CN1952106B - Avirulent binucleate rhizoctonia BNR-1 agricultural agent and its uses - Google Patents
Avirulent binucleate rhizoctonia BNR-1 agricultural agent and its uses Download PDFInfo
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- CN1952106B CN1952106B CN200610070048A CN200610070048A CN1952106B CN 1952106 B CN1952106 B CN 1952106B CN 200610070048 A CN200610070048 A CN 200610070048A CN 200610070048 A CN200610070048 A CN 200610070048A CN 1952106 B CN1952106 B CN 1952106B
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Abstract
The invention relates to a nonpathogenic dicaryon threads pyrenomycetes as well as its preparing process and the application of its preparations, which has something to do with the agricultural biotechnology field. The preserving number of the nonpathogenic dicaryon threads pyrenomycetes BNR-1 is CGMCC NO.1844. The preparing process comprises the following steps: cultivating the strain for 3-4d oninclined plane; cultivating the strain in shaking flask for 2-3d; cultivating the strain on solid medium for 5-7d. The granules mixed by culture on the solid medium, glucose and/or calcium carbonatecan be used to prevent and treat cotton's instant-drying disease and the wheat's stripe-drying disease.
Description
Technical field
Dry thread Pyrenomycetes) and formulation preparation and application the present invention relates to a kind of Avirulent binucleate rhizoctonia species (Rhizoctonia solani) BNR-1 (classification name:, belong to agricultural biological technical field.
Background technology
Enter 21 century, along with improving constantly of expanding economy and living standards of the people, people's health perception and environmental consciousness are strengthened greatly, reduce the use of chemical pesticide, adopt green bio pesticide control disease pest and weed to become the inexorable trend of agricultural chemicals development, the Ministry of Agriculture and national petrochemical industry office have also all put into effect corresponding support policy to the development of biological husbantry.Although biological pesticide has had certain development, still there are some outstanding problems in research and development and aspect such as application, as the quantity of present China biocontrol fungicide and of less types, prevention effect is lower or unstable.Therefore must enlarge the germ plasm resource of biocontrol strain, wherein this deficiency has been alleviated in the research and development utilization of Avirulent binucleate rhizoctonia species.
About the research of Avirulent binucleate rhizoctonia species, existing both at home and abroad report.Since 1984; respectively from Israel; the U.S.; Canada; Japan; Avirulent binucleate rhizoctonia species bacterial strain (the B.SNEH that is used for plant protection is isolated on ground such as Australia; E.YAMOAH and A.STEWART.HYPOVIRULENT RHIZOCTONIA SPP.ISOLATES FROM NEW ZEALAND SOILS PROTECT RADISH SEEDLINGS AGAINSTDAMPING-OFF CAUSED BYR.SOLANI.New Zealand Plant Protection; 2004 (57): 54～58.) (B.SNEH; E.YAMOAH and A. utilization isolating no pathogenicity rhizoctonia from New Zealand's soil is prevented and treated by the microbial radish seedling canker of miliary damping-off; New Zealand's plant protection, 2004 (57): 54～58).Some Avirulent binucleate rhizoctonia species bacterial strain can inducing cotton under greenhouse test system's resistance; and effect is than remarkable (the Suha Jabaji-Hare and Stephen M.Neate.Nonpathogenic Binucleate Rhizoctonia spp.andBenzothiadiazole Protect Cotton Seedlings Against Rhizoctonia Damping-Off and AlternariaLeaf Spot in Cotton.PHYTOPATHOLOGY of chemical inducer diazosulfide; 2005; 95 (9): 1031～1036) (Soviet Union breathes out; Si Difen. the canker that the opposing of Avirulent binucleate rhizoctonia species and diazosulfide protection cotton seedling is caused by rhizoctonia and by the full microbial spot disease of chain lattice; Plant Pathology; 2005,95 (9): 1031～1036).The minority bacterial strain can promote plant-growth, increases output (Sneh, B.; Zeidan, M.; Ichielevich-Auster, M.et al.Increased growth responses induced by anon-pathogenic Rhizoctonia solani.Can.J.Bot.1986,64:2372～2378) (Sneh, B.; Zeidan, M.; Ichielevich-Auster, the growth-promoting functions of M.et al. no pathogenicity dry thread Pyrenomycetes, Canada, 1986,64:2372～2378).
