CN102876622B - Biocontrol bacterium and application of biocontrol bacterium to preventing and controlling cotton blight and cotton greensickness - Google Patents
Biocontrol bacterium and application of biocontrol bacterium to preventing and controlling cotton blight and cotton greensickness Download PDFInfo
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Abstract
The invention discloses Streptomyces rochei 42561 which is preserved in the general microorganism center of the China Committee for Culture Collection of Microorganisms (CCCCM), with the preservation number of CGMCC (China General Microbiological Culture Collection Center) NO.6556. The bacteriostatic and prophylactic actions of the biocontrol bacterium 42561 are comprehensively tested according to a cup-plate method, a mycelial growth rate inhibition method, a spore germination inhibition method, greenhouse pot culture and field tests; and the testing results show that the strain has a remarkable prevention effect on cotton blight and cotton greensickness, can be used as an effective biological pesticide preparation agent and has excellent application and development prospects.
Description
Technical field
The present invention relates to biological control field, be specifically related to Lou Che Shi streptomycete (
streptomyces rochei) and biological and ecological methods to prevent plant disease, pests, and erosion purposes.
Background technology
Cotton wilt and cotton verticillium wilt are the main diseases of harm Cotton Production, and cotton wilt and cotton verticillium wilt are on the rise in China cotton region in recent years.Yet, also do not prevent and treat the beneficial agents of cotton wilt and cotton verticillium wilt at present.From Microbial resources, finding biological pesticide is one of effective way, and the agricultural antibiotic that actinomycetes produce is most important source wherein.The development of the biological pesticides such as jingganmycin, Avrmectin and Qinlingmycin popularization have fully proved reasonableness and the superiority of screening economical and efficient agricultural antibiotic from actinomycetic meta-bolites.The biocontrol microorganisms of the withered and yellow bacterium that withers of control cotton of registering at present have not been reported, but the biological prevention and control agent commodity of existing other crop blights of control.For example, the trichoderma harziarum of the commodity F-stop by name that Harman in 1993 etc. register, for the control of vegetables reaping hook root rot; Germany biological prevention and control agent PROPHYTA, for champac shape bacterium (
talaromyces flavus), be mainly used in the control of vegetables verticillium.
Therefore, the efficient biocontrol microorganisms by screening control cotton wilt and cotton verticillium wilt from actinomycetes is object of the present invention.
Summary of the invention
Technical problem to be solved by this invention is: a kind of effective biocontrol microorganisms is provided, specifically Lou Che Shi streptomycete (
streptomyces rochei), on September 11st, 2012 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preserving number is CGMCC No.6556.
Technical scheme provided by the invention is: a kind of Lou Che Shi streptomycete (
streptomyces rochei), (preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City), preserving number is CGMCC No.6556, after testing survival on September 11st, 2012, to be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Lou Che Shi streptomycete described in the present invention also provides is as the purposes of biocontrol microorganisms, especially for control cotton wilt and cotton verticillium wilt.
The present invention has following beneficial effect: result shows that Lou Che Shi streptomycete (biocontrol microorganisms 42561) fermented liquid is respectively 23.96mm and 32.67mm to cotton-wilt fusarium (Fusarium oxysporum) and verticillium dahliae (verticillium dahliae) antibacterial circle diameter, its mycelial growth inhibition rate is respectively to 89.41% and 83.75%, and inhibition of germination is respectively 73.56% and 68.75%; Results from pot experiment test shows, biocontrol microorganisms 42561 fermented liquids all show good preventive and therapeutic effect to cotton wilt and cotton verticillium wilt, and prevention effect is respectively 36.73% and 44.82%; Field test shows that biocontrol microorganisms 42561 fermented liquids have certain effect to the field control of cotton spoting verticillium wilt, after dispenser for the third time the 15th and 30 days, its prevention effect reaches respectively 33.06% and 41.65%.Therefore, 42561 pairs of cotton wilts of biocontrol microorganisms and cotton verticillium wilt preventive effect are remarkable, can be used as effective biological pesticide preparation, have good application and development prospect.
