CN106135285B - Application of the Rhodopseudomonas palustris biocontrol agent in prevention and control rice blast - Google Patents
Application of the Rhodopseudomonas palustris biocontrol agent in prevention and control rice blast Download PDFInfo
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- CN106135285B CN106135285B CN201610333888.XA CN201610333888A CN106135285B CN 106135285 B CN106135285 B CN 106135285B CN 201610333888 A CN201610333888 A CN 201610333888A CN 106135285 B CN106135285 B CN 106135285B
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- rhodopseudomonas palustris
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- biocontrol agent
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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 a kind of application of Rhodopseudomonas palustris biocontrol agent in prevention and control rice blast, and Rhodopseudomonas palustris biocontrol agent is by Rhodopseudomonas palustris LY 6（Rhodopseudomonas palustris LY‑6）Culture obtains.The present invention for the first time using Rhodopseudomonas palustris be applied to prevention and control rice blast, have the characteristics that it is environmental-friendly, nontoxic to people and animals, to crop without poisoning, using simple and convenient.
Description
Technical field
The present invention relates to biological pesticide technical field more particularly to a kind of Rhodopseudomonas palustris biocontrol agent in prevention and control rice
Application in seasonal febrile diseases.
Background technology
Rice blast is one of important disease of rice, and the disease is widely distributed, and there are about more than 80 countries and regions hairs in the whole world
It is sick this, also there are generation in China South And North Rice Regions.Plant disease epidemic area, the general underproduction 10%~20%, when serious up to 40%~
50%, especially even No kernels or seeds are gathered, as in a year of scarcity for heavy field.Prevention for rice blast, at present or based on chemical prevention, but due to anti-
Pharmacological property enhances and finds that applications of pesticide pole is not scientific and reasonable not in time, and applications of pesticide dosage is severely exceeded with number, causes 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 to be reduced still
It is so the major issue faced in current China's Rice Production.
Biological control with its is nontoxic, harmless, pollution-free, be not likely to produce the resistance to the action of a drug and be efficient the advantages that, in plant pest
It is increasingly valued by people in prevention.Photosynthetic bacteria（Photosynthetic Bacteria, PBS）Being can be in anaerobism item
The general name of photosynthetic a kind of bacterium of oxygen is not put under part, is occurred earliest on the earth with the synthesis of original luminous energy
The prokaryotes of system.Its species is various, widely distributed, generally existing Yu Haiyang, rivers, lake, paddy field, activated sludge and soil
In earth, it is known that photosynthetic bacteria share Chlorobiaceae（Chlorobiaceae）, red Thiobacteriaceae（Chromatiaceae）, it is red
Spirillaceae（Rhodospirillaceae）3 sections.The metabolic function of photosynthetic bacteria complexity and unique physiological and ecological characteristic make it
It is used widely in fields such as farming animals industry, feed, environmental protection, food processing, bioenergy and medicines and health protections.In agricultural production side
Face, with processing, crop quality improvement, soil remediation and the getting fat of photosynthetic bacteria progress seed seedling, the prevention and control of plant diseases, pest control, waste water
Processing and guarantee agricultural product security are current research hotspots.Photosynthetic bacteria itself is nontoxic, has fixed nitrogen, carbon sequestration, promotes sulphur
Ringing, and rich in physiology such as B family vitamin, pantothenic acid, folic acid, carotenoid, ubiquinone, proline, uracil, cytimidines
Active material is to increasing soil fertility, stimulating plant cell activity, root system development, raising crop photosynthesis efficiency and reproduction is promoted to give birth to
Long ability has very strong promotion functions.Although photosynthetic bacteria obtains going deep into expansion in the application study of agriculture field, and is agriculture
The high-quality of product, safety, health provide strong guarantee.
Certain progress is had been achieved in terms of the biological control research of rice blast at present.In microbial source biological pesticide
In, bacillus and fungus Trichoderma can effectively inhibit the nutrient growth of Pyricularia oryzae.At present, the red vacation in photosynthetic bacteria marsh
Unit cell bacteria agent does not appear in the newspapers also in the prevention and control of rice blast.
The content of the invention
The technical problem to be solved by the present invention is to overcome the deficiencies in the prior art, provide a kind of Rhodopseudomonas palustris biological and ecological methods to prevent plant disease, pests, and erosion
Application of the microbial inoculum in prevention and control rice blast, using microbial inoculum made of Rhodopseudomonas palustris bacterial strain, production cost is low, to rice blast
Control effect is notable.
