CN106135285A - The application in prevention and control rice blast of the Rhodopseudomonas palustris biocontrol agent - Google Patents
The application in prevention and control rice blast of the Rhodopseudomonas palustris biocontrol agent Download PDFInfo
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- CN106135285A CN106135285A CN201610333888.XA CN201610333888A CN106135285A CN 106135285 A CN106135285 A CN 106135285A CN 201610333888 A CN201610333888 A CN 201610333888A CN 106135285 A CN106135285 A CN 106135285A
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- rhodopseudomonas palustris
- application
- biocontrol agent
- rice
- prevention
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
Abstract
The invention discloses the application in prevention and control rice blast of a kind of Rhodopseudomonas palustris biocontrol agent, Rhodopseudomonas palustris biocontrol agent is by Rhodopseudomonas palustris LY 6(Rhodopseudomonas palustris LY 6) cultivate obtain.The present invention utilizes Rhodopseudomonas palustris to be applied to prevention and control rice blast first, have environmental friendliness, nontoxic to people and animals, to crop without poisoning, use the features such as simple and convenient.
Description
Technical field
The present invention relates to biological pesticide technical field, particularly relate to a kind of Rhodopseudomonas palustris biocontrol agent prevention and control rice
Application in pestilence.
Background technology
Rice blast is one of important disease of Oryza sativa L., and this disease is widely distributed, and the whole world there are about more than 80 countries and regions and sends out
Sick this, the most all there is generation China South And North Rice Regions.Plant disease epidemic area, the general underproduction 10%～20%, time serious up to 40%～
50%, even No kernels or seeds are gathered, as in a year of scarcity for field heavy especially.For the preventing and treating of rice blast, the most still based on chemical prevention, but due to anti-
The property of medicine strengthens and finds that applications of pesticide pole is the most scientific and reasonable not in time, and applications of pesticide dosage and number of times severe overweight cause environment
The significant problems such as pollution and agricultural product quality and safety.Therefore, pesticide dosage and green composite prevention and control rice blast how are reduced still
It it is so the major issue faced in current China Rice Production.
Biological control is nontoxic with it, harmless, pollution-free, be not likely to produce Drug resistance and the advantage such as efficiently, at plant pest
Preventing and treating is increasingly subject to people's attention.Photosynthetic bacteria (Photosynthetic Bacteria, PBS) is can be at anaerobism bar
Do not put the general name of a photosynthetic bacterioid of oxygen under part, be that the original luminous energy that has occurred the earliest on the earth synthesizes
The prokaryote of system.It is of a great variety, widely distributed, is prevalent in ocean, rivers, lake, paddy field, activated sludge and soil
In earth, it is known that photosynthetic bacteria have Chlorobiaceae (Chlorobiaceae), red Thiobacteriaceae
(Chromatiaceae), Rhodospirillaceae (Rhodospirillaceae) 3 sections.Photosynthetic bacteria complexity
Metabolic function and unique physiological and ecological characteristic make it protect at farming animals industry, feedstuff, environmental protection, food processing, bioenergy and medicine
It is used widely in the field such as strong.In terms of agricultural production, photosynthetic bacteria carries out the process of seed seedling, crop quality changes in utilization
Good, soil remediation processes with weight gaining, the prevention and control of plant diseases, pest control, waste water and ensures that agricultural product security is current study hotspot.Photosynthetic
Antibacterial itself is nontoxic, has fixed nitrogen, carbon sequestration, promotion sulfur cycle effect, and rich in vitamin B group, pantothenic acid, folic acid, carotenoids
The biological active substanceies such as element, ubiquinone, proline, uracil, cytosine are to increasing soil fertility, stimulate plant cell activity, rush
Enter root system development, raising crop photosynthesis efficiency and reproductive growth ability and there is the strongest promotion functions.Although, photosynthetic bacteria is in agriculture
The applied research in industry field obtains deep expansion, and provides strong guarantee for the high-quality of agricultural product, safety, health.
Certain progress is had been achieved at present in terms of the Biological control research of rice blast.At microbial source biological pesticide
In, bacillus cereus and fungus Trichoderma can effectively suppress nourishing and growing of Pyricularia oryzae.At present, the red vacation in photosynthetic bacteria marsh
Zymomonas mobilis microbial inoculum did not the most appear in the newspapers in the prevention and control of rice blast.
Summary of the invention
The technical problem to be solved in the present invention is to overcome the deficiencies in the prior art, it is provided that a kind of Rhodopseudomonas palustris biological and ecological methods to prevent plant disease, pests, and erosion
Microbial inoculum application in prevention and control rice blast, utilizes the microbial inoculum that Rhodopseudomonas palustris bacterial strain is made, and production cost is low, to rice blast
Control effect is notable.
For solving above-mentioned technical problem, it is provided that a kind of Rhodopseudomonas palustris biocontrol agent answering at prevention and control rice blast
With, described Rhodopseudomonas palustris biocontrol agent is by Rhodopseudomonas palustris LY-6(Rhodopseudomonas palustris LY-6) cultivation obtains.Preferably, described cultural method include activation, seed culture, productive culture,
The steps such as NaCl solution is resuspended.
Above-mentioned application, it is preferred that described rice blast is leaf blast in rice, described Rhodopseudomonas palustris biocontrol agent is anti-
Application process in control leaf blast in rice is: spray described Rhodopseudomonas palustris biocontrol agent in the rice leaf in tiller later stage
On.
Above-mentioned application, it is preferred that described rice blast is Pyricularia Oryzae, described Rhodopseudomonas palustris biocontrol agent is anti-
Application process in control Pyricularia Oryzae is: described Rhodopseudomonas palustris biocontrol agent is sprayed the rice leaf heading stage.
Above-mentioned application, it is preferred that the concentration of described Rhodopseudomonas palustris biocontrol agent is 5 × 109Cfu/mL～5
×1010Cfu/mL, the amount of spraying is 15 L/ mus～20 L/ mus, sprays once every 10 d～15 d.
Above-mentioned application, it is preferred that the preparation method of described Rhodopseudomonas palustris biocontrol agent specifically includes following step
Rapid:
(1) activation: the bacterial strain of Rhodopseudomonas palustris LY-6 is pressed the cultivation of double-layer plate culture method and occurs to red single bacterium colony；
(2) seed culture: red single bacterium colony step (1) cultivated accesses in serum bottle, uses photosynthetic bacterial liquid
Culture medium culturing obtains bacterium solution to exponential phase；
(3) productive culture: the bacterium solution obtained after step (2) seed culture is inoculated in production bottle, produces training with photosynthetic bacteria
Foster base carries out productive culture to exponential phase, is biocontrol agent.
Above-mentioned application, it is preferred that in described step (2), seed culture temperature be 30 DEG C～32 DEG C, illumination condition be
2500 Lx～4000 Lx, pH are to carry out under the conditions of 7.0.
Above-mentioned application, it is preferred that in described step (3), productive culture temperature be 30 DEG C～32 DEG C, illumination condition be
Carry out under the conditions of 2500 Lx～4000 Lx, pH=7.0.
Above-mentioned application, it is preferred that the inoculum concentration of bacterium solution described in described step (3) is that described Photosynthetic bacterium strain produces
5 %～10 % of culture medium cumulative volume.
Rhodopseudomonas palustris LY-6 that the application is used (Rhodopseudomonas palustris
LY-6), being preserved in China typical culture collection center, deposit number is the red false unit cell in CCTCC NO: M 2014525(marsh
Bacterium LY-6 patent applied for is protected, number of patent application: 201410750073.2), it is big that depositary institution address is positioned at Wuhan, China
Learning, preservation date is on November 5th, 2014, and this bacterial strain is named as Rhodopseudomonas palustris LY-6
(Rhodopseudomonas palustris LY-6)。
Described Rhodopseudomonas palustris bacterial strain LY-6, has a following principal character: Gram-negative, identifies through Physiology and biochemistry
Being Rhodopseudomonas palustris with molecular biology identification, main biological property has: cultivate 4 d on bilayer solid plating medium
～8 d, forming red circular colonies, colony edge is the most smooth, colony diameter 0.2 mm～0.60 mm；Fluid medium
Middle cultivation 7 d is peony；Well-grown under anaerobism and micro-oxygen conditions；Physiology characteristic is V-P reaction and clark and Lubsreaction
Negative, H2S reaction, gelatin liquefaction, urease test, indole test are the most positive, optimum growth temperature 30 DEG C～32 DEG C, pH=7.
Compared with prior art, it is an advantage of the current invention that:
(1) the invention provides a kind of Rhodopseudomonas palustris LY-6 application at prevention and control rice blast, Rhodopseudomonas palustris LY-
Containing secreting albumen and metabolite outward in the biocontrol agent of 6, can effectively prevent and treat rice blast, such as leaf blast in rice, Pyricularia Oryzae；Have
Environmental friendliness, nontoxic to people and animals, to crop without poisoning, use the features such as simple and convenient.