Accompanying drawing explanation
The taxonomy of Fig. 1 biocontrol microorganisms 42561.
The dull and stereotyped face-off method of Fig. 2 biocontrol microorganisms 42561 fungistatic effect.Wherein, a left side: verticillium dahliae is target bacterium; Right: Fusarium oxysporum is target bacterium.A:42561; B: culture block.
Fig. 3 biocontrol microorganisms 42561 fermented liquid cup-plate method fungistatic effects, a left side: verticillium dahliae is target bacterium; Right: Fusarium oxysporum is target bacterium.A:42561 fermented liquid; B: liquid nutrient medium.
The inhibition of Fig. 4 biocontrol microorganisms 42561 fermented liquids to verticillium dahliae mycelial growth.A: biocontrol microorganisms 42561 fermentation liquor treatment groups; B: liquid nutrient medium control group.
The inhibition of Fig. 5 biocontrol microorganisms 42561 fermented liquids to Fusarium oxysporum mycelial growth.A: biocontrol microorganisms 42561 fermentation liquor treatment groups; B: liquid nutrient medium control group.
The restraining effect of Fig. 6 biocontrol microorganisms 42561 fermented liquids to 2 kind of plant pathogenic fungi spore germinations.A: verticillium dahliae---42561 fermentation liquor treatment groups; B: verticillium dahliae---liquid nutrient medium control group); C: Fusarium oxysporum---42561 fermentation liquor treatment groups; D: Fusarium oxysporum---liquid nutrient medium control group.
The results from pot experiment test of Fig. 7 biocontrol microorganisms 42561 fermented liquids to cotton wilt.A:42561 fermentation liquor treatment group; B: derosal treatment group C: clear water control group.
The results from pot experiment test of Fig. 8 biocontrol microorganisms 42561 fermented liquids to cotton verticillium wilt.A:42561 fermentation liquor treatment group; B: derosal treatment group C: clear water control group.
Fig. 9 field efficiency test cotton cane sectional view.A: morbidity field 42561 fermentation liquor treatment groups; B: morbidity field blank group; C: morbidity field derosal treatment group; D: healthy cotton plant.
Embodiment
Detailed description below by embodiment is further illustrated the present invention, but is not limitation of the present invention, only does example explanation.
the separation of embodiment mono-biocontrol microorganisms
Adopt heptangle well salt lake, Xinjiang (height above sea level 1584.49m, 43.48 ° of north latitude, 91.48 ° of east longitudes) pedotheque of 0～30cm, with 8 kinds of conventional mediums (GW1 substratum, GPY substratum, chitosan asparagin culture, glycerine arginine substratum, the glycerine minimal medium containing 5%, 10%, 15%, 20% and 25% sodium-chlor, N.F,USP MANNITOL minimal medium, ISP-4 substratum and PY substratum), adopt dilution-plate method (to take the aseptic aqueous solution that 1 g pedotheque is put into 9 mL, fully vibration, makes 10
-1suspension liquid.Draw 0.1 mL suspension liquid, evenly coat on the isolation medium flat board preparing in advance, be inverted for 37 ℃ and cultivate.Culture dish adopts preservative film sealing, reduces substratum moisture loss, the Extending culture time.), isolate altogether 7 genus totally 35 strain actinomycetes, be distributed in streptomyces and Nocardiopsis more, the glycerine arginine substratum of actinomycetes 42561 separated self-contained 5% sodium-chlor, its culture medium prescription is: glycerine 5 g/L, arginine 1 g/L, dipotassium hydrogen phosphate 1 g/L, magnesium sulfate 0.5 g/L, calcium carbonate 0.5 g/L, sodium-chlor 50 g/L, agar powder 17 g/L, pH 7.5.By stand facing each other method primary dcreening operation and cup-plate method of flat board, sieve again, final Isolation and screening has gone out the best biocontrol microorganisms 42561 of fungistatic effect.
the evaluation of embodiment bis-biocontrol microorganisms
For further determining that the taxonomy of biocontrol microorganisms 42561, the present invention adopt traditional classification method and molecular classification method to carry out classification to this bacterial strain and identify.
