CN103695350B - One strain streptomyces griseus CCCQ171 and application thereof - Google Patents
One strain streptomyces griseus CCCQ171 and application thereof Download PDFInfo
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Abstract
Do you the invention discloses a strain streptomyces griseus (Streptomyces? griseochromogenes) CCCQ171 and application thereof, does is described its deposit number of streptomyces griseus CCCQ171: CGMCC? No.6227, according to place of production called after Jiangling bacterium.The invention also discloses the application of described streptomyces griseus CCCQ171 in agricultural microbial agent and agricultural microbial agent thereof, containing streptomyces griseus CCCQ171 in described agricultural microbial agent, described agricultural microbial agent is for preventing and treating tobacco diseases or evoking tobacco produces resistance to tobacco diseases.Described streptomyces griseus CCCQ171 is separated to obtain from vega soil, with natural, ecological, there is natural harmonious blending, compare with chemical pesticide that soil ecology has no side effect, noresidue, therefore, in the biological control practice of disease, there is potential business development and using value.
Description
Technical field
The present invention relates to microorganism and microbe application tobacco diseases prevention and control field, be specifically related to that there is bacterial strain one strain streptomyces griseus (Streptomycesgriseochromogenes) and the application thereof that tobacco diseases prevents and treats function.
Background technology
Nature, in long-term evolution process, exists between plant and ambient microorganisms and generally makes phenomenon mutually.There is the plant-microorganism symbiosis that two classes are useful: a class is nutritious function, as nitrogen-fixing bacteria or mycorrhizal fungi (mycorrhizalfungi); One class has defense function, as suppressed endophyte and the epiphyte of pathogenic bacteria.The symbiotic microorganism that this two class is useful can improve the adaptive faculty of plant.Agrochemicals (chemical fertilizer and agricultural chemicals) and the planting type of " intensive cultivation " play an important role in agriculture production.Under this planting type, in raise crop, this equilibrium system of plant-microorganism is broken, and lacks useful plant-microorganism symbiosis, makes crop production height depend on chemical fertilizer and agricultural chemicals.And the frequent use of fertilizer and pesticide causes the pollution of global environment, and pollutent enters human body eventually through food chain.Therefore exogenous application beneficial microorganism, rebuilds ecological friendly, useful plant-microorganism symbiote and receives much attention, and show the potentiality that this microbial preparation of application carries out biological control.
The integrated control of tobacco diseases is the important step in leaf tobacco production always, and chemical prevention for a long time occupies very important status in tobacco diseases control strategy.On tobacco leaf production, a large amount of and long-term use continuously of chemical pesticide and the unreasonable of operation technique, black root of tobacco bacterium is developed immunity to drugs, and the resistance of the former bacterial strain of root Black Rotten of different sources also also exists difference.The appearance of endurance strain not only increases the cost of controlling disease, and increases the weight of to use agricultural chemicals to the pollution of environment, and the destruction of the ecosystem, also directly affects quality of tobacco simultaneously.Overcome Plant diseases resistance, reduce chemical bactericide to the pollution of environment, ecological destruction and agricultural byproducts in chemical pesticide residual in, Biocontrol microorganism has unique advantage, the biological pesticide utilizing actinomycetic secondary metabolite to prepare, there is pollution-free, noresidue, not easily harmful organism developed immunity to drugs, high with Environmental compatibility, the main body of nuisanceless green agricultural chemicals and the developing direction of pesticide in future have been become to features such as person poultry safeties.Therefore, the biological control research of Efforts To Develop Biocontrol microorganism is the important channel of agricultural sustainable development, has important production practical significance.
Summary of the invention
In order to solve the problems of the technologies described above, the invention provides strain streptomyces griseus (Streptomycesgriseochromogenes) CCCQ171 and an application thereof.
One strain streptomyces griseus (Streptomycesgriseochromogenes) CCCQ171, its deposit number is: CGMCCNo.6227, depositary institution is: China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, and preservation day is: on June 15th, 2012.
The 16SrDNA sequence of described streptomyces griseus CCCQ171 is as shown in SEQIDNo:1.
The application of described streptomyces griseus CCCQ171 in agricultural microbial agent.
The application of streptomyces griseus CCCQ171 described above in agricultural microbial agent, described agricultural microbial agent is for preventing and treating tobacco diseases or evoking tobacco produces resistance to tobacco diseases.
