CN102433285A - Bacterium Paenibacillus polymyma SHL-1 and application thereof in prevention and treatment of stalk rot of cymbidium sinense - Google Patents
Bacterium Paenibacillus polymyma SHL-1 and application thereof in prevention and treatment of stalk rot of cymbidium sinense Download PDFInfo
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- 241000179039 Paenibacillus Species 0.000 title claims abstract description 31
- 241000173252 Cymbidium sinense Species 0.000 title abstract 3
- 230000002265 prevention Effects 0.000 title abstract 2
- 238000002360 preparation method Methods 0.000 claims abstract description 8
- 238000004321 preservation Methods 0.000 claims abstract description 4
- 241000732800 Cymbidium Species 0.000 claims description 22
- 241000894006 Bacteria Species 0.000 claims description 21
- 239000007788 liquid Substances 0.000 claims description 20
- 230000001580 bacterial effect Effects 0.000 claims description 18
- 238000009472 formulation Methods 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 10
- 235000015097 nutrients Nutrition 0.000 claims description 10
- 239000002207 metabolite Substances 0.000 claims description 2
- 238000000855 fermentation Methods 0.000 claims 1
- 230000004151 fermentation Effects 0.000 claims 1
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 7
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- 239000003242 anti bacterial agent Substances 0.000 abstract 1
- 231100000252 nontoxic Toxicity 0.000 abstract 1
- 230000003000 nontoxic effect Effects 0.000 abstract 1
- 241000194105 Paenibacillus polymyxa Species 0.000 description 7
- 201000010099 disease Diseases 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
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- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
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- 241001530056 Athelia rolfsii Species 0.000 description 1
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
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Abstract
The invention discloses a bacterium Paenibacillus polymymaSHL-1, a biological agent prepared from the same and application of the bacterium Paenibacillus polymymaSHL-1 in prevention and treatment of stem rot of cymbidium sinense. The bacterium Paenibacillus polymymaSHL-1 is preserved in China center for type culture Collection at 2011, 12 and 2 days, with the preservation number of CCTCCM 2011441. The preparation method comprises the steps of inoculating Paenibacillus polymymaSHL-1 into an LB liquid culture medium with the pH value of 7.0, and carrying out shake culture at the temperature of 28 ℃ for 48 hours at the speed of 200r/min to obtain the biological agent. The biological agent has strong inhibition effect on cymbidium sinense, is environment-friendly and nontoxic in source, and has small influence on the ecological environment; in addition, the culture condition requirement is low, and the antibacterial agent has an antibacterial effect on various pathogenic fungi and has a good development and application prospect.
Description
Technical field
The invention belongs to field of biological pesticide, be specifically related to a strain bacterium
Paenibacillus polymymaThe biotechnological formulation of SHL-1 and preparation thereof and the application in control Chinese cymbidium stem rot.
Background technology
Chinese cymbidium (
Cymbidium. sinense) have long cultivation history in China, mainly be distributed in ground such as Guangdong, Guangxi, Fujian, Zhejiang, Sichuan, Yunnan.In the industry of flowers and plants, Chinese cymbidium belongs to high-grade potted flower main breed, has high economic worth and ornamental value.In recent years, along with the expansion of Chinese cymbidium cultivated area, various disease and pests also constantly occur, and become one of limiting factor that influences Chinese cymbidium production.The Chinese cymbidium stem rot (
Fusarium oxysporum), be blight again, be important disease during Chinese cymbidium is produced, causing harm can cause plant withered when serious, causes serious threat for the production of Chinese cymbidium.Each city, county (district) investigation find that this disease takes place general in the Chinese cymbidium planting base in Guangdong Province, and it is serious to cause harm, and a part flower sickness rate is up to 30%, and production causes and has a strong impact on to Chinese cymbidium.
