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Mixtures and Compositions Comprising Paenibacillus Strains Or Fusaricidins and Chemical Pesticides

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Mixtures and Compositions Comprising Paenibacillus Strains Or Fusaricidins and Chemical Pesticides (19) TZZ¥ Z Z_T (11) EP 3 205 208 A1 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 16.08.2017 Bulletin 2017/33 A01N 37/46 (2006.01) A01N 63/02 (2006.01) A01P 7/00 (2006.01) A01P 3/00 (2006.01) (21) Application number: 16154807.8 (22) Date of filing: 09.02.2016 (84) Designated Contracting States: (71) Applicant: BASF SE AL AT BE BG CH CY CZ DE DK EE ES FI FR GB 67056 Ludwigshafen am Rhein (DE) GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR (72) Inventors: Designated Extension States: • Siebe, Isabella BA ME 69221 Dossenheim (DE) Designated Validation States: • Liebmann, Burghard MA MD 64625 Bensheim (DE) • Jabs, Thorsten (83) Declaration under Rule 32(1) EPC (expert 67454 Hassloch (DE) solution) • Schuster, Annette 67105 Schifferstadt (DE) (54) MIXTURES AND COMPOSITIONS COMPRISING PAENIBACILLUS STRAINS OR FUSARICIDINS AND CHEMICAL PESTICIDES (57) The present invention relates to mixtures com- tabolite thereof, and at least one chemical pesticide. The prising, as active components, at least one isolated bac- present invention also relates to a method of controlling terial strain, which is a member of the genus Paenibacil- or suppressing plant pathogens or preventing plant path- lus, or a cell-free extract thereof or at least one metabolite ogen infections by applying such composition. The thereof, and at least one chemical pesticide. The present present invention also relates to mixtures of fusaricidins invention also relates to compositions comprising at least which are pesticidal metabolites produced by the above- one of such bacterial strains, whole culture broth or a mentioned strains, and chemical pesticides. cell-free extract or a fraction thereof or at least one me- EP 3 205 208 A1 Printed by Jouve, 75001 PARIS (FR) EP 3 205 208 A1 Description Field of the Invention 5 [0001] The present invention relates to novel mixtures comprising, as active components, at least one isolated bacterial strain, which is a member of the genus Paenibacillus, or a cell-free extract thereof or at least one metabolite thereof, and at least one chemical pesticide. The present invention also relates to compositions comprising at least one of such bacterial strains, whole culture broth or a cell-free extract or a fraction thereof or at least one metabolite thereof, and at least one chemical pesticide. The present invention also relates to a method of controlling or suppressing plant pathogens 10 or preventing plant pathogen infections by applying such composition. The present invention also relates to mixtures of fusaricidins which are pesticidal metabolites produced by the abovementioned strains, and chemical pesticides. Background of the Invention 15 [0002] In the technical field of controlling phytopathogenic fungi affecting plants or crops it is well known to apply biopesticides, for example selected from bacteria, like spore-forming bacteria, or fungi which are not detrimental to the plant or crop to be treated and which biological control agents may be further combined with classical organic chemical antagonists of plant pathogens. [0003] Biopesticides have been defined as a form of pesticides based on micro-organisms (bacteria, fungi, viruses, 20 nematodes, etc.) or natural products (compounds or extracts from biological sources) (U.S. Environmental Protection Agency: http://www.epa.gov/pesticides/biopesticides/). [0004] Biopesticides are typically created by growing and concentrating naturally occurring organisms and/or their metabolites including bacteria and other microbes, fungi, viruses, nematodes, proteins, etc. They are often considered to be important components of integrated pest management (IPM) programmes, and have received much practical 25 attention as substitutes to synthetic chemical plant protection products (PPPs). [0005] Biopesticides fall into two major classes, microbial and biochemical pesticides: (1) Microbial pesticides consist of bacteria, fungi or viruses (and often include the metabolites that bacteria and fungi produce). Entomopathogenic nematodes are also classed as microbial pesticides, even though they are multi- 30 cellular. (2) Biochemical pesticides are naturally occurring substances that control pests or provide other crop protection uses as defined below, but are relatively non-toxic to mammals. [0006] For controlling phytopathogenic fungi several microbial pesticides comprising spore-forming bacteria such as 35 Bacillus subtilis have been described earlier, see e. g. WO 1998/050422; WO 2000/029426; WO 1998/50422 and WO 2000/58442. [0007] WO 2009/0126473 discloses agriculturally acceptable aqueous compositions comprising bacterial or fungal spores contained in an aqueous/organic solvent and which may further comprise insect control agents, pesticides, fungicides or combinations thereof. Spores of bacteria of the genus Bacillus are a preferred species. 