In order to solve the above technical problems, provide a kind of Rhodopseudomonas palustris biocontrol agent answering in prevention and control rice blast
With the Rhodopseudomonas palustris biocontrol agent is by Rhodopseudomonas palustris LY-6（Rhodopseudomonas palustris
LY-6）Culture obtains.Preferably, the cultural method includes the steps such as activation, seed culture, productive culture, NaCl solution resuspension
Suddenly.
Above-mentioned application, it is preferred that the rice blast is leaf blast in rice, and the Rhodopseudomonas palustris biocontrol agent is anti-
Control leaf blast in rice in application process be：The Rhodopseudomonas palustris biocontrol agent is sprayed on to the rice leaf in tiller later stage
On.
Above-mentioned application, it is preferred that the rice blast is Pyricularia Oryzae, and the Rhodopseudomonas palustris biocontrol agent is anti-
Control Pyricularia Oryzae in application process be：The Rhodopseudomonas palustris biocontrol agent is sprayed on to the rice leaf at heading stage.
Above-mentioned application, it is preferred that the concentration of the Rhodopseudomonas palustris biocontrol agent is 5 × 109Cfu/mL~5
×1010Cfu/mL, the amount of spraying are 15 L/ mus~20 L/ mus, are sprayed once every the d of 10 d~15.
Above-mentioned application, it is preferred that the preparation method of the Rhodopseudomonas palustris biocontrol agent specifically includes following step
Suddenly：
（1）Activation：The bacterial strain of Rhodopseudomonas palustris LY-6 is dropped out by double-layer plate cultivation culture to red single bacterium
It is existing；
（2）Seed culture：By step（1）The red single bacterium colony of culture is accessed in serum bottle, uses photosynthetic bacteria
Fluid nutrient medium culture to exponential phase obtains bacterium solution；
（3）Productive culture：By step（2）The bacterium solution obtained after seed culture is inoculated into production bottle, is given birth to photosynthetic bacteria
It produces culture medium and carries out productive culture to exponential phase, be biocontrol agent.
Above-mentioned application, it is preferred that the step（2）In, seed culture temperature be 30 DEG C~32 DEG C, illumination condition is
2500 Lx~4000 Lx, pH is carry out under the conditions of 7.0.
Above-mentioned application, it is preferred that the step（3）In, productive culture temperature be 30 DEG C~32 DEG C, illumination condition is
It is carried out under the conditions of 2500 Lx~4000 Lx, pH=7.0.
Above-mentioned application, it is preferred that the step（3）Described in bacterium solution inoculum concentration be the Photosynthetic bacterium strain production
The % of 5 % of culture medium total volume~10.
Rhodopseudomonas palustris LY-6 used by the application (Rhodopseudomonas palustris LY-6), protect
China typical culture collection center is hidden in, deposit number is CCTCC NO: M 2014525（Rhodopseudomonas palustris LY-6 is
Patent protection, number of patent application：201410750073.2）, depositary institution address is located at Wuhan, China university, preservation date
For on November 5th, 2014, the bacterial strain be named as Rhodopseudomonas palustris LY-6 (Rhodopseudomonas palustris
LY-6)。
The Rhodopseudomonas palustris bacterial strain LY-6 has following main feature：Gram-negative is identified through Physiology and biochemistry
It is Rhodopseudomonas palustris with molecular biology identification, main biological property has：4 d are cultivated on bilayer solid plating medium
~8 d, form red circular colonies, the mm of neat smooth, 0.2 mm of colony diameter of colony edge~0.60；Fluid nutrient medium
7 d of middle culture are in peony；The well-grown under anaerobism and micro-oxygen conditions；Physiology characteristic is reacted for V-P and clark and Lubsreaction
Negative, H2S reactions, gelatin liquefaction, urease test, indole test are positive, 30 DEG C~32 DEG C of optimum growth temperature, pH=7.
Compared with prior art, the advantage of the invention is that：
（1）The present invention provides a kind of Rhodopseudomonas palustris LY-6 in the application of prevention and control rice blast, the red false unit cell in marsh
Containing excretion albumen and metabolite in the biocontrol agent of bacterium LY-6, rice blast can be effectively prevented, such as leaf blast in rice, Rice Panicle
Pest；Have the characteristics that it is environmental-friendly, nontoxic to people and animals, to crop without poisoning, using simple and convenient.
