to a method for combating pests by treating transgenic plants, preferably insect-resistant transgenic plant such as Bt-plants or Vip-plants with a compound of formula (I), preferably with a compound of formula (I-5).
For the purpose of the invention, genetically modified organisms (GMOs), e.g. plants or seeds, are genetically modified plants (or transgenic plants) are plants of which a heterologous gene has been stably integrated into genome. The expression “heterologous gene” essentially means a gene which is provided or assembled outside the plant and when introduced in the nuclear, chloroplastic or mitochondrial genome gives the transformed plant new or improved agronomic or other properties by expressing a protein or polypeptide of interest or by downregulating or silencing other gene(s) which are present in the plant (using for example, antisense technology, cosuppression technology, RNA interference—RNAi—technology or microRNA—miRNA—technology). A heterologous gene that is located in the genome is also called a transgene. A transgene that is defined by its particular location in the plant genome is called a transformation or transgenic event.
Depending on the plant species or plant cultivars, their location and growth conditions (soils, climate, vegetation period, diet), the treatment according to the invention may also result in superadditive (“synergistic”) effects. Thus, for example, reduced application rates and/or a widening of the activity spectrum and/or an increase in the activity of the active compounds and compositions which can be used according to the invention, better plant growth, increased tolerance to high or low temperatures, increased tolerance to drought or to water or soil salt content, increased flowering performance, easier harvesting, accelerated maturation, higher harvest yields, bigger fruits, larger plant height, greener leaf color, earlier flowering, higher quality and/or a higher nutritional value of the harvested products, higher sugar concentration within the fruits, better storage stability, increased combating of pests, especially nematodes and insects and/or processability of the harvested products are possible, which exceed the effects which were actually to be expected.
At certain application rates, the active compound combinations according to the invention may also have a strengthening effect in plants. Accordingly, they are also suitable for mobilizing the defense system of the plant against attack by unwanted microorganisms. This may, if appropriate, be one of the reasons of the enhanced activity of the combinations according to the invention, for example against fungi. Plant-strengthening (resistance-inducing) substances are to be understood as meaning, in the present context, those substances or combinations of substances which are capable of stimulating the defense system of plants in such a way that, when subsequently inoculated with unwanted microorganisms, the treated plants display a substantial degree of resistance to these microorganisms. In the present case, unwanted microorganisms are to be understood as meaning phytopathogenic fungi, bacteria and viruses. Thus, the substances according to the invention can be employed for protecting plants against attack by the abovementioned pathogens within a certain period of time after the treatment. The period of time within which protection is effected generally extends from 1 to 10 days, preferably 1 to 7 days, after the treatment of the plants with the active compounds.
Plants and plant cultivars which are preferably to be treated according to the invention include all plants which have genetic modified material which impart particularly advantageous, useful traits to these plants (whether obtained by breeding and/or biotechnological means).
Plants and plant cultivars which are also preferably to be treated according to the invention are resistant against one or more biotic stresses, i.e. said plants show a better defense against animal and microbial pests, such as against nematodes, insects, mites, phytopathogenic fungi, bacteria, viruses and/or viroids.
Examples of nematode or insect resistant plants are described in e.g. U.S. patent application Ser. Nos. 11/765,491, 11/765,494, 10/926,819, 10/782,020, 12/032,479, 10/783,417, 10/782,096, 11/657,964, 12/192,904, 11/396,808, 12/166,253, 12/166,239, 12/166,124, 12/166,209, 11/762,886, 12/364,335, 11/763,947, 12/252,453, 12/209,354, 12/491,396, 12/497,221, 12/644,632, 12/646,004, 12/701,058, 12/718,059, 12/721,595, 12/638,591, and in WO 11/002992, WO 11/014749, WO 11/103247, WO 11/103248, WO 12/135436, WO 12/135501.
Examples of plants resistant to other types of pathogens are described in e.g. WO13/050410.
Plants and plant cultivars which may also be treated according to the invention are those plants which are resistant to one or more abiotic stresses. Abiotic stress conditions may include, for example, drought, cold temperature exposure, heat exposure, osmotic stress, flooding, increased soil salinity, increased mineral exposure, ozone exposure, high light exposure, limited availability of nitrogen nutrients, limited availability of phosphorus nutrients, shade avoidance.
Plants and plant cultivars which may also be treated according to the invention, are those plants characterized by enhanced yield characteristics. Increased yield in said plants can be the result of, for example, improved plant physiology, growth and development, such as water use efficiency, water retention efficiency, improved nitrogen use, enhanced carbon assimilation, improved photosynthesis, increased germination efficiency, improved combating of insects and accelerated maturation. Yield can furthermore be affected by improved plant architecture (under stress and non-stress conditions), including but not limited to, early flowering, flowering control for hybrid seed production, seedling vigor, plant size, internode number and distance, root growth, seed size, fruit size, pod size, pod or ear number, seed number per pod or ear, seed mass, enhanced seed filling, reduced seed dispersal, reduced pod dehiscence and lodging resistance. Further yield traits include seed composition, such as carbohydrate content, protein content, oil content and composition, nutritional value, reduction in anti-nutritional compounds, improved processability and better storage stability.
Examples of plants with the above-mentioned traits are non-exhaustively listed in Table A.
Agrostis
stolonifera
Brassica
napus
anthropi.
Lycopersicon
esculentum
Lycopersicon
esculentum
Medicago
sativa (Alfalfa)
Agrobacterium tumefaciens.
Nicotiana
tabacum
Nicotiana
tabacum
Oryza
sativa (Rice)
Oryza
sativa (Rice)
Oryza
sativa (Rice)
Oryza
sativa (Rice)
Oryza
sativa (Rice)
Streptomyces hygroscopicus).
Brassica
napus
anthropi.
Oryza
sativa (Rice)
Streptomyces hygroscopicus).
Oryza
sativa (Rice)
Oryza
sativa (Rice)
Oryza
sativa (Rice)
Prunus
domestica
Solanum
tuberosum
Bacillus thuringiensis (subsp. Tenebrionis).
Solanum
tuberosum
Bacillus thuringiensis (subsp. Tenebrionis).
Solanum
tuberosum
thuringiensis (subsp. Tenebrionis) and the
Solanum
tuberosum
thuringiensis (subsp. Tenebrionis) and the
Brassica
napus
Triticum
aestivum
Triticum
aestivum
Triticum
aestivum
Triticum
aestivum
Fusarium resistance (trichothecene 3-O-
Triticum
aestivum
Triticum
aestivum
Triticum
aestivum
Triticum
aestivum
Triticum
aestivum
Brassica
napus
viridochromogenes, an aerobic soil bacteria.
coli and Streptomyces viridochromogenes,
Bacillus thuringiensis subsp. kurstaki, and
viridochromogenes.
viridochromogenes. Resistance to other
frugiperda, A. ipsilon, and S. albicosta, is
Bacillus thuringiensis Cry1Ab delta-
Bacillus thuringiensis Vip3Aa20 insecticidal
Brassica
napus
amyloliquefaciens, RF lines contained the
Streptomyces hygroscopicus.
viridochromogenes. Corn rootworm-
thuringiensis.
thuringiensis subsp. kurstaki, and the
Bacillus thuringiensis var aizawai and the
viridochromogenes was introduced as a
Brassica
napus
amyloliquefaciens, RF lines contained the
Streptomyces hygroscopicus.
Brassica
napus
amyloliquefaciens, RF lines contained the
Streptomyces hygroscopicus.
E.coli was used as a selectable marker;
Bacillus thuringiensis and the 5-
nubilalis) by introduction of a synthetic
Bacillus thuringiensis subsp. kurstaki HD-1.
Brassica
napus
thuringiensis subsp. kurstaki HD-1 present
Bacillus thuringiensis subspecies
kumamotoensis strain EG4691 present in
Agrobacterium tumefaciens strain CP4
thuringiensis subsp. kumamotoensis.
thuringiensis subspecies kumamotoensis
thuringiensis providing resistance to number
Brassica
napus
amyloliquefaciens; PPT resistance was via
amyloliquefaciens; PPT resistance was via
Brassica
napus
viridochromogenes.
viridochromogenes.