1 morphologic observation substratum
Inorganic salt Starch Agar (ISP-4) substratum: starch 10.00 g, (NH
4)
2sO
44.00 g, MgSO
42.00 g, K
2hPO
42.00 g, CaCO
31.00 g, Nacl 40.00 g, agar 18.00 g, distilled water are dissolved to 1000ml, pH 7.2-7.4,121 ℃ of high pressure moist heat sterilization 30min.
2 primers
Adopt actinomycetes universal primer You Saiyin biotechnology (Shanghai) Co., Ltd. synthetic.Its sequence is respectively: 27F(5 '-AGAGTTTGATCCTGGCTC-3 ') and 1492R(5 '-CGGCTACCTTGTTACGACTT-3 ')
3 test methods
3.1 morphological observation
Simple microscope is observed: adopt inserted sheet method, cultivate biocontrol microorganisms 42561 for 37 ℃.With observation by light microscope, record mycelia form, aerial hyphae and substrate mycelium growing state, whether mycelium produces arrangement mode, the shape of fibrillae of spores and fibrillae of spores; Spore shape and size; The having or not of spore, shape, size and generation type etc.
The molecular biology identification of 3.2 biocontrol microorganisms 42561
(A) extraction of biocontrol microorganisms 42561 genomic dnas
42561 thalline on collection culture plate are put into the aseptic centrifuge tube of 1.5 mL, add 480 μ L's
1 * TE damping fluid.Add 20 μ L N,O-Diacetylmuramidases (50 mg/mL), put into 37 ℃ of shaking tables, 200rpm shaken overnight.Every pipe adds the SDS of 50 μ L 20%, adds the Proteinase K of 5 μ L 20 mg/mL, puts into 60 ℃ of shaking tables, 200 r/min, 1 h that vibrates.Phenol-chloroform-the primary isoamyl alcohol (25:24:1) that adds 550 μ L, centrifugal 10 min of 12000rpm, get supernatant and move in another centrifuge tube, and extracting is 2-3 time repeatedly.Get supernatant, add isopyknic dehydrated alcohol, add the sodium acetate (3mol/L) of 0.1 times of volume, put into 4 ℃ of refrigerators and precipitate the about 0.5h of DNA.The centrifugal 10min of 12000rpm, abandons supernatant.With 70% ethanol of 200 μ L, clean centrifugal product 2 times, centrifugal 5 min of 12000rpm, abandon supernatant, by ethanol volatilization completely.With the aseptic ultrapure water of 50 μ L, fully dissolve the DNA of bottom, 1% agarose gel electrophoresis detects DNA extraction quality, the DNA of extraction is put into-20 ℃ of refrigerators and save backup.
(B) amplification of biocontrol microorganisms 42561 16S rDNA genes
With actinomycetes 16S rDNA gene universal primer 27F(5 '-AGAGTTTGATCCTGGCTC-3 ') and 1492R(5 '-CGGCTACCTTGTTACGACTT-3 ') 16S rDNA gene fragment in amplification actinomycetes genomic dna.
The PCR reaction system of 25 μ L is: ddH
2o 20.4 μ L, 10 * Buffer(damping fluid is containing Mg
2+) 2.5 μ L, dNTPs 0.5 μ L, primer 2 7F(10 μ mol/L) 0.5 μ L, primer 1492R(10 μ mol/L) 0.5 μ L, Taq archaeal dna polymerase 0.1 μ L, template DNA 0.5 μ L.
PCR reaction conditions is: 94 ℃ of 4 min of denaturation; 94 ℃ of 1min of sex change, 56 ℃ of 1 min that anneal, extends 72 ℃ of 2 min, 30 circulations; 72 ℃ of 8min of total elongation.Having reacted rear use 1% agarose gel electrophoresis detects.Qualified PCR product carries out sequencing.
