JPH06506345A - bactericidal microorganisms - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention]

bactericidal microorganisms Technical field The present invention relates to plant protection products for agricultural use. More specifically, the present invention Concerning its use for protecting plants from attack by microorganisms, especially novel strains of fungi. Background technology Fluorescence of microorganisms ii Strains consisting of Pseudomonas fluorescens is known to be a useful agricultural fungicide. The antifungal activity of countless strains of microorganisms It changes drastically. Agriculturally useful bacterial strains target specific fungi (fungi). It is a strain that has a useful level of activity against. One such goal is to make the crop In addition, damping-off disease, which is particularly troublesome under certain climatic conditions, ) is a group of mosquitoes with a disease known as J. Living things to control diseases chemical control agents (BCAs) or biological biopesticides s) is more effective or more preferred than the use of standard chemical control agents. obtain. One object of the present invention is to provide an agriculturally useful fungicide. Disclosure of invention According to the present invention, with the accession number NCl8401119 dated September 1, 1989, National Collection of Industrial and Marine Bacteria ( Fluorescent image of a new bacterial strain (Pseudomonas fluo) deposited at NCIB) rescens) Biovar I is provided. The present invention also provides the use of fluorophores as the active ingredient in admixture with an agriculturally acceptable carrier composition. Pseudomonas fluorescens biovarue, strain NC It consists of an agricultural fungicidal composition containing IB 40189. Agronomic compositions that can be used Examples of types of products are seed coating compositions, liquids for soaking roots or soil, and granular or powdered forms. It is a composition. Base materials for these compositions are well known to those skilled in the art. Furthermore, the fluorescent image of the present invention Pseudomonas fluorescens A bactericidal effective amount of Opal I, strain NCIB 40189 was added to plants, roots or seeds thereof, or Protection of crops from mosquito and fungal infections, consisting of application to the growing medium of plants provide a method. This treatment involves applying a preparation containing microorganisms of the strain of the present invention. This can be done by The bactericidal microorganisms of the present invention include banquet disease fungus Rh1zoctonia, Pythium hdanoesu, Molds related to "damping-off" such as those of the genus Fusarium The microorganism of the present invention is particularly effective against infections caused by take- Geramanomyces graminis (Gaeumannom), which causes disease of all It is also active against heather 1. The strain of the present invention (NCIB 40189) is from Chiba, Swindon, Wiltshire, UK. It was isolated from wheat roots collected from wild wheat grown in Seldon. The strain is On September 1, 1989, the National Co11 ctions of Industrial and Marine Bact area (23rd Street, Macha Drive, Aberdeen, Scotland, A B2IRY). The following describes the activity of bacterial isolates against some mosquitoes and fungi that infect agricultural crops. Describes the isolation, characterization, and selection of. Kodohiki Nikohe The bacterium was developed by Dray Cotto, Chiseldon, Swindon, Wiltshire, UK. cott) isolated from the roots of native wheat excavated from the fields of farms known as Ta. Wash the roots of the native wheat to remove all attached soil and place it in a glass bead-containing flask. into sterile distilled water. The wheat roots were shaken with glass beads for 15 minutes, then the washings were mixed with 10 = 1 and 10: Kings B agar (Proteose Bebutton No!) at a dilution rate of 3. 