Brassica
napus
thuringiensis var.aizawai and the
viridochromogenes; Insect resistance
Bacillus thuringiensis var. aizawai. Corn
Streptomyces viridochromogenes.
Brassica
napus
amyloliquefaciens; fertility restoration by
Brassica
napus
amyloliquefaciens; fertility restoration by
Brassica
napus
Brassica
napus
viridochromogenes, an aerobic soil bacteria.
Brassica
rapa (Polish
napus line T45. This trait is mediated by the
Brassica
rapa (Polish
Carica
papaya
Carica
papaya
Beta vulgaris
Cichorium
intybus
amyloliquefaciens; PPT resistance was via
Cucumis
melo (Melon)
Cucurbita
pepo (Squash)
Cucurbita
pepo (Squash)
Dianthus
caryophyllus
Dianthus
caryophyllus
Dianthus
caryophyllus
Beta vulgaris
viridochromogenes, an aerobic soil bacteria.
Streptomyces viridochromogenes.
Streptomyces viridochromogenes.
Beta vulgaris
Agrobacterium tumefaciens CP4;
Streptomyces hygroscopicus.
Gossypium
hirsutum
thuringiensis subsp. kurstaki.
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
thuringiensisvar. aizawai. The PAT
viridochromogenes was introduced as a
Beta vulgaris
Gossypium
hirsutum
thuringiensissubsp. kurstaki. The PAT
viridochromogenes was introduced as a
Gossypium
hirsutum
pneumoniae.
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
thuringiensisAB88. The APH4 encoding
Gossypium
hirsutum
Gossypium
hirsutum
thuringiensis. The APH4 encoding gene
Brassica
napus
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Brassica
napus
Gossypium
hirsutum
Streptomyces hygroscopicus; WO
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Agrobacterium tumefaciens. Insect
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Brassica
napus
Gossypium
hirsutum
Gossypium
hirsutum
Gossypium
hirsutum
Helianthus
annuus
Lens
culinaris
Linum
usitatissimum
Lycopersicon
esculentum
Bacillus thuringiensis subsp. Kurstaki.
Lycopersicon
esculentum
Oryza
sativa (Rice)
Oryza
sativa (Rice)
Beta vulgaris
Gossypium
hirsutum
Brassica (A
Brassica
Brassica
Oryza
sativa (Rice)
mays) coding sequence > fertility restoration
sativum) 3′-untranslated region. A CP4
Agrobacterium sp. strain CP4 and encoding
Agrobacterium tumefaciens .
Arabidopsis thaliana, containing an internal
annuus (sunflower) > hppdPf W336: the
Pseudomonas fluorescens strain A32
tumefaciens. 2) Ph4a748: sequence
acidovorans) encodes the aryloxyalkanoate
Brassica napus plant.
ing a corn
Plants that may be treated according to the invention are hybrid plants that already express the characteristic of heterosis or hybrid vigor which results in generally higher yield, vigor, health and resistance towards biotic and abiotic stresses). Such plants are typically made by crossing an inbred male-sterile parent line (the female parent) with another inbred male-fertile parent line (the male parent). Hybrid seed is typically harvested from the male sterile plants and sold to growers. Male sterile plants can sometimes (e.g. in corn) be produced by detasseling, i.e. the mechanical removal of the male reproductive organs (or males flowers) but, more typically, male sterility is the result of genetic determinants in the plant genome. In that case, and especially when seed is the desired product to be harvested from the hybrid plants it is typically useful to ensure that male fertility in the hybrid plants is fully restored. This can be accomplished by ensuring that the male parents have appropriate fertility restorer genes which are capable of restoring the male fertility in hybrid plants that contain the genetic determinants responsible for male-sterility. Genetic determinants for male sterility may be located in the cytoplasm. Examples of cytoplasmic male sterility (CMS) were for instance described in Brassica species (WO 92/05251, WO 95/09910, WO 98/27806, WO 05/002324, WO 06/021972 and U.S. Pat. No. 6,229,072). However, genetic determinants for male sterility can also be located in the nuclear genome. Male sterile plants can also be obtained by plant biotechnology methods such as genetic engineering. A particularly useful means of obtaining male-sterile plants is described in WO 89/10396 in which, for example, a ribonuclease such as barnase is selectively expressed in the tapetum cells in the stamens. Fertility can then be restored by expression in the tapetum cells of a ribonuclease inhibitor such as barstar (e.g. WO 91/02069).
Plants or plant cultivars (obtained by plant biotechnology methods such as genetic engineering) which may be treated according to the invention are herbicide-tolerant plants, i.e. plants made tolerant to one or more given herbicides. Such plants can be obtained either by genetic transformation, or by selection of plants containing a mutation imparting such herbicide tolerance.
Herbicide-resistant plants are for example glyphosate-tolerant plants, i.e. plants made tolerant to the herbicide glyphosate or salts thereof. Plants can be made tolerant to glyphosate through different means. For example, glyphosate-tolerant plants can be obtained by transforming the plant with a gene encoding the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Examples of such EPSPS genes are the AroA gene (mutant CT7) of the bacterium Salmonella typhimurium (Science 1983, 221, 370-371), the CP4 gene of the bacterium Agrobacterium sp. (Curr. Topics Plant Physiol. 1992, 7, 139-145), the genes encoding a Petunia EPSPS (Science 1986, 233, 478-481), a Tomato EPSPS (J. Biol. Chem. 1988, 263, 4280-4289), or an Eleusine EPSPS (WO 01/66704). It can also be a mutated EPSPS as described in for example EP 0837944, WO 00/66746, WO 00/66747 or WO 02/26995, WO 11/000498. Glyphosate-tolerant plants can also be obtained by expressing a gene that encodes a glyphosate oxido-reductase enzyme as described in U.S. Pat. No. 5,776,760 and U.S. Pat. No. 5,463,175. Glyphosate-tolerant plants can also be obtained by expressing a gene that encodes a glyphosate acetyl transferase enzyme as described in for example WO 02/036782, WO 03/092360, WO 05/012515 and WO 07/024782. Glyphosate-tolerant plants can also be obtained by selecting plants containing naturally-occurring mutations of the above-mentioned genes, as described in for example WO 01/024615 or WO 03/013226. Plants expressing EPSPS genes that confer glyphosate tolerance are described in e.g. U.S. patent application Ser. Nos. 11/517,991, 10/739,610, 12/139,408, 12/352,532, 11/312,866, 11/315,678, 12/421,292, 11/400,598, 11/651,752, 11/681,285, 11/605,824, 12/468,205, 11/760,570, 11/762,526, 11/769,327, 11/769,255, 11/943,801 or 12/362,774. Plants comprising other genes that confer glyphosate tolerance, such as decarboxylase genes, are described in e.g. U.S. patent application Ser. Nos. 11/588,811, 11/185,342, 12/364,724, 11/185,560 or 12/423,926.
Other herbicide resistant plants are for example plants that are made tolerant to herbicides inhibiting the enzyme glutamine synthase, such as bialaphos, phosphinothricin or glufosinate. Such plants can be obtained by expressing an enzyme detoxifying the herbicide or a mutant glutamine synthase enzyme that is resistant to inhibition, e.g. described in U.S. patent application Ser. No. 11/760,602. One such efficient detoxifying enzyme is an enzyme encoding a phosphinothricin acetyltransferase (such as the bar or pat protein from Streptomyces species). Plants expressing an exogenous phosphinothricin acetyltransferase are for example described in U.S. Pat. Nos. 5,561,236; 5,648,477; 5,646,024; 5,273,894; 5,637,489; 5,276,268; 5,739,082; 5,908,810 and 7,112,665.