Up to the present, all worlds, the Plant diseases biological bactericide of having registered on domestic mainly contain bacterium and fungi two big classes, are the microbial inoculum of biocontrol strain but also do not have a kind of with non-virulent double-core rhizoctonia (Rhizoctonia solani).
Summary of the invention
The purpose of this invention is to provide a kind of novel biocontrol microorganisms, and utilize this biocontrol microorganisms to develop a kind of Agrotechnical formulation, to enlarge the biocontrol microorganisms kind, effective controlling plant diseases.
One of purpose of the present invention provides a kind of Avirulent binucleate rhizoctonia species (Rhizoctoniasolani) BNR-1 with biological control function, classification name: dry thread Pyrenomycetes (Rhizoctonia solani).
Two of purpose of the present invention provides the solid fermentation method of a kind of Avirulent binucleate rhizoctonia species BNR-1.
Three of purpose of the present invention provides the Agrotechnical formulation of a kind of Avirulent binucleate rhizoctonia species BNR-1.
Four of purpose of the present invention provides the application of Avirulent binucleate rhizoctonia species BNR-1 Agrotechnical formulation.
Below technical scheme of the present invention is described respectively.
1, Avirulent binucleate rhizoctonia species BNR-1.
Avirulent binucleate rhizoctonia species BNR-1 (Rhizoctonia solani), classification name: dry thread Pyrenomycetes (Rhizoctoniasolani), preserving number: CGMCC NO.1844.
The feature description of this bacterial strain is as follows:
Mycelia is dendroid branch, and is more sparse, light brown, slightly hangs in bifurcation and contracts, and the place of being not far from here forms barrier film, and mycelia is nearly right angle branch.Sclerotium is from mycelia, and particle is flat, grows in stromal surface, inside and outside solid colour, and the sclerotium superficial cell is slightly little, does not have obvious boundary with inside, and each cell contains 2 nucleus.
This strains separation, can be grown rapidly at plant rhizosphere the plant no pathogenicity surely from plant rhizosphere, by with pathogenic bacteria competition nutrition and inducing plant system's resistance of pathogenic bacteria being reached the disease effect that prevents and treats.
2, the preparation of the solid fermentation culture of Avirulent binucleate rhizoctonia species BNR-1CGMCC NO.1844.
The solid fermentation method of a kind of Avirulent binucleate rhizoctonia species BNR-1CGMCC NO.1844 carries out according to the following steps, all is weight percentage:
(1) slant strains: adopt solid PDA substratum or water agar, Avirulent binucleate rhizoctonia species BNR-1 classification is named: dry thread Pyrenomycetes (Rhizoctonia solani) CGMCC NO.1844 is seeded on the slant tube, cultivates 3～4 days for 25～28 ℃.
(2) shake a bottle bacterial classification: adopt liquid PDB substratum, above-mentioned slant strains is seeded in the liquid shaking bottle, placed on the shaking table 28 ℃ of shaking culture 2～3 days.
(3) solid spawn: adopt solid medium, water content is 45～55%wt, sterilizes 50～65 minutes for 120～121 ℃, and liquid spawn and solid medium are mixed, and inoculum size is 5～10%wt, and inoculation finishes and transfers to the solid culture indoor cultivation.
(4) cultivate: shallow-layer substratum thickness is 4～6cm, and the material temperature control is at 28 ± 2 ℃, and room temperature is controlled at 23～32 ℃, and the relative water content of air is controlled at 45～85%wt, and incubation time is 5～7 days, and it is The more the better that sclerotium produces.
3, the Agrotechnical formulation of Avirulent binucleate rhizoctonia species BNR-1.
The Agrotechnical formulation of a kind of Avirulent binucleate rhizoctonia species BNR-1 is mixed and made into granule by solid fermentation culture and starch, glucose and/or the lime carbonate of Avirulent binucleate rhizoctonia species BNR-1.The weight ratio of the solid fermentation culture of Avirulent binucleate rhizoctonia species BNR-1 and other composition total amounts is 1: 1～1.5.
The Agrotechnical formulation preparation method of Avirulent binucleate rhizoctonia species BNR-1 is: with solid fermentation culture natural air drying above-mentioned 2, (4) preparation, add starch, glucose and/or light calcium carbonate mix, mist projection granulating on rolling disc.The diameter of granule is 1～2mm.