The application of streptomyces griseus CCCQ171 described above in agricultural microbial agent, described tobacco diseases is black root of tobacco, black shank or Alternaria alternate.
A kind of agricultural microbial agent, containing streptomyces griseus CCCQ171.
Agricultural microbial agent as above, described agricultural microbial agent is for preventing and treating tobacco diseases or evoking tobacco produces resistance to tobacco diseases.
Agricultural microbial agent as above, described tobacco diseases is black root of tobacco, black shank or Alternaria alternate.
The streptomyces griseus CCCQ171 obtained is separated from cigarette strain rhizosphere soil, be Late Cambrian and a new strains of qualification, itself has very strong restraining effect to various plants pathogenic bacterias such as black root of tobacco germs, and its fermentation broth on tobacco root Black Rotten has good control effect.According to bacterial strain bacterium colony cultural colony, morphological specificity and Physiological-biochemical Characters and 16SrDNA sequence alignment analysis, determine that CCCQ171 bacterial strain is streptomyces griseus, according to place of production called after Jiangling bacterium.Because this bacterial strain is separated to obtain from vega soil, with natural, ecological, there is natural harmonious blending, compare with chemical pesticide that soil ecology has no side effect, noresidue, therefore, in the biological control practice of disease, there is potential business development and using value.
Accompanying drawing explanation
Fig. 1 be in the embodiment of the present invention 2 CCCQ171 bacterial strain to the dull and stereotyped antagonistic effect figure of black root of tobacco germ.
Fig. 2 be in the embodiment of the present invention 3 CCCQ171 bacterial strain to the greenhouse Controlling effect figure of black root of tobacco.
Fig. 3 be in the embodiment of the present invention 3 CCCQ171 bacterial strain to the direct inhibition figure of black root of tobacco germ mycelia.
Embodiment
For a better understanding of the present invention, below in conjunction with drawings and Examples, the present invention is described further.
If no special instructions, the routine operation that the operation adopted in following examples is well known to the skilled person, reagent, bacterial classification etc. are commercially available prod.
The selection systems of embodiment 1CCCQ171 bacterial strain
Actinomycetes 17 strain that the multiple bacterial strains obtained obtain bacteriostatic activity after primary dcreening operation and multiple sieve is separated from the vega soil of Chongqing.Primary dcreening operation adopts dull and stereotyped face-off method, and multiple sieve adopts chloroform fumigation and steaming method.Eventually pass the sick efficiency test of control determine to black root of tobacco have better preventive and therapeutic effect for streptomyces griseus of the present invention (Streptomycesgriseochromogenes) CCCQ171 bacterial strain.
1.1 colony characteristicses are observed
Solid plate substratum is made with yeastex malt-agar culture (ISP-2), oatmeal nutrient agar (ISP-3), inorganic salt starch agar medium (ISP-4), glycerine asparagine nutrient agar (ISP-5) and potato leaching juice substratum, Cha Shi (Czapck) substratum and improvement Gao Shi No. 1 substratum.By the streak inoculation of CCCQ171 bacterial strain on above 7 kinds of culture medium flat plates, 28 DEG C of constant temperature culture 7-14d, after its mycelial growth is stable, observe single colonial morphology, aerial hyphae, substrate mycelium, fibrillae of spores color respectively with anatomical lens.
In different culture media, CCCQ171 colonial morphology is respectively:
ISP-2 substratum: bacterium colony is circular, centre has fold protruding, and colony edge has haloing, aerial hyphae white, down shape, fibrillae of spores canescence, substrate mycelium Vandyke brown.
ISP-3 substratum: bacterium colony is circular, and edge mycelia be down shape, the cotton-shaped or powdery of aerial hyphae, becomes grey, fibrillae of spores grey, the deep red brown of substrate mycelium, chromogenesis after first white.
ISP-4 substratum: colony edge is irregular, slightly circular, aerial hyphae white long wool hair shape, fibrillae of spores white, substrate mycelium is faint yellow, does not produce pigment.
ISP-5 substratum: bacterium colony is circular, and edge mycelia is radially, sparse, aerial hyphae grey long wool hair shape, fibrillae of spores beige, substrate mycelium from faint yellow to isabelline, chromogenesis.