Further herbicide-tolerant plants are also plants that are made tolerant to the herbicides inhibiting the enzyme hydroxyphenylpyruvatedioxygenase (HPPD). HPPD is an enzyme that catalyze the reaction in which para-hydroxyphenylpyruvate (HPP) is transformed into homogentisate. Plants tolerant to HPPD-inhibitors can be transformed with a gene encoding a naturally-occurring resistant HPPD enzyme, or a gene encoding a mutated or chimeric HPPD enzyme as described in WO 96/38567, WO 99/24585, WO 99/24586, WO 09/144079, WO 02/046387, U.S. Pat. No. 6,768,044, WO 11/076877, WO 11/076882, WO 11/076885, WO 11/076889. WO 11/076892. WO13/026740, WO13/092552, WO13/092551 or WO12/092555. Tolerance to HPPD-inhibitors can also be obtained by transforming plants with genes encoding certain enzymes enabling the formation of homogentisate despite the inhibition of the native HPPD enzyme by the HPPD-inhibitor. Such plants and genes are described in WO 99/34008 and WO 02/36787. Tolerance of plants to HPPD inhibitors can also be improved by transforming plants with a gene encoding an enzyme having prephenate deshydrogenase (PDH) activity in addition to a gene encoding an HPPD-tolerant enzyme, as described in WO 04/024928. Further, plants can be made more tolerant to HPPD-inhibitor herbicides by adding into their genome a gene encoding an enzyme capable of metabolizing or degrading HPPD inhibitors, such as the CYP450 enzymes shown in WO 07/103567 and WO 08/150473.
Still further herbicide resistant plants are plants that are made tolerant to acetolactate synthase (ALS) inhibitors. Known ALS-inhibitors include, for example, sulfonylurea, imidazolinone, triazolo-pyrimidines, pyrimidinyoxy(thio)benzoates, and/or sulfonylaminocarbonyltriazolinone herbicides. Different mutations in the ALS enzyme (also known as acetohydroxyacid synthase, AHAS) are known to confer tolerance to different herbicides and groups of herbicides, as described for example in Tranel and Wright (Weed Science 2002, 50, 700-712), but also, in U.S. Pat. Nos. 5,605,011, 5,378,824, 5,141,870, and 5,013,659. The production of sulfonylurea-tolerant plants and imidazolinone-tolerant plants is described in U.S. Pat. Nos. 5,605,011; 5,013,659; 5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107; 5,928,937; and 5,378,824; and WO 96/33270. Other imidazolinone-tolerant plants are also described in for example WO 04/040012, WO 04/106529, WO 05/020673, WO 05/093093, WO 06/007373, WO 06/015376, WO 06/024351, and WO 06/060634. Further sulfonylurea- and imidazolinone-tolerant plants are also described in for example WO 07/024782, WO 2011/076345, WO 2012058223, WO 2012150335 and U.S. Patent Application 61/288,958.
Other plants tolerant to imidazolinone and/or sulfonylurea can be obtained by induced mutagenesis, selection in cell cultures in the presence of the herbicide or mutation breeding as described for example for soybeans in U.S. Pat. No. 5,084,082, for rice in WO 97/41218, for sugar beet in U.S. Pat. No. 5,773,702 and WO 99/057965, for lettuce in U.S. Pat. No. 5,198,599, or for sunflower in WO 01/065922.
Plants tolerant to 2,4 D or dicamba are for example described in U.S. Pat. No. 6,153,401.
Plants or plant cultivars (obtained by plant biotechnology methods such as genetic engineering) which may also be treated according to the invention are insect-resistant transgenic plants, i.e. plants made resistant to attack by certain target insects. Such plants can be obtained by genetic transformation, or by selection of plants containing a mutation imparting such insect resistance.
An “insect-resistant transgenic plant”, as used herein, includes any plant containing at least one transgene comprising a coding sequence encoding:
Of course, an insect-resistant transgenic plant, as used herein, also includes any plant comprising a combination of genes encoding the proteins of any one of the above classes 1 to 10. In one embodiment, an insect-resistant plant contains more than one transgene encoding a protein of any one of the above classes 1 to 10, to expand the range of target insect species affected when using different proteins directed at different target insect species, or to delay insect resistance development to the plants by using different proteins insecticidal to the same target insect species but having a different mode of action, such as binding to different receptor binding sites in the insect.
An “insect-resistant transgenic plant”, as used herein, further includes any plant containing at least one transgene comprising a sequence producing upon expression a double-stranded RNA which upon ingestion by a plant insect pest inhibits the growth of this insect pest, as described e.g. in WO 07/080126, WO 06/129204, WO 07/074405, WO 07/080127 and WO 07/035650.
Plants or plant cultivars (obtained by plant biotechnology methods such as genetic engineering) which may also be treated according to the invention are tolerant to abiotic stresses. Such plants can be obtained by genetic transformation, or by selection of plants containing a mutation imparting such stress resistance. Particularly useful stress tolerance plants include:
Plants or plant cultivars (obtained by plant biotechnology methods such as genetic engineering) which may also be treated according to the invention show altered quantity, quality and/or storage-stability of the harvested product and/or altered properties of specific ingredients of the harvested product such as:
Plants or plant cultivars (that can be obtained by plant biotechnology methods such as genetic engineering) which may also be treated according to the invention are plants, such as oilseed rape or related Brassica plants, with altered seed shattering characteristics. Such plants can be obtained by genetic transformation, or by selection of plants contain a mutation imparting such altered seed shattering characteristics and include plants such as oilseed rape plants with delayed or reduced seed shattering as described in WO 2009/068313 and WO 2010/006732, WO 2012090499.
Plants or plant cultivars (that can be obtained by plant biotechnology methods such as genetic engineering) which may also be treated according to the invention are plants, such as Tobacco plants, with altered post-translational protein modification patterns, for example as described in WO 10/121818 and WO 10/145846.
Particularly useful transgenic plants which may be treated according to the invention are plants containing transformation events, or combination of transformation events, that are the subject of petitions for non-regulated status, in the United States of America, to the Animal and Plant Health Inspection Service (APHIS) of the United States Department of Agriculture (USDA) whether such petitions are granted or are still pending. At any time this information is readily available from APHIS (4700 River Road, Riverdale, Md. 20737, USA), for instance on its internet site (URL http://www.aphis.usda.gov/brs/not_reg.html). On the filing date of this application the petitions for nonregulated status that were pending with APHIS or granted by APHIS were those listed in Table B which contains the following information:
Additional particularly useful plants containing single transformation events or combinations of transformation events are listed for example in the databases from various national or regional regulatory agencies (see for example http://gmoinfo.jrc.it/gmp_browse.aspx and http://www.cera-gmc.org/?action=gm_crop_database).
Further particularly transgenic plants include plants containing a transgene in an agronomically neutral or beneficial position as described in any of the patent publications listed in Table C.
Additional particularly useful plants containing single transformation events or combinations of transformation events are listed for example in the databases from various national or regional regulatory agencies (see for example http://gmoinfo.jrc.it/gmp_browse.aspx and http://www.cera-gmc.org/?action=gm_crop_database).
