CN107251909B - Application of the Pseudomonas fluorescens pf27 in anti-Bemisia tabaci and prevention and treatment potato disease - Google Patents
Application of the Pseudomonas fluorescens pf27 in anti-Bemisia tabaci and prevention and treatment potato disease Download PDFInfo
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Abstract
The invention discloses application of the Pseudomonas fluorescens pf27 in anti-Bemisia tabaci and prevention and treatment potato disease.Pseudomonas fluorescens pf27 or its microbial inoculum of the invention or its fermentation liquid or its be metabolized liquid and have a good control efficiency to the Bemisia tabaci for endangering numerous crop pests such as the vegetables such as tomato, cucumber and cotton, and being capable of the main potato disease such as wide spectrum, efficient inhibition of potato tar spot, late blight and potato scab.The effect that the anti-Bemisia tabaci insect pest of crops not only can be improved, also solves the problems such as pesticide residue and environmental pollution, may advantageously facilitate environmental protection and agricultural sustainable development, with good application prospect.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to Pseudomonas fluorescens pf27 is in anti-Bemisia tabaci and prevention and treatment potato
Application in disease.
Background technique
Potato is the biggish crops of current plantation amount, since its growth adaptability is strong, delicious and full of nutrition etc.
Feature, planting range is larger, and the fourth-largest staple food grain being gradually identified as after corn, wheat, rice, people are to potato
Demand is also increasing.Some diseases are easy to appear during potato growth, such as late blight, early blight, shot hole and disease
The multiple diseases such as viral disease have seriously affected the growing state of potato, result even in potato tubers rot etc..And agriculture
Industry pest Bemisia tabaci is commonly called as little Bai moth, is a kind of insect pest of kainogenesis in recent years, seriously endangers the vegetables such as tomato, cucumber, capsicum
And numerous crops such as cotton.Currently, both at home and abroad to anti-Zhiduo County of pest and disease damage from cultural control and chemical prevention, control efficiency
It is unobvious, and cause a degree of environmental pollution.
Biological control is more and more paid attention to due to starting the advantages of its own.Pseudomonad usually has plant rush
Raw effect, while plant can be enhanced for the resistance of pathogen and abiotic stress, it is referred to as plant-growth promoting rhizobacteria (Plant
Growth-promoting bacteria, PGPB) or plant growth-promoting rhizobacteria, Pseudomonas fluorescens (Psedomonas
It fluorescens, can be by inhibiting the generation of pathogen, inducible system resistance, generating plant) as a kind of important biocontrol bacteria
Object hormone promotes the growth and development of plant with plant growth amount is improved, and is most common in microbial manure and biological prevention and control agent production
It is also one of most important strain.Shi Jingying etc. (2011) research discovery pseudomonas aeruginosa (P.aeruginosa) MGP3 fermentation
Liquid respectively reaches 50% and 71% to the control efficiency for adopting rear papaya anthracnose and phytophthora root rot, and MGP3 bacterium solution can be significant
Reduce the latent infection rate of fruit anthrax-bacilus and the disease index of anthracnose in addition to seedling stage.Wei Qiaojie etc. (2013) research discovery is short of money
Antibacterial Bacillus sp.B and organic fertilizer fermentation, which are used in combination, has significant preventive and therapeutic effect to cucumber fusarium axysporum, and disease incidence reduces
66.7%, disease index has dropped 67%, control efficiency reaches 80.7%；And simple manure use disease incidence not only cannot
It reduces, but also is risen.
Therefore, develop new, highly efficient and safe microbial bactericide control disease be improve society
The needs of production, while the needs of even more agricultural safety sustainable development, meet social development demand.
Summary of the invention
It is an object of the present invention to provide Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its
Fermentation liquid or its new application for being metabolized liquid or its bacteria suspension or its culture solution.
The present invention provides Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its fermentation liquid or its
It is metabolized the application of liquid or its bacteria suspension or its culture solution in prevention and treatment potato disease.
The present invention also provides Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its fermentation liquid or
It is metