Patent Application
20230098202
The present invention relates in general to compounds having fungicidal and bactericidal properties for agricultural uses.
Plant pests and diseases represent major challenges to productivity in modern agriculture. Rusts are a diverse group of plant pathogens with tens of genera and thousands of species. They have huge economic importance and may cause tens of percent's loss in yield in cereals, maize and soybean (Gessese 2019; Groth et al., 1998; Hershman et al., 2011).
Puccinia spp. is an obligatory pathogenic fungus and a major genus in plant rusts belonging to phylogenetic lineage of Basidiomycetes. Puccinia spp. causes a wide range of commercially significant plant diseases in cereals (such as yellow rust in wheat) and maize (common rust)—(Gessese 2019; Groth et al., 1998).
Soil-borne plant pathogens cause crucial damage to agricultural crops. The phytopathogenic fungus Rhizoctonia spp. belongs to phylogenetic lineage of Basidiomycetes. It causes a wide range of commercially significant plant diseases, such as brown patch, damping off in seedlings, root rot and belly rot in vegetable crops and sheath blight in rice. All Rhizoctonia diseases, and subsequent secondary infections in plants are difficult to control (Erlacher et al., 2014).
Pythium spp. is phytopathogenic fungus-like organism which belongs to phylogenetic lineage of eukaryotic microorganisms called Oomycetes which causes the widespread “damping off” disease of tobacco, tomato, mustard, chilies and cress seedlings (Martin & Loper, 2010).
Phytophthora spp. is an obligatory plant fungal like pathogen which belongs to phylogenetic lineage of eukaryotic microorganisms called Oomycetes. Phytophthora infestans is a serious potato disease known as potato blight resulting in foliage blight and rot of tubers. The disease can cause complete loss of a potato harvest (Sedláková et al., 2012). Phytophthora attacks the aerial parts of many plant species and it is the major cause of leaf blight, canker fruit rot diseases in tomato, pumpkins and other crops.
Botrytis spp. is a ubiquitous filamentous fungal pathogen of a wide range of plant species belonging to phylogenetic lineage of Ascomycetes. Botrytis can infect all aerial parts of its host plants to a certain extent. Botrytis causes a disease called grey mold in diverse array of agronomically important crops and commodity plants, such as grapevine, tomato, strawberry, cucumber, bulb flowers, cut flowers and ornamentals (J. A. L. van Kan, 2005).
Fusarium spp. is a large genus of filamentous fungi belonging to phylogenetic lineage of Ascomycetes. Many species of Fusarium are pathogenic to plants and cause serious diseases like wilt or ‘rot’ of economically important plants, mostly vegetables. In addition, Fusarium species infects cereals causing head blight and ear rot in maize and cause to mycotoxins accumulation under certain conditions (J. E. E. Jenkins, Y. S. Clark and A. E. Buckle, 1998).
Alternaria spp. is a ubiquitous fungal genus with numerous species that cause significant damage to agricultural products including cereal grains, fruits and vegetables—apples, potatoes, tomatoes and others (Patriarca, A., & Fernández Pinto, V. 2018).
Pseudomonas spp. is a plant pathogenic bacterial genus which is virulent in the diverse arrays of crop plants and causes to significant leaf and stem lesions. Pseudomonas spp. causes the following diseases in economically significant crops plants and orchards such as: pith necrosis in parsnip and tomato, brown blotch and leaf sheath brown rot in rice, bacterial canker in almonds and olive knot disease in olives (Moore L. W., 1988; Hofte M. and De Vos P., 2006).
A variety of methods have been tested for the management of Pseudomonas spp. in crop plants. They include cultural management, host resistance, biological control with microbial antagonists and chemical control. None of them gives full control.
The number of available active ingredients for crop protection purposes against these diseases is diminishing from year to year due to increasing pest resistance, erratic climatic conditions and mounting regulatory pressure. New active ingredients are urgently needed for development of novel environmentally sustainable crop protection solutions.
In one aspect of the present invention, a method for controlling, preventing, reducing or eradicating instances of plant-pathogen infestation on a plant, plant organ, plant part, or plant propagation material, the method comprises: applying to a plant, plant part, plant organ or plant propagation material, or to soil surrounding said plant, a pesticidally effective amount of at least one compound of formula (I):
wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, methyl, hydroxy and methoxy group, and halogen atom (F, Cl, Br, I); R5 and R6 are independently selected from hydrogen, methyl and ethyl; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy; or stereoisomers, or agriculturally acceptable salts thereof.
In a specific embodiment, the compounds of formula (I) which are applied in the method of the present invention are:
Compound 3: (1S,4R)-4-(3,4-dichlorophenyl)-N-methyl-1,2,3,4-tetrahydronaphthalen-1-aminium chloride,
Compound 1: (5R,6R,7R)-5-(3,4-dihydroxyphenyl)-6,7-dimethyl-5,6,7,8-tetrahydronaphthalene-2,3-diol, and
Compound 2: 4-((1R,2R,3R)-7-hydroxy-6-methoxy-2,3-dimethyl-1,2,3,4-tetrahydronaphthalen-1-yl)benzene-1,2-diol.
In some embodiments, Compound 3 is applied to a plant-pathogen which is a member selected from: a Basidomycete of the class Pucciniomycetes or the genus Rhizoctonia; an Ascomycota of the class Dothideomycetes or a genus selected from Botrytis and Fusarium; and a Heterokontophyta of the class Oomycota.
In other embodiments, Compound 1 is applied to a plant-pathogen which is a member selected from: a Basidomycete of the class Pucciniomycetes or the genus Rhizoctonia; a Heterokontophyta of the class Oomycota; and a protobacterium of the order Pseudomonadales.
In still other embodiments, Compound 2 is applied to a plant-pathogen which is a member selected from: a Basidomycete of the class Pucciniomycetes or the genus Rhizoctonia; a Heterokontophyta of the family Pythiaceae; and a protobacterium of the order Pseudomonadales.
In another aspect of the present invention, a pesticide composition comprises at least one compound of formula (I),
wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, methyl, hydroxy and methoxy group, and halogen atom (F, Cl, Br, I); R5 and R6 are independently selected from hydrogen, methyl and ethyl; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy group; stereoisomers or agriculturally acceptable salts thereof.
In a specific embodiment, the compounds of formula (I) of the composition of the present invention are:
Compound 3: (1S,4R)-4-(3,4-dichlorophenyl)-N-methyl-1,2,3,4-tetrahydronaphthalen-1-aminium chloride,
Compound 1: (5R,6R,7R)-5-(3,4-dihydroxyphenyl)-6,7-dimethyl-5,6,7,8-tetrahydronaphthalene-2,3-diol, and
Compound 2: 4-((1R,2R,3R)-7-hydroxy-6-methoxy-2,3-dimethyl-1,2,3,4-tetrahydronaphthalen-1-yl)benzene-1,2-diol.
It has been found in accordance with the present invention that 1-phenyl-tetralin derivatives of the following formula (I), stereoisomers or agriculturally acceptable salts thereof are potent pesticides against several Basidomyceta, Ascomycota and Heterokontophyta fungi as well as protobacteria of the genus Pseudomonas:
wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, methyl, hydroxy and methoxy group, and halogen atom (F, Cl, Br, I);
R5 and R6 are independently selected from hydrogen, methyl and ethyl; and
R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy.
In a particular embodiment, the compounds of the present invention are the compounds of formula (I), wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, methyl, hydroxy and methoxy group, and halogen atom (F, Cl, Br, I); R5 and R6 are methyl; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy group.
In a more particular embodiment, the compounds of the present invention are the compounds of formula (I), wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, hydroxy, and methoxy group; R5 and R6 are methyl; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy group.
In a specific embodiment, the compounds of the present invention are the compounds of formula (I), wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, hydroxy, and methoxy; R5 and R6 are methyl; and R7 is hydrogen.
In the specific embodiment of the present invention, the compounds are:
In a further particular embodiment, the compounds of the present invention are the compounds of formula (I), wherein R1a, R1b, R2 are independently selected from hydrogen and halogen atom (F, Cl, Br, I); R3, R4, R5 and R6 are hydrogen; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy group.
In yet further particular embodiment, the compounds of the present invention are the compounds of formula (I), wherein R1a, R1b, R2 are independently selected from hydrogen and chlorine atom; R3, R4, R5 and R6 are hydrogen; and R7 is methylamino group.
The specific compound of the present invention according to the above embodiment is:
In general, Compound 1 is a 1-phenyl-tetralin derivative, which is a member of the class of 1-aryl tetralin lignans. Compound 2 is another 1-phenyl-tetralin derivative, which is also a member of the class of 1-aryl tetralin lignans. Compound 3 is known as sertraline hydrochloride and it is a selective serotonin reuptake inhibitor (SSRI) anti-depressant drug. These three specific compounds are stereoisomeric 1-phenyl-tetralin derivatives of formula (I).
The present invention provides in one aspect a method for controlling, preventing, reducing or eradicating plant-pathogen infestation or instances thereof, on a plant, plant organ, plant part, or plant propagation material, the method comprising: applying to a plant, plant organ or plant propagation material, or to soil surrounding said plant, a pesticidally effective amount of at least one compound of Compound 3: (1S,4R)-4-(3,4-dichlorophenyl)-N-methyl-1,2,3,4-tetrahydronaphthalen-1-aminium chloride or stereoisomers or agriculturally acceptable salts thereof as an active pesticidal ingredient, or a pesticide composition of compound 3, wherein said plant-pathogen is a member selected from: a Basidomycete of the class Pucciniomycetes or the genus Rhizoctonia; an Ascomycota of the class Dothideomycetes or a genus selected from Botrytis and Fusarium; and; a Heterokontophyta of the class Oomycota.
In another aspect, the present invention provides a method for controlling, preventing, reducing or eradicating instances of plant-pathogen infestation on a plant, plant organ, plant part, or plant propagation material, the method comprising: applying to a plant, plant part, plant organ or plant propagation material, or to soil surrounding said plant, a pesticidally effective amount of Compound 1: (5R,6R,7R)-5-(3,4-dihydroxyphenyl)-6,7-dimethyl-5,6,7,8-tetrahydronaphthalene-2,3-diol, or stereoisomers, or agriculturally acceptable salts thereof, wherein said plant-pathogen is a member selected from: a Basidomycete of the class Pucciniomycetes or the genus Rhizoctonia; a Heterokontophyta of the class Oomycota; and a protobacterium of the order Pseudomonadales.
In additional aspect, the present invention provided a method for controlling, preventing, reducing or eradicating instances of plant-pathogen infestation on a plant, plant organ, plant part, or plant propagation material, the method comprising: applying to a plant, plant part, plant organ or plant propagation material, or to soil surrounding said plant, a pesticidally effective amount of Compound 2: 4-((1R,2R,3R)-7-hydroxy-6-methoxy-2,3-dimethyl-1,2,3,4-tetrahydronaphthalen-1-yl)benzene-1,2-diol, or stereoisomers, or agriculturally acceptable salts thereof, wherein said plant-pathogen is a member selected from: a Basidomycete of the class Pucciniomycetes or the genus Rhizoctonia; a Heterokontophyta of the family Pythiaceae; and a protobacterium of the order Pseudomonadales.
The method of treatment of the present invention according to anyone of the embodiments disclosed herein is useful for example against the following diseases: common rust in corn; crown rust of oats and ryegrass; stem rust of wheat and Kentucky bluegrass, or black rust of cereals; daylily rust; wheat rust in grains; brown or red rust; ‘yellow rust’ in cereals; ‘brown rust’ or ‘orange rust’ in sugarcane; or coffee rust; leaf and stem rust in barley; potato blight, Phytophthora palmivora in cacao, canker fruit rot diseases in tomato and pumpkins; Phytophthora spp. crown and collar rot in pome and stone fruit; “damping off” disease caused by Pythium spp. in tobacco, tomato, cucumbers, mustard, chilies and cress seedlings; gray mold (Botrytis cinerea) in table and wine grapes, strawberries and vegetable crops; Fusarium spp. causing wilt or ‘rot’ of vegetables, bananas; Fusarium spp. head and ear rot in maize; Fusarium graminearum head blight in small grains; Rhizoctonia spp. causing brown patch, damping off in seedlings, root rot and belly rot in vegetables and sheath blight in rice; Alternaria spp. causing spots, rots and blights on leaves and fruits.
In certain embodiments, the plant-pathogen is a member of the class Pucciniomycetes of an order selected from Helicobasidiales, Pachnocybales, Platygloeales, Pucciniales, and Septobasidiales. In specific embodiments, the plant-pathogen is a member of the order Pucciniales.
In some embodiments, the Pucciniales plant-pathogen is a member of a family selected from Chaconiaceae, Coleosporiaceae, Cronartiaceae, Melampsoraceae, Mikronegeriaceae, Phakopsoraceae, Phragmidiaceae, Pileolariaceae, Pucciniaceae, Pucciniosiraceae, Pucciniastraceae, Raveneliaceae, Sphaerophragmiaceae, Uncolaceae, Uropyxidaceae, mitosporic Pucciniales and Pucciniales incertae sedis. In particular embodiments, the Pucciniales plant-pathogen is a member of the family Pucciniaceae.
In other embodiments, the Pucciniaceae plant-pathogen is a member of the genus Puccinia, such as Puccinia sorghi, Puccinia coronate, Puccinia graminis, Puccinia hemerocallidis, Puccinia hemerocallidis, Puccinia persistens subsp. Triticina, Puccinia striiformis, Puccinia melanocephala, Puccinia kuehnii and Hemileia vastatrix. In specific embodiments, the Puccinia plant-pathogen is selected from Puccinia sorghi and Puccinia triticina.
In further embodiments, the method of the invention is useful for controlling, preventing, reducing or eradicating any one of the Pucciniomycetes plant-pathogens described above, and in particular Puccinia sorghi and Puccinia triticina, by applying as described above any one of compounds 3, 1 or 2, or any combination thereof.
In some embodiments, the plant-pathogen is a member of the genus Rhizoctonia (which is in the Ceratobasidiaceae family of the order Cantharellales), such as Rhizoctonia solani, Rhizoctonia bataticola also known as Macrophomina phaseolina, Rhizoctonia carotae also known as Fibulorhizoctonia carotae, Rhizoctonia cerealis, Rhizoctonia crocorum also known as Thanatophytum crocorum, Rhizoctonia fragariae, Rhizoctonia goodyerae-repentis also known as Ceratobasidium cornigerum, Rhizoctonia oryzae also known as Waitea circinate, and Rhizoctonia ramicola also known as Ceratorhiza ramicola. In particular embodiments, the plant-pathogen is Rhizoctonia solani.
In further embodiments, the method of the invention is useful for controlling, preventing, reducing or eradicating any one of the Rhizoctonia plant-pathogens described above, and in particular Rhizoctonia solani, by applying as described above any one of compounds 3, 1 or 2, or any combination thereof.
In yet further embodiments, the plant-pathogen is a member of the class Dothideomycetes of an order selected from Capnodiales, Dothideales, Myriangiales, Hysteriales, Jahnulales, Mytilinidiales, Pleosporales, Botryosphaeriales, Microthyriales, Patellariales, and Trypetheliales. In specific embodiments, the plant-pathogen is a member of the order Pleosporales.
In other embodiments, the Pleosporales plant-pathogen is a member of a family selected from Aigialaceae, Amniculicolaceae, Cucurbitariaceae, Delitschiaceae, Diademaceae, Didymellaceae, Didymosphaeriaceae, Halojulellaceae, Lentitheciaceae, Leptosphaeriaceae, Lindgomycetaceae, Lop hiostomataceae, Massariaceae, Massarinaceae, Melanommataceae, Montagnulaceae, Phaeosphaeriaceae, Phaeotrichaceae, Pleomassariaceae, Pleosporaceae, Sporormiaceae, Venturiaceae, Teichosporaceae, Tetraplosphaeriaceae, Testudinaceae, Trematosphaeriaceae, and Zopfiaceae. In particular embodiments, the Pleosporales plant-pathogen is a member of the family Pleosporaceae.
In some embodiments, the Pleosporaceae plant-pathogen is a member of a genus selected from Alternaria, Bipolaris, Cochliobolus, Crivellia, Decorospora, Exserohilum, Falciformispora, Kriegeriella, Lewia, Macrospora, Monascostroma, Pithomyces, Platysporoides, Pleospora, Pseudoyuconia, Pyrenophora, Setosphaeria, and Zeuctomorpha. In specific embodiments, the Pleosporaceae plant-pathogen is a member of the genus Alternaria.
In still other embodiments, the Alternaria plant-pathogen is selected from Alternaria alternata, Alternaria alternantherae, Alternaria arborescens, Alternaria arbusti, Alternaria blumeae, Alternaria brassicae, Alternaria brassicicola, Alternaria burnsii, Alternaria carotiincultae, Alternaria carthami, Alternaria celosiae, Alternaria cinerariae, Alternaria citri, Alternaria conjuncta, Alternaria cucumerina—grows on various cucurbits, Alternaria dauci, Alternaria dianthi, Alternaria dianthicola, Alternaria eichhorniae, Alternaria euphorbiicola, Alternaria gaisen, Alternaria helianthin, Alternaria helianthicola, Alternaria hungarica, Alternaria infectoria, Alternaria japonica, Alternaria limicola, Alternaria linicola, Alternaria longipes, Alternaria mali, Alternaria molesta, Alternaria panax, Alternaria perpunctulata, Alternaria petroselini, Alternaria porri, Alternaria quercicola, Alternaria radicina, Alternaria raphanin, Alternaria saponariae, Alternaria selini, Alternaria senecionis, Alternaria solani, Alternaria smyrnii, Alternaria tenuissima, Alternaria triticina, Alternaria ventricosa, and Alternaria zinnia. In further specific embodiments, the Alternaria plant-pathogen is selected from Alternaria alternata and Alternaria solani.
In other embodiments, the method of the invention is useful for controlling, preventing, reducing or eradicating any one of the Dothideomycetes plant-pathogens described above, and in particular Alternaria alternata by applying compound 3 as described above; and Alternaria solani, by applying as described above any one of compounds 3, 1 or 2, or any combination thereof.
In still other embodiments, the plant-pathogen is a member of the of the class Leotiomycetes of an order selected from Cyttariales, Erysiphales, Helotiales, Leotiales, and Rhytismatales, Thelebolales.
In further embodiments, the plant-pathogen is a member of the order Helotiales. In yet further embodiments, the Helotiales plant-pathogen is a member of a family selected from Ascocorticiaceae, Dermateaceae, Helotiaceae, Hemiphacidiaceae, Hyaloscyphaceae, Loramycetaceae, Phacidiaceae, Rutstroemiaceae, Sclerotinaceae, and Vibrisseaceae. In particular embodiments, the Helotiales plant-pathogen is a member of the family Sclerotiniaceae. In other particular embodiments, the Sclerotiniaceae plant-pathogen is a member of the genus Botrytis.
In certain embodiments, the Botrytis plant-pathogen is selected from Botrytis aclada, Botrytis allii, Botrytis allii-fistulosi, Botrytis ampelophila, Botrytis anacardii, Botrytis anthophila, Botrytis argillacea, Botrytis arisaemae, Botrytis artocarpi, Botrytis bifurcata, Botrytis bryi, Botrytis capsularum, Botrytis carnea, Botrytis caroliniana, Botrytis carthami, Botrytis cercosporaecola, Botrytis cercosporicola, Botrytis cinerea, Botrytis citricola, Botrytis citrina, Botrytis convallarias, Botrytis croci, Botrytis cryptomeriae, Botrytis densa, Botrytis diospyri, Botrytis elliptica, Botrytis fabae, Botrytis fabiopsis, Botrytis galanthina, Botrytis gladioli, Botrytis gossypina, Botrytis hormini, Botrytis hyacinthi, Botrytis isabellina, Botrytis latebricola, Botrytis liliorum, Botrytis limacidae, Botrytis luteobrunnea, Botrytis lutescens, Botrytis mali, Botrytis monilioides, Botrytis necans, Botrytis paeoniae, Botrytis peronosporoides, Botrytis pistiae, Botrytis platensis, Botrytis pruinosa, Botrytis pseudocinerea, Botrytis pyramidalis, Botrytis rivoltae, Botrytis rosea, Botrytis rubescens, Botrytis rudiculoides, Botrytis sekimotoi, Botrytis septospora, Botrytis setuligera, Botrytis sinoallii, Botrytis sonchina, Botrytis splendida, Botrytis squamosa, Botrytis taxi, Botrytis terrestris, Botrytis tracheiphila, Botrytis trifolii, Botrytis tulipae, Botrytis viciae-hirsutae, and Botrytis yuae. In some embodiments, the plant-pathogen is Botrytis cinerea.
In other embodiments, the plant-pathogen is a member of the class Sordariomycetes of an order selected from Coronophorales, Glomerellales, Hypocreales, Melanosporales, Microascales, Boliniales, Calosphaeriales, Chaetosphaeriales, Coniochaetales, Diaporthales, Magnaporthales, Ophiostomatales, Sordariales, Xylariales, Koralionastetales, Lulworthiales, Meliolales, Phyllachorales, and Trichosphaeriales.
In still other embodiments, the plant-pathogen is a member of the order Hypocreales. In certain embodiments, the Hypocreales plant-pathogen is a member a family selected from Bionectriaceae, Cordycipitaceae, Clavicipitaceae, Hypocreaceae, Nectriaceae, Niessliaceae, Ophiocordycipitaceae, and Stachybotryaceae. In particular embodiments, the Hypocreales plant-pathogen is a member of the family Nectriaceae.
In further embodiments, the Nectriaceae plant-pathogen is a member of the genus Fusarium. In certain embodiments, the Fusarium plant-pathogen is selected from Fusarium acaciae, Fusarium acaciae-mearnsii, Fusarium acutatum, Fusarium aderholdii, Fusarium acremoniopsis, Fusarium affine, Fusarium arthrosporioides, Fusarium avenaceum, Fusarium bubigeum, Fusarium circinatum, Fusarium crookwellense, Fusarium culmorum, Fusarium graminearum, Fusarium incarnatum, Fusarium langsethiae, Fusarium mangiferae, Fusarium merismoides, Fusarium oxysporum, Fusarium pallidoroseum, Fusarium poae, Fusarium proliferatum, Fusarium pseudograminearum, Fusarium redolens, Fusarium sacchari, Fusarium solani, Fusarium sporotrichioides, Fusarium sterilihyphosum, Fusarium subglutinans, Fusarium sulphureum, Fusarium tricinctum, Fusarium venenatum, Fusarium verticillioides, and Fusarium virguliforme. In some embodiments, the plant-pathogen is the species Fusarium oxysporum.
In yet further embodiments, the plant-pathogen is a member of the class Oomycota of an order selected from Lagenidiales, Leptomitales, Peronosporales, Rhipidiales, and Saprolegniales. In certain embodiments, the plant-pathogen is a member of the class Oomycota of the order Peronosporales.
In some embodiments, the Peronosporales plant-pathogen is a member of a family selected from Lagenidiaceae, Olpidiosidaceae, Sirolpidiaceae, Leptomitaceae, Albuginaceae, Peronosporaceae, Pythiaceae, Rhipidaceae, Ectrogellaceae, Haliphthoraceae, Leptolegniellaceae, and Saprolegniaceae. In particular embodiments, the plant-pathogen is a member of the family Peronosporaceae or Pythiaceae.
In certain embodiments, the Peronosporaceae plant-pathogen is a member of a genus selected from Baobabopsis, Basidiophora, Benua, Bremia, Calycofera, Eraphthora, Graminivora, Hyaloperonospora, Nothophytophthora, Novotelnova, Paraperonospora, Perofascia, Peronosclerospora, Peronospora, Phytophthora, Plasmopara, Plasmoverna, Protobremia, Pseudoperonospora, Sclerophthora, Sclerospora, and Viennotia.
In certain embodiments, the Peronosporaceae plant-pathogen is a member of the genus Phytophthora. In specific embodiments, the Phytophthora plant-pathogen is selected from Phytophthora acerina, Phytophthora agathidicida, Phytophthora alni, Phytophthora x alni, Phytophthora alticola, Phytophthora amaranthi, Phytophthora amnicola, Phytophthora amnicola x moyootj, Phytophthora andina, Phytophthora aquimorbida, Phytophthora arecae, Phytophthora arenaria, Phytophthora cf. arenaria, Phytophthora aff arenaria, Phytophthora asiatica, Phytophthora asparagi, Phytophthora aff asparagi, Phytophthora attenuata, Phytophthora austrocedrae, Phytophthora balyanboodja, Phytophthora batemanensis, Phytophthora bilorbang, Phytophthora bisheria, Phytophthora bishii, Phytophthora boehmeriae, Phytophthora boodjera, Phytophthora borealis, Phytophthora botryosa, Phytophthora cf. botryosa, Phytophthora aff. botryosa, Phytophthora brassicae, Phytophthora cactorum, Phytophthora cactorum var. applanata, Phytophthora cactorum x hedraiandra, Phytophthora cajani, Phytophthora cambivora, Phytophthora capensis, Phytophthora capsici, Phytophthora aff. capsici, Phytophthora captiosa, Phytophthora castaneae, Phytophthora castanetorum, Phytophthora chlamydospora, Phytophthora chrysanthemi, Phytophthora cichorii, Phytophthora aff. cichorii, Phytophthora cinnamomi, Phytophthora cinnamomi var. cinnamomi, Phytophthora cinnamomi var. parvispora, Phytophthora cinnamomi var. robiniae, Phytophthora citricola, Phytophthora aff citricola, Phytophthora citrophthora, Phytophthora citrophthora var. clementina, Phytophthora aff citrophthora, Phytophthora clandestina, Phytophthora cocois, Phytophthora colocasiae, Phytophthora condilina, Phytophthora constricta, Phytophthora cooljarloo, Phytophthora crassamura, Phytophthora cryptogea, Phytophthora aff cryptogea, Phytophthora cuyabensis, Phytophthora cyperi, Phytophthora dauci, Phytophthora aff dauci, Phytophthora drechsleri, Phytophthora drechsleri var. cajani, Phytophthora elongata, Phytophthora cf. elongata, Phytophthora erythroseptica, Phytophthora erythroseptica var. pisi, Phytophthora aff erythroseptica, Phytophthora estuarina, Phytophthora europaea, Phytophthora fallax, Phytophthora flexuosa, Phytophthora fluvialis, Phytophthora fluvialis x moyootj, Phytophthora foliorum, Phytophthora formosa, Phytophthora formosana, Phytophthora fragariae, Phytophthora fragariaefolia, Phytophthora frigida, Phytophthora gallica, Phytophthora gemini, Phytophthora gibbosa, Phytophthora glovera, Phytophthora gonapodyides, Phytophthora gondwanensis, Phytophthora gregata, Phytophthora cf. gregata, Phytophthora hedraiandra, Phytophthora aff hedraiandra, Phytophthora x heterohybrida, Phytophthora heveae, Phytophthora hibernalis, Phytophthora himalayensis, Phytophthora himalsilva, Phytophthora aff himalsilva, Phytophthora humicola, Phytophthora aff humicola, Phytophthora hydrogena, Phytophthora hydropathica, Phytophthora idaei, Phytophthora ilicis, Phytophthora x incrassata, Phytophthora infestans, Phytophthora aff infestans, Phytophthora inflata, Phytophthora insolita, Phytophthora cf. insolita, Phytophthora intercalaris, Phytophthora intricata, Phytophthora inundata, Phytophthora ipomoeae, Phytophthora iranica, Phytophthora irrigata, Phytophthora katsurae, Phytophthora kelmania, Phytophthora kernoviae, Phytophthora kwongonina, Phytophthora lactucae, Phytophthora lacustris, Phytophthora lacustris x riparia, Phytophthora lateralis, Phytophthora lilii, Phytophthora litchii, Phytophthora litoralis, Phytophthora litoralis x moyootj, Phytophthora macilentosa, Phytophthora macrochlamydospora, Phytophthora meadii, Phytophthora aff meadii, Phytophthora medicaginis, Phytophthora medicaginis x cryptogea, Phytophthora megakarya, Phytophthora megasperma, Phytophthora melonis, Phytophthora mengei, Phytophthora mexicana, Phytophthora cf. mexicana, Phytophthora mirabilis, Phytophthora mississippiae, Phytophthora morindae, Phytophthora moyootj, Phytophthora moyootj x fluvialis, Phytophthora moyootj x litoralis, Phytophthora moyootj x thermophila, Phytophthora x multiformis, Phytophthora multivesiculata, Phytophthora multivora, Phytophthora nagaii, Phytophthora nemorosa [11], Phytophthora nicotianae, Phytophthora nicotianae var. parasitica, Phytophthora nicotianae x cactorum, Phytophthora niederhauserii, Phytophthora cf. niederhauserii, Phytophthora obscura, Phytophthora occultans, Phytophthora oleae, Phytophthora ornamentata, Phytophthora pachypleura, Phytophthora palmivora, Phytophthora palmivora var. palmivora, Phytophthora parasitica, Phytophthora parasitica var. nicotianae, Phytophthora parasitica var. piperina, Phytophthora parsiana, Phytophthora aff. parsiana, Phytophthora parvispora, Phytophthora x pelgrandis, Phytophthora phaseoli, Phytophthora pini, Phytophthora pinifolia, Phytophthora pisi, Phytophthora pistaciae, Phytophthora plurivora, Phytophthora pluvialis, Phytophthora polonica, Phytophthora porri, Phytophthora primulae, Phytophthora aff. primulae, Phytophthora pseudocryptogea, Phytophthora pseudolactucae, Phytophthora pseudorosacearum, Phytophthora pseudosyringae, Phytophthora pseudotsugae, Phytophthora aff. pseudotsugae, Phytophthora psychrophila, Phytophthora quercetorum, Phytophthora quercina, Phytophthora quininea, Phytophthora ramorum, Phytophthora rhizophorae, Phytophthora richardiae, Phytophthora riparia, Phytophthora rosacearum, Phytophthora aff. rosacearum, Phytophthora rubi, Phytophthora sansomea, Phytophthora sansomeana, Phytophthora aff. sansomeana, Phytophthora x serendipita, Phytophthora sinensis, Phytophthora siskyouensis, Phytophthora sojae, Phytophthora stricta, Phytophthora sulawesiensis, Phytophthora syringae, Phytophthora tabaci, Phytophthora tentaculata, Phytophthora terminalis, Phytophthora thermophila, Phytophthora thermophila x amnicola, Phytophthora thermophila x moyootj, Phytophthora trifolii, Phytophthora tropicalis, Phytophthora cf. tropicalis, Phytophthora tubulina, Phytophthora tyrrhenica, Phytophthora uliginosa, Phytophthora undulata, Phytophthora uniformis, Phytophthora vignae, Phytophthora vignae f. sp. adzukicola, Phytophthora virginiana, and Phytophthora vulcanica. In other specific embodiments, the said plant-pathogen is the species Phytophthora infestans.
In still other embodiments, the Peronosporaceae plant-pathogen is a member of the family Pythiaceae. In certain embodiments, the Pythiaceae plant-pathogen is a member of a genus selected from Cystosiphon, Diasporangium, Globisporangium, Lagenidium, Myzocytium, Phytophthora, Pythium, and Trachysphaera.
In further embodiments, the Pythiaceae plant-pathogen is a member of the genus Pythium. In specific embodiments, the Pythium plant-pathogen is a species selected from Pythium aphanidermatum, Pythium acanthicum, Pythium acanthophoron, Pythium acrogynum, Pythium adhaerens, Pythium amasculinum, Pythium anandrum, Pythium angustatum, Pythium apleroticum, Pythium aquatile, Pythium aristosporum, Pythium arrhenomanes, Pythium attrantheridium, Pythium bifurcatum, Pythium boreale, Pythium buismaniae, Pythium butleri, Pythium camurandrum, Pythium campanulatum, Pythium canariense, Pythium capillosum, Pythium carbonicum, Pythium carolinianum, Pythium catenulatum, Pythium chamaehyphon, Pythium chondricola, Pythium citrinum, Pythium coloratura, Pythium conidiophorum, Pythium contiguanum, Pythium cryptoirregulare, Pythium cucurbitacearum, Pythium cylindrosporum, Pythium cystogenes, Pythium debaryanum, Pythium delicense, Pythium destruens, Pythium diclinum, Pythium dimorphum, Pythium dissimile, Pythium dissotocum, Pythium echinulatum, Pythium emineosum, Pythium erinaceum, Pythium flevoense, Pythium folliculosum, Pythium glomeratum, Pythium graminicola, Pythium grandisporangium, Pythium guiyangense, Pythium helicandrum, Pythium helicoides, Pythium heterothallicum, Pythium hydnosporum, Pythium hypogynum, Pythium indigoferae, Pythium inflatum, Pythium insidiosum, Pythium intermedium, Pythium irregulare, Pythium iwayamae, Pythium jasmonium, Pythium kunmingense, Pythium Morale, Pythium longandrum, Pythium longisporangium, Pythium lutarium, Pythium macrosporum, Pythium mamillatum, Pythium marinum, Pythium marsupium, Pythium mastophorum, Pythium megacarpum, Pythium middletonii, Pythium minus, Pythium monospermum, Pythium montanum, Pythium multisporum, Pythium myriotylum, Pythium nagaii, Pythium nodosum, Pythium nunn, Pythium oedochilum, Pythium okanoganense, Pythium oligandrum, Pythium oopapillum, Pythium ornacarpum, Pythium orthogonon, Pythium ostracodes, Pythium pachycaule, Pythium pachycaule, Pythium paddicum, Pythium paroecandrum, Pythium parvum, Pythium pectinolyticum, Pythium periilum, Pythium periplocum, Pythium perniciosum, Pythium perplexum, Pythium phragmitis, Pythium pleroticum, Pythium plurisporium, Pythium polare, Pythium polymastum, Pythium porphyrae, Pythium prolatum, Pythium proliferatum, Pythium pulchrum, Pythium pyrilobum, Pythium quercum, Pythium radiosum, Pythium ramificatum, Pythium regulare, Pythium rhizo-oryzae, Pythium rhizosaccharum, Pythium rostratifingens, Pythium rostratum, Pythium salpingophorum, Pythium scleroteichum, Pythium segnitium, Pythium speculum, Pythium spinosum, Pythium splendens, Pythium sterilum, Pythium stipitatum, Pythium sulcatum, Pythium terrestris, Pythium torulosum, Pythium tracheiphilum, Pythium ultimum, Pythium ultimum var. ultimum, Pythium uncinulatum, Pythium undulatum, Pythium vanterpoolii, Pythium viniferum, Pythium violae, Pythium volutum, Pythium zingiberis, and Pythium zingiberum. In other specific embodiments, the plant-pathogen is the species Pythium aphanidermatum.
In other embodiments of the present invention, the method of the invention is useful for controlling, preventing, reducing or eradicating any one of the Oomycotaplant-pathogens described above, and in particular Phytophthora infestans by applying compound 3 as described above; and Pythium aphanidermatum, by applying as described above compound 3 or 1, or a combination thereof.
In further embodiments, the plant-pathogen is a member of the genus Pseudomonas, such as Pseudomonas aeruginosa and Pseudomonas syringae. In particular embodiments, the plant-pathogen is the species Pseudomonas syringae. In yet further embodiments, the method of the invention is useful for controlling, preventing, reducing or eradicating any one of the Pseudomonas pathogens described above, and in particular Pseudomonas syringae, by applying as described above compound 1 or 2, or a combination thereof.
In another aspect, the present invention provides a pesticide composition comprising a pesticidally effective amount of at least one compound of formula (I),
wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, methyl, hydroxy and methoxy group, and halogen atom (F, Cl, Br, I); R5 and R6 are independently selected from hydrogen, methyl and ethyl; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy; stereoisomers or agriculturally acceptable salts thereof.
In a further embodiment, the present invention provides a pesticide composition comprising a compound of formula (I), wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, methyl, hydroxy and methoxy group, and halogen atom (F, Cl, Br, I); R5 and R6 are methyl; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy group.
In yet further embodiment, the present invention provides a pesticide composition comprising a compound of formula (I), wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, hydroxy, and methoxy group; R5 and R6 are methyl; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy group.
In a particular embodiment, the present invention provides a pesticide composition comprising a compound of formula (I), wherein R1a, R1b, R2, R3 and R4 are independently selected from hydrogen, hydroxy, and methoxy; R5 and R6 are methyl; and R7 is hydrogen.
In a specific embodiment, the pesticide composition of the present invention comprises:
In another embodiment, the present invention provides a pesticide composition comprising a compound of formula (I), wherein R1a, R1b, R2 are independently selected from hydrogen and halogen atom (F, Cl, Br, I); R3, R4, R5 and R6 are hydrogen; and R7 is selected from hydrogen, methyl, amino, methylamino, dimethylamino, hydroxy and methoxy group.
In a specific embodiment, the present invention provides a pesticide composition comprising a compound of formula (I), wherein R1a, R1b, R2 are independently selected from hydrogen and chlorine atom; R3, R4, R5 and R6 are hydrogen; and R7 is methylamino group.
In another specific embodiment, the pesticide composition of the present invention comprises:
In certain embodiments, the pesticide composition or formulation of any one of the above embodiments further comprises an agriculturally suitable or acceptable solvent or solubilising agent. In other certain embodiments, the agriculturally acceptable solvent or solubilising agent is a water-miscible solvent capable of dissolving or solubilising 1-phenyl-tetralin compounds.
In some embodiments, the water-miscible solvent capable of dissolving or solubilising 1-phenyl-tetralin compounds is a polar solvent, such as an alcohol, a ketone, a lactone, a keto-alcohol, a glycol, a glycoether, an amide, an alkanolamine, a sulfoxide and a pyrrolidone. In particular embodiments, the composition of any one of the above embodiments comprises a solvent selected from dimethyl-sulfoxide or ethanol. In specific embodiments, the composition further comprises a polysorbate-type non-ionic surfactant, such as polysorbate 20.
The pesticide composition of the present invention may be formulated into a formulation to facilitate application of the active pesticidal ingredient. The formulation may be a water-miscible formulation, such as a suspension concentrate (SC), a capsule suspension (CS), water-dispersible granules (WG), an emulsifiable concentrate (EC), a wettable powder (WP), a soluble (liquid) concentrate (SL), or a soluble powder (SP).
The composition or formulation of the present invention may further comprise at least one adjuvant, carrier, diluent, and/or surfactant. Non-limiting examples of adjuvants are activator adjuvants, such as cationic, anionic or non-ionic surfactants, oils and nitrogen-based fertilisers capable of improving activity of the pesticide product. Oils may be crop oils, such as paraffin or naphtha-based petroleum oil, crop oil concentrates based on emulsifiable petroleum-based oil, and vegetable oil concentrates derived from seed oil, usually cotton, linseed, soybean, or sunflower oil, used to control grassy weeds. Nitrogen-based fertilisers may be ammonium sulphate or urea-ammonium nitrate.
A non-limiting example of a polysaccharide adjuvant used also as a thixotropic agent in the compositions of the present embodiments, is Xanthan gum (commercially available under trademark KELZAN® by CP Kelco), which is produced from simple sugars using a fermentation process, and derives its name from the species of bacteria used, Xanthomonas campestris. Oils used as adjuvants may be crop oils, such as paraffin or naphtha-based petroleum oil, crop oil concentrates based on emulsifiable petroleum-based oil, and vegetable oil concentrates derived from seed oil, usually cotton, linseed, soybean, or sunflower oil, used to control grassy weeds. Nitrogen-based fertilisers may be ammonium sulphate or urea-ammonium nitrate.
Non-limiting examples of solubilising agents or solvents are petroleum-based solvents, the aforementioned oils, liquid mixtures of fatty acids, ethanol, glycerol and dimethyl sulfoxide. The agriculturally acceptable solvent or solubilising agent may be a water-miscible solvent capable of dissolving or solubilising 1-phenyl-tetralin compounds, such as a polar solvent, e.g., an alcohol, a ketone, a lactone, a keto-alcohol, a glycol, a glycoether, an amide, an alkanolamine, a sulfoxide and a pyrrolidone. Non-limiting examples of carriers are precipitated silica, colloidal silica, attapulgite, china clay, talc, kaolin and combinations thereof.
The pesticide composition or formulation of the present invention may further comprise a diluent, such as lactose, starch, urea, water soluble inorganic salts and combination thereof. The pesticide composition or formulation may further comprise one or more surfactants, such as polysorbate-type non-ionic surfactant, for example Polysorbate 20 or trisiloxane non-ionic surfactant, styrene acrylic dispersant polymers, acid resin copolymer based dispersing agents, potassium polycarboxylate, sodium alkyl naphthalene sulphonate blend, sodium diisopropyl naphthalene sulphonate, sodium salt of naphthalene sulphonate condensate, lignin sulfonate salts and combinations thereof.
Trisiloxane non-ionic surfactants or polyether dimethyl siloxanes (PEMS), often referred to as super-spreaders or super-wetters, are added to pesticides to enhance their activity and the rain fastness of the active substance by promoting rapid spreading over the hydrophobic surfaces of leaves. Some spreaders of the modified trisiloxane type combine a very low molecular weight trisiloxane with a polyether group and capable of reducing surface tension and rapidly spreading on difficult to wet surfaces.
The active agent, composition, or formulation comprising it, is applied in the method of any one of the above embodiments to the plant or part, organ or plant propagation material thereof by spraying, immersing, dressing, coating, pelleting or soaking.
In certain embodiments, the concentration of the 1-phenyl-tetralin compounds of the present invention, in the composition or formulation comprising it may be in the range of 10-2000, 10-1500, 10-1000, 10-900, 10-800, 10-700, 10-600, 10-500, 10-400, 10-300, 10-200, 10-900, 20-800, 20-700, 20-600, 20-500, 20-400, 20-300, 20-200, 20-100, 20-90, 20-80, 20-70, 20-60, 20-50, 20-40, 20-30, 20-20. 30-2000, 30-1500, 30-1000, 30-900, 30-800, 30-700, 30-600, 30-500, 30-400, 30-300, 30-200, 30-100, 30-0, 30-100, 30-90, 30-80, 30-70, 30-60, 30-50, 30-40, 40-2000, 40-1500, 40-1000, 40-900, 40-800, 40-700, 40-600, 40-500, 40-400, 40-300, 40-200, 40-100, 40-90, 40-80, 40-70, 40-60, 40-50, 50-2000, 50-1500, 50-1000, 50-900, 50-800, 50-700, 50-600, 50-500, 50-400, 50-300, 50-200, 50-100, 50-90, 50-80, 50-70, 50-60, 60-2000, 60-1500, 60-1000, 60-900, 60-800, 60-700, 60-600, 60-500, 60-400, 60-300, 60-200, 60-100, 60-90, 60-80, 60-70, 70-2000, 70-1500, 70-1000, 70-900, 70-800, 70-700, 70-600, 70-500, 70-400, 70-300, 70-200, 70-100, 70-90, 70-80, 80-2000, 80-1500, 80-1000, 80-900, 80-800, 80-700, 80-600, 80-500, 80-400, 80-300, 80-200, 80-100, 80-90, 90-2000, 90-1500, 90-1000, 90-900, 90-800, 90-700, 90-600, 90-500, 90-400, 90-300, 90-200, 90-100, 100-2000, 100-1500, 100-1000, 100-900, 100-800, 100-700, 100-600, 100-500, 100-400, 100-300, 100-200, 200-2000, 200-1500, 200-1000, 200-900, 200-800, 200-700, 200-600, 200-500, 200-400, 200-300, 300-2000, 300-1500, 300-1000, 300-900, 300-800, 300-700, 300-600, 300-500, 300-400, 400-2000, 400-1500, 400-1000, 400-900, 400-800, 400-700, 400-600, 400-500, 500-2000, 500-1500, 500-1000, 500-900, 500-800, 500-700, 500-600, 600-2000, 600-1500, 600-1000, 600-900, 600-800, 600-700, 700-2000, 700-1500, 700-1000, 700-900, 700-800, 800-2000, 800-1500, 800-1000, 800-900, 900-2000, 900-1500, 900-1000, 1000-2000, or 1000-1500 ppm.
In particular, the concentration of 1-phenyl-tetralin compounds in the composition or formulation comprising it may be 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 1000, 1500 or 2000 ppm.
Any one of the above concentration ranges or concentrations can be used in accordance with any one of the above embodiments of the method of the present invention, including against any one of the aforementioned pathogens and by means of any one of the above mentioned means of applying the composition or formulation.
The term “plant organ” as used herein refers to the leaf, stem, root, and reproductive structures. The term “plant part” as used herein refers to a vegetative plant material such as a cutting or a tuber; a leaf, flower, bark or a stem. The term “plant propagation material” as used herein refers to a seed, root, fruit, tuber, bulb, rhizome, or part of a plant. The term “pesticidal effective amount” as used herein refers to an amount of the pesticide that is able to bring about death to at least one pest, or to noticeably reduce pest growth, feeding, or normal physiological development. The terms “class”, “order”, “family”, “genus”, and “species” are used herein according to Art 3.1 of the International Code of Nomenclature for algae, fungi, and plants.
The term “comprising”, used in the claims, is “open ended” and means the elements recited, or their equivalent in structure or function, plus any other element or elements which are not recited. It should not be interpreted as being restricted to the means listed thereafter; it does not exclude other elements or steps. It needs to be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps or components, or groups thereof. Thus, the scope of the expression “a composition comprising x and z” should not be limited to compositions consisting only of components x and z. Also, the scope of the expression “a method comprising the steps x and z” should not be limited to methods consisting only of these steps.
Unless otherwise indicated, all numbers used in this specification are to be understood as being modified in all instances by the term “about”. Unless specifically stated, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within two standard deviations of the mean. In one embodiment, the term “about” means within 10% of the reported numerical value of the number with which it is being used, preferably within 5% of the reported numerical value. For example, the term “about” can be immediately understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. In other embodiments, the term “about” can mean a higher tolerance of variation depending on for instance the experimental technique used. Said variations of a specified value are understood by the skilled person and are within the context of the present invention. As an illustration, a numerical range of “about 1 to about 5” should be interpreted to include not only the explicitly recited values of about 1 to about 5, but also include individual values and sub-ranges within the indicated range. Thus, included in this numerical range are individual values such as 2, 3, and 4 and sub-ranges, for example from 1-3, from 2-4, and from 3-5, as well as 1, 2, 3, 4, 5, or 6, individually. This same principle applies to ranges reciting only one numerical value as a minimum or a maximum.
Unless otherwise clear from context, all numerical values provided herein are modified by the term “about”. Other similar terms, such as “substantially”, “generally”, “up to” and the like are to be construed as modifying a term or value such that it is not an absolute. Such terms will be defined by the circumstances and the terms that they modify as those terms are understood by those of skilled in the art. This includes, at very least, the degree of expected experimental error, technical error and instrumental error for a given experiment, technique or an instrument used to measure a value.
As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items. Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. Well-known functions or constructions may not be described in detail for brevity and/or clarity.
The invention will now be illustrated by the following non-limiting Examples.
RPM—Revolutions per minute
RCF—Relative centrifugal force
CFU—Colony forming unit
PDBC—Potato dextrose broth with 20 μg/ml chloramphenicol
PDAC—Potato dextrose agar with 20 μg/ml chloramphenicol
PDAT—Potato dextrose agar with 12 μg/ml tetracycline
DMSO—Dimethyl sulfoxide
LB—LB broth
LBA—LB agar
SCH—Schmittner medium
2:PDBC—PDBC diluted 2 fold by sterile distilled water
PDA—Potato dextrose agar
PDBT—Potato dextrose broth with 12 μg/ml tetracycline
Background: Puccinia sorghi is a fungus of belonging to the Basidiomycetes and it is an air borne pathogen. Puccinia spores were grown on corn plants in a growth room and fresh spore suspension is prepared from the infected corn leaves for each experiment. Since Puccinia sorghi is an obligatory pathogen and does not grow on synthetic medium, the germination of the spores was monitored as indication for 1-phenyl-tetralin compounds bioactivity.
Summary: Diluted in DMSO 1-phenyl-tetralin compounds (Compound 1: (5R,6R,7R)-5-(3,4-dihydroxyphenyl)-6,7-dimethyl-5,6,7,8-tetrahydronaphthalene-2,3-diol, Compound 2: 4-((1R,2R,3R)-7-hydroxy-6-methoxy-2,3-dimethyl-1,2,3,4-tetrahydronaphthalen-1-yl)benzene-1,2-diol, and Compound 3: (1S,4R)-4-(3,4-dichlorophenyl)-N-methyl-1,2,3,4-tetrahydronaphthalen-1-aminium chloride) were added separately to microplate wells and mixed with freshly prepared spore suspensions. The germination of the spores was monitored by visual inspection under the microscope.
The following materials, methods and equipment were used:
Materials: Tween® 20 (Tidea Company INC) non-ionic detergent, DMSO—dimethyl-sulfoxide (J. T. Baker—Poland) solvent, chloramphenicol (Alfa Aesar—UK)
Equipment: Centrifuge, Shaker, Incubator, Microscope, Filtration system
See results in Example 9.
Summary: Diluted in DMSO 1-phenyl-tetralin compounds (Compound 1, 2 or 3) were added separately to microplate wells and mixed with 50 μl of hyphae suspension and growth of the fungus, starting from blended hyphae, was monitored by plate reader and visual inspection.
The following Materials, methods and equipment were used:
Equipment: Plate reader, Centrifuge, Shaker, Incubator.
A. Inoculum Preparation of Rhizoctonia solani Hyphae
See results in Example 9.
Summary: Diluted in DMSO 1-phenyl-tetralin compounds (Compound 1, 2 or 3) were added separately to microplate wells and mixed with 50 μl of zoospores in PDBC suspension and the growth of the fungus, starting from zoospores, was monitored by plate reader and visual inspection.
The following Materials, methods and equipment were used:
Equipment: Plate reader, Centrifuge, Shaker, Incubator.
See results in Example 9.
Summary: Microplates with Compound 1, 2 or 3 diluted in DMSO was mixed with frozen spore suspension and the growth of the fungus was monitored, starting from frozen spores by visual inspection.
Background: Botrytis cinerea is a fungus belonging to Ascomycetes and it is an air-borne pathogen. It is quite easy to produce large amounts of Botrytis spores, which survive in liquid 60% glycerol at −20° C. Thus, we used frozen spores' stocks in the bioactivity screening experiments rather than prepare fresh spores for each experiment.
Aim: To determine the effect of a 1-phenyl-tetralin compound on the survival and growth of Botrytis.
The following Materials, methods and equipment were used:
Equipment: Centrifuge—Eppendorf 5810R; Shaker—Scientific Industries, Multi Microplate Genie; Incubator—Pol-Eco Aparatura; Plate reader.
See results in Example 9.
Summary: Compound 1, 2 or 3 diluted in DMSO was added to microplate wells and mixed with freshly prepared spore suspension and growth of the fungus, starting from frozen spores, was monitored using the plate reader and by visual inspection.
Background: Fusarium is a fungus of belonging to the Ascomycetes, and it is a soil borne pathogen. It is quite easy to produce large amounts of spores of Fusarium and they survive in liquid 60% glycerol at −20° C. Thus, we used frozen spores' stock in the bioactivity screening experiments rather than prepare fresh spores for each experiment.
Aim: To determine the effect of 1-phenyl-tetralin compounds on the survival and growth of Fusarium.
The following materials, methods and equipment were used:
Equipment: Plate reader, Centrifuge, Shaker, Incubator.
Collect the readout of the plate at 3 more dates: 3 d, 7 d, 14 d and 21 d following the assay start
See results in Example 9.
Background: Phytophthora infestans is an obligatory pathogen from Oomycetes which is very difficult to grow on synthetic medium. Therefore, the bioactivity screening system based on leaf discs prepared from detached tomato leaves were used.
Summary: Compounds 1, 2 or 3 diluted in DMSO were added to tomato leaf discs infected with Phytophthora and the disease progress was monitored by visual inspection.
General description: Inoculation and maintenance on tomato leaves, preparation of spore suspension, their growth on leaf discs in microplates and inspection by magnifying glass of Phytophthora infestans severity of infection.
The following materials, methods and equipment were used:
See results in Example 9.
Background: Pseudomonas is a rod-shaped Gram-negative bacterium. Frozen bacterial stock in 60% glycerol was used as an inoculum for the bioactivity screening experiments.
Summary: Compounds 1 or 2 diluted in DMSO were added to microplate wells and mixed with frozen bacteria suspension and growth of the Pseudomonas was monitored by visual inspection. The following materials, methods and equipment were used:
See results in Example 9.
Background: Alternaria alternata is a major plant pathogen and cause large damage to many agricultural crops. Alternaria alternata is a fungus of belonging to the Ascomycetes, and it is an air borne pathogen. It is quite easy to produce large amounts of spores of Alternaria alternata, and they survive in liquid 60% glycerol at −20° C., that led to decision to use frozen spore stock in this screening rather than prepare fresh spores for each experiment.
Summary: Compound 1, 2 or 3 diluted in DMSO was added to microplate wells and mixed with frozen bacteria suspension and growth of Alternaria was monitored by visual inspection.
The following materials, methods and equipment were used:
See results in Example 9.
1-phenyl-tetralin compounds were screened against selected agricultural pests (as indicated in the tables below). The bioactivity values are in % and reflect the potential of eradicating the target pests.