Potato leaching juice substratum: bacterium colony intermediate projections, there is haloing at edge, and aerial hyphae white long wool hair shape, fibrillae of spores grey, substrate mycelium, from dark reddish purple look to grey, slightly produces filbert pigment.
Czapck substratum: radially, aerial hyphae white long wool hair shape, edge is sparse, and mesospore silk is cotton-shaped, grey, and substrate mycelium is colourless, does not produce pigment for colony edge.
Improvement Gao Shi No. 1 substratum: bacterium colony is comparatively large, rounded, intermediate projections, aerial hyphae white, fibrillae of spores canescence, substrate mycelium is dark yellow to Vandyke brown, chromogenesis.
1.2 mycelium and spore observation of characteristics
Inserted sheet method is adopted to observe hypha form and the spore feature of CCCQ171 bacterial strain.Get CCCQ171 bacterial strain bacteria suspension to coat on improvement Gao Shi No. 1 substratum, with the sterilized cover glass of tweezers gripping, enter in substratum with about 45 ° of oblique cuttings, every dull and stereotyped slotting 4-5 sheet.Flat-plate inverted after inserted sheet is placed in 28 DEG C of cultivations, after 3-5 days, carefully takes out cover glass with tweezers, then will have a cover of bacterium on slide glass, directly with the aerial hyphae of microscopic examination CCCQ171 bacterial strain, spore chain and spore raw situation.The mycelium of CCCQ171 bacterial strain and spore feature: spore chain length and straight, cluster, has minority spore chain spirrillum, and spore is subsphaeroidal.Form, tentatively determine that the classification of CCCQ171 bacterial strain belongs to actinomycetes thus, Streptomycetaceae streptomyces.
1.3 molecular biology identification
By extracting CCCQ171 strain gene group DNA and 16SrDNAPCR amplification, the 16SrDNA fragment length that CCCQ171 bacterial strain amplifies is 1487bp, and sequence is as shown in SEQIDNo.1, close in heredity with streptomyces (Streptomycessp.).
Streptomyces griseus CCCQ171 is Late Cambrian and a new strains of qualification, binding molecule biological assay, cultural characters, morphological specificity, gramstaining and physiological and biochemical test result, CCCQ171 bacterial strain is defined as streptomyces griseus (Streptomycesgriseochromogenes), according to place of production called after Jiangling bacterium.The live body pure culture of this bacterial classification is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 15th, 2012, deposit number: CGMCCNo.6227, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica.
Embodiment 2CCCQ171 bacterial strain antimicrobial spectrum is tested
Adopt dull and stereotyped face-off method, get different germ bacterial strain to activate, with sterilizing punch tool (diameter 6mm) colony edge region punching make bacterium sheet, be forwarded to the dull and stereotyped central authorities of PDA, cultivate 48h for 28 DEG C, after it is grown surely, at both sides, distance center 30mm place access CCCQ171 bacterial strain, often process repetition 3 times.28 DEG C of constant temperature culture 7d, measure antibacterial bandwidth, measure the colony diameter of germ, calculate bacteriostasis rate.The antagonistic effect of CCCQ171 bacterial strain to tobacco germ is as shown in table 1.
The screening of table 1CCCQ171 bacterial strain antimicrobial spectrum
Note: in table 1, "-" represents do not have inhibition zone.
Measure the antagonistic activity of CCCQ171 bacterial strain to 9 kinds of germs to screen, measure inhibition zone respectively, calculate bacteriostasis rate.Result shows that CCCQ171 bacterial strain has obvious fungistatic effect to Tobacco Root maize ear rot germ, red-star like disease germ, balck shank germ, bacteriostasis rate reaches more than 50%, wherein CCCQ171 bacterial strain can reach 80.01% to tobacco pine root fungus bacteriostasis rate, and antibacterial bandwidth reaches 17.09mm(as shown in Figure 1).Microscopic examination finds, CCCQ171 is that mycelia branch increases to germ mycelia effect main manifestations, and front end is elongated, and vesicle is not obvious.
The application of embodiment 3CCCQ171 bacterial strain
3.1CCCQ171 bacterial strain fermentation liquor is to the restraining effect of spore germination
After black root of tobacco bacterium cultivates 10d on potato dextrose agar, scraping spore, adds sterilized water adjustment spore concentration and is about 1 × 10
5individual/mL, for subsequent use.
