CN105794853A - Application of rhodopseudomonas palustris fermentation liquor to rice blast prevention and control - Google Patents
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Abstract
The invention discloses application of rhodopseudomonas palustris fermentation liquor to rice blast prevention and control.The rhodopseudomonas palustris fermentation liquor is obtained by culturing rhodopseudomonas palustris LY-6.Rhodopseudomonas palustris is used for preventing and controlling rice blast for the first time, and the rhodopseudomonas palustris fermentation liquor has the advantages of being environmentally friendly, nontoxic to human and livestock, free of phytotoxicity to crops, easy and convenient to apply and the like.
Description
Technical field
The present invention relates to biological pesticide technical field, particularly relate to a kind of Rhodopseudomonas palustris fermentation liquid at prevention and control rice blast
Application in disease.
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), red
3 sections of Spirillaceae (Rhodospirillaceae).Photosynthetic bacteria can secrete substantial amounts of albumen and secondary metabolism in incubation
Product, the metabolic function of its complexity and unique physiological and ecological characteristic make it at farming animals industry, feedstuff, environmental protection, food processing, biology
It is used widely in the field such as the energy and medicines and health protection.
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, and actinomycetes fermentation liquor is to rice blast
The preventing and treating of sick leaf pestilence also has certain effect.Photosynthetic bacteria Rhodopseudomonas palustris fermentation liquid in the prevention and control of rice blast the most not
Appear in the newspapers.
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 fermentation
Liquid application in prevention and control rice blast, utilizes the microbial inoculum that Rhodopseudomonas palustris bacterial strain is made, and production cost is low, prevents rice blast
Control effect is notable.
For solve above-mentioned technical problem, it is provided that a kind of Rhodopseudomonas palustris fermentation liquid in the application of prevention and control rice blast,
Described Rhodopseudomonas palustris fermentation liquid is by Rhodopseudomonas palustris LY-6(Rhodopseudomonas palustris LY-6)
Cultivation obtains.Preferably, described cultural method includes the steps such as activation, seed culture, productive culture, centrifugal filtration.
Above-mentioned application, it is preferred that described rice blast is leaf blast in rice, described Rhodopseudomonas palustris fermentation liquid is in prevention and control
Application process in leaf blast in rice is: sprayed on the rice leaf in tiller later stage by described Rhodopseudomonas palustris fermentation liquid.
Above-mentioned application, it is preferred that described rice blast is Pyricularia Oryzae, described Rhodopseudomonas palustris fermentation liquid is in prevention and control
Application process in Pyricularia Oryzae is: described Rhodopseudomonas palustris fermentation liquid is sprayed the rice leaf heading stage.Above-mentioned
Application, it is preferred that the concentration of described Rhodopseudomonas palustris fermentation liquid is 5 × 109Cfu/mL～5 × 1010Cfu/mL, spray
The amount of executing 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 fermentation liquid 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: 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；
(4) prepared by fermentation liquid: the bacterium solution centrifugal filtration obtained by step (3) productive culture, collects filtrate, is marsh red false single
Born of the same parents' fermented liquid.
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.
Above-mentioned application, it is preferred that in described step (4), described centrifugal rotating speed is 8000 rpm～12000 rpm,
Temperature is 4 DEG C, centrifugation time is 15 min～20 min.
Above-mentioned application, it is preferred that in described step (4), described centrifugal filtration specially takes the supernatant after being centrifuged, uses
0.22 μm filter membrane carries out sucking filtration.
The Rhodopseudomonas palustris LY-6 (Rhodopseudomonas palustris LY-6) that the application is used, protects
Being 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 positioned at Wuhan, China university, preservation day
Phase 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 fermentation liquid of 6, can effectively prevent and treat rice blast, such as leaf blast in rice, Pyricularia Oryzae；There is ring
Border is friendly, 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 fermentation liquid 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 fermentation liquid in land for growing field crops prevention effect comparison diagram to leaf blast in rice.
Fig. 2 be in present example 3 fermentation liquid 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), is preserved in China typical culture collection center, and deposit number is
CCTCC NO:M 2014525, depositary institution address is positioned at Wuhan, China university, and preservation date is on November 5th, 2014.
Embodiment 1:
The preparation method of the Rhodopseudomonas palustris LY-6 fermentation liquid 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 5 % of Photosynthetic bacterium strain production medium cumulative volume, temperature be 30 DEG C～32 DEG C, illumination condition be 2500 Lx～
Under the conditions of 4000 Lx, pH=7.0, cultivate to exponential phase.
(4) prepared by fermentation liquid: the bacterium solution that step (3) productive culture obtains is carried out 8000 rpm, 4 DEG C, 15 min be centrifuged
After, collect supernatant, then carry out sucking filtration with 0.22 μm filter membrane and remove the thalline of residual in supernatant, collect filtrate, i.e. prepare marsh
Rhodopseudomonas fermentation liquid.
Comparative example 1:
The fermentation liquid obtained in embodiment 1 is put in autoclave, inactivates 30 min at 121 DEG C and i.e. obtain the fermentation liquid of inactivation.