(2) production technology of the Rhodopseudomonas palustris LY-6 of the present invention is simple, incubation time is short, low cost, and this bacterium
Strain can prepare the biocontrol agent for rice blast prevention and control by simple, quick, the operational approach of low cost.
Accompanying drawing explanation
For making the purpose of the embodiment of the present invention, technical scheme and advantage clearer, below in conjunction with the embodiment of the present invention
In accompanying drawing, the technical scheme in the embodiment of the present invention is carried out clear, complete description.
Fig. 1 be in present example 2 biocontrol agent in land for growing field crops prevention effect comparison diagram to leaf blast in rice.
Fig. 2 be in present example 3 biocontrol agent in land for growing field crops prevention effect comparison diagram to Pyricularia Oryzae.
Detailed description of the invention
Below in conjunction with Figure of description and concrete preferred embodiment, the invention will be further described, but the most therefore and
Limit the scope of the invention.
Material and instrument employed in following example are commercially available.The Rhodopseudomonas palustris LY-6 of the present invention
(Rhodopseudomonas palustris LY-6), it is preserved in China typical culture collection center,
Deposit number is CCTCC NO:M 2014525, and depositary institution address is positioned at Wuhan, China university, and preservation date is 2014 11
The moon 5.
Embodiment 1:
The preparation method of the Rhodopseudomonas palustris LY-6 biocontrol agent of a kind of present invention, comprises the following steps:
(1) activation: cultivated by double-layer plate culture method by the conserving species of Rhodopseudomonas palustris LY-6 bacterial strain, cultivates extremely
Red single bacterium colony occurs；The culture medium used is photosynthetic bacteria solid medium, and formula is: (NH4)2SO4 0.1 wt%、Mg
SO4 0.02 wt%、NaHCO3 0.5 wt%、K2HPO4 0.05 wt%, NaCl 0.02 wt%, yeast extract 0.15 wt% and agar
1.5 wt%, pH=7.0.
(2) seed culture: step (1) is turned out red single bacterium colony access in 120 mL serum bottle, in temperature be
30 DEG C～32 DEG C, illumination condition be 2500 Lx～under the conditions of 4000 Lx, pH=7.0, with photosynthetic bacterial liquid culture medium culturing extremely
Exponential phase；The culture medium used is photosynthetic bacteria liquid culture medium, and formula is: (NH4)2SO4 0.1 wt%、MgSO4 0.02
wt%、NaHCO3 0.5 wt%、K2HPO4 0.05 wt%, NaCl 0.02 wt%, yeast extract 0.15 wt%(are mass fraction),
pH=7.0。
(3) productive culture: the bacterium solution obtained after step (2) seed culture is inoculated in production bottle, raw with photosynthetic bacteria
Producing culture medium (medium component is consistent with above-mentioned photosynthetic bacteria liquid culture medium) and carry out productive culture, the inoculum concentration of bacterium solution is institute
State the 5% of Photosynthetic bacterium strain production medium cumulative volume, temperature be 30 DEG C～32 DEG C, illumination condition be 2500 Lx～4000
Under the conditions of Lx, pH=7.0, cultivate to exponential phase, i.e. prepare biocontrol agent.
Comparative example 1:
The biocontrol agent obtained in embodiment 1 is put in autoclave, inactivates 30 min at 121 DEG C and i.e. obtain the biocontrol microorganisms of inactivation
Agent.
Embodiment 2: the Rhodopseudomonas palustris LY-6 biocontrol agent field prevention and control to leaf pestilence
Biocontrol agent (the concentration 5 × 10 respectively embodiment 1 prepared9Cfu/mL), the biological and ecological methods to prevent plant disease, pests, and erosion of the inactivation prepared in comparative example 1
Microbial inoculum, chemical pesticide (Isoprothiolane, 120 mL/ mus) spray in the rice leaf in tiller later stage, and each process sets 3 repetitions, often
Mu 20 L, spray once every 15 d, use 3 times altogether.After last dispenser 15 d, calculate rice leaf by 5 method samplings
Disease index, and calculate respective prevention effect.