20g; Glycerol 10mR; Potassium sulfate 1.5g; Magnesium sulfate 1.5 g of heptahydrate; 12 g of industrial agar and 1 drop of distilled water at pH 7.2 and cyclohexahydrate. Cymid 754/mQ; Chloromycetin 12.547mQi and ampicillin (containing 5J@/mQ). Agar plates were incubated at 20°C for 3 days. Bacteria were randomly taken out from the agar plate and placed on 1/10) Ribsyn soybean juice agar (TSA ) (3 g tryptic soybean juice; 15 g agar; IQ water) . Bacteria were maintained on 1/10 TSA and stored at 4°C until required. 】”J1 Bon Seal 1u Nine Envelopes Fluorescence image (Pseudomonas fluorescens) strain NCIB The morphological characteristics of 40189 are from the National Co., Ltd. Determined by Industrial and Marine Bacteria and are shown in Table 1 below. Table 1 Morphology of P fluorescens strain NCIB 40189 Hoshiri: Gram-negative rods (rods) 1ヱ:　+ve Presentation 111J stop n Nitrate reduction reaction - Production of indole - Acid from glucose (+) Arginine dehydrolase + Urease - Niscrin hydrolysis - Gelatin hydrolysis 10 β-galactosidase - Assimilation of glucose Assimilation of arabinose 10 Mannose assimilation 10 Assimilation of mannitol 10 Assimilation of N-acetylglucosamine Assimilation of maltose 10 Assimilation of gluconate Caprete Assimilation 10 Assimilation of adipate - Malay assimilation 10 Assimilation of citrate 10 Assimilation of phenylacetate - Cytochrome oxidase + Biocyanin - Fluorescence 10 Gas glucose - Acid glucose PNS + 0NPG - Nitrate ~ Nitrite - Nitrite - Nitrogen - Residual nitrogen 10 Propylene glycol - Gel puncture 20℃ + Gel flat plate 10 Casein (+) Starch - Phenylalanine deaminase - Necessary growth factors - Tetracycline 10 Novobiocin - Polymyxin B + Leban + (The test is RE Buchanan, N E Gibbons (ads) 1 974゜Bergey's Manual of Determinative Bacteriology. 8th edition; ST Covan, KJ 5teel, 1974. Manu al for Identification of Medical Bact area). -nobe 1 plate Fluorescence imaging (Pseudomonas fluoresce) using Petri dish soil sorting ns) strain MCl8401119 was used as a damping-off fungus (h. It was tested for activity against. Fill a Petri dish (5 cm) with 10 mΩ potato dextrose agar (PDA). Filled. 7-day-old culture of damping-off fungus (h' Eshi ultimate) grown at 20°C An agar plug (5 mm) from the plant was centered on the PDA. Agar plate at 20℃ During this period, the damping-off fungus (P ultimum) colonizes on the agar plate. It turned into a knee. Minster Mendip loam soil at 120℃ for 60 minutes Autoclaved. Add sterile wheat germ seeds (1 wt/wt%) to this loam soil. and mixed thoroughly. Approximately 8g of soil was placed in each Petri dish. coated. The following treatment agent was used (2m12 were applied per one bird dish). 1) Fluorescent image (Pseudomonas fluorescens) strain NCI B 401119 (in 1/10 tryptic soybean juice (TSB) for 48 hours at 12°C culture grown in ); 2) Metalaxyl (100 ppm active ingredient AI); 3) Control: 1/10 TSB only. Duplicate agar plates were prepared and incubated at 10°C for 4 days. The results are shown in Table 2 below. Sex was recorded on a scale of O-3. A grade of zero means that there has been considerable growth of H Junjiang 4. A grade of 3 means no aerial mycelial growth in the Petri dish. Table 2 Active treatment agent of MCl840189 against damping-off fungus (P ultimum) activity grade NCIB 40189 3 Metalaxyl 3 Control O Results show that strain NCIB 40189 inhibits the growth of mosquitoes and fungi in soil. did. Its activity can be compared to the standard chemical fungicide metalaxyl. Any J Shimaji 2 Yuj

【skin Fluorescence image (Pseudomonas fluorescens) strain NCIB The efficacy of 40189 was tested against damping-off fungi (Mu'1 ultimum) and white group fungi (Rh). izoctonia 5 olani). Where the results show If the strain NCIB 40189 is grown under greenhouse conditions, P ultimum and Dan5ol Develops activity against ani. Activity against ヱULTIMUH: Cultures of P. ultimum were grown on PDA flat roots for 7 days at 20°C. 20 0g silver sand, 5g cornmeal and 4g Bemax wheat germ and 4011IQ water was autoclaved at 120°C for 20 minutes and then and charge it into the flask. 