CN103382451B - Streptomyces griseochromogenes YN-6 and application thereof to club root control - Google Patents
Streptomyces griseochromogenes YN-6 and application thereof to club root control Download PDFInfo
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- CN103382451B CN103382451B CN201310341746.4A CN201310341746A CN103382451B CN 103382451 B CN103382451 B CN 103382451B CN 201310341746 A CN201310341746 A CN 201310341746A CN 103382451 B CN103382451 B CN 103382451B
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Abstract
The invention provides a Streptomyces griseochromogenes YN-6 separated from germ-carrying soil of cruciferae crops infected with club root, and the CCTCC No is M2013187. The strain YN-6 has good control effect on the club root of the cruciferae crops through verification of club root germ resting spore germination inhabitation and greenhouse pot experiments. The strain is high in adaptability and colonization capacity under field natural conditions, not prone to antagonistic reaction with complicated microorganism species in field soil, the disease control effect is stable, defects of chemical control, disease-resistant variety and agricultural control methods are effectively overcome, and the strain is an ideal anti-biological inoculant applicable to field club root control.
Description
Technical field
The present invention relates to biological control field, specifically, relate to a strain grey and produce look streptomycete YN-6 and the application in preventing and treating club root thereof.
Background technology
Crop in cruciferae club root is by rape plasmodiophora brassicae (Plasmodiophora brassicae), to infect the worldwide soil causing to pass Plant diseases.Rape plasmodiophora brassicae is obligate parasite, with its statospore, in the soil with invalid body and the barnyard manure that do not become thoroughly decomposed, survives the winter, and survival ability is extremely strong, generally can survive at least 8 years, and condition is suitable survives more than 15 years.Main prophylactico-therapeutic measures for crop in cruciferae club root at present has chemical prevention, disease-resistant variety utilization, cultural control and biological control etc.
Chemical prevention is the more general method of preventing and treating adopting, derosal, quintozene, m-tetrachlorophthalodinitrile, handsome (fluazinam), the section's good (cyanogen frost) etc. of obtaining of good fortune all have certain prevention effect to club root, effectively preventing disease in a short time, the output of guaranteeing crop can not be greatly affected, but the cost of this kind of method control is relatively high, and most medicament can leave residually on crop, the edible safety of crop is had a huge impact.
Breeding resistant variety is the Perfected process of preventing and treating club root.But because disease-resistant variety cultivate to need of long durationly, the cycle is long, and there is Physiological Race Differentiation in pathogen plasmodiophora, after disease-resistant variety is applied the several years, with regard to susceptibleization, therefore, control club root completely by disease-resistant variety also very difficult.
Cultural control method is as hoted water treatment of seeds, enrich white lime or plant ash etc., and these treatment processs also have certain limitation, as executes white lime and changed soil ph, but life-time service can destroy Soil structure, is not suitable for life-time service.
Biological control utilizes beneficial microorganism to kill or suppresses the growth of pathogenic micro-organism and reduce the quantity of pathogenic micro-organism, thereby control, Plant diseases occurs, a class measure of development.From controlling the sustainability of Plant diseases, consider, biological control is a class green safety effectively preventing method.
Summary of the invention
The object of this invention is to provide a strain grey and produce look streptomycete YN-6 and the application in control crop in cruciferae club root thereof.
In order to realize the object of the invention, the invention provides a strain grey and produce look streptomycete (Streptomyces griseochromogenes) YN-6, this bacterium be from dye have the crop in cruciferae of club root with the isolated streptomycete that crop in cruciferae club root is had to better prevention effect soil bacteria, now be preserved in Chinese Typical Representative culture collection center, address: Wuhan, China, Wuhan University, postcode 430072, deposit number CCTCC No:M2013187, preservation date on May 8th, 2013.
Above-mentioned grey is produced the cultural method of look streptomycete YN-6: grey is produced to look streptomycete YN-6 and be inoculated in the Gause I substratum of pH value for 7.2-7.4, at 24-28 ℃, under dark condition, cultivate 5-6d.
Adopt plate inserted sheet method to observe the form that grey is produced look streptomycete YN-6: the elongated multi-branched of substrate mycelium, straight shape or curve, do not rupture; Aerial hyphae spiral, relatively sturdy, easy fracture; Spore is partially cylindrical, and there is depression (Fig. 1 and 2) on surface.
The physio-biochemical characteristics of bacterial strain YN-6: produce melanochrome and H
2s, can utilize D-Fructose, glucose, sucrose, wood sugar, PEARLITOL 25C, can not make gelatine liquefication, can not make Starch Hydrolysis (not producing amylase), can not decomposition of cellulose (not producing cellulase), can not utilize L-rhamnosyl, L-arabinose, inositol.
Grey is produced look streptomycete YN-616S rDNA sequencing result as shown in Seq ID No.1.
The present invention also provides by grey and produces biocontrol fungicide prepared by look streptomycete YN-6.