(C) compare of analysis of sequencing result
DNAMAN5.2 software splicing for sequencing result, sequence is compared by the bacterial strain sequence of the effective publication in BLAST and GenBank database, download the 16S rDNA gene order of the bacterial strain of effective publication that similarity is higher, with MEGA 4.1 softwares, sequence is entered to build the row structure of phylogenetic tree, determine actinomycetic taxonomy.
4 test-results
The morphological observation result of 4.1 biocontrol microorganisms 42561
The molecular biology identification result of 4.2 biocontrol microorganisms 42561
4.2.1 the 16S rDNA sequencing result of biocontrol microorganisms 42561
DNAMAN5.2 software splicing for sequencing result, determines that this fragment is by 1348 based compositions, obtains the 16S rDNA sequence of biocontrol microorganisms 42561 as shown in SEQ ID NO.1
.
4.2.2 homology evolutionary tree builds
By BLAST, compare, in GenBank database, download the sequence nearer with actinomycetes 16S rDNA gene order similarity to be measured, with MEGA5.1 software, actinomycetes 16S rDNA gene order is carried out to multiple sequence comparison, phylogenetic tree construction.Biocontrol microorganisms 42561 evolutionary trees see Fig. 1, biocontrol microorganisms 42561 and Lou Che Shi streptomycete (
streptomyces rochei) 16S rDNA gene order (similarity is greater than 99.9%) gather in same phyletic evolution branch, therefore by this biocontrol microorganisms be accredited as Lou Che Shi streptomycete (
streptomyces rochei) 42561.
embodiment tri-biocontrol microorganisms 42561 bacteriostatic activities
1 for examination pathogenic bacteria
Verticillium Dahliae Infecting Cotton: verticillium dahliae (
verticillium dahliae), the former bacterium of cotton wilt: Fusarium oxysporum cotton specialized form (
fusarium oxysporumsp.
vasinfectum) by Production and Construction Corps of Xinjiang's Tarim Basin living resources conservation, utilize key lab to provide.
2 test plants and medicament
Test plant: cotton (new land in 36)
Treatment agent: biocontrol microorganisms 42561 fermented liquids
Contrast medicament: carbendazol wettable powder (Shandong Eastcom biological pesticide company limited)
3 substratum
Potato dextrose agar (PDA): get 200g peeling potato, boiling water boiling 30min after being cut into small pieces, four layers of filtered through gauze, filtrate adds glucose 20g, agar 18g, distilled water is settled to 1000mL, pH nature.After Erlenmeyer flask packing, 121 ℃ of high pressure moist heat sterilization 30min.
Activation medium: inorganic salt Starch Agar (ISP-4) substratum: starch 10.00 g, (NH
4)
2sO
44.00 g, MgSO
42.00 g, K
2hPO
42.00 g, CaCO
31.00 g, Nacl 40.00 g, agar 18.00 g, distilled water are dissolved to 1000ml, pH 7.2-7.4,121 ℃ of high pressure moist heat sterilization 30min.
Seed culture medium: inorganic salt Starch Agar (ISP-4) substratum
Fermention medium: Semen Maydis powder 15g, peptone 5g, glucose 5g, CaCO
31g, NaCl 40g, tap water are dissolved to 1000 ml, pH 7.2-7.4,121 ℃ of high pressure moist heat sterilization 30min.
4 experimental techniques
4.1 strain fermentations are cultivated
4.2 fermentation liquor pretreatment
Fermented liquid, with the solids such as mycelium in the centrifugal 10min removal of the speed fermented liquid of 8000rmp, is got to supernatant liquor as potted plant and treatment agent field experiment; Millipore filtration (0.45 μ m) removes by filter after the fine impurities suspending in supernatant liquor as cup-plate method, suppresses mycelial growth method and suppress the treatment agent of spore germination method.