Particularly useful transgenic plants which may be treated according to the invention are plants containing transformation events, or a combination of transformation events, and that are listed for example in the databases for various national or regional regulatory agencies including Event 531/PV-GHBK04 (cotton, insect control, described in WO 2002/040677), Event 1143-14A (cotton, insect control, not deposited, described in WO 06/128569); Event 1143-51B (cotton, insect control, not deposited, described in WO 06/128570); Event 1445 (cotton, herbicide tolerance, not deposited, described in US-A 2002-120964 or WO 02/034946Event 17053 (rice, herbicide tolerance, deposited as PTA-9843, described in WO 10/117737); Event 17314 (rice, herbicide tolerance, deposited as PTA-9844, described in WO 10/117735); Event 281-24-236 (cotton, insect control—herbicide tolerance, deposited as PTA-6233, described in WO 05/103266 or US-A 2005-216969); Event 3006-210-23 (cotton, insect control—herbicide tolerance, deposited as PTA-6233, described in US-A 2007-143876 or WO 05/103266); Event 3272 (corn, quality trait, deposited as PTA-9972, described in WO 06/098952 or US-A 2006-230473); Event 33391 (wheat, herbicide tolerance, deposited as PTA-2347, described in WO 2002/027004), Event 40416 (corn, insect control—herbicide tolerance, deposited as ATCC PTA-11508, described in WO 11/075593); Event 43A47 (corn, insect control—herbicide tolerance, deposited as ATCC PTA-11509, described in WO 11/075595); Event 5307 (corn, insect control, deposited as ATCC PTA-9561, described in WO 10/077816); Event ASR-368 (bent grass, herbicide tolerance, deposited as ATCC PTA-4816, described in US-A 2006-162007 or WO 04/053062); Event B16 (corn, herbicide tolerance, not deposited, described in US-A 2003-126634); Event BPS-CV127-9 (soybean, herbicide tolerance, deposited as NCIMB No. 41603, described in WO 10/080829); Event BLR1 (oilseed rape, restoration of male sterility, deposited as NCIMB 41193, described in WO 2005/074671), Event CE43-67B (cotton, insect control, deposited as DSM ACC2724, described in US-A 2009-217423 or WO 06/128573); Event CE44-69D (cotton, insect control, not deposited, described in US-A 2010-0024077); Event CE44-69D (cotton, insect control, not deposited, described in WO 06/128571); Event CE46-02A (cotton, insect control, not deposited, described in WO 06/128572); Event COT102 (cotton, insect control, not deposited, described in US-A 2006-130175 or WO 04/039986); Event COT202 (cotton, insect control, not deposited, described in US-A 2007-067868 or WO 05/054479); Event COT203 (cotton, insect control, not deposited, described in WO 05/054480); Event DAS21606-3/1606 (soybean, herbicide tolerance, deposited as PTA-11028, described in WO 012/033794), Event DAS40278 (corn, herbicide tolerance, deposited as ATCC PTA-10244, described in WO 11/022469); Event DAS-44406-6/pDAB8264.44.06.1 (soybean, herbicide tolerance, deposited as PTA-11336, described in WO 2012/075426), Event DAS-14536-7/pDAB8291.45.36.2 (soybean, herbicide tolerance, deposited as PTA-11335, described in WO 2012/075429), Event DAS-59122-7 (corn, insect control—herbicide tolerance, deposited as ATCC PTA 11384, described in US-A 2006-070139); Event DAS-59132 (corn, insect control—herbicide tolerance, not deposited, described in WO 09/100188); Event DAS68416 (soybean, herbicide tolerance, deposited as ATCC PTA-10442, described in WO 11/066384 or WO 11/066360); Event DP-098140-6 (corn, herbicide tolerance, deposited as ATCC PTA-8296, described in US-A 2009-137395 or WO 08/112019); Event DP-305423-1 (soybean, quality trait, not deposited, described in US-A 2008-312082 or WO 08/054747); Event DP-32138-1 (corn, hybridization system, deposited as ATCC PTA-9158, described in US-A 2009-0210970 or WO 09/103049); Event DP-356043-5 (soybean, herbicide tolerance, deposited as ATCC PTA-8287, described in US-A 2010-0184079 or WO 08/002872); Event EE-1 (brinjal, insect control, not deposited, described in WO 07/091277); Event FI117 (corn, herbicide tolerance, deposited as ATCC 209031, described in US-A 2006-059581 or WO 98/044140); Event FG72 (soybean, herbicide tolerance, deposited as PTA-11041, described in WO 2011/063413), Event GA21 (corn, herbicide tolerance, deposited as ATCC 209033, described in US-A 2005-086719 or WO 98/044140); Event GG25 (corn, herbicide tolerance, deposited as ATCC 209032, described in US-A 2005-188434 or WO 98/044140); Event GHB119 (cotton, insect control—herbicide tolerance, deposited as ATCC PTA-8398, described in WO 08/151780); Event GHB614 (cotton, herbicide tolerance, deposited as ATCC PTA-6878, described in US-A 2010-050282 or WO 07/017186); Event GJ11 (corn, herbicide tolerance, deposited as ATCC 209030, described in US-A 2005-188434 or WO 98/044140); Event GM RZ13 (sugar beet, virus resistance, deposited as NCIMB-41601, described in WO 10/076212); Event H7-1 (sugar beet, herbicide tolerance, deposited as NCIMB 41158 or NCIMB 41159, described in US-A 2004-172669 or WO 04/074492); Event JOPLIN1 (wheat, disease tolerance, not deposited, described in US-A 2008-064032); Event LL27 (soybean, herbicide tolerance, deposited as NCIMB41658, described in WO 06/108674 or US-A 2008-320616); Event LL55 (soybean, herbicide tolerance, deposited as NCIMB 41660, described in WO 06/108675 or US-A 2008-196127); Event LLcotton25 (cotton, herbicide tolerance, deposited as ATCC PTA-3343, described in WO 03/013224 or US-A 2003-097687); Event LLRICE06 (rice, herbicide tolerance, deposited as ATCC 203353, described in U.S. Pat. No. 6,468,747 or WO 00/026345); Event LLRice62 (rice, herbicide tolerance, deposited as ATCC 203352, described in WO 2000/026345), Event LLRICE601 (rice, herbicide tolerance, deposited as ATCC PTA-2600, described in US-A 2008-2289060 or WO 00/026356); Event LY038 (corn, quality trait, deposited as ATCC PTA-5623, described in US-A 2007-028322 or WO 05/061720); Event MIR162 (corn, insect control, deposited as PTA-8166, described in US-A 2009-300784 or WO 07/142840); Event MIR604 (corn, insect control, not deposited, described in US-A 2008-167456 or WO 05/103301); Event MON15985 (cotton, insect control, deposited as ATCC PTA-2516, described in US-A 2004-250317 or WO 02/100163); Event MON810 (corn, insect control, not deposited, described in US-A 2002-102582); Event MON863 (corn, insect control, deposited as ATCC PTA-2605, described in WO 04/011601 or US-A 2006-095986); Event MON87427 (corn, pollination control, deposited as ATCC PTA-7899, described in WO 11/062904); Event MON87460 (corn, stress tolerance, deposited as ATCC PTA-8910, described in WO 09/111263 or US-A 2011-0138504); Event MON87701 (soybean, insect control, deposited as ATCC PTA-8194, described in US-A 2009-130071 or WO 09/064652); Event MON87705 (soybean, quality trait—herbicide tolerance, deposited as ATCC PTA-9241, described in US-A 2010-0080887 or WO 10/037016); Event MON87708 (soybean, herbicide tolerance, deposited as ATCC PTA-9670, described in WO 11/034704); Event MON87712 (soybean, yield, deposited as PTA-10296, described in WO 2012/051199), Event MON87754 (soybean, quality trait, deposited as ATCC PTA-9385, described in WO 10/024976); Event MON87769 (soybean, quality trait, deposited as ATCC PTA-8911, described in US-A 2011-0067141 or WO 09/102873); Event MON88017 (corn, insect control—herbicide tolerance, deposited as ATCC PTA-5582, described in US-A 2008-028482 or WO 05/059103); Event MON88913 (cotton, herbicide tolerance, deposited as ATCC PTA-4854, described in WO 04/072235 or US-A 2006-059590); Event MON88302 (oilseed rape, herbicide tolerance, deposited as PTA-10955, described in WO 2011/153186), Event MON88701 (cotton, herbicide tolerance, deposited as PTA-11754, described in WO 2012/134808), Event MON89034 (corn, insect control, deposited as ATCC PTA-7455, described in WO 07/140256 or US-A 2008-260932); Event MON89788 (soybean, herbicide tolerance, deposited as ATCC PTA-6708, described in