The present invention also provides grey to produce the application in control crop in cruciferae (as Chinese cabbage, four seasons cream Plantula Brassicae chinensis, stone row cabbage heart etc.) club root of look streptomycete YN-6 or described biocontrol fungicide.
After streptomycete YN-6 activates on Gause I liquid nutrient medium flat board, 2 bacterium cake accesses of beating cut-off footpath 5mm with punch tool are equipped with in the 250mL triangular flask of 150mL Gause I liquid nutrient medium, under 28 ℃ of 180r/min conditions, shaking culture 6d in shaking table, with sterile gauze (4 layers), filter, obtain streptomycete YN-6 nutrient solution.
Carry out greenhouse pot culture test: adopt bacterium soil inoculation method to infect club root by crop, respectively at seeding time, after planting 7d, after planting 14d employing root-pouring method impose on streptomycete YN-6 nutrient solution or described biocontrol fungicide the root of crop, the preferred 20mL of each every alms bowl 15-25mL() 20mL, the preferred 30mL of every 3-5d cast Hoagland nutritive medium 25-35mL().
The present invention from dye have a crop in cruciferae of club root with the grey that is separated to soil bacteria, produce look streptomycete YN-6, by pathogen plasmodiophora statospore Germination suppression and greenhouse pot culture verification experimental verification bacterial strain YN-6 are had to good prevention effect to crop in cruciferae club root.This bacterial strain under the natural condition of field adaptive faculty and colonazition strong, be difficult for field soil in there is antagonistic action between the microbe species of complexity, prevent and treat disease effect stability, effectively having evaded the deficiency existing in the methods such as chemical prevention, disease-resistant variety and cultural control, is the desirable biocontrol fungicide that is suitable for land for growing field crops club root control.
Accompanying drawing explanation
Fig. 1 is that in the embodiment of the present invention 2, grey is produced the form (* 600) of look streptomycete YN-6 on Gause I substratum.
Fig. 2 is the spore chain form (* 30000) that in the embodiment of the present invention 2, grey is produced look streptomycete YN-6.
Fig. 3 is that in the embodiment of the present invention 2, grey is produced the cultivation results of look streptomycete YN-6 on different culture media flat board.
Fig. 4 adopts dull and stereotyped face-off culture method to identify the inhibition of streptomycete YN-6 to various pathogenic bacteria in the embodiment of the present invention 2.
Embodiment
Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.If do not specialize, the conventional means that in embodiment, technique means used is well known to those skilled in the art, the raw materials used commercial goods that is.
Embodiment 1 grey is produced the preparation of look streptomycete YN-6 nutrient solution
By from dye have a crop in cruciferae of club root with the grey that is separated to soil bacteria, produce look streptomycete YN-6(CCTCC No:M2013187) after the activation of Gause I liquid nutrient medium flat board, 2 bacterium cake accesses of beating cut-off footpath 5mm with punch tool are equipped with in the 250mL triangular flask of 150mL Gause I liquid nutrient medium, under 28 ℃ of 180r/min dark conditions, shaking culture 6d in shaking table, with sterile gauze (4 layers), filter, obtain streptomycete YN-6 nutrient solution.
Embodiment 2 is by testing and the prevention effect of greenhouse pot culture test evaluation bacterial strain YN-6 to crop in cruciferae pathogen plasmodiophora pathogen plasmodiophora statospore Germination suppression
The indicator strain using in the present embodiment is orange anthrax bacteria (Colletotrichum geloeosporiolides), ustilaginoidea virens (Ustiloginoidea virens), Phytophthora capsici germ (Phytophthorac apsici), capsicum point spore anthrax bacteria (Colletotrichum spp.), Pyricularia oryzae (Pyricularia oryzae).
1 pair of pathogen plasmodiophora statospore Germination suppression experiment
Bacterial strain YN-6 nutrient solution, pathogen plasmodiophora statospore suspension, Collina juice are added in the triangular flask of sterilizing with the amount of 2mL, 2mL, 20mL respectively, it is contrast that the sterilized water of take replaces Antagonistic Fungi nutrient solution, pH value 6.3,24 ℃ of dark culturing, after 5d, with lichens red colouring 3min microscopy statospore, sprout situation, repeat 3 times, it is 61.75% to pathogen plasmodiophora statospore Germination suppression rate that result shows.
2 greenhouse pot culture tests
Adopt bacterium soil inoculation method to infect club root by crop, respectively at seeding time, after planting 7d, after planting 14d adopts root-pouring method streptomycete YN-6 nutrient solution to be imposed on to the root of crop, each every alms bowl waters 20mL.Clear water, 75% m-tetrachlorophthalodinitrile (* 600 times of liquid) are set for contrasting, totally 6 processing, each processes 6 alms bowls, every alms bowl 5 strains.The about 30mL of every 3-5d cast Hoagland nutritive medium, a situation arises (table 1 and table 2) to plant about 30d " Invest, Then Investigate " club root.