4.3 Antibacterial Activity methods
(1) isolated activity is measured
Isolated activity is measured and is adopted dull and stereotyped face-off method, cup-plate method, inhibition mycelial growth rate method, inhibition spore germination method to carry out, and concrete grammar is as follows:
(A) dull and stereotyped face-off method: first by biocontrol microorganisms 42561 with carry out respectively flat board for examination pathogenic bacteria and activate, cultivated the hypha,hyphae of 5 days to being equipped with accordingly in the bottle of 5mL sterilized water by the inoculating needle scraping after calcination sterilizing respectively, made 10
6～10
7the fungi bacteria suspension of individual spore/mL, draws the bacteria suspension of 60 μ L to PDA flat board, with sterilized spreading rod, by after bacteria suspension coating evenly, on Bechtop, dries up a little, makes fungi indication flat board.With aseptic punch tool, from the actinomycetes flat board of cultivating 5 days, laying bacterium cake (Φ=7mm) is put on the target bacterium flat board having prepared, with ISP-4 culture block, compare, then being placed in 28 ℃ of constant incubators cultivates after 4～5d, utilize right-angled intersection method to measure antibacterial circle diameter (antibacterial circle diameter=antibacterial circle diameter observed value-7mm), every processing repeats 3 times.
(B) cup-plate method:
By sterilized water wash-out from culture plate for cultured pathogenic fungi spore, make 10
6～10
7the spore suspension of individual spore/mL, draws the spore suspension of 60 μ L to PDA flat board, with sterilized spreading rod by bacteria suspension coating evenly, on Bechtop, dries up a little.Equidistant placement Oxford cup on the flat board that carries disease germs, adds 200 μ L tunnings in each Oxford cup, take fermention medium as contrast, cultivates 4～5d for 28 ℃, and right-angled intersection method is measured antibacterial circle diameter, and every processing repeats 3 times.
(C) suppress mycelial growth rate method:
By pretreated fermented liquid with melt after be cooled to 45 ℃ of left and right PDA substratum pour in sterile petri dish after mixing with the ratio of 1:9, make flat board, then pathogenic bacteria bacterium cake (Φ=7mm) be placed in to dull and stereotyped central authorities, repeat 3 times, in 28 ℃ of constant temperature culture.When the mycelia of pathogenic fungi is about to be paved with full ware in contrasting ware, by right-angled intersection method, measure for examination fungal colony growth diameter, calculate as follows mycelial growth inhibition rate:
(D) suppress spore germination method:
Adopt depression slide method to measure the restraining effect of strain fermentation product to the spore germination of test plant pathogenic fungi.By sterilized water wash-out from culture plate for cultured pathogenic fungi spore, by multilayer filtered through gauze, to remove mycelia, make spore suspension.Spore concentration is with under 10 * 10 low power lens, and each visual field has 20～30 spores to be advisable.Fermented liquid is mixed with the ratio of 1:1 with spore suspension, get one after another drop of being added on depression slide, every processing repeats for 3 times.At 25 ℃, moisturizing is cultivated, after the germination rate of control group reaches 90%, the spore germination rate of check processing, every processing repeats 3 above visuals field of random observation, the sum of investigation spore is no less than 200, and the short radius that spore germ tube length is greater than spore is considered as sprouting.According to the following public inhibition of germination that calculates.
(2) greenhouse pot culture determination of activity
The control test of cotton spoting verticillium wilt: adopt root-pouring method to carry out, Growth of Potted Cotton, after cotton emerges, every basin stays 3 of seedlings, starts to inoculate pathogenic bacteria when cotton grows 3～4 true leaves.During inoculation, it is 1 * 10 that every basin pours into concentration
7the suspension 30mL of individual spore/mL.25 ℃, under the above condition of relative humidity 90%, cultivate.After stable disease, the every basin of fermented liquid is carried out to root irrigation with 30mL to the cotton seedling of having fallen ill.With derosal 10mg ﹒ mL
-1(every basin 30mL) be medicament in contrast, to use clear water, makes blank, every processing 10 basins, and experiment repeats 3 times.Every 7d, be administered once, administration is 3 times altogether, and the 15d after administration and 30d investigate respectively cotton seedling growth and incidence the last time.By following criterion calculation disease index and prevention effect.