The series bacillus genus (
Paenibacillus) be to equal the new genus set up in 1993 by Ash, comprise 11 kinds that belong to bacillus originally, its type species be Paenibacillus polymyxa (
Paenibacillus polymyxa).Research shows, Paenibacillus polymyxa can produce the multiple meta-bolitess that anti-microbial effect is arranged such as peptide class, protein-based, ucleosides, and plurality of plant diseases is had preventive and therapeutic effect.Kavitha etc. (2005) are separated to a kind of molecular weight from Paenibacillus polymyxa VLB16 fermented liquid be 37KDa and resistant to elevated temperatures protein, this albumen can suppress Pyricularia oryzae (
Magnaporthe oryzae), Rhizoctonia solani Kuhn (
Rhizoctonia solani) mycelial growth.Sun Changpo discovers,
P. polymymaCp_S316 to comprise cucumber fusarium axysporum (
Fusarium oxysporumF. sp.
Cucumerinum) all have the intensive antagonistic action interior 8 kinds of pathogenic fungies and 4 kinds of pathogenetic bacterias.Khan etc. have reported
P. polymymaCan also act as host plant through fixed nitrogen and molten phosphorus nutritive ingredient is provided, promote plant strain growth, improve the disease resistance of plant etc.It is thus clear that, Paenibacillus polymyxa (
P. polymyma) has a broad antifungal spectrum, effectively meta-bolites is various, has the potentiality that are developed to biological pesticide.
Summary of the invention
The objective of the invention is to deficiency, the bacterium of the strong bacteriostatic action of a kind of tool is provided to existing biological control Chinese cymbidium stem rot technology
Paenibacillus polymymaSHL-1.
Another object of the present invention is to provide by above-mentioned bacterium
Paenibacillus polymymaThe preparation of the fermented liquid of SHL-1 and biotechnological formulation.
Another object of the present invention is to provide the preparation method of above-mentioned biotechnological formulation.
Another object of the present invention is to provide above-mentioned bacterium
Paenibacillus polymymaThe application in the control of Chinese cymbidium stem rot of SHL-1 and fermented liquid thereof.
Above-mentioned purpose of the present invention is achieved through following scheme:
The SHL-1 bacterial strain is to separate from the dendrobium blade through conventional tissue isolation.This bacterial strain is observed after cultivating 48 h on the LB substratum, and its single bacterium colony is circular, white, and smooth surface, opaque, slightly to swell, neat in edge is non-wrinkled.This bacterium in the LB of pH7.0 nutrient solution, 28 ℃, 200 r/min shaking culture 24h, fermented liquid is muddy shape, the thalline adhesion becomes cotton-shaped, 24h ~ 48h fermented liquid begins to become clarification, cotton-shaped thalline scatters.This bacterial strain is through 16S rDNA sequential analysis, and the number of landing is HQ317186.1's among its sequence and the GenBank
Paenibacillus polymyxaHomology reaches 99%; Show through BIOLOG Bacteria Identification system software analysis, the SHL-1 bacterial strain only with DB in Paenibacillus polymyxa (
Paenibacillus polymyma) mate well, its similarity Similarity (SIM) is 0.834, and distance of positions Distance (DIS) is 2.48, and possibility Probability (PROB) is 99%.In on December 2nd, 2011 in China's typical culture collection center preservation, preserving number is CCTCC M 2011441.
A kind of biotechnological formulation is by the antagonistic bacterium bacterial strain
Paenibacillus polymymaSHL-1 gets through the liquid fermenting preparation, and its preparation method is: will
Paenibacillus polymymaSHL-1 is seeded to the LB liquid nutrient medium of pH7.0, and 200 r/min shaking culture, 48 h under 28 ℃ can obtain.