By following formulated improvement Gause I substratum, then autoclaving is for subsequent use: KNO
31.0g; K
2hPO
40.5g; MgSO
4gS
2o0.5g; NaCl0.5g; FeSO
4eS
2o0.01g; Zulkovsky starch 20.0g; Agar 17.0g; PH:7.2 ~ 7.4.By CCCQ171 bacterial strain 28 DEG C of constant temperature culture 7 ~ 10d on improvement Gao Shi No. 1 slant medium, treat that it produces enough spores, sterilized water is poured into along inclined-plane, after inclination, did not just have inclined-plane, make bacteria suspension with asepsis ring scraping surface thalline.In the triangular flask of 250mL, load 100mL improve Gao Shi No. 1 nutrient solution, after sterilizing cooling, access above-mentioned bacteria suspension, inoculum size is 100mL/L, and shaking speed is 180rpm, collects fermented liquid after 28 DEG C of shaking culture 8d, fermented liquid is in 4 DEG C, the centrifugal 20min of 15000rpm, gets supernatant liquor, and obtaining concentration is 4 × 10
8the CCCQ171 bacterial strain fermentation liquor stoste of individual/mL.
With sterilized water by CCCQ171 bacterial strain fermentation liquor stoste (4 × 10
8individual/mL) be diluted to 25%, 50%, the concentration of 75%, with liquid-transfering gun from lower concentration to high density, the CCCQ171 bacterial strain fermentation liquor drawing 0.5mL successively adds in small test tube, then draws 1 × 10
5individual/mL black root of tobacco bacterium spore suspension 0.5mL, fully mixes, if sterilized water is contrast.Drawing above-mentioned mixed solution with micro sample adding appliance drips on depression slide, is then put in the culture dish with shallow seated groundwater, adds a cover.48h is cultivated in 15 ~ 20 DEG C of moisturizings.Often process and repeat for 3 times.Examining under a microscope statistics, being greater than spore short radius person for sprouting with spore germ tube length.
Be calculated as follows suppression spore germination rate:
Relative inhibition of germination (%)=[(contrast spore germination rate-process spore germination rate)/contrast spore germination rate] × 100
……………………………………（1）
Table 2CCCQ171 bacterial strain fermentation liquor is to the restraining effect of Tobacco Root maize ear rot spore germination
Test-results is as shown in table 2, and CCCQ171 bacterial strain fermentation liquor different concns treatment group all has significant restraining effect to the sprouting of Tobacco Root maize ear rot Spores; With comparing, after 75%, 100% concentration C CCQ171 bacterial strain fermentation liquor process, Tobacco Root maize ear rot Spores is not all sprouted, and Germination suppression rate reaches 100%.
3.2CCCQ171 bacterial strain is without the restraining effect of fermented liquid to mycelial growth
Adopt containing bacterium flat band method, with sterilized water, CCCQ171 bacterial strain fermentation liquor is diluted to the concentration of 50%, after the CCCQ171 bacterial strain fermentation liquor drawing 1mL mixes with 9mLPDA substratum, pouring culture dish into, is contrast with sterilized water.Be that the punch tool of 6mm cuts pure culture biscuits involvng inoculation in the above-mentioned plate center containing CCCQ171 fermented liquid cultivating the black root of tobacco colony edge of 7d with diameter, mycelia faces down, cultivate 7 ~ 8d for 28 DEG C, measure bacterium colony expansion diameter by right-angled intersection method, be calculated as follows mycelial growth inhibition rate.
Mycelial growth inhibition rate (%)=[(contrast bacterium colony expansion diameter-process bacterium colony expansion diameter)/contrast bacterium colony expansion diameter] × 100
……………………………………（2）
As shown in Figure 3, black root of tobacco bacterium is containing CCCQ171 bacterial strain without the substratum of fermented liquid showing mycelia front end growth retardation, bacterium colony expansion slowly, stoste inhibition reaches 57.22%, shows that the secondary metabolite in CCCQ171 bacterial strain fermentation liquor is inhibited to germ mycelial growth.