US-A 2006-282915 or WO 06/130436); Event MS11 (oilseed rape, pollination control—herbicide tolerance, deposited as ATCC PTA-850 or PTA-2485, described in WO 01/031042); Event MS8 (oilseed rape, pollination control—herbicide tolerance, deposited as ATCC PTA-730, described in WO 01/041558 or US-A 2003-188347); Event NK603 (corn, herbicide tolerance, deposited as ATCC PTA-2478, described in US-A 2007-292854); Event PE-7 (rice, insect control, not deposited, described in WO 08/114282); Event RF3 (oilseed rape, pollination control—herbicide tolerance, deposited as ATCC PTA-730, described in WO 01/041558 or US-A 2003-188347); Event RT73 (oilseed rape, herbicide tolerance, not deposited, described in WO 02/036831 or US-A 2008-070260); Event SYHT0H2/SYN-000H2-5 (soybean, herbicide tolerance, deposited as PTA-11226, described in WO 2012/082548), Event T227-1 (sugar beet, herbicide tolerance, not deposited, described in WO 02/44407 or US-A 2009-265817); Event T25 (corn, herbicide tolerance, not deposited, described in US-A 2001-029014 or WO 01/051654); Event T304-40 (cotton, insect control—herbicide tolerance, deposited as ATCC PTA-8171, described in US-A 2010-077501 or WO 08/122406); Event T342-142 (cotton, insect control, not deposited, described in WO 06/128568); Event TC1507 (corn, insect control—herbicide tolerance, not deposited, described in US-A 2005-039226 or WO 04/099447); Event VIP1034 (corn, insect control—herbicide tolerance, deposited as ATCC PTA-3925, described in WO 03/052073), Event 32316 (corn, insect control-herbicide tolerance, deposited as PTA-11507, described in WO 11/084632), Event 4114 (corn, insect control-herbicide tolerance, deposited as PTA-11506, described in WO 11/084621), EE-GM3/FG72 (soybean, herbicide tolerance, ATCC Accession No PTA-11041, WO 2011/063413A2), event DAS-68416-4 (soybean, herbicide tolerance, ATCC Accession No PTA-10442, WO2 011/066360A1), event DAS-68416-4 (soybean, herbicide tolerance, ATCC Accession No PTA-10442, WO 2011/066384A1), event DP-040416-8 (corn, insect control, ATCC Accession No PTA-11508, WO 2011/075593A1), event DP-043A47-3 (corn, insect control, ATCC Accession No PTA-11509, WO 2011/075595A1), event DP-004114-3 (corn, insect control, ATCC Accession No PTA-11506, WO 2011/084621A1), event DP-032316-8 (corn, insect control, ATCC Accession No PTA-11507, WO 2011/084632A1), event MON-88302-9 (oilseed rape, herbicide tolerance, ATCC Accession No PTA-10955, WO 2011/153186A1), event DAS-21606-3 (soybean, herbicide tolerance, ATCC Accession No. PTA-11028, WO 2012/033794A2), event MON-87712-4 (soybean, quality trait, ATCC Accession No. PTA-10296, WO 2012/051199A2), event DAS-44406-6 (soybean, stacked herbicide tolerance, ATCC Accession No. PTA-11336, WO 2012/075426A1), event DAS-14536-7 (soybean, stacked herbicide tolerance, ATCC Accession No. PTA-11335, WO 2012/075429A1), event SYN-000H2-5 (soybean, herbicide tolerance, ATCC Accession No. PTA-11226, WO 2012/082548A2), event DP-061061-7 (oilseed rape, herbicide tolerance, no deposit No available, WO 2012071039A1), event DP-073496-4 (oilseed rape, herbicide tolerance, no deposit No available, US2012131692), event 8264.44.06.1 (soybean, stacked herbicide tolerance, Accession No PTA-11336, WO 2012075426A2), event 8291.45.36.2 (soybean, stacked herbicide tolerance, Accession No. PTA-11335, WO 2012075429A2), event SYHT0H2 (soybean, ATCC Accession No. PTA-11226, WO 2012/082548A2), event MON88701 (cotton, ATCC Accession No PTA-11754, WO 2012/134808A1), event KK179-2 (alfalfa, ATCC Accession No PTA-11833, WO2013003558A1), event pDAB8264.42.32.1 (soybean, stacked herbicide tolerance, ATCC Accession No PTA-11993, WO 2013010094A1), event MZDT09Y (corn, ATCC Accession No PTA-13025, WO 2013012775A1), event KK179-2 (alfalfa, ATCC Accession No PTA-11833), WO2013003558A1, event pDAB8264.42.32.1 (soybean, stacked herbicide tolerance, ATCC Accession No PTA-1 1993), WO2013010094A1, event MZDT09Y (corn, ATCC Accession No PTA-13025), WO2013012775A1, event VCO-01981-5 (corn, herbicide tolerance, NCIMB Accession No 41842), WO2013014241A1, event DAS-81419-2 X DAS-68416-4 (soybean stacked insect resistance and herbicide tolerance, ATCC Accession No PTA-10442), WO2013016516A1, event DAS-81419-2 (soybean stacked insect resistance and herbicide tolerance, ATCC Accession No PTA-12006), WO2013016527A1, event HCEM485 (corn, herbicide tolerance, ATCC Accession No PTA-12014), WO2013025400A1, event pDAB4468.18.07.1 (cotton, herbicide tolerance, ATCC Accession No PTA-12456), WO2013112525A2, event pDAB4468.19.10.3 (cotton, herbicide tolerance, ATCC Accession No PTA-12457), WO2013112527A1.
In an advantageous embodiment, the compounds of the formula (I) are used for treating transgenic plants comprising at least one gene or gene fragment coding for a Bt toxin or Vip-related toxin.
Preferably, the compounds of the formula (I) are used for treating transgenic plants comprising at least one gene or gene fragment coding for a Bt toxin. A Bt toxin is a protein originating from or derived from the soil bacterium Bacillus thuringiensis which either belongs to the group of the crystal toxins (Cry) or the cytolytic toxins (Cyt). In the bacterium, they are originally formed as protoxins and are only metabolized in alkaline medium—for example in the digestive tract of certain feed insects—to their active form. There, the active toxin then binds to certain hydrocarbon structures at cell surfaces causing pores to be formed which destroy the osmotic potential of the cell, which may effect cell lysis. The result is the death of the insects. Bt toxins are active in particular against certain harmful species from the orders of the Lepidoptera (butterflies), Homoptera, Diptera and Coleoptera (beetles) in all their development stages; i.e. from the egg larva via their juvenile forms to their adult forms.
It has been known for a long time that gene sequences coding for Bt toxins, parts thereof or else peptides or proteins derived from Bt toxins can be cloned with the aid of genetic engineering into agriculturally useful plants to generate transgenic plants having endogenous resistance to pests sensitive to Bt toxins. For the purpose of the invention, the transgenic plants coding for at least one Bt toxin or proteins derived therefrom are defined as “Bt plants”.
The “first generation” of such Bt plants generally only comprise the genes enabling the formation of a certain toxin, thus only providing resistance to one group of pathogens. An example of a commercially available maize variety comprising the gene for forming the Cry1Ab toxin is “YieldGard®” from Monsanto which is resistant to the European corn borer. In contrast, in the Bt cotton variety (Bollgard®), resistance to other pathogens from the family of the Lepidoptera is generated by introduction by cloning of the genes for forming the Cry1Ac toxin. Other transgenic crop plants, in turn, express genes for forming Bt toxins with activity against pathogens from the order of the Coleoptera. Examples that may be mentioned are the Bt potato variety “NewLeaf®” (Monsanto) capable of forming the Cry3A toxin, which is thus resistant to the Colorado potato beetle, and the transgenic maize variety “YieldGard®” (Monsanto) which is capable of forming the Cry 3Bb1 toxin and is thus protected against various species of the Western corn rootworm.
In a “second generation”, the multiply transgenic plants, already described above, expressing or comprising at least two foreign genes were generated.
Preference according to the invention is given to transgenic plants with Bt toxins from the group of the Cry family (see, for example, http://www.lifesci.susx.ac.uk/home/Neil_Crickmore/Bt/.
Preferred are transgenic plants with Bt toxins from the group of the
Particular preference is given to the genes or gene sections of the subfamilies cry1, cry2, cry3, cry5 and cry9; especially preferred are members of the subfamily cry1A such as cry1Aa, cry1Ac, cry2Ab.