The experiment proof: above-mentioned biotechnological formulation has the intensive bacteriostatic action to Chinese cymbidium stem rot germ, and average antibacterial band is 12.0mm; The fermented liquid dialogue palm parasitica of this bacterial strain (
PhytophthoraSp.), the poinsettia parasitica (
PhytophthoraSp.), African chrysanthemum southern blight bacterium (
Sclerotium rolfsii), Da Ye agaric nuclear germ (
Sclerotinia sclerotiorum) and butterfly orchid bennet rotten pathogenic bacteria (
Fusarium fujikuroi) fungistatic effect arranged, wherein the average antibacterial band of dialogue palm eqpidemic disease reaches 9.3mm, and the average antibacterial band of butterfly orchid bennet rotten pathogenic bacteria is reached 9.8mm, has the bacteriostatic action of wide spectrum.
Therefore, bacterium of the present invention
Paenibacillus polymymaSHL-1 and fermented liquid thereof can be used for the control of Chinese cymbidium stem rot.
Compared with prior art, the present invention has following beneficial effect: 1. biotechnological formulation of the present invention has had strong inhibitory effects to Chinese cymbidium stem rot germ, and this biotechnological formulation source environment-protecting asepsis property, and is little to eco-environmental impact; 2. the present invention adopts
Paenibacillus polymymaThe SHL-1 bacterial strain, its culture condition requires low, and the several diseases fungal pathogens is had bacteriostatic action, has good development prospect.
Description of drawings
Fig. 1 does
Paenibacillus polymymaThe single bacterium colony back view of SHL-1 on the LB substratum;
Fig. 2 does
Paenibacillus polymymaThe SHL-1 fermented liquid is to the restraining effect of Chinese cymbidium stem rot germ.
Embodiment
Various culture medium prescriptions:
The PDA substratum: take by weighing the 200g yam, clean the peeling chopping, add water 1000ml and boil half hour, filtered through gauze adds 10～20g glucose and 17～20g agar again, abundant filtered through gauze while hot after the dissolving, and 121 ℃ of sterilizations are after 20 minutes, and the cooling back is stored subsequent use.
The LB substratum: Tryptones 10 g, yeast extract powder 5 g, Nacl 10 g, agar powder 15 g are settled to 1000 mL, and pH 7.2.Above material does not add agar powder and is the LB liquid nutrient medium.
Embodiment 1
Paenibacillus polymymaThe SHL-1 nutrient solution is active to the inhibition of Chinese cymbidium stem rot germ
The SHL-1 bacterial strain is to separate from the dendrobium blade through conventional tissue isolation.This bacterial strain is observed after cultivating 48 h on the LB substratum, and its single bacterium colony is circular, white, and smooth surface, opaque, slightly to swell, neat in edge is non-wrinkled.This bacterium is through 16S rDNA sequential analysis, and the number of landing is HQ317186.1's among its sequence and the GenBank
Paenibacillus polymyxaHomology reaches 99%; Show through BIOLOG Bacteria Identification system software analysis, the SHL-1 bacterial strain only with DB in Paenibacillus polymyxa (
Paenibacillus polymyma) mate well, its similarity Similarity (SIM) is 0.834, and distance of positions Distance (DIS) is 2.48, and possibility Probability (PROB) is 99%.
Adopt inhibition zone method to measure the activity of this bacterial strain live body bacterium liquid.Bacterial isolates is inoculated in the LB nutrient solution, in 28 ℃, 200 r/min shaking culture, 48 h.The use internal diameter is that the punch tool of 0.6 mm punches on the PDA substratum, adds bacterium bacterium liquid 80 μ L in the hole, is inoculating Chinese cymbidium stem rot germ bacterium piece apart from these 3 cm places, hole.To add the LB liquid nutrient medium is contrast, and each handles repetition 3 times, cultivates in 25 ℃ of constant incubators, when the contrast colony diameter reaches 6 cm, measures antibacterial bandwidth.Right through measuring SHL-1 bacterial strain live body bacterium liquid
F. oxysporumHave obvious restraining effect, producing average bandwidth is the antibacterial band of 12.0 mm, makes near antagonism bacterium one side
F. oxysporumMycelia stops growing.Therefore, the strain in the present embodiment has the bacterial strain SHL-1 of antagonistic activity, called after to Chinese cymbidium stem rot germ
Paenibacillus polymymaSHL-1, and on December 2nd, 2011 in China's typical culture collection center preservation, preserving number is CCTCC M 2011441.