4, the application of Avirulent binucleate rhizoctonia species BNR-1 Agrotechnical formulation.
The application of Avirulent binucleate rhizoctonia species BNR-1 Agrotechnical formulation is used to prevent and treat cotton seedling blight or wheat hypochnus.
The effect of Avirulent binucleate rhizoctonia species BNR-1 Agrotechnical formulation is as follows:
(1) materials and methods: the test plant pathogenic fungi is cotton rhizoctonia solani (Rhizoctonia solani) and rhizoctonia cerealis (Rhizoctonia cerealis), purified back inoculation millet substratum is cultivated for 25 ℃ and was covered with small rice grain to pathogenic fungi in 4～5 days.For the examination wheat breed is good star 99, and cotton variety is Shandong 9154, with 0.1% mercuric chloride water surface sterilization 1min, and aseptic water washing, vernalization is to showing money or valuables one carries unintentionally under 30 ℃ of conditions.Seed adopts jingganmycin (80 μ g/mL), triazolone (80 μ g/mL), clear water seed soaking to handle 30min respectively, is directly used in following test after the presprouting of seeds of Agrotechnical formulation BNR-1 granule treatment group.
Insert 20 in the seed of above-mentioned different treatment in every basin respectively, be covered with 2 of the millets of pathogenic fungi, (diameter is 1～2mm) to add 2 of Avirulent binucleate rhizoctonia species BNR-1 granules of the present invention in addition for examination Agrotechnical formulation treatment group.3 repetitions are established in every processing, and district's group is arranged at random, hot-house culture, and temperature is 18～25 ℃, and humidity is 40～70%, regularly quantitatively waters, and trial period is 30 days.
The investigation of the cotton seedling blight state of an illness is calculated with prevention effect:
0 grade: anosis; 1: basal part of stem has little scab, accounts for below 1/4 of stem girth; 2 grades: the basal part of stem scab is bigger, accounts for the 1/4-1/2 of stem girth; 3 grades: the basal part of stem scab accounts for more than 1/2 of stem girth, but does not destroy whole stem girth as yet; 4 grades: the basal part of stem scab occupies whole stem girths, plant death.
The investigation of the wheat hypochnus state of an illness is calculated with prevention effect:
0 grade: leaf sheath, stem stalk are not all infected; 1 grade: scab is only limited on the leaf sheath, does not have scab on the stem stalk; 2 grades: though on the stem stalk scab is arranged, scab is grown crosswise and is not reached stem stalk girth and have 1/3; 3 grades: scab is grown crosswise and is reached 1/3 of stem stalk girth, but does not reach 2/3 of stem stalk girth; 4 grades: scab is grown crosswise and is reached 2/3 of stem stalk girth, until scab around the stem stalk.
Total strain number of diseased plant number * 100%/each processing of each processing of diseased plant rate %=
Disease index %=∑ (each sick level * strain numbers at different levels) * 100%/(the highest sick grade * always investigate strain number)
Prevention effect %=(contrast disease index-processing disease index) * 100%/contrast disease index
(2) result: Agrotechnical formulation BNR-1 granule of the present invention under greenhouse experiment in various degree preventive and therapeutic effect all being arranged for examination two kind of plant pathogenic fungies, significant difference on 0.05 level compared with the control, prevention effect all is better than contrast medicament (table 1).
Table 1 Agrotechnical formulation BNR-1 of the present invention granule is to the greenhouse prevention effect of two kinds of crop pests
(annotate: data are three multiple mean values, adopt the Duncan method in the SPSS10.0 statistical software.Be illustrated in significant difference on 0.05 level between different lowercase marks.)
Avirulent binucleate rhizoctonia species BNR-1 (Rhizoctonia solani), classification name: dry thread Pyrenomycetes (Rhizoctoniasolani).The preservation time: on October 20th, 2006, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, address: No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, preserving number: CGMCC NO.1844.
Description of drawings
Fig. 1 is the nucleus dyeing (picture of sarranine-O) of double-core rhizoctonia BNR-1 (dry thread Pyrenomycetes) (Rhizoctonia solani) CGMCC NO.1844.
Embodiment
The present invention will be further described below in conjunction with embodiment.
The embodiment bacterial strain uses therefor is: Avirulent binucleate rhizoctonia species BNR-1 (Rhizoctonia solani), classification name: dry thread Pyrenomycetes (Rhizoctonia solani), the preservation time: on October 20th, 2006, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, address: No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, preserving number: CGMCC NO.1844.