174 of the h Huangshan plate were inoculated. The flask was cultured at 20°C for 7 days, and then The contents of each flask were then mixed with fine sand to a final weight of 300 g. Add one flask containing byxium to 8Q Mendip Minestera. Mixed with soil to give standard dilution (X). Add this to clean Mendip loam soil. Further dilution to give a x/16 dilution used in the following tests. The test was conducted using a pot (diameter 7.62 cm) filled to 3/4 with soil infested with disease bacteria. I did it inside. Five peas were sown in each pot and covered with clean soil. . The following treatments were used (35 mQ added as soaking liquid to each pot): 1) Fluorescent image (Pseudomonas fluorescens) strain NCI B 40189 (in 1/10 tryptic soybean juice (TSB) for 48 hours at 20°C grown culture); 2) Metalaxyl (AI of 0.1-5.0 ppro); 3) Control; 1/10 TSB only Five replicates were performed for each treatment. Peas were grown for 14 days at 15°C and seedling germination was monitored weekly. Ento Water beans daily; pathogens promote damping-off in cold, damp conditions. Special care was taken to ensure that the sea urchin soil remained moist during the test. The results are shown in Table 3 below. Table 3 Strain NCIB 4011 for establishment and maintenance of pea in the presence of D. ultimum 19 effects NCIB 40189 76 metalaxyl 0, lppm 64 0.5ppm 114 1.0ppm 84 5, Oppm 96 Control 28 Results for strain NCIB 40189 and untreated control are shown. According to some sources, a strain of Pseudomonas fluorescens is applied. If you do this, you can germinate a large number of pea beans. Activity of strain NCIB 40189 The activity can be comparable to that of the chemical fungicide metalaxyl in terms of the level of disease developed. Activity against Dan5 OLANI: A culture of 1.5 olani was grown on PDA plates for 7 days at 20°C. 200g A mixture of silver sand, 4g Bemax wheat germ and 4On+1 water was heated to 120℃ for 20 minutes. Autoclaved at ℃ and then charged to a flask, 174 plates were inoculated. The flasks were cultured at 20°C for 7 days, and each flask was The contents of the Sco were mixed with fine sand to a final weight of 300 g Rh1zoct 60 Mendip Minster 374 in flask with onia inoculation tube A standard dilution (Y) was obtained by mixing with a-m soil. Add this to an even cleaner version of Mendipro. A Y/8 dilution was obtained for use in the following tests by diluting with soil. The test was conducted using a pot (diameter 7.62 cm) filled to 3/4 of the soil with disease-infested soil. I did it inside. Five peas were sown in each pot and covered with clean soil. . To use the following treatment agent, 35 mΩ was added as an immersion liquid to each pot): 1) Fluorescence image Pseudomonas fluorescens strain NCIB 401!19 (grown in 1710 tryptic soybean juice (TSB) for 48 hours at 20°C culture); 2) Bendiculone (monseren of 1o or 1100pp AI: Moncer) en) ; 3) Control with disease bacteria; 1/10 TSB only 4) Healthy control; Rh1zoc 1/10 TSB fleas on soil not infested with tonfa. Five replicates were performed for each treatment. Peas were grown for 14 days at 24°C and seedling germination was monitored weekly. Ento I watered the beans every day. The results are shown in Table 4 below. Table 4 Strain NCIB 4018 for establishment and maintenance of pea in the presence of R5olani 9 effects Treatment agent Germination rate of peas (%) NCI840189 118 montserene 10ppm 4g 100 ppm 60 Control with pathogens 2゜ Healthy contrast 92 Pseudomonas fluorescens strain NCIB Application of 40189 significantly increased the Peas can be sprouted. In fact, applying bacterial strain NCIB 40189 results in a healthy peas at levels comparable to the full control (treatment without fi5olani). Can be established and maintained. The activity of BCA is also higher than that of the chemical fungicide monselen. Seems expensive. の　・　　：】L stage】λνhairyoshi Pseudomonas fluorescens strain NCIB 40189t- Its activity against white MJPf bacteria (Nori Niyama Aim fa 5 olani) It was further tested for its properties and ability to protect peas from pathogens. The test consists of half-filling with disease-infested soil (prepared by the method described in the previous part). The test was carried out in a shallow rectangular box (tray) (165 mm x 95 mm). 