CN105794853B - Application of the Rhodopseudomonas palustris zymotic fluid in prevention and control rice blast - Google Patents
Application of the Rhodopseudomonas palustris zymotic fluid in prevention and control rice blast Download PDFInfo
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- CN105794853B CN105794853B CN201610333926.1A CN201610333926A CN105794853B CN 105794853 B CN105794853 B CN 105794853B CN 201610333926 A CN201610333926 A CN 201610333926A CN 105794853 B CN105794853 B CN 105794853B
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- rhodopseudomonas palustris
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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
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
Abstract
The invention discloses a kind of application of Rhodopseudomonas palustris zymotic fluid in prevention and control rice blast, and Rhodopseudomonas palustris zymotic fluid is by Rhodopseudomonas palustris LY 6（Rhodopseudomonas palustris LY‑6）Culture obtains.The present invention for the first time utilize 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 fields more particularly to a kind of Rhodopseudomonas palustris zymotic fluid in prevention and control rice blast
Application in disease.
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 is nontoxic with its, it is harmless, pollution-free, be not likely to produce drug resistance and it is 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 for not put photosynthetic a kind of bacterium of oxygen under part is that occur earliest on the earth with the synthesis of original luminous energy
The prokaryotes of system.Its type 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.Photosynthetic bacteria can secrete a large amount of albumen and secondary metabolism in incubation
Product, complicated metabolic function and unique physiological and ecological characteristic make it in farming animals industry, feed, environmental protection, food processing, biology
It is used widely in the fields such as the energy and medicines and health protection.
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, and actinomycetes fermentation liquor is to rice blast
The prevention of sick leaf pest also has certain effect.Photosynthetic bacteria Rhodopseudomonas palustris zymotic fluid in the prevention and control of rice blast also not
It appears in the newspapers.
Invention content
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 fermentation
Application of the liquid in prevention and control rice blast, using microbial inoculum made of Rhodopseudomonas palustris bacterial strain, production cost is low, anti-to rice blast
Control significant effect.
In order to solve the above technical problems, a kind of application of Rhodopseudomonas palustris zymotic fluid in prevention and control rice blast is provided,
The Rhodopseudomonas palustris zymotic fluid is by Rhodopseudomonas palustris LY-6（Rhodopseudomonas palustris LY-6）
Culture obtains.Preferably, the cultural method includes activation, seed culture, productive culture, centrifugal filtration.
Above-mentioned application, it is preferred that the rice blast is leaf blast in rice, and the Rhodopseudomonas palustris zymotic fluid is in prevention and control
Application process in leaf blast in rice is：The Rhodopseudomonas palustris zymotic fluid is sprayed on the rice leaf in tiller later stage.
Above-mentioned application, it is preferred that the rice blast is Pyricularia Oryzae, and the Rhodopseudomonas palustris zymotic fluid is in prevention and control
Application process in Pyricularia Oryzae is：The Rhodopseudomonas palustris zymotic fluid is sprayed on to the rice leaf at heading stage.It is above-mentioned
Application, it is preferred that the amount of spraying of the Rhodopseudomonas palustris zymotic fluid is 15 L/ mus~20 L/ mus, every 10 d~15
D is sprayed once.
Above-mentioned application, it is preferred that the preparation method of the Rhodopseudomonas palustris zymotic fluid specifically includes following steps：
（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 accesses 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；
（4）It is prepared by zymotic fluid：By step（3）The bacterium solution centrifugal filtration that productive culture obtains, collects filtrate, and as marsh is red
Pseudomonad zymotic fluid.
Above-mentioned application, it is preferred that the step（2）In, seed culture is temperature is 30 DEG C~32 DEG C, illumination condition is
2500 Lx~4000 Lx, pH are carried out under the conditions of being 7.0.
Above-mentioned application, it is preferred that the step（3）In, productive culture is temperature is 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 %~10 of culture medium total volume.
Above-mentioned application, it is preferred that the step（4）In, the rotating speed of the centrifugation be the rpm of 8000 rpm~12000,
Temperature is 4 DEG C, centrifugation time is the min of 15 min~20.