Embodiment 2: the Rhodopseudomonas palustris LY-6 fermentation liquid field prevention and control to leaf pestilence
Fermentation liquid (the concentration 5 × 10 respectively embodiment 1 prepared9Cfu/mL), in comparative example 1 prepare inactivation fermentation liquid,
Chemical pesticide (Isoprothiolane, 120 mL/ mus) sprays in the rice leaf in tiller later stage, and each process sets 3 repetitions, every mu 20
L, sprays once every 15 d, uses 3 times altogether.After last dispenser 15 d, calculate the rice leaf state of an illness by 5 method samplings and refer to
Number, and calculate respective prevention effect.
Fig. 1 is that leaf blast in rice is prevented by the fermentation liquid of the inactivation of comparative example 1, the fermentation liquid of embodiment 1 and chemical pesticide
Controlling effect contrast figure, A, B represent significant difference (P < 0.01).As can be known from Fig. 1: use the fermentation liquid of inactivation, fermentation liquid with
And the respective prevention effect of pesticide be 8.7 %, 47.4 % and 44.4 %, and found by statistical analysis, spray fermentation liquid and
The prevention effect of chemical pesticide does not has significant difference.
Embodiment 3: the Rhodopseudomonas palustris LY-6 fermentation liquid field prevention and control to fringe pestilence
Fermentation liquid (the concentration 5 × 10 respectively embodiment 1 prepared9Cfu/mL), the fermentation liquid of the inactivation prepared in comparative example 1
Spraying the rice leaf heading stage with chemical pesticide (Isoprothiolane, 120 mL/ mus), each process sets 3 repetitions, every mu 20
L, sprays once every 15 d, uses 2 times altogether.After 15 d, calculate Oryza sativa L. tassel disease index by 5 method samplings, and calculate each
From prevention effect.
Fig. 2 is that Pyricularia Oryzae is prevented by the fermentation liquid of the inactivation of comparative example 1, the fermentation liquid of embodiment 1 and chemical pesticide
Controlling effect contrast figure, A, B represent significant difference (P < 0.01).Use the fermentation liquid of inactivation, fermentation liquid and pesticide respective
Prevention effect is 11.4 %, 34.7 % and 46.3 %, and is found by statistical analysis, sprays the anti-of fermentation liquid and chemical pesticide
Control effect and there is no 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 (10)
1. Rhodopseudomonas palustris fermentation liquid application in prevention and control rice blast, it is characterised in that described Rhodopseudomonas palustris
Fermentation liquid is by Rhodopseudomonas palustris LY-6(Rhodopseudomonas palustris LY-6) cultivate obtain.
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' fermented liquid application process in prevention and control leaf blast in rice is: sprayed after tiller by described Rhodopseudomonas palustris fermentation liquid
On the rice leaf of phase.
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' fermented liquid application process in prevention and control Pyricularia Oryzae is: spray described Rhodopseudomonas palustris fermentation liquid heading stage
Rice leaf.
Application the most according to any one of claim 1 to 3, it is characterised in that described Rhodopseudomonas palustris fermentation liquid
Concentration be 5 × 109Cfu/mL～5 × 1010Cfu/mL, the amount of spraying is 15 L/ mus～20 L/ mus, every 10 d～15 d
Spray once.
Application the most according to any one of claim 1 to 3, it is characterised in that described Rhodopseudomonas palustris fermentation liquid
Preparation method specifically include 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；
(4) prepared by fermentation liquid: the bacterium solution centrifugal filtration obtained by step (3) productive culture, collects filtrate, is marsh red false single
Born of the same parents' fermented liquid.
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.
Application the most according to claim 5, it is characterised in that in described step (4), described centrifugal rotating speed is 8000
Rpm～12000 rpm, temperature are 4 DEG C, centrifugation time is 15 min～20 min.
Application the most according to claim 5, it is characterised in that in described step (4), described centrifugal filtration specially takes
Supernatant after Li Xin, carries out sucking filtration with 0.22 μm filter membrane.
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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 |
| 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 |
| 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 |
| 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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| CN102876615B (en) * | 2012-10-17 | 2014-04-16 | 湖南省植物保护研究所 | Rhodopseudomonas palustris strains, application of rhodopseudomonas palustris strains, fungicide pesticide as well as preparation method and application of fungicide pesticide |
| CN103224904B (en) * | 2013-05-24 | 2014-05-21 | 湖南省植物保护研究所 | Rhodopseudomonas strain, biocontrol microbial inoculum and biocontrol fermentation liquid, and corresponding preparation methods and application thereof in controlling phytophthora blight of pepper |
| CN104403978B (en) * | 2014-12-10 | 2018-01-12 | 湖南省植物保护研究所 | Rhodopseudomonas palustris bacterial strain, the preparation method of microbial inoculum and microbial inoculum, extracellular protein and its extracting method and application |
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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 |
| 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 |
| 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 |
| 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 |
| 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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