The used substratum of embodiment is as follows, and each components in proportions of solid medium is weight ratio:
PDA substratum: potato 200g, glucose 10g, agar 15g, tap water 1000mL.
Water agar: agar 15g, tap water 1000mL.
Solid medium 1: wheat bran: rice husk: Semen Maydis powder: chitin powder=7: 1: 1.5: 0.5.
Solid medium 2: wheat bran: cotton seed hull: Semen Maydis powder: chitin powder=7: 1: 1.5: 0.5.
Solid medium 3: wheat bran: cotton seed hull: starch=7: 1: 2.
Solid medium 4: wheat bran: cotton seed hull=8: 2.
Embodiment 1: Avirulent binucleate rhizoctonia species BNR-1.
Avirulent binucleate rhizoctonia species BNR-1 (Rhizoctonia solani), classification name: dry thread Pyrenomycetes (Rhizoctoniasolani), the preservation time: on October 20th, 2006, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, address: No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, preserving number: CGMCC NO.1844.This strains separation, can be grown rapidly at plant rhizosphere the plant no pathogenicity surely from plant rhizosphere, by with pathogenic bacteria competition nutrition and inducing plant system's resistance of pathogenic bacteria being reached the disease effect that prevents and treats.This bacterial strain mycelia is dendroid, and is more sparse, light brown, slightly hangs in bifurcation and contracts, and the place of being not far from here forms barrier film, and mycelia is nearly right angle branch.Sclerotium is from mycelia, and particle is flat, grows in stromal surface, inside and outside solid colour, and the sclerotium superficial cell is slightly little, does not have obvious boundary with inside.Each cell contains 2 nucleus (as shown in Figure 1).
Embodiment 2: the preparation of the solid fermentation culture of Avirulent binucleate rhizoctonia species BNR-1CGMCC NO.1844.
(1) slant strains: adopt solid PDA substratum, with Avirulent binucleate rhizoctonia species BNR-1CGMCCNO.1844 (classification name: dry thread Pyrenomycetes (Rhizoctonia solani)) be seeded on the test tube substratum, cultivated 3 days for 28 ℃.
(2) eggplant bottle bacterial classification: adopt liquid PDA substratum, test tube strains is seeded in the liquid eggplant bottle, placed on the shaking table 28 ℃ of shaking culture 2 days.
(3) solid spawn: adopt solid medium 1, wheat bran: rice husk: Semen Maydis powder: chitin powder=7: 1: 1.5: 0.5, water content is 50%wt, sterilized 60 minutes for 121 ℃, liquid spawn and solid medium are mixed, inoculum size is 6%wt, and inoculation finishes and transfers to the solid culture indoor cultivation.
(4) cultivate: substratum thickness is 5cm, and the material temperature control is at 28 ± 2 ℃, and room temperature is controlled at 25～30 ℃, and the relative water content of air is controlled at 45～85%wt, and incubation time is 7 days.Get the solid fermentation culture of Avirulent binucleate rhizoctonia species BNR-1.
Embodiment 3: the preparation of the solid fermentation culture of Avirulent binucleate rhizoctonia species BNR-1CGMCC NO.1844.As described in embodiment 1, different is that step (3) adopts solid medium 2, wheat bran: cotton seed hull: Semen Maydis powder: chitin powder=7: 1: 1.5: 0.5.
Embodiment 4: the preparation of the solid fermentation culture of Avirulent binucleate rhizoctonia species BNR-1CGMCC NO.1844.
As described in embodiment 1, different is that step (3) adopts solid medium 3, wheat bran: cotton seed hull: starch=7: 1: 2.
Embodiment 5: the preparation of the solid fermentation culture of Avirulent binucleate rhizoctonia species BNR-1CGMCC NO.1844.
As described in embodiment 1, different is that step (3) adopts solid medium 4, wheat bran: cotton seed hull=8: 2.
Embodiment 6: the Agrotechnical formulation of Avirulent binucleate rhizoctonia species BNR-1, and component is as follows:
Double-core rhizoctonia BNR-1 solid fermentation culture (air-dry) 60g
Starch 30g
Glucose 10g
Light calcium carbonate 30g
The preparation method: with product (the double-core rhizoctonia BNR-1 solid fermentation culture) natural air drying of embodiment 1, adding starch, glucose, light calcium carbonate mix, and mist projection granulating on rolling disc, the diameter of granule are 1～2mm.