Fig. 1 is that the biocontrol agent of the inactivation of comparative example 1, the biocontrol agent of embodiment 1 and chemical pesticide are to leaf blast in rice
Prevention effect comparison diagram, A, B represent significant difference (P < 0.01).As can be known from Fig. 1: use the biocontrol agent of inactivation, life
Anti-microbial inoculum and the respective prevention effect of pesticide are 8.7%, 30.9% and 44.4%, and are found by statistical analysis, spray biological and ecological methods to prevent plant disease, pests, and erosion
The prevention effect of microbial inoculum and chemical pesticide does not has significant difference.
Embodiment 3: the Rhodopseudomonas palustris LY-6 biocontrol agent field prevention and control to fringe pestilence
Biocontrol agent (the concentration 5 × 10 respectively embodiment 1 prepared9Cfu/mL), the biological and ecological methods to prevent plant disease, pests, and erosion of the inactivation prepared in comparative example 1
Microbial inoculum and chemical pesticide (Isoprothiolane, 120 mL/ mus) spray the rice leaf heading stage, and each process sets 3 repetitions, every mu
20 L, spray once every 15 d, use 2 times altogether.After 15 d, calculate Oryza sativa L. tassel disease index by 5 method samplings, and calculate
Respective prevention effect.
Fig. 2 is that the biocontrol agent of the inactivation of comparative example 1, the biocontrol agent of embodiment 1 and chemical pesticide are to Pyricularia Oryzae
Prevention effect comparison diagram, A, B represent significant difference (P < 0.01).Use the biocontrol agent of inactivation, biocontrol agent and agriculture
The respective prevention effect of medicine, it is 11.4%, 34.4% and 46.3%, and is found by statistical analysis, spray biocontrol agent and chemistry
The prevention effect of pesticide does not has significant difference.
The above, be only presently preferred embodiments of the present invention, and the present invention not makees any pro forma restriction.Though
So the present invention discloses as above with preferred embodiment, but is not limited to the present invention.Any it is familiar with those skilled in the art
Member, in the case of without departing from the spirit of the present invention and technical scheme, may utilize in method and the technology of the disclosure above
Hold and technical solution of the present invention is made many possible variations and modification, or be revised as the Equivalent embodiments of equivalent variations.Therefore,
Every content without departing from technical solution of the present invention, according to the present invention technical spirit to made for any of the above embodiments any simply
Amendment, equivalent, equivalence change and modification, all still fall within the range of technical solution of the present invention protection.
Claims (8)
1. Rhodopseudomonas palustris biocontrol agent application in prevention and control rice blast, it is characterised in that the red false unit cell in described marsh
Bacterium biocontrol agent is by Rhodopseudomonas palustris LY-6(Rhodopseudomonas palustris LY-
6) cultivation obtains.
Application the most according to claim 1, it is characterised in that described rice blast is leaf blast in rice, described marsh is red false single
Born of the same parents' bacterium biocontrol agent application process in prevention and control leaf blast in rice is: sprayed by described Rhodopseudomonas palustris biocontrol agent in dividing
On the rice leaf in tiller later stage.
Application the most according to claim 1, it is characterised in that described rice blast is Pyricularia Oryzae, described marsh is red false single
Born of the same parents' bacterium biocontrol agent application process in prevention and control Pyricularia Oryzae is: spray described Rhodopseudomonas palustris biocontrol agent in taking out
The rice leaf of fringe phase.
Application the most according to any one of claim 1 to 3, it is characterised in that described Rhodopseudomonas palustris biocontrol microorganisms
The concentration of agent is 5 × 109Cfu/mL, the amount of spraying is 20 L/ mus, sprays once every 15 d.
Application the most according to any one of claim 1 to 3, it is characterised in that described Rhodopseudomonas palustris biocontrol microorganisms
The preparation method of agent specifically includes following steps:
(1) activation: the bacterial strain of Rhodopseudomonas palustris LY-6 is pressed the cultivation of double-layer plate culture method and occurs to red single bacterium colony；
(2) seed culture: described red single bacterium colony step (1) cultivated accesses to, in serum bottle, use photosynthetic bacteria
Fluid medium cultivation obtains bacterium solution to exponential phase；
(3) productive culture: the described bacterium solution obtained after step (2) seed culture is inoculated in production bottle, raw with photosynthetic bacteria
Produce culture medium and carry out productive culture to exponential phase, be biocontrol agent.
Application the most according to claim 5, it is characterised in that in described step (2), seed culture temperature be 30 DEG C～
32 DEG C, illumination condition is 2500 Lx～4000 Lx, pH are to carry out under the conditions of 7.0.
Application the most according to claim 5, it is characterised in that in described step (3), productive culture temperature be 30 DEG C～
32 DEG C, illumination condition is 2500 Lx～carries out under the conditions of 4000 Lx, pH=7.0.
Application the most according to claim 5, it is characterised in that described in described step (3), the inoculum concentration of bacterium solution is described
5 %～10 % of Photosynthetic bacterium strain production medium cumulative volume.