Furthermore, it is preferred to use plants which, in addition to the genes for one or more Bt toxins, express or contain, if appropriate, also genes for expressing, for example, a protease or peptidase inhibitor (such as in WO-A 95/35031), of herbicide resistances (for example to glufosinate or glyphosate by expression of the pat gene or bar gene) or for becoming resistant to nematodes, fungi or viruses (for example by expressing a gluconase, chitinase). However, they may also be genetically modified in their metabolic properties, so that they show a qualitative and/or quantitative change of ingredients (for example by modification of the energy, carbohydrate, fatty acid or nitrogen metabolism or by metabolite currents influencing these (see above).
In one preferred embodiment, a Bt-plant, preferably a Bt-soybean, comprises event MON87701 which is described in, e.g., WO2009/064652. Thus, in one preferred embodiment, a Bt-soybean seeds comprising said event of which a representative sample was deposited at the ATCC under Accession No. PTA-8194 are treated with a ryanodine receptor modulator according to the present invention.
In another preferred embodiment, a Bt-soybean comprises event pDAB9582.814.19.1 and/or event pDAB4468.04.16.1 which are described in, e.g., WO 2013/016516. This breeding stacks comprise cry1F, cry1Ac and pat and aad-12 and pat, as described in WO 2012/075426. Thus, in one preferred embodiment, a Bt-soybean seeds of which comprising said events were deposited at the ATCC under Accession No. PTA-10442 (pDAB4468.04.16.1) are treated with a ryanodine receptor modulator according to the present invention.
In one preferred embodiment, the method of the invention is characterized in that the Bt-plant, preferably a Bt-soybean plant, comprises at least one cry-gene or a cry-gene fragment coding for a Bt toxin.
In one preferred embodiment, said method is characterized in that the Bt-plant, preferably Bt-soybean plant, comprises at least one cry1A-gene or cry1A-gene fragment coding for a Bt toxin.
In one preferred embodiment, said method is characterized in that said Bt-plant, preferably Bt-soybean plant, further comprising a cryF gene or cryF-gene fragment coding for a Bt toxin.
In another preferred embodiment, said method is characterized in that said plant, preferably said soybean plant, comprises event MON87701.
In a more preferred embodiment, said soybean plant comprises event MON87701 and event MON89788, e.g. Intacta™ Roundup Ready™ 2 Pro.
In another preferred embodiment, said method is characterized in that said soybean plant comprising DNA that comprises a first sequence selected from the group consisting of bp 1385-1415 of SEQ ID NO: 1; bp 1350-1450 of SEQ ID NO: 1; bp 1300-1500 of SEQ ID NO: 1; bp 1200-1600 of SEQ ID NO: 1; bp 137-168 of SEQ ID NO:2; bp 103-203 of SEQ ID NO:2; and bp 3-303 of SEQ ID NO:2; and a second sequence selected from the group consisting bp 2680-2780 of SEQ ID NO: 3; bp 2630-2830 of SEQ ID NO: 15; bp 2530-2930 of SEQ ID NO: 15; bp 9071-9171 of SEQ ID NO: 15; bp 9021-9221 of SEQ ID NO: 15; and, bp 8921-9321 of SEQ ID NO: 15 said first and second sequences being diagnostic for the presence of soybean event pDAB9582.814.19.1::pDAB4468.04.16.1. pDAB9582.814.19.1::pDAB4468.04.16.1 are disclosed in WO 2013/016516.
In one preferred embodiment, said method is characterized in that said soybean plant comprising a nucleotide sequence of SEQ ID NO: 4, SEQ ID NO:5, or complement thereof.
In one preferred embodiment, said method is characterized in that said soybean plant comprising a nucleotide sequence of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:9 or complement thereof.
In one preferred embodiment, said method is characterized in that said soybean plant comprising a nucleotide sequence of SEQ ID NO:6 from positions 1 to 5757, the nucleotide sequence of SEQ ID NO:8 from positions 1 to 6426, and the nucleotide sequence of SEQ ID NO:7 from positions 379 to 2611, or complement thereof.
In one preferred embodiment, said method is characterized in that said soybean plant comprising a nucleotide sequence essentially of the nucleotide sequence of SEQ ID NO: 9 or complement thereof.
In another preferred embodiment, said method is characterized in that said pest is selected from the group consisting of Pseudoplusia includens (soybean looper), Anticarsia gemmatalis (velvet bean caterpillar) and Spodoptera frugiperda (fall armyworm).
In another preferred embodiment, said method is characterized in that the use form of the ryanodine receptor modulator is present in a mixture with at least one mixing partner.
A second aspect refers to a method for improving the utilization of the production potential of transgenic soybean plants in the absent of a pest. Preferred embodiments of this aspect are identical to the preferred embodiments disclosed for the first aspect of the present invention.
A third aspect refers to a synergistic composition comprising Bt toxins encoded by a nucleotide sequence that comprises
A fourth aspect refers to a Bt-soybean plant, characterized in that at least 0.00001 g of a ryanodine receptor modulator as described herein is attached to it.
SEQ ID No: 1 (disclosed in WO 2013/016516) is the 5′ DNA flanking border sequence for soybean event pDAB9582.814.19.1. Nucleotides 1-1400 are genomic sequence. Nucleotides 1401-1535 are a rearranged sequence from pDAB9582. Nucleotides 1536-1836 are insert sequence.
SEQ ID No: 2 (disclosed in WO 2013/016516) is the 3′ DNA flanking border sequence for soybean event pDAB9582.814.19.1. Nucleotides 1-152 are insert sequence. Nucleotides 153-1550 are genomic sequence.
SEQ ID No: 3 (disclosed in WO 2013/016516) is the confirmed sequence of soybean event pDAB4468.04.16.1. Including the 5′ genomic flanking sequence, pDAB4468 T-strand insert, and 3′ genomic flanking sequence.
SEQ ID No:4 (disclosed in WO 2009/064652) is a A 20 nucleotide sequence representing the junction between the soybean genomic DNA and an integrated expression cassette. This sequence corresponds to positions 5748 to 5767 of SEQ ID NO:9. In addition, SEQ ID NO: 1 is a nucleotide sequence corresponding to positions 5748 through 5757 of SEQ ID NO:6 and the integrated right border of the TIC 107 expression cassette corresponding to positions 1 through 10 of SEQ ID NO:8. SEQ ID NO:1 also corresponds to positions 5748 to 5767 of the 5′ flanking sequence, SEQ ID NO:6.
SEQ ID No: 5 (disclosed in WO 2009/064652) is a 20 nucleotide sequence representing the junction between an integrated expression cassette and the soybean genomic DNA. This sequence corresponds to positions 12174 to 12193 of SEQ ID NO:9. In addition, SEQ ID NO:2 is a nucleotide sequence corresponding positions 6417 through 6426 of SEQ ID NO:8 and the 3′ flanking sequence corresponding to positions 379 through 388 of SEQ ED NO:7.
SEQ ID No: 6 (disclosed in WO 2009/064652) is the 5′ sequence flanking the inserted DNA of MON87701 up to and including a region of transformation DNA (T-DNA) insertion.
SEQ ID No: 7 (disclosed in WO 2009/064652) is the 3′ sequence flanking the inserted DNA of MON87701 up to and including a region of T-DNA insertion.
SEQ ID No: 8 (disclosed in WO 2009/064652) is the sequence of the integrated TIC 107 expression cassette, including right and left border sequence after integration.
SEQ ID No: 9 (disclosed in WO 2009/064652) is a 14,416 bp nucleotide sequence representing the contig of the 5′ sequence flanking the inserted DNA of MON87701 (SEQ ID NO:6), the sequence of the integrated expression cassette (SEQ ID NO:8) and the 3′ sequence flanking the inserted DNA of MON87701 (SEQ ID NO: 7).