Embodiment 7: the Agrotechnical formulation of Avirulent binucleate rhizoctonia species BNR-1, and component is as follows:
Double-core rhizoctonia BNR-1 solid fermentation culture (air-dry) 60g
Starch 30g
Light calcium carbonate 30g
The preparation method is with the product of embodiment 4 (double-core rhizoctonia BNR-1 solid fermentation culture) natural air drying, adds starch and light calcium carbonate and mixes, and mist projection granulating on rolling disc, the diameter of granule are 1～2mm.
Claims (2)
1. the Agrotechnical formulation of an Avirulent binucleate rhizoctonia species BNR-1, it is characterized in that it being the granule that solid fermentation culture and starch, glucose and/or lime carbonate by Avirulent binucleate rhizoctonia species BNR-1 are mixed and made into, the weight ratio of the solid fermentation culture of Avirulent binucleate rhizoctonia species BNR-1 and other composition total amounts is 1: 1～1.5;
The solid fermentation culture of described Avirulent binucleate rhizoctonia species BNR-1 is made by following method, and the per-cent in each step all is weight percentage:
(1) slant strains: adopt solid PDA substratum or water agar, Avirulent binucleate rhizoctonia species BNR-1CGMCC NO.1844 is seeded on the slant tube, cultivated 3～4 days for 25～28 ℃;
(2) shake a bottle bacterial classification: adopt liquid PDB substratum, above-mentioned slant strains is seeded in the liquid shaking bottle, placed on the shaking table 28 ℃ of shaking culture 2～3 days;
(3) solid spawn: adopt solid medium, water content is 45～55%wt, sterilizes 50～65 minutes for 120～121 ℃, and liquid spawn and solid medium are mixed, and inoculum size is 5～10%wt, and inoculation finishes and transfers to the solid culture indoor cultivation;
(4) cultivate: shallow-layer substratum thickness is 4～6cm, and the material temperature control is at 28 ± 2 ℃, and room temperature is controlled at 23～32C, and the relative water content of air is controlled at 45～85%wt, and incubation time is 5～7 days.
2. the application of the described Avirulent binucleate rhizoctonia species BNR-1 of claim 1 Agrotechnical formulation is used to prevent and treat wheat hypochnus or cotton seedling blight.
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| CN101914457B (en) * | 2008-04-24 | 2012-07-25 | 中国医学科学院药用植物研究所 | Fungus for promoting germination of bletilla striata seeds |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5948671A (en) * | 1998-07-24 | 1999-09-07 | The United States Of America As Represented By The Secretary Of Agriculture | Control of replant disease of tree fruits with Pseudomonas putida |
| WO2000004779A1 (en) * | 1998-07-24 | 2000-02-03 | Instituto Tecnico Agronomico Provincial, S.A. | Protection of plants using rhizoctonia |
| CN1766091A (en) * | 2005-09-23 | 2006-05-03 | 中国农业大学 | Bacillus subtilis and its uses |
-
2006
- 2006-11-08 CN CN200610070048A patent/CN1952106B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5948671A (en) * | 1998-07-24 | 1999-09-07 | The United States Of America As Represented By The Secretary Of Agriculture | Control of replant disease of tree fruits with Pseudomonas putida |
| WO2000004779A1 (en) * | 1998-07-24 | 2000-02-03 | Instituto Tecnico Agronomico Provincial, S.A. | Protection of plants using rhizoctonia |
| CN1766091A (en) * | 2005-09-23 | 2006-05-03 | 中国农业大学 | Bacillus subtilis and its uses |
Non-Patent Citations (2)
| Title |
|---|
| Suha Jabaji-Hare等.nonpathogenic binucleate rhizoctonia spp. and benzothiadiazole protect cotton seedlings against rhizoctonia damping-off and alternaria leaf spot in cotton.phytopathology95 9.2005,95(9),1030-1036. |
| Suha Jabaji-Hare等.nonpathogenic binucleate rhizoctonia spp. and benzothiadiazole protect cotton seedlings against rhizoctonia damping-off and alternaria leaf spot in cotton.phytopathology95 9.2005,95(9),1030-1036. * |
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| CN1952106A (en) | 2007-04-25 |
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