Blight stage division:
0 grade: healthy tree;
1 grade: diseased plant blade has the classical symptom of performance below 10%;
3 grades: diseased plant blade has 11%-25% performance classical symptom;
5 grades: diseased plant blade has 26%-50% to show symptom, plant type is downgraded;
7 grades: diseased plant blade has 51%-90% to show symptom, and plant type is obviously downgraded;
9 grades: diseased plant blade is whole aobvious symptom almost, dried-up coming off even, branch stem is withered, and there is acute wilting death in whole strain sometimes.
Verticillium stage division:
0 grade: healthy tree;
1 grade: diseased plant blade has aobvious symptom below 10%, the faint yellow irregular scab of blade performance;
3 grades: diseased plant blade has 11%25% aobvious symptom, produces faint yellow irregular scab between blade master pulse;
5 grades: diseased plant blade has 26%-50% to show symptom, the most of yellowing of color and the tawny of scab, blade edge has volume withered;
7 grades: diseased plant blade has more than 51% aobvious symptom, the most digital display tawny of scab, blade edge volume is withered, has minority blade to wither and fall;
9 grades: diseased plant leaf abscission becomes polished rod and plant death, sometimes there is the dead symptom of acute wilting.
Drug effect method of calculation
(3) field control effectiveness test
Agriculture one teacher of the field control effectiveness test Shi Production and Construction Corps of Xinjiang repeatedly kicks into row for 12 28, selects to occur verticillium and the heavier plot of blight throughout the year.Test minute 3 processing, are specifically treated to: (1) biocontrol microorganisms 42561 fermented liquid groups, (2) derosal group, (3) clear water control group.Repeat 3 times, community area is 30m
2, random district group is arranged, and control time is cotton buds and bolls phase and the term of opening bolls.Sowing time is on April 28th, 2012, and cotton variety is in new land 36, and soil is silty loam, middle fertility, and irrigation conditions is better.When cotton grows to 5～6 true leaves, respectively cotton seedling is carried out to pathogenic fungi root irrigation once.When the state of an illness starts to manifest, every 7d dispenser once, dispenser is three times altogether, and 15d and 30d after last dispenser investigate respectively growth rate of cotton plant and incidence.Application method and investigation and method of calculation are with greenhouse pot experiment.
5 results and analysis
The indoor Antifungal Activity in Vitro measurement result of 5.1 biocontrol microorganisms 42561
5.1.1 the antagonistic activity measurement result of biocontrol microorganisms 42561
Adopt dull and stereotyped face-off method, detect biocontrol microorganisms 42561 to verticillium dahliae and Fusarium oxysporum antagonistic activity, as shown in Figure 2, wherein the antibacterial circle diameter of 42561 pairs of verticillium dahliaes of biocontrol microorganisms and Fusarium oxysporum reaches respectively 34.75 and 24.20mm.
5.1.2 biocontrol microorganisms 42561 fermented liquid antagonistic activity measurement results
Cup-plate method is measured the restraining effect of biocontrol microorganisms 42561 fermented liquids to verticillium dahliae and Fusarium oxysporum, as shown in Figure 3.Fermented liquid has strong restraining effect to verticillium dahliae and Fusarium oxysporum, and antibacterial circle diameter is respectively 32.67 and 23. 96mm.
5.1.3 biocontrol microorganisms 42561 fermented liquids are to the restraining effect for examination pathogenic fungi mycelial growth
From Fig. 4, Fig. 5,42561 bacterial strain fermentation liquors all have good restraining effect to verticillium dahliae and Fusarium oxysporum mycelial growth, and its inhibiting rate is all greater than 80%.Wherein the inhibiting rate of verticillium dahliae and Fusarium oxysporum is respectively to 83. 41% and 89.75%.