（2）The simple production process of the Rhodopseudomonas palustris LY-6 of the present invention, incubation time are short, at low cost, and the bacterium
The biocontrol agent for rice blast prevention and control can be prepared in strain by simple, quick, inexpensive operating method.
Description of the drawings
To make the purpose, technical scheme and advantage of the embodiment of the present invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, clear, complete description is carried out to the technical solution in the embodiment of the present invention.
Fig. 1 is for biocontrol agent in present example 2 in crop field to the control effect comparison diagram of leaf blast in rice.
Fig. 2 is for biocontrol agent in present example 3 in crop field to the control effect comparison diagram of Pyricularia Oryzae.
Specific embodiment
Below in conjunction with Figure of description and specific preferred embodiment, the invention will be further described, but not therefore and
It limits the scope of the invention.
Material and instrument employed in following embodiment are commercially available.The Rhodopseudomonas palustris LY-6 of the present invention
（Rhodopseudomonas palustris LY-6）, China typical culture collection center is preserved in, deposit number is
CCTCC NO：M 2014525, depositary institution address are located at Wuhan, China university, and preservation date is on November 5th, 2014.
Embodiment 1：
A kind of preparation method of the Rhodopseudomonas palustris LY-6 biocontrol agents of present invention, comprises the following steps：
（1）Activation：The preservation kind of Rhodopseudomonas palustris LY-6 bacterial strains by double-layer plate cultivation is cultivated, is trained
It supports to red single bacterium colony and occurs；The culture medium used is formulated for photosynthetic bacteria solid medium and is：（NH4）2SO4 0.1 wt%、
Mg SO4 0.02 wt%、NaHCO3 0.5 wt%、K2HPO4 0.05 0.02 wt% of wt%, NaCl, 0.15 wt% of yeast extract and fine jade
1.5 wt% of fat, pH=7.0.
（2）Seed culture：By step（1）Turn out red single bacterium colony is accessed in 120 mL serum bottles, be in temperature
30 DEG C~32 DEG C, illumination condition under the conditions of 2500 Lx~4000 Lx, pH=7.0, with photosynthetic bacterial liquid medium culture extremely
Exponential phase；The culture medium used is formulated for photosynthetic bacteria liquid culture medium and is：（NH4）2SO4 0.1 wt%、MgSO4 0.02
wt%、NaHCO3 0.5 wt%、K2HPO4 0.05 0.02 wt% of wt%, NaCl, 0.15 wt% of yeast extract（It is mass fraction）,
pH=7.0。
（3）Productive culture：By step（2）The bacterium solution obtained after seed culture is inoculated into production bottle, is given birth to photosynthetic bacteria
Produce culture medium（Medium component is consistent with above-mentioned photosynthetic bacteria liquid culture medium）Productive culture is carried out, the inoculum concentration of bacterium solution is institute
State the 5% of Photosynthetic bacterium strain production medium total volume, temperature be 30 DEG C~32 DEG C, illumination condition is 2500 Lx~4000
Under the conditions of Lx, pH=7.0, cultivate to exponential phase, that is, biocontrol agent is made.
Comparative example 1：
The biocontrol agent obtained in embodiment 1 is put into autoclave, the life that 30 min obtain inactivation is inactivated at 121 DEG C
Anti- microbial inoculum.
Embodiment 2：Prevention and control of the Rhodopseudomonas palustris LY-6 biocontrol agents field to leaf pest
Respectively by biocontrol agent made from embodiment 1（Concentration 5 × 109cfu/mL）, inactivation obtained in comparative example 1
Biocontrol agent, chemical pesticide（Isoprothiolane, 120 mL/ mus）The rice leaf in tiller later stage is sprayed on, each processing sets 3 weights
Multiple, 20 L, sprays once every 15 d per acre, co-administers 3 times.After 15 d of last time dispenser, rice is calculated by 5 method samplings
Blade disease index, and calculate respective control effect.
Fig. 1 is the biocontrol agent of inactivation, the biocontrol agent of embodiment 1 and the chemical pesticide of comparative example 1 to leaf blast in rice
Control effect comparison diagram, A, B represent significant difference（P<0.01）.As can be known from Fig. 1：Using the biocontrol agent of inactivation, life
Anti- microbial inoculum and the respective control 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 control effect of microbial inoculum and chemical pesticide does not have significant difference.