3.3CCCQ171 the greenhouse control disease test of bacterial strain
Choose 4 ~ 6 leaf cigarette seedlings grown fine, every strain is filled with and is executed 10mLCCCQ171 bacterial strain fermentation liquor stoste (1 × 10
8individual spore/mL), co-processing 30 strain cigarette seedling, with fill with execute equivalent sterilized water 30 strain cigarette seedlings in contrast.Bacterium cake method is adopted to inoculate black root of tobacco bacterium after 72h.Observe incidence, after 8d, cigarette seedling is extracted, by following standard survey diseased plant number and severity etc. after rinsing well, be calculated as follows sickness rate, disease index and relative control effect, carry out statistical study with SPSS software.
Sickness rate (%)=disease plant/total strain number × 100 ... (3)
Disease index (DI)=[∑ (disease index × with sick level strain number)]/[the highest sick progression × total strain number] × 100 ... (4)
Relative control effect (%)=[(CK disease index-process disease index)/CK disease index] × 100 ... (5)
Black root of tobacco severity Scaling standard (in units of whole strain) is as follows:
Table 3CCCQ171 bacterial strain is to the test of Tobacco Root maize ear rot greenhouse control disease
As shown in Fig. 2 and table 3, CCCQ171 bacterial strain greenhouse pot culture prevention effect is better, and after bacterium liquid process tobacco seedling, experimental group root system color is normal, the blackening of control group root system.Sickness rate and the disease index of experimental group plant obviously reduce, and prevention effect reaches 70.53%, compared with the control, α=0.05 level reach remarkable, α=0.0l level reach extremely remarkable.
Streptomyces griseus CCCQ171 itself has very strong restraining effect to various plants pathogenic bacterias such as black root of tobacco germs, and its fermentation broth on tobacco root Black Rotten has good control effect.Therefore, described streptomyces griseus CCCQ171 may be used for the agricultural microbial agent preparing control tobacco diseases, can containing conventional sterilant, sterilant in described agricultural microbial agent, described agricultural microbial agent can use with other conventional sterilant, pesticide compositional.Described agricultural microbial agent can be obtained by fermentation mode.Because this bacterial strain is separated to obtain from vega soil, with natural, ecological, there is natural harmonious blending, compare with chemical pesticide that soil ecology has no side effect, noresidue, therefore, in the biological control practice of disease, there is potential business development and using value.
The effect of embodiment 4CCCQ171 bacterial strain inducing Tobacco resistance
4.1 inoculation process
Choose the cigarette seedling (4 ~ 6 leaf age) grown fine, fill with and execute bacterium liquid, every strain is filled with and is executed 10mLCCCQ171 bacterial strain fermentation liquor stoste (1 × 10
8individual spore/mL), execute equivalent sterilized water for contrast to fill with.After 0d, 2d, 4d, 6d, 8d, gather blade (avoiding damaged blade) respectively, often process 3 times and repeat.Measure phenylalanine ammonia lyase PAL, peroxidase POD and polyphenoloxidase PPO respectively active.
4.2 enzyme assay
(1) extraction of phenylalanine ammonia lyase PAL, determination of activity:
Take tobacco leaf 2.0g, add 5mMpH8.8 mercaptoethanol borate buffer 10.0mL, PVP0.5g and a small amount of quartz sand, ice bath grinds, four layers of filtered through gauze, 10000rpm frozen centrifugation 10min.Get enzyme liquid 1.0mL, add 0.02ML-phenylalanine 1.0mL, distilled water 2.0mL, cumulative volume is 4.0mL.Contrast does not add substrate, adding distil water 1.0mL, and reaction solution is placed in 30 DEG C of waters bath with thermostatic control and is incubated, and steams ultraviolet spectrophotometer measure light absorption value (OD value) at 290nm place after 0.5h with UV-8500.
(2) extraction of peroxidase POD, determination of activity:
Get tobacco leaf 0.2g, add 0.01MpH5.9 phosphoric acid buffer 2.0mL and a small amount of quartz sand, ice bath grinds, 10000rpm frozen centrifugation 10min.Get enzyme liquid 0.1mL, add pH5.9 phosphoric acid buffer to 4.0mL, add 0.02M more wound order phenol 1.0mL, mixing.During mensuration, add 0.06mol/LH
2o
2, under 420nm wavelength, after 5min, measure light absorption value (OD value).Contrast does not add H
2o
2, add 1.0mL phosphoric acid buffer.