5.1.4 the restraining effect of biocontrol microorganisms 42561 fermented liquids to 2 kind of plant pathogenic fungi spore germinations
As seen from Figure 6, to test plant pathogenic fungi, spore germination all has good restraining effect to biocontrol microorganisms 42561 fermented liquids, and verticillium dahliae and Fusarium oxysporum inhibition of germination are respectively to 68.75% and 73. 56%.
5.2.2 pot-culture method is measured the result of biocontrol microorganisms 42561 fermented liquid bacteriostatic activities
5.2.2.1 the prevention effect of biocontrol microorganisms 42561 fermented liquids to cotton spoting verticillium wilt
By Fig. 7, Fig. 8 and table 1, can be found out: biocontrol microorganisms 42561 fermented liquids have certain prevention effect to cotton wilt and cotton verticillium wilt.Prevention effect is respectively 36.73% and 44.82%, far above 10.00% and 9.51% of contrast medicament derosal.
The potted plant prevention effect of table 1 biocontrol microorganisms 42561 fermented liquids to cotton spoting verticillium wilt
5.2.2.3 field control effectiveness test result
On indoor in vitro and potted plant basis, biocontrol microorganisms 42561 is carried out to the test of field control cotton spoting verticillium wilt.As can be seen from Table 1, biocontrol microorganisms 42561 fermented liquids have certain effect to the field control of cotton spoting verticillium wilt on the one hand, the 15th and 30d after dispenser for the third time, and its prevention effect reaches respectively 33.06% and 41.65%; Another aspect is that effect is slow, although 42561 fermented liquids can not fundamentally be blocked the propagation of pathogenic fungi, it has effectively delayed the further deterioration of the state of an illness.
The field control effect of table 2 biocontrol microorganisms 42561 fermented liquids to cotton spoting verticillium wilt
Claims (3)
- A Lou Che Shi streptomycete ( streptomyces rochei), on September 11st, 2012, being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No.6556.
- 2. the purposes as biocontrol microorganisms according to Lou Che Shi streptomycete claimed in claim 1, is characterized in that it is for preventing and treating cotton wilt or cotton verticillium wilt.
- 3. a method of utilizing Lou Che Shi streptomycete described in claim 1 to prepare biocontrol fungicide, it is characterized in that described Lou Che Shi streptomycete is fermented on fermention medium obtains fermented liquid as biocontrol fungicide, wherein fermention medium is: Semen Maydis powder 15g/L, peptone 5g/L, glucose 5g/L, CaCO 31g/L, NaCl 40 g/L, adjusting pH is 7.2-7.4; Inoculum size is: the inoculum size by 6% is inoculated in fermention medium; Culture condition is: under 37 ℃, 180rpm condition, shaking table is cultivated 4 days.
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| CN103789222A (en) * | 2013-07-11 | 2014-05-14 | 青岛农业大学 | Biocontrol bacterium streptomyces rochei A-1 for controlling apple fruit ring spot and biocontrol agent thereof |
| CN103966083B (en) * | 2014-05-26 | 2015-10-28 | 丁海铭 | Cup-plate method microorganism detection system calibration calibration standard model |
| CN105961439B (en) * | 2016-05-26 | 2019-06-14 | 陕西博秦生物工程有限公司 | Crop seed soaking agent based on actinomyces and preparation method thereof and seed-soaking method |
| CN105994379B (en) * | 2016-05-26 | 2019-06-25 | 陕西博秦生物工程有限公司 | Coating agent for seed based on actinomyces and preparation method thereof and coating method |
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| CN109874808B (en) * | 2019-01-25 | 2021-02-02 | 青岛农业大学 | Application of arbuscular mycorrhizal fungi and/or plant symbiotic actinomycetes in preparation of biocontrol preparation for hot pepper and solanaceae vegetables |
| CN114456973B (en) * | 2022-01-11 | 2022-08-05 | 湖北省生物农药工程研究中心 | Streptomyces rochei in tobacco and application thereof in prevention and control of tobacco diseases |
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