Embodiment 3：Prevention and control of the Rhodopseudomonas palustris LY-6 biocontrol agents field to fringe pest
Respectively by biocontrol agent made from embodiment 1（Concentration 5 × 109cfu/mL）, inactivation obtained in comparative example 1
Biocontrol agent and chemical pesticide（Isoprothiolane, 120 mL/ mus）The rice leaf at heading stage is sprayed on, each processing sets 3 repetitions,
20 L per acre is sprayed once every 15 d, is co-administered 2 times.After 15 d, rice tassel disease index is calculated by 5 method samplings, and
Calculate respective control effect.
Fig. 2 is the biocontrol agent of inactivation, the biocontrol agent of embodiment 1 and the chemical pesticide of comparative example 1 to Pyricularia Oryzae
Control effect comparison diagram, A, B represent significant difference（P<0.01）.Using the biocontrol agent, biocontrol agent and agriculture of inactivation
The respective control effect of medicine is 11.4%, 34.4% and 46.3%, and is found by statistical analysis, sprays biocontrol agent and chemistry
The control effect of pesticide does not have significant difference.
The above described is only a preferred embodiment of the present invention, not make limitation in any form to the present invention.Though
So the present invention is disclosed as above with preferred embodiment, however is not limited to the present invention.It is any to be familiar with those skilled in the art
Member, in the case where not departing from the Spirit Essence of the present invention and technical solution, all using in the methods and techniques of the disclosure above
Appearance makes technical solution of the present invention many possible changes and modifications or is revised as the equivalent embodiment of equivalent variations.Therefore,
Every content without departing from technical solution of the present invention, technical spirit according to the invention is to made for any of the above embodiments any simple
Modification, equivalent substitution, equivalence changes and modification, still fall within technical solution of the present invention protection in the range of.
Claims (8)
1. application of the Rhodopseudomonas palustris biocontrol agent in prevention and control rice blast, which is characterized in that the red false unit cell in the marsh
Bacterium biocontrol agent is by Rhodopseudomonas palustris LY-6（Rhodopseudomonas palustris LY-6）Culture obtains.
2. application according to claim 1, which is characterized in that the rice blast is leaf blast in rice, and the marsh is red false single
Application process of the born of the same parents bacterium biocontrol agent in prevention and control leaf blast in rice be：The Rhodopseudomonas palustris biocontrol agent is sprayed on point
On the rice leaf in tiller later stage.
3. application according to claim 1, which is characterized in that the rice blast is Pyricularia Oryzae, and the marsh is red false single
Application process of the born of the same parents bacterium biocontrol agent in prevention and control Pyricularia Oryzae be：The Rhodopseudomonas palustris biocontrol agent is sprayed on pumping
The rice leaf of ear period.
4. application according to any one of claim 1 to 3, which is characterized in that the Rhodopseudomonas palustris biocontrol microorganisms
The concentration of agent is 5 × 109Cfu/mL, the amount of spraying are 20 L/ mus, are sprayed once every 15 d.
5. application according to any one of claim 1 to 3, which is characterized in that the Rhodopseudomonas palustris biocontrol microorganisms
The preparation method of agent specifically includes following steps：
（1）Activation：The bacterial strain of Rhodopseudomonas palustris LY-6 is occurred by double-layer plate cultivation culture to red single bacterium colony；
（2）Seed culture：By step（1）The red single bacterium colony of culture is accessed in serum bottle, is trained with photosynthetic bacterial liquid
It supports base culture to exponential phase and obtains bacterium solution；
（3）Productive culture：By step（2）The bacterium solution obtained after seed culture is inoculated into production bottle, is given birth to photosynthetic bacteria
It produces culture medium and carries out productive culture to exponential phase, be biocontrol agent.
6. application according to claim 5, which is characterized in that the step（2）In, seed culture temperature for 30 DEG C~
32 DEG C, illumination condition be to be carried out under the conditions of 2500 Lx~4000 Lx, pH are 7.0.
7. application according to claim 5, which is characterized in that the step（3）In, productive culture temperature for 30 DEG C~
32 DEG C, illumination condition is carries out under the conditions of 2500 Lx~4000 Lx, pH=7.0.
8. application according to claim 5, which is characterized in that the step（3）Described in the inoculum concentration of bacterium solution be described
The % of 5 % of Photosynthetic bacterium strain production medium total volume~10.
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| CN108192849B (en) * | 2018-03-13 | 2021-03-19 | 长沙艾格里生物肥料技术开发有限公司 | Rhodopseudomonas palustris strain, Rhodopseudomonas palustris microbial inoculum and application thereof |
| 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 |
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| 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 |
| 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 |
| 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 |
| 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 |
| 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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