A nucleic acid molecule is said to be the “complement” of another nucleic acid molecule if they exhibit complete complementarity. As used herein, molecules are said to exhibit “complete complementarity” when every nucleotide of one of the molecules is complementary to a nucleotide of the other. Two molecules are said to be “minimally complementary” if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under at least conventional “low-stringency” conditions. Similarly, the molecules are said to be “complementary” if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under conventional “high-stringency” conditions. Conventional stringency conditions are described by Sambrook et al, 1989, and by Haymes et al, In: Nucleic Acid Hybridization, A Practical Approach, IRL Press, Washington, D.C. (1985), Departures from complete complementarity are therefore permissible, as long as such departures do not completely preclude the capacity of the molecules to form a double-stranded structure. In order for a nucleic acid molecule to serve as a primer or probe it need only be sufficiently complementary in sequence to be able to form a stable double-stranded structure under the particular solvent and salt concentrations employed.
As used herein, a “substantially homologous sequence” is a nucleic acid sequence that will specifically hybridize to the complement of the nucleic acid sequence to which it is being compared under high stringency conditions. Appropriate stringency conditions which promote DNA hybridization, for example, 6.0× sodium chloride/sodium citrate (SSC) at about 45<0>C, followed by a wash of 2.0×SSC at 50<0>C, are known to those skilled in the art or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. For example, the salt concentration in the wash step can be selected from a low stringency of about 2.0×SSC at 50<0>C to a high stringency of about 0.2×SSC at 50<0>C. In addition, the temperature in the wash step can be increased from low stringency conditions at room temperature, about 22<0>C, to high stringency conditions at about 65<0>C. Both temperature and salt may be varied, or either the temperature or the salt concentration may be held constant while the other variable is changed. In a preferred embodiment, a nucleic acid of the present invention will specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 and 2 or complements thereof or fragments of either under moderately stringent conditions, for example at about 2.0×SSC and about 65<0>C. In a particularly preferred embodiment, a nucleic acid of the present invention will specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 and SEQ ID NO:2 or complements or fragments of either under high stringency conditions. In one aspect of the present invention, a preferred marker nucleic acid molecule of the present invention has the nucleic acid sequence set forth in SEQ ID NO:1 and SEQ ID NO:2 or complements thereof or fragments of either. In another aspect of the present invention, a preferred marker nucleic acid molecule of the present invention shares 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and 100% sequence identity with the nucleic acid sequence set forth in SEQ ID NO:1 and SEQ ID NO:2 or complement thereof or fragments of either. In a further aspect of the present invention, a preferred marker nucleic acid molecule of the present invention shares 95% 96%, 97%, 98%, 99% and 100% sequence identity with the sequence set forth in SEQ ID NO:1 and SEQ ID NO: 2 or complement thereof or fragments of either. SEQ ID NO:1 and SEQ ID NO:2 may be used as markers in plant breeding methods to identify the progeny of genetic crosses similar to the methods described for simple sequence repeat DNA marker analysis, in “DNA markers: Protocols, applications, and overviews: (1997) 173-185, Cregan, et al., eds., Wiley-Liss NY”; all of which is herein incorporated by reference. The hybridization of the probe to the target DNA molecule can be detected by any number of methods known to those skilled in the art, these can include, but are not limited to, fluorescent tags, radioactive tags, antibody based tags, and chemilluminescent tags.
Regarding the amplification of a target nucleic acid sequence (e.g., by PCR) using a particular amplification primer pair, “stringent conditions” are conditions that permit the primer pair to hybridize only to the target nucleic-acid sequence to which a primer having the corresponding wild-type sequence (or its complement) would bind and preferably to produce a unique amplification product, the amplicon, in a DNA thermal amplification reaction.
The term “specific for (a target sequence)” indicates that a probe or primer hybridizes under stringent hybridization conditions only to the target sequence in a sample comprising the target sequence.
In a particularly preferred variant, the process according to the invention is used for treating transgenic vegetable, maize, soya bean, cotton, tobacco, rice, potato and sugar beet varieties. These are preferably Bt plants.
The vegetable plants or varieties are, for example, the following useful plants:
Bt vegetables including exemplary methods for preparing them are described in detail, for example, in Barton et al., 1987, Plant Physiol. 85: 1103-1109; Vaeck et al., 1987, Nature 328: 33-37; Fischhoff et al., 1987, Bio/Technology 5: 807-813. In addition, Bt vegetable plants are already known as commercial varieties, for example the potato cultivar NewLeaf® (Monsanto). The preparation of Bt vegetables is also described in U.S. Pat. No. 6,072,105.
Likewise, Bt cotton is already known in principle, for example from U.S. Pat. No. 5,322,938. In the context of the present invention, particular preference is given to Bt cotton with the trade names NuCOTN33® and NuCOTN33B®.
The use and preparation of Bt maize has likewise already been known for a long time, for example from Ishida, Y., Saito, H., Ohta, S., Hiei, Y., Komari, T., and Kumashiro, T. (1996). High efficiency transformation of maize (Zea mayz L.) mediated by Agrobacterium tumefaciens, Nature Biotechnology 4: 745-750. EP-B-0485506, too, describes the preparation of Bt maize plants. Furthermore, different varieties of Bt maize are commercially available, for example under the following names (company/companies is/are in each case given in brackets): KnockOut® (Novartis Seeds), NaturGard® (Mycogen Seeds), Yieldgard® (Novartis Seeds, Monsanto, Cargill, Golden Harvest, Pioneer, DeKalb inter alia), Bt-Xtra® (DeKalb) and StarLink® (Aventis CropScience, Garst inter alia). For the purpose of the present invention, particular preference is given especially to the following maize cultivars: KnockOut®, NaturGard®, Yieldgard®, Bt-Xtra® and StarLink®.
For soya beans, too, Roundup®Ready cultivar or cultivars resistant to the herbicide Liberty Link® are available and can be treated according to the invention. In the case of rice, a large number of “Golden Rice” lines are available which are likewise characterized in that, by virtue of a transgenic modification, they have an increased content of provitamin A. They, too, are examples of plants which can be treated by the method according to the invention, with the advantages described.
The method according to the invention is suitable for controlling a large number of harmful organisms which occur in particular in vegetables, maize and cotton, in particular insects and arachnids, very particularly preferably insects. The pests mentioned include:
The method according to the invention for the treatment of Bt vegetables, Bt maize, Bt cotton, Bt soya beans, Bt tobacco and also Bt rice, Bt sugar beets or Bt potatoes is particularly suitable for controlling aphids (Aphidina), whiteflies (Trialeurodes), thrips (Thysanoptera), spider mites (Arachnida), soft scale insects or mealy bugs (Coccoidae and Pseudococcoidae, respectively).
The active compounds which can be used according to the invention can be employed in customary formulations, such as solutions, emulsions, wettable powders, water- and oil-based suspensions, powders, dusts, pastes, soluble powders, soluble granules, granules for broadcasting, suspoemulsion concentrates, natural compounds impregnated with active compound, synthetic substances impregnated with active compound, fertilizers and also microencapsulations in polymeric substances.
These formulations are prepared in a known manner, for example by mixing the active compounds with extenders, i.e. liquid solvents and/or solid carriers, if appropriate using surfactants, i.e. emulsifiers and/or dispersants and/or foam-formers. The formulations are prepared either in suitable plants or else before or during application.
Wettable powders are preparations which can be dispersed homogeneously in water and which, in addition to the active compound and beside a diluent or inert substance, also comprise wetting agents, for example polyethoxylated alkylphenols, polyethoxylated fatty alcohols, alkylsulphonates or alkylphenylsulphonates and dispersants, for example sodium lignosulphonate, sodium 2,2′-dinaphthylmethane-6,6′-disulphonate.
Dusts are obtained by grinding the active compound with finely distributed solid substances, for example talc, natural clays, such as kaolin, bentonite, pyrophillite or diatomaceous earth. Granules can be prepared either by spraying the active compound onto granular inert material capable of adsorption or by applying active compound concentrates to the surface of carrier substances, such as sand, kaolinites or granular inert material, by means of adhesives, for example polyvinyl alcohol, sodium polyacrylate or mineral oils. Suitable active compounds can also be granulated in the manner customary for the preparation of fertilizer granules—if desired as a mixture with fertilizers.