Embodiment 2
Paenibacillus polymymaThe antimicrobial spectrum of SHL-1 nutrient solution
With several kinds of flowers fungal disease pathogenic bacterias is object, measures through inhibition zone method
Paenibacillus polymymaThe antimicrobial spectrum of SHL-1 bacterial strain.Bacterial isolates is inoculated in the LB nutrient solution, in 28 ℃, 200 r/min shaking culture, 48 h.The use internal diameter is that the punch tool of 0.6 mm punches on the PDA substratum; Add bacterium bacterium liquid 80 μ L in the hole, inoculating white palm parasitica, poinsettia parasitica, African chrysanthemum southern blight bacterium, Da Ye agaric nuclear germ and butterfly orchid bennet rotten pathogenic bacteria bacterium piece apart from these 3 cm places, hole respectively.To add the LB liquid nutrient medium is contrast, and each handles repetition 3 times, cultivates in 25 ℃ of constant incubators, when the contrast colony diameter reaches 6 cm, measures antibacterial bandwidth.
Table 1
Paenibacillus polymymaThe SHL-1 fermented liquid is to the restraining effect of 5 kinds of pathogenic bacterias
Can find out by table 1,
Paenibacillus polymymaSHL-1 has bacteriostatic action to the pathogenic bacteria of these several kinds of diseases, and this bacterial strain has a broad antifungal spectrum is described.
Claims (4)
1. bacterium
Paenibacillus polymymaSHL-1, this bacterial strain on December 2nd, 2011 in China's typical culture collection center preservation, preserving number is CCTCC M 2011441.
2. a biotechnological formulation is characterized in that this biotechnological formulation is by bacterium
Paenibacillus polymymaSHL-1 gets through the liquid fermentation and culture preparation.
3. the preparation method of the said biotechnological formulation of claim 2, it is characterized in that comprising the steps: with
Paenibacillus polymymaSHL-1 is seeded to the LB liquid nutrient medium of pH7.0, and 200 r/min shaking culture, 48 h under 28 ℃ can obtain.
4. the said bacterium of claim 1
Paenibacillus polymymaThe application in control Chinese cymbidium stem rot of SHL-1 or its meta-bolites.
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| CN 201110437834 CN102433285B (en) | 2011-12-23 | 2011-12-23 | Bacterium Paenibacillus polymyxa SHL-1 and application thereof in prevention and treatment of stalk rot of cymbidium sinense |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103173386A (en) * | 2013-03-12 | 2013-06-26 | 山东农业大学 | Bio-control strain G1 for preventing and controlling pepper phytophthora blights and applications thereof |
| CN103621560A (en) * | 2013-11-28 | 2014-03-12 | 仲恺农业工程学院 | Paenibacillus polymyxa and difenoconazole compound bactericide and application thereof |
| CN112063555A (en) * | 2020-09-10 | 2020-12-11 | 仲恺农业工程学院 | Paenibacillus polymyxa BFP-1 strain, biological agent and application |
-
2011
- 2011-12-23 CN CN 201110437834 patent/CN102433285B/en active Active
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103173386A (en) * | 2013-03-12 | 2013-06-26 | 山东农业大学 | Bio-control strain G1 for preventing and controlling pepper phytophthora blights and applications thereof |
| CN103173386B (en) * | 2013-03-12 | 2015-01-28 | 山东农业大学 | Bio-control strain G1 for preventing and controlling pepper phytophthora blights and applications thereof |
| CN103621560A (en) * | 2013-11-28 | 2014-03-12 | 仲恺农业工程学院 | Paenibacillus polymyxa and difenoconazole compound bactericide and application thereof |
| CN112063555A (en) * | 2020-09-10 | 2020-12-11 | 仲恺农业工程学院 | Paenibacillus polymyxa BFP-1 strain, biological agent and application |
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