40 [0008] WO 2006/017361 discloses compositions for controlling plant pathogens and comprising at least one beneficial bacterium, at least one beneficial fungus, at least on nutrient and at least one compound which extends the effective lifetime of such a composition. The group of beneficial bacteria e.a. comprises bacteria of Paenibacillus polymyxa and Paenibacillus durum. [0009] WO 1999/059412 discloses a Paenibacillus polymyxastrain PKB1 (bearing ATCC accession no. 202127) active 45 against several phytopathogenic fungi. [0010] WO 2011/069227 discloses a P. polymyxa strain JB05-01-1 (bearing ATCC accession no. PTA-10436) having a highly inhibitory effect against pathogenic bacteria, pre-dominantly food-borne human pathogenic bacteria. [0011] Raza et al. (BrazilianArch. Biol. Techol. 53,1145-1154, 2010; Eur. J.Plant Pathol.125: 471-483, 2009) described a fusaricidin-producing Paenibacillus polymyxastrain SQR-21 effective against Fusarium oxysporum. 50 [0012] Another Paenibacillus polymyxa strain called AC-1 is known from Microbial Research 2016 (in press doi:10.1016/j.micres.2016.01.004) and product Topseed from Green Biotech Co., Ltd. 45-70 Yadang-ri, Gyoha-Eup Paju Kyungki-Do, Korea (South) 413 -830. [0013] A further Paenibacillus polymyxa strain presumably called HY96-2 is known from Biocontrol Science and Tech- nology 24 (4), 426-435 (2014) and shall be marketed under the name KangDiLeiDe by Shanghai Zeyuan Marine Bio- 55 technology Co.,Ltd. [0014] The genome of several Paenibacillus polymyxa strains has been published so far: inter alia for strain M-1 (NCBI acc. no. N_017542; J. Bacteriol. 193 (29), 5862-63, 2011; BMC Microbiol. 13, 137, 2013), strain CR1 (GenBank acc. no. CP006941; Genome Announcements 2 (1), 1, 2014) and strain SC2 (GenBank acc. nos. CP002213 and CP002214; 2 EP 3 205 208 A1 NCBI acc. no. N_014622; J. Bacteriol. 193 (1), 311-312, 2011), for further strains see legend of Figure 12 herein. The P. polymyxa strain M-1 has been deposited in China General Microbiological Culture Collection Center (CGMCC) under acc. no. CGMCC 7581. [0015] In the unpublished PCT application PCT/EP2015/067925 new strains of the genusPaenibacillus have been 5 characterized. Said bacterial strains Lu16774, Lu 17007 and Lu17015 had been isolated from crop acreage in Germany and deposited under the Budapest Treaty with the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) on February 20th, 2013 by BASF SE, Germany: 1) Paenibacillus strain Lu16774 deposited under Accession No. DSM 26969, 10 2) Paenibacillus strain Lu 17007 deposited under Accession No. DSM 26970, and 3) Paenibacillus strain Lu17015 deposited under Accession No. DSM 26971. [0016] As used herein, the term Paenibacillus strain is identical to the termPaenibacillus sp. strain and means a bacterial strain form the genus Paenibacillus. The genus Paenibacillus includes all species Paenibacillus spp.. 15 [0017] In abovementioned PCT/EP2015/067925, the strains Lu16774, Lu17007 and Lu17015 were determined to belong to the genus Paenibacillus on the following morphological and physiological observations (see Example 2.3 in PCT/EP2015/067925 and herein): - rod-shaped cells 20 - ellipsoidal spores - swollen sporangium - anaerobic growth - fermentation of a variety of sugars including glucose, arabinose, xylose, mannit, fructose, raffinose, trehalose and glycerol with acid formation 25 - gas production from glucose - arginine dihydrolase negative - no utilization of citrate - no growth in presence of 5% or more sodium chloride - production of extracellular hydrolytic enzymes degrading starch, gelatine, casein and esculin. 30 [0018] Further, these strains Lu16774, Lu17007 and Lu17015 were also determined to belong to the genus Paeniba- cillus by 16S rDNA analysis by having the Paenibacillu-specific 22-base sequence in 16S rDNA (5’ to 3’): 5’-TCGATACCCTTGGTGCCGAAGT-3’ (see SEQ ID NO:1 (nucleotides 840-861), SEQ ID NO:2 (840-861), SEQ 35 ID NO:3 (844-865) and SEQ ID NO:4 (840-861) in sequence listings of PCT/EP2015/067925 and herein). [0019] Further, sequencing of the complete 16S rDNA in comparison to 24 different Paenibacillus strains resulted in clustering of the strains Lu16774, Lu17007 and Lu17015 with the type strains of Paenibacillus brasiliensis, P. kribbensis, P. jamilae, P. peoriae, and P. polymyxa, more preferably to P. peoriae, in particular Paenibacillus peoriae strain BD-62 40 (see Figs. 1 and 2 in PCT/EP2015/067925 and herein). It is known thatP. polymyxa and P. peoriae have 16S rDNA sequence identity values of 99.6 to 99.7% (J. Gen. Appl. Microbiol. 48, 281-285 (2002)). [0020] "Percent Identity" or "percent similarity" between
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