Suitable for use as auxiliaries are substances which are suitable for imparting to the composition itself and/or to preparations derived therefrom (for example spray liquors, seed dressings) particular properties such as certain technical properties and/or also particular biological properties. Typical suitable auxiliaries are: extenders, solvents and carriers.
Suitable extenders are, for example, water, polar and nonpolar organic chemical liquids, for example from the classes of the aromatic and non-aromatic hydrocarbons (such as paraffins, alkylbenzenes, alkylnaphthalenes, chlorobenzenes), the alcohols and polyols (which, if appropriate, may also be substituted, etherified and/or esterified), the ketones (such as acetone, cyclohexanone), esters (including fats and oils) and (poly)ethers, the unsubstituted and substituted amines, amides, lactams (such as N-alkylpyrrolidones) and lactones, the sulphones and sulphoxides (such as dimethyl sulphoxide).
If the extender used is water, it is also possible to employ, for example, organic solvents as auxiliary solvents. Essentially, suitable liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics and chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example petroleum fractions, mineral and vegetable oils, alcohols such as butanol or glycol and also their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethyl sulphoxide, and also water.
Suitable solid carriers are for example, ammonium salts and ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals, such as finely divided silica, alumina and silicates; suitable solid carriers for granules are: for example, crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, and also synthetic granules of inorganic and organic meals, and granules of organic material such as paper, sawdust, coconut shells, maize cobs and tobacco stalks; suitable emulsifiers and/or foam-formers are: for example, nonionic and anionic emulsifiers, such as polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, for example alkylaryl polyglycol ethers, alkylsulphonates, alkyl sulphates, arylsulphonates and also protein hydrolysates; suitable dispersants are nonionic and/or ionic substances, for example from the classes of the alcohol-POE and/or -POP ethers, acid and/or POP POE esters, alkylaryl and/or POP POE ethers, fat and/or POP POE adducts, POE- and/or POP-polyol derivatives, POE- and/or POP-sorbitan or -sugar adducts, alkyl or aryl sulphates, alkyl- or arylsulphonates and alkyl or aryl phosphates or the corresponding PO-ether adducts. Furthermore, suitable oligo- or polymers, for example those derived from vinylic monomers, from acrylic acid, from EO and/or PO alone or in combination with, for example, (poly)alcohols or (poly)amines. It is also possible to employ lignin and its sulphonic acid derivatives, unmodified and modified celluloses, aromatic and/or aliphatic sulphonic acids and their adducts with formaldehyde.
Tackifiers such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules or lattices, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, as well as natural phospholipids such as cephalins and lecithins, and synthetic phospholipids, can be used in the formulations.
It is possible to use colorants such as inorganic pigments, for example iron oxide, titanium oxide and Prussian Blue, and organic dyestuffs, such as alizarin dyestuffs, azo dyestuffs and metal phthalocyanine dyestuffs, and trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.
Other possible additives are perfumes, mineral or vegetable, optionally modified oils, waxes and nutrients (including trace nutrients), such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.
Stabilizers, such as low-temperature stabilizers, preservatives, antioxidants, light stabilizers or other agents which improve chemical and/or physical stability may also be present.
These individual types of formulation are known in principle and are described, for example, in: “Pesticides Formulations”, 2nd Ed., Marcel Dekker N.Y.; Martens, 1979, “Spray Drying Handbook”, 3rd Ed., G. Goodwin Ltd. London.
Based on his general expert knowledge, the person skilled in the art is able to choose suitable formulation auxiliaries (in this context, see, for example, Watkins, “Handbook of Insecticide Dust Diluents and Carriers”, 2nd Ed., Darland Books, Caldwell N.J.).
In a preferred embodiment, the plants or plant parts are treated according to the invention with an oil-based suspension concentrate. An advantageous suspension concentrate is known from WO 2005/084435 (EP 1 725 104 A2). It consists of at least one room-temperature-solid active agrochemical substance, at least one “closed” penetrant, at least one vegetable oil or mineral oil, at least one nonionic surfactant and/or at least one anionic surfactant, and optionally one or more additives from the groups of the emulsifiers, foam inhibitors, preservatives, antioxidants, colorants and/or inert filler materials. Preferred embodiments of the suspension concentrate are described in the above-mentioned WO 2005/084435. For the purpose of the disclosure, both documents are incorporated herein in their entirety by way of reference.
In a further preferred embodiment, the plants or plant parts are treated according to the invention with compositions comprising ammonium or phosphonium salts and, if appropriate, penetrants. Advantageous compositions are known from WO2007/068355 and from the not prior-published EP 07109732.3. They consist of at least one compound of the formula (I) and at least one ammonium or phosphonium salt and, if appropriate, penetrants. Preferred embodiments are described in WO2007/068355 and the not prior-published EP 07109732.3. For the purpose of the disclosure, these documents are incorporated herein in their entirety by way of reference.
In general, the formulations comprise from 0.01 to 98% by weight of active compound, preferably from 0.5 to 90%. In wettable powders, the active compound concentration is, for example, from about 10 to 90% by weight, the remainder to 100% by weight consisting of customary formulation components. In the case of emulsifiable concentrates, the active compound concentration can be from about 5 to 80% by weight. In most cases, formulations in the form of dusts comprise from 5 to 20% by weight of active compound, sprayable solutions comprise about 2 to 20% by weight. In the case of granules, the active compound content depends partially on whether the active compound is present in liquid or solid form and on which granulation auxiliaries, fillers, etc., are used.
The required application rate may also vary with external conditions such as, inter alia, temperature and humidity. It may vary within wide limits, for example between 0.1 g/h and 5.0 kg/ha or more of active substance. However, they are preferably between 0.1 g/ha and 1.0 kg/ha. Owing to the synergistic effects between Bt vegetables and the insecticide, particular preference is given to application rates of from 0.1 to 500 g/ha.
For compounds of the formula (I), preference is given to application rates of from 10 to 500 g/ha; particularly preferred are from 10 to 200 g/ha.
In a particular embodiment of the method according to the invention, the compound of the formula (I) is employed in an application rate of from 0.1 g/ha to 5.0 kg/ha, preferably from 0.1 to 500 g/ha and particularly preferably from 50 to 500 g/ha and especially preferably from 50 to 200 g/ha.
In their commercial formulations and in the use forms prepared from these formulations, the active compounds according to the invention may be present as mixtures with other active compounds, such as insecticides, attractants, sterilants, acaricides, nematicides, fungicides, growth-regulating substances or herbicides.
A mixture with other known compounds, such as herbicides, fertilizers, growth regulators, safeners, semiochemicals, or else with agents for improving plant properties is also possible.
The active compound content of the use forms prepared from the commercial formulations can be from 0.00000001 to 95% by weight, preferably between 0.00001 and 1% by weight, of active compound.
For preparing the stock solution, 20 mg of active compound is solved in 200 μl of dimethylformamide and filled-up with 9.78 ml SC blank formulation of Belt. The final test concentrations are prepared by dilution with water.
The test is conducted with conventional soybean plants (Glycine max; non-transgenic) and transgenic soybean plants containing a Cry1Ac gene (Intacta from Monsanto). When the plants are in stage V2 (3 nodes with 2 unfolded trifoliolates), they are treated by spray application with the active compound preparation. After application, clip-cages with 5-6 L2 larvae of the fall army worm (Spodoptera frugiperda) are placed on the leaves.
After the specified period of time, feeding damage (white holes on leaves) of Spodoptera frugiperda on conventional soybean,
According to the present application in this test the following combinations of transgenic plant and compound shows a superior effect compared to the treated, non-transgenic plant respectively the non-treated, transgenic plant:
Results of the experiments 1, 2 and 9 to 18 of Table A are shown in
| Number | Date | Country | Kind |
|---|---|---|---|
| 13164466.8 | Apr 2013 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2014/057667 | 4/15/2014 | WO | 00 |