Above-mentioned application, it is preferred that the step（4）In, the centrifugal filtration is specially the supernatant taken after centrifugation, is used
0.22 μm of filter membrane is filtered.
Rhodopseudomonas palustris LY-6 used by the application (Rhodopseudomonas palustris LY-6), protect
It is hidden in China typical culture collection center, deposit number is CCTCC NO: M 2014525（Rhodopseudomonas palustris LY-6
Patent applied for is protected, number of patent application：201410750073.2）, depositary institution address is located at Wuhan, China university, preservation day
Phase is on November 5th, 2014, which is 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, neat smooth, colony diameter 0.2 mm~0.60 mm of colony edge；Fluid nutrient medium
7 d of middle culture are in peony；The well-grown under anaerobism and micro-oxygen conditions；Physiology characteristic is V-P reactions 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 the prior art, the advantages of the present invention are as follows：
（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
Contain excretion albumen and metabolite in the zymotic fluid of bacterium LY-6, rice blast can be effectively prevented, such as leaf blast in rice, Pyricularia Oryzae；
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 zymotic fluid for rice blast prevention and control can be prepared in strain by simple, quick, inexpensive operating method.
Description of the drawings
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical solution in the embodiment of the present invention carries out clear, complete description.
Fig. 1 is for zymotic fluid in present example 2 in crop field to the control effect comparison diagram of leaf blast in rice.
Fig. 2 is for zymotic fluid in present example 3 in crop field to the control effect comparison diagram of Pyricularia Oryzae.
Specific implementation mode
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）, it is preserved in China typical culture collection center, deposit number is
CCTCC NO：M 2014525, depositary institution address are located at Wuhan, China university, and the deposit date is on November 5th, 2014.
Embodiment 1：
A kind of preparation method of the Rhodopseudomonas palustris LY-6 zymotic fluids of the present invention, includes the following steps：
（1）Activation：The preservation kind of Rhodopseudomonas palustris LY-6 bacterial strains is cultivated by double-layer plate cultivation, 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 accesses in 120 mL serum bottles, be in temperature
30 DEG C~32 DEG C, illumination condition be 2500 Lx~4000 Lx, pH=7.0 under the conditions of, extremely with photosynthetic bacterial liquid medium culture
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
5 % for stating Photosynthetic bacterium strain production medium total volume, temperature be 30 DEG C~32 DEG C, illumination condition be 2500 Lx~
Under the conditions of 4000 Lx, pH=7.0, culture to exponential phase.By step（3）Bacterium solution after productive culture, bacterial concentration
It is 5 × 109 cfu/mL。
（4）It is prepared by zymotic fluid：By step（3）The bacterium solution that productive culture obtains carries out 8000 rpm, 4 DEG C, 15 min centrifugations
Afterwards, supernatant is collected, is then carried out filtering remaining thalline in removal supernatant with 0.22 μm of filter membrane, collects filtrate, obtain marsh
Red pseudomonas zymotic fluid.
Comparative example 1：
The zymotic fluid obtained in embodiment 1 is put into autoclave, the fermentation that 30 min obtain inactivation is inactivated at 121 DEG C
Liquid.
Embodiment 2：Prevention and control of the Rhodopseudomonas palustris LY-6 zymotic fluids field to leaf pest
Respectively by zymotic fluid, the chemical pesticide of inactivation obtained in zymotic fluid made from embodiment 1, comparative example 1（Rice blast
Spirit, 120 mL/ mus）It is sprayed on the rice leaf in tiller later stage, each processing sets 3 repetitions, and 20 L, sprays every 15 d per acre
Once, it co-administers 3 times.After 15 d of last time dispenser, rice leaf disease index is calculated by 5 method samplings, and calculate respective
Control effect.
Fig. 1 is that the zymotic fluid of inactivation, the zymotic fluid of embodiment 1 and the chemical pesticide of comparative example 1 prevent leaf blast in rice
Effect contrast figure is controlled, A, B represent significant difference（P<0.01）.As can be known from Fig. 1：Using the zymotic fluid of inactivation, zymotic fluid with
And the respective control effect of pesticide is 8.7 %, 47.4 % and 44.4 %, and is found by statistical analysis, spray zymotic fluid and
The control effect of chemical pesticide does not have significant difference.