The prevention effect of table 1YN-6 to susceptible kind four seasons cream Plantula Brassicae chinensis club root
The prevention effect of table 2.YN-6 to the special pure breeding stone row of susceptible kind cabbage heart club root
Cabbage clubroot is by Protozoa rape plasmodiophora brassicae (P.brassicae), to infect the worldwide soil causing to pass Plant diseases, main harm root, form tumour not of uniform size, when serious, cause that plant butt rot and even complete stool are withered, cause heavy economic losses.The major measure that control crop in cruciferae club root is taked at present has the chemical prevention of comprising, disease-resistant variety utilization, and cultural control etc., because soil-borne disease chemical control consumption is large, exist hidden danger to brassicaceous vegetable edible safety; Disease-resistant variety is few at present, can not meet need of production; Cultural control only can be as assisting in preventing and treating measure.It is a kind of safe, effective, free of contamination method of preventing and treating that application antagonistic microbe carries out this disease of biological control.From the angle of Sustainable development, consider, screening biocontrol microorganisms controlling plant diseases is considered to safe and effective, the free of contamination method of preventing and treating.
The present invention is separated to bacterial strain YN-6 from dye the band soil bacteria of the crop in cruciferae that has club root, by the flat board culture experiment that stands facing each other, find that this bacterial strain is to multiple cause of disease indicator, as orange anthrax bacteria (Colletotrichum geloeosporiolides), ustilaginoidea virens (Ustiloginoidea virens), Phytophthora capsici germ (Phytophthora capsici), capsicum point spore anthrax bacteria (Colletotrichum spp.), Pyricularia oryzae (Pyricularia oryzae) etc. all have good antagonistic effect.
Fig. 3 is for being used the dull and stereotyped result of cultivating grey product look streptomycete YN-6 of different culture media.
Fig. 4 is for adopting dull and stereotyped face-off culture method to identify the inhibition of streptomycete YN-6 to various pathogenic bacteria.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements, all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (6)
1. grey is produced look streptomycete (Streptomyces griseochromogenes) YN-6, and its preserving number is CCTCC No:M2013187.
2. the cultural method of grey product look streptomycete YN-6 described in claim 1, is characterized in that, grey is produced to look streptomycete YN-6 and be inoculated in the Gause I substratum that pH value is 7.2-7.4, at 24-28 ℃, under dark condition, cultivates 5-6d.
3. by grey described in claim 1, produce biocontrol fungicide prepared by look streptomycete YN-6.
4. the application of biocontrol fungicide in control crop club root described in grey product look streptomycete YN-6 or claim 3 described in claim 1, wherein, described crop is crop in cruciferae Chinese cabbage, stone row cabbage heart.
5. application according to claim 4, is characterized in that, adopts root-pouring method that grey is produced to the root that look streptomycete YN-6 nutrient solution or described biocontrol fungicide impose on the crop that catches an illness, each every alms bowl 15-25mL, every 3-5d cast Hoagland nutritive medium 25-35mL.
6. application according to claim 5, is characterized in that, adopts root-pouring method that grey is produced to the root that look streptomycete YN-6 nutrient solution or described biocontrol fungicide impose on the crop that catches an illness, each every alms bowl 20mL, every 3-5d cast Hoagland nutritive medium 30mL.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101311262A (en) * | 2007-05-22 | 2008-11-26 | 上海市农药研究所 | Streptomyces griseus and uses thereof |
| CN101416641A (en) * | 2008-09-12 | 2009-04-29 | 云南农业大学 | Biological preparation capable of preventing and treating cruciferae club root and use thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101311262A (en) * | 2007-05-22 | 2008-11-26 | 上海市农药研究所 | Streptomyces griseus and uses thereof |
| CN101416641A (en) * | 2008-09-12 | 2009-04-29 | 云南农业大学 | Biological preparation capable of preventing and treating cruciferae club root and use thereof |
Non-Patent Citations (5)
| Title |
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| Isolation and characterization of endophytic actinomycetes from Chinese cabbage roots as antagonists to Plasmodiophora brassicae;Lee S O 等;《J Microbiol Biotechnol》;20081231;第18卷(第11期);第1741-1746页 * |
| L.H. CHEAH 等.BIOLOGICAL CONTROL OF CLUBROOT ON CAULIFLOWER WITH TRICHODERMA AND STREPTOMYCES SPP..《New Zealand Plant Protection》.2000,第53卷第18-21页. * |
| Lee S O 等.Isolation and characterization of endophytic actinomycetes from Chinese cabbage roots as antagonists to Plasmodiophora brassicae.《J Microbiol Biotechnol》.2008,第18卷(第11期),第1741-1746页. * |
| 十字花科根肿病研究进展;王靖 等;《植物保护》;20111231;第37卷(第6期);153-158 * |
| 王靖 等.十字花科根肿病研究进展.《植物保护》.2011,第37卷(第6期),153-158. * |
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