(3) extraction of polyphenoloxidase PPO, determination of activity:
Get tobacco leaf 0.2g, add 0.051MpH6.8 phosphoric acid buffer 0.4mL and a small amount of quartz sand, ice bath grinds, 10000rpm frozen centrifugation 10min.Get enzyme liquid 0.01mL, add 0.05MpH7.8 phosphoric acid buffer 1.0mL, 0.02M pyrocatechol 1.5mL.Contrast not enzyme-added liquid.Under 389nm wavelength, light absorption value (OD value) is measured after reaction 30min.
Cigarette seedling is after the process of CCCQ171 bacterial strain, in blade, PAL activity rises gradually, after 4d, reach a significant peak value, 6d, 8d start to decline, but in blade after the process of CCCQ171 bacterium liquid, PAL activity is all significantly higher than contrast in the level of α=0.05; The process of CCCQ171 bacterial strain is little on the impact of Tobacco Peroxidase activity change, and 4,6d is lower than contrast; In CCCQ171 bacterial strain process cigarette seedling rear blade, polyphenol oxidase activity rises gradually, reaches a peak value after 2d, and 4, start after 6d to decline, there is another peak value after 8d, and high compared with the peak value after 2d.Occur declining to when impinging upon 2d, rise afterwards always, but all lower than the activity for the treatment of group, show that CCCQ171 bacterial strain all can produce relevant to polyphenol oxidase activity to the resistance of root Black Rotten by evoking tobacco.Result shows that CCCQ171 bacterial strain can produce Resistant enzymes by evoking tobacco plant to a certain extent, thus improves the resistance to root Black Rotten.
Streptomyces griseus CCCQ171 itself has evoking tobacco produces resistance effect to tobacco diseases.Therefore, described streptomyces griseus CCCQ171 may be used for preparing evoking tobacco produces resistance agricultural microbial agent to tobacco diseases, can containing conventional sterilant, sterilant in described agricultural microbial agent, described agricultural microbial agent can use with other conventional sterilant, pesticide compositional.Described agricultural microbial agent can be obtained by fermentation mode.Because this bacterial strain is separated to obtain from vega soil, with natural, ecological, there is natural harmonious blending, compare with chemical pesticide that soil ecology has no side effect, noresidue, therefore, in the biological control practice of disease, there is potential business development and using value.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (7)
1. strain streptomyces griseus (Streptomycesgriseochromogenes) CCCQ171, it is characterized in that, its deposit number is: CGMCCNo.6227, and the 16SrDNA sequence of this bacterial strain is as shown in SEQIDNo:1.
2. the application of streptomyces griseus according to claim 1 (Streptomycesgriseochromogenes) CCCQ171 in agricultural microbial agent.
3. the application of streptomyces griseus according to claim 2 (Streptomycesgriseochromogenes) CCCQ171 in agricultural microbial agent, it is characterized in that, described agricultural microbial agent is for preventing and treating tobacco diseases or evoking tobacco produces resistance to tobacco diseases.
4. the application of streptomyces griseus according to claim 3 (Streptomycesgriseochromogenes) CCCQ171 in agricultural microbial agent, it is characterized in that, described tobacco diseases is black root of tobacco, black shank or Alternaria alternate.
5. an agricultural microbial agent, is characterized in that, containing bacterial strain according to claim 1 in described agricultural microbial agent.
6. agricultural microbial agent according to claim 5, is characterized in that, described agricultural microbial agent is for preventing and treating tobacco diseases or evoking tobacco produces resistance to tobacco diseases.
7. agricultural microbial agent according to claim 6, is characterized in that, described tobacco diseases is black root of tobacco, black shank or Alternaria alternate.
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Effective date of registration: 20160822 Address after: 510000, 88, 6 Hui Jin Street, five mountain road, Guangdong, Guangzhou, Tianhe District, 8, 601, one room Patentee after: GUANGZHOU ZHENGE AGRICULTURAL SCIENCE & TECHNOLOGY Co.,Ltd. Patentee after: GUANGDONG ZHENGE BIOTECHNOLOGY Co.,Ltd. Address before: 526000 District, Gaoyao Industrial Park, Gaoyao District, Guangdong, Zhaoqing Patentee before: GUANGDONG ZHENGE BIOTECHNOLOGY Co.,Ltd. |