Embodiment 3：Prevention and control of the Rhodopseudomonas palustris LY-6 zymotic fluids field to fringe pest
Respectively by the zymotic fluid and chemical pesticide of inactivation obtained in zymotic fluid made from embodiment 1, comparative example 1（Rice blast
Spirit, 120 mL/ mus）It is sprayed on the rice leaf at heading stage, each processing sets 3 repetitions, and 20 L, one is sprayed every 15 d per acre
It is secondary, it co-administers 2 times.After 15 d, rice tassel disease index is calculated by 5 method samplings, and calculate respective control effect.
Fig. 2 is that the zymotic fluid of inactivation, the zymotic fluid of embodiment 1 and the chemical pesticide of comparative example 1 prevent Pyricularia Oryzae
Effect contrast figure is controlled, A, B represent significant difference（P<0.01）.It is respective using the zymotic fluid, zymotic fluid and pesticide of inactivation
Control effect is 11.4 %, 34.7 % and 46.3 %, and is found by statistical analysis, and the anti-of zymotic fluid and chemical pesticide is sprayed
Controlling effect does not have significant difference.
The above described is only a preferred embodiment of the present invention, being not intended to limit the present invention in any form.Though
So the present invention has been disclosed with preferred embodiment as above, and however, it is not intended to limit the invention.It is any to be familiar with those skilled in the art
Member, in the case where not departing from the Spirit Essence and technical solution of the present invention, all using in the methods and techniques of the disclosure above
Appearance makes many possible changes and modifications to technical solution of the present invention, or is revised as the equivalent embodiment of equivalent variations.Therefore,
Every content without departing from technical solution of the present invention is made to the above embodiment any simple according to the technical essence of the invention
Modification, equivalent replacement, equivalence changes and modification, still fall within technical solution of the present invention protection in the range of.
Claims (10)
1. application of the Rhodopseudomonas palustris zymotic fluid in prevention and control rice blast, which is characterized in that the Rhodopseudomonas palustris
Zymotic fluid is by Rhodopseudomonas palustris LY-6（Rhodopseudomonas palustris LY-6）Activated, seed culture, life
Then bacterium solution is collected filtrate and obtained by production culture to exponential phase through centrifugal filtration.
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 born of the same parents' fermented liquid in prevention and control leaf blast in rice be：After the Rhodopseudomonas palustris zymotic fluid is sprayed on tiller
On the rice leaf of phase.
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 born of the same parents' fermented liquid in prevention and control Pyricularia Oryzae be：The Rhodopseudomonas palustris zymotic fluid is sprayed on heading stage
Rice leaf.
4. application according to any one of claim 1 to 3, which is characterized in that the Rhodopseudomonas palustris zymotic fluid
The amount of spraying be 15 L/ mus~20 L/ mus, sprayed once every the d of 10 d~15.
5. application according to any one of claim 1 to 3, which is characterized in that the Rhodopseudomonas palustris zymotic fluid
Preparation method specifically include 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 accesses 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；
（4）It is prepared by zymotic fluid：By step（3）Filtrate is collected in the bacterium solution centrifugal filtration that productive culture obtains, and as marsh is red false single
Born of the same parents' fermented liquid.
6. application according to claim 5, which is characterized in that the step（2）In, seed culture temperature be 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 be 30 DEG C~
32 DEG C, illumination condition be 2500 Lx~4000 Lx, pH=7.0 under the conditions of carry out.
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 %~10 of Photosynthetic bacterium strain production medium total volume.
9. application according to claim 5, which is characterized in that the step（4）In, the rotating speed of the centrifugation is 8000
The rpm of rpm~12000, temperature are 4 DEG C, centrifugation time is the min of 15 min~20.
10. application according to claim 5, which is characterized in that the step（4）In, the centrifugal filtration is specially to take
Supernatant after centrifugation is filtered with 0.22 μm of filter membrane.
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN110294794A (en) * | 2019-07-09 | 2019-10-01 | 湖南省植物保护研究所 | A kind of Rhodopseudomonas palustris GroEL albumen and its preparation method and application |
| 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 |
| 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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