Rules for Bioactivity Relative Value Calculation (Expressed in % from Maximal Value)
a. Puccinia sorghi, Phytophthora infestans—activity grade (1/2/3) X repeats #/12 (maximal value 3×4=12)×100
b. Alternaria alternata, Botrytis cinerea, Rhizoctonia solani, Sclerotinia sclerotiorum, Fusarium oxysporum, Pythium aphanidermatum—activity grade (1/2/3) X repeats #X days of activity/252 (maximal value 3×4×21=252)×100
c. Pseudomonas syringae, Pectobacterium caratovorum—activity grade (1/2/3) X repeats # X days of activity/168 (maximal value 3×4×14=168)×100
Puccinia sorghi
Phytophthora infestans
Rhizoctonia solani
Pythium aphanidermatum
Pseudomonas syringae
Puccinia sorghi
Rhizoctonia solani
Pythium aphanidermatum
Pseudomonas syringae
Puccinia sorghi
Phytophthora infestans
Botrytis cinerea
Alternaria alternata
Rhizoctonia solani
Pythium aphanidermatum
Fusarium oxysporum
In summary, 1-phenyl-tetralin compounds are demonstrated to be effective pesticides against the following pests: Puccinia sorghi (positive results are provided in in-planta results section below), Phytophthora infestans (positive results in tomato detached leaves validation experiments and greenhouse in-vivo validation experiments provided), Rhizoctonia solani, Pythium aphanidermatum, Alternaria alternata, Botrytis cinerea (positive results in in-vivo tomato validation experiments under greenhouse conditions provided), Fusarium oxysporum and Pseudomonas syringae.
To evaluate the effect of a tested compounds in infected plants compared to control plants (infected but not treated) the data was analysed by Student's t-test and the p-value is calculated. The minimum number of repeats in each experiment was 3. Results were considered significant if p<0.05. The data presented as mean with standard error mean from biological replicates. * means that p-value <0.05, ** means that p-value is <0.01, *** means that p-value is <0.001, # means that p-value <0.1, n.s.—means non-significant effect vs. control.
Three types of stock solutions were used for final 1-phenyl-tetralin compound formulation preparation at 400 ppm (Formulation 1):
(A) 1-phenyl-tetralin compound solution in water and acetic acid.
The final formulated 1-phenyl-tetralin compound was applied as 400 ppm or diluted to the required concentrations and applied to plants.
Three types of stock solutions were used for final 1-phenyl-tetralin compound formulation preparation at 400 ppm:
(A) 1-phenyl-tetralin compound suspension in water.
The final formulation which was applied to wheat plants is composed of:
4% of stock solution (A), 10% of stock solutions (B), 3.3% of stock (C), and 82.7% of water.
The final formulated 1-phenyl-tetralin compound was applied as 400 ppm or diluted to the required concentrations and applied to plants.
Protocol name: Puccinia sorghi infection of corn seedlings test
General description: Inoculation on corn, collection, Puccinia sorghi spores' suspension preparation and 1-phenyl-tetralin compounds 1 or 3 bioactivity evaluation against Puccinia sorghi.
The following materials, methods and equipment were used:
Compound 1 was dissolved in dimethyl-sulfoxide solvent with 1:9 weight to weight ratio and then brought up to the final volume used for the validation with double distilled water. Before spraying, the non-ionic detergent Tween® 20 was added to final concentration of 0.05%.
Compound 3 Formulations 1-5 for Exps. 270, 284, 294 (with Reference to
Compound 3 was dissolved in absolute ethanol with 1:36 weight to weight ratio or in dimethyl-sulfoxide with 1:17 weight to weight ratio, sonicated for 5 mins and then another part of non-ionic detergent either Tween® 20 with weight to weight 1:4.5 ratio to Compound 3 or Silwet® in 1:1 weight to weight ratio to Compound 3 was added for formulation finalisation. In some cases, Na2CO3 was used to adjust pH to 6.
Several experiments were conducted under controlled environment in growth rooms where the potential of Compound 1 and Compound 3 to prevent and control Puccinia sorghi in corn plants was estimated (
General description: Inoculation of wheat with leaf rust, spraying of potentially bioactive compounds to control the infection, and procedure for evaluation of the infection level.
Two experiments were conducted under controlled environment in growth rooms where the bioactivity potential of Compound 1 to prevent and control Puccinia sorghi in corn plants was estimated (
General description: Detached leaves of tomato were treated by 1-phenyl-tetralin compound and infected by spores of Phytophthora infestans.
Phytophthora spore suspension preparation: Prepare spores according to Example 6 and dilute by water to 1000 spores/ml.
Compound 3 was dissolved in absolute ethanol with 1:36 weight to weight ratio or in dimethyl-sulfoxide with 1:17 weight to weight ratio, sonicated for 5 mins and then another part of non-ionic detergent either Tween® 20 with weight to weight 1:4.5 ratio to Compound 3 or Silwet® in 1:1 weight to weight ratio to Compound 3 was added for formulation finalisation. In some cases, sodium carbonate (Na2CO3) was used to adjust pH to 6.
Three independent experiments were conducted in detached tomato leaves where the bioactivity potential of Compound 3 to prevent and control Phytophthora infestans was estimated (see the results in Tables 4-6 below). The severity of infection by Phytophthora was evaluated using the following grades which expresses the leaf area covered by fungus: 0=clear; 1=low coverage; 2=medium coverage; 3=high coverage.
Compound 3 controlled the Phytophthora infection with the efficacies between 73% to 83% at 200 ppm.
Phytophthora
Phytophthora inoculated, not treated
Phytophthora
Phytophthora inoculated, not treated
Phytophthora inoculated, not
General description: Leaves of wheat, with colonies of leaf rust were sprayed by formulated Compound 3 (treatment) and the percentage of spore germination was evaluated in three time points following the treatment (1, 7 and 14 d).
Sub protocol: 1) Pustules germination; 2) Inoculation of wheat with Puccinia.
See formulation 2 preparation in Formulation section in Example 9.
Three experiments were conducted under greenhouse conditions where the bioactivity potential of Compound 3 to inhibit Puccinia triticina spore's germination was estimated (
General description: Severity of late blight disease caused by Phytophtora infestans was evaluated following treatment with 1-phenyl-tetralin derivatives. Sporangium was used to infect 3-4 weeks-old tomato young plants following curative treatment with the 1-phenyl-tetralin derivatives.
In the preventative approach, inoculation was applied following two repeating treatments with Compound 3:
See formulation preparation in Formulation section in Example 9.
Seven independent experiments were conducted in tomato plants infected with Phytophthora, where the potential of Compound 3 to prevent and control Phytophthora infestans (
Compound 3 controlled the Phytophthora infection with the efficacy up to 100%.
General description: Severity of early blight disease caused by Alternaria solani was evaluated following treatment with Compound 3. Spores isolates were used to infect leaves of 3-4 weeks old tomato young plants following preventative treatment by 1-phenyl-tertralin compounds.
4) Early blight symptoms (Alternaria) were observed as yellow-brownish spots that appeared on the infected spot. The yellow-brown diameter of the decay on each labelled leaflet was measured. The total leaflet size was measured as well.
See Formulation 2 preparation in Formulation section in Example 9.
Experiments were conducted in tomato plants infected with Alternaria, where the bioactivity potential of Compound 3 to prevent and control Alternaria solani was estimated (see
See Formulations 1 and 2 preparation in Formulations section in Example 9.
Two independent experiments were conducted in tomato plants infected with Botrytis where the potential of Compound 3 to prevent and control Botrytis cinerea was estimated (see
The present application claims the benefit of priority to U.S. Patent Application No. 62/969,111 filed Feb. 2, 2020, the contents of which are incorporated by reference herein in their entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/IL2021/050110 | 1/31/2021 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 62969111 | Feb 2020 | US |