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Cited By (9)
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| CN108179129A (en) * | 2018-03-13 | 2018-06-19 | 长沙艾格里生物肥料技术开发有限公司 | A kind of application of Rhodopseudomonas palustris microbial inoculum in prevention and control false smut |
| CN108192849A (en) * | 2018-03-13 | 2018-06-22 | 长沙艾格里生物肥料技术开发有限公司 | Rhodopseudomonas palustris bacterial strain, Rhodopseudomonas palustris bacteria agent and its application |
| CN108893418A (en) * | 2018-05-10 | 2018-11-27 | 湖南省植物保护研究所 | A kind of Rhodopseudomonas palustris LY-6 microbial inoculum and preparation and its application to the prevention and treatment of tomato southern blight |
| CN110731345A (en) * | 2019-08-28 | 2020-01-31 | 湖南省植物保护研究所 | Pesticide composition and application thereof in preventing and treating potato scab |
| CN110982750A (en) * | 2019-12-26 | 2020-04-10 | 河北大河生物科技有限公司 | High-density fermentation method for rhodopseudomonas palustris and application of high-density fermentation method |
| CN111363056A (en) * | 2020-03-17 | 2020-07-03 | 湖南省植物保护研究所 | Rhodopseudomonas palustris exopolysaccharide and preparation method and application thereof |
| CN113862179A (en) * | 2021-09-15 | 2021-12-31 | 上海农乐生物制品股份有限公司 | Rhodopseudomonas palustris, application and method for preparing 5-ALA by using rhodopseudomonas palustris |
| CN115067362A (en) * | 2022-08-15 | 2022-09-20 | 湖南省植物保护研究所 | Sterilization composition containing rhodopseudomonas palustris and tricyclazole |
| CN115191444A (en) * | 2022-08-15 | 2022-10-18 | 湖南省植物保护研究所 | Bactericidal composition containing rhodopseudomonas palustris and isoprothiolane |
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| CN108192849A (en) * | 2018-03-13 | 2018-06-22 | 长沙艾格里生物肥料技术开发有限公司 | Rhodopseudomonas palustris bacterial strain, Rhodopseudomonas palustris bacteria agent and its application |
| CN108179129A (en) * | 2018-03-13 | 2018-06-19 | 长沙艾格里生物肥料技术开发有限公司 | A kind of application of Rhodopseudomonas palustris microbial inoculum in prevention and control false smut |
| CN108893418A (en) * | 2018-05-10 | 2018-11-27 | 湖南省植物保护研究所 | A kind of Rhodopseudomonas palustris LY-6 microbial inoculum and preparation and its application to the prevention and treatment of tomato southern blight |
| CN110731345A (en) * | 2019-08-28 | 2020-01-31 | 湖南省植物保护研究所 | Pesticide composition and application thereof in preventing and treating potato scab |
| CN110982750A (en) * | 2019-12-26 | 2020-04-10 | 河北大河生物科技有限公司 | High-density fermentation method for rhodopseudomonas palustris and application of high-density fermentation method |
| CN111363056A (en) * | 2020-03-17 | 2020-07-03 | 湖南省植物保护研究所 | Rhodopseudomonas palustris exopolysaccharide and preparation method and application thereof |
| CN111363056B (en) * | 2020-03-17 | 2021-07-09 | 湖南省植物保护研究所 | Rhodopseudomonas palustris exopolysaccharide and preparation method and application thereof |
| US11473115B2 (en) | 2020-03-17 | 2022-10-18 | Hunan Plant Protection Institute | Exopolysaccharide from Rhodopseudomonas palustris and method for preparing and use thereof |
| CN113862179A (en) * | 2021-09-15 | 2021-12-31 | 上海农乐生物制品股份有限公司 | Rhodopseudomonas palustris, application and method for preparing 5-ALA by using rhodopseudomonas palustris |
| CN115067362A (en) * | 2022-08-15 | 2022-09-20 | 湖南省植物保护研究所 | Sterilization composition containing rhodopseudomonas palustris and tricyclazole |
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| CN115067362B (en) * | 2022-08-15 | 2023-08-11 | 湖南省植物保护研究所 | Sterilization composition containing rhodopseudomonas palustris and tricyclazole |
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