25 Pea grains were sown in each shallow box and covered with clean soil. The treatments for BCA (strain NCIB 40189) and control were as follows. 1) Immersion solution: 100-bacterial suspension added to each shallow box (109 cells/m U 2)! I Granules 10g of preformed granules added to each shallow box; carrying bacterial cells The granules (model A, B, C) are BCA* thick maximum capacity crab 4.5 x 101 inoculated at 01 m cells/g; 3) 100 mQ of untreated water was added as an immersion liquid to each shallow box as a control. Three replicates were performed for each treatment. Peas were grown for 7-14 days at 24°C. Water the peas daily and keep them young. Seedling germination was monitored weekly. The test results were collated from all experiments and shown in Table 5 below. Active soaking solution of strain NCIB 40189 against D. olani 73 Granule A 65 Granule B 74 Granule C, 66 Unprocessed 31 Application of strain NCIB 40189 resulted in a reduction in peas when compared to the untreated control. The germination rate (%) of beans was significantly increased. Obtained with immersion liquid or granule BCA treatment agent There is no statistically significant difference between the results. ! Kantoyo Oka Fluorescent i1 (Pseudomonas fluorescens) strain NCIB 40189 against the disease "damping off" in peas and corn. used in outdoor tests as a biological control agent. Strain NCIB 40189 outdoors Tests were conducted on its ability to inhibit diseases of the genus Mudandetan and Fusarium. It was. The results are compared to standard chemical treatments and are reported below. In summary, damping-off Caused by the fungus Fusarium ultimum or Fusarium 5olani. With the application of strain NCIB 401119, the disease caused by standard chemical treatments can be eliminated. These results show that the reduction to levels comparable to those obtained by are doing. Effective against four swordfish crabs The test was conducted in France using peas as the test crop. Pathogenic bacteria are as follows. It was added at various ratios as an inoculum to wheat grains prepared in different manners. Batch amount of wheat seeds ( 1 kg'') was soaked in water overnight, drained and autoclaved at 121°C. The grown fungal spores were allowed to germinate and grow for two days, and then the seeds were incubated at 121°C. Reprocessed by autoclaving. 7-day-old plate of the damping-off fungus Rikaie Ultimate ( grown on potato dextrose agar at 20°C) was added to wheat seeds and Grow for 10 days, shake to redistribute mycelium, and leave for another 11 days of growth. The inoculum was then ready for use within the next 14 days. Strain NCIB 40189 was applied as a rubber/peat composition (107 cells 7 seeds), APRON (trade name) was used as a chemical control, 1 Application was made at a rate of 30 g of active ingredient AI per 00 kg of seeds. Compare two types of plants Used: untreated pea seeds and blank pea seeds (BCA or chemical (seeds were treated without the addition of agents). The results are shown in Table 6 below, and the "number of germinated seeds" is 200 seeds on a 5m long strip. The average number of pea plants that germinated 22 days after seeding is reported. " The letters that appear after the number of germination indicate statistical significance; common letters are There is no appreciable difference between comrades who have a probability value of 5%. Table 6 Outdoor activity of NCl340189 against P. ultimum: pea germination 0 100 ZOO NCI840189 1g0,25AB 153.00CDG 134.50F GH Ablon 171.50AC169,50ACB 157.25BCDP not yet Processing 172.50 AC130,25GHI 111.25 I blank test 17 6.00 AC144j4 DEH119,00HI The first column of Table 6 lists the pathogenic bacteria. The results of a control experiment in which the test was conducted in normal field soil without the addition of soil are shown. Plants grew normally in the absence of pathogens in the soil. Nurturing among fellow treatment agents There is no significant difference, and this means that adding strain NCIB 401119 will not cause drug damage. It is shown that. The other numbers in Table 6 give the results for the two treatments tested at various disease MWi application rates. . The presence of pathogens in the soil increases the Reduced the number of pea plants that germinated from the test seeds. This is a disease This indicates that the disease has a negative effect on plant growth. Of course . When strain NCIB 40189 is present, the number of plants that germinate even in the presence of pathogenic bacteria This significantly increases the Effects on FUSARIUM An outdoor test was conducted in the United States (Mississippi) using corn as a test crop. Ta. The pathogen (Dan5olani) was used as an inoculum of wheat grain prepared as described above. It was added at various application rates. Bacterial strain NCIB 40189 was applied as a rubber/peat composition (cells on the 107 side /seed), the chemical control agent is CAPTAN (trade name), Used as a seed treatment at recommended application rates. Use untreated seeds as a plant control there was. The results are shown in Table 7 below. 0 germination indicates the germination of corn plants 15 days after sowing. It was expressed as bud rate (%). The letters after the germination percentage (%) indicate statistical significance. common There is no acceptable difference between comrades with a probability value of 5%. germination of plants NCIB 40189 65,20! I C74,583AC Cabtan 69 ,5113 BC69,375BC unprocessed 65.833 C51,250D table The first column of 7 shows the results of a control test in which no test pathogen was added. In the absence of fungi, the plants were successfully hosted. There are no real differences between treatment agents. This indicates that adding bacterial strain NCIB 40189 does not cause drug damage. There is. The presence of the pathogenic fungus, Fusarium 5olani, in the soil was compared to the control experiment. reduced the number of corn plants that germinated from untreated seeds when compared to child This is a natural sign that a disease is present and is having a negative impact on plant growth. However, when strain NCIB 40189 is present, germination occurs even in the presence of pathogens. Increase the proportion (%) of plants that grow. In fact, strain MCl8401+1 What is the process in 9? Plants should be germinated so that germination rates are not appreciably different from germination rates in the absence of pathogens. Improved bud performance. Continuation of front page (81) Designated countries EP (AT, BE, CH, DE. DK, ES, FR, GB, GR, IT, LU, MC, NL, SE), 0A (BF , BJ, CF, CG, CI, CM, GA, GN, ML, MR, SN, TD, TG. ), AU, BB, BG, BR, CA, C3, FI, HU, JP. KP, KR, LK, MG, MN, MW, No, PL, R○, R U, SD, US (72) Inventor: Powell, Case, Adrian, United Kingdom. berkshire a Lucy 12.5 Enis, Nyar, Bracknell. Panfield, Emmettu Park, Bits Cross, 6 (72) Inventor: Campbell, Richard, United Kingdom, Bristol Binis 8 ・1 Nicey, Woodland Road, University of Bristol, DE Part of Batany (no house number)

Claims (10)

[Claims] 1. Contains the microorganism Pseudomonas fluorescens. A fungicide made of National Collection of India with accession number NCIB40189 on the 1st. Strial and Marine Bacteria (National Collection s of Industrial and Marine Bacteria Fungicides deposited with 2. Claim 1 as the active ingredient in admixture with an agriculturally acceptable carrier composition. A fungicidal agricultural composition containing a fungicide according to. 3. Applying an effective amount of the fungicide according to claim 1 to the plants or seeds. A method to prevent fungal attacks on plants. 4. 4. The method according to claim 3, wherein the fungicide is applied to plants. 5. 4. The method according to claim 3, wherein the fungicide is applied to the seeds before sowing. 6. Applying the fungicide to the growing medium in which the plants are growing or intended to grow. The method according to claim 3. 7. The claim that the fungus is from the genus Rhizoctonia The method described in Box 3. 8. Claim 3, wherein the mold is from the genus Pythium. the method of. 9. Claims that the fungus is from the genus Fusarium The method described in 3. 10. The fungus is Gaeumannomyces graminis. 4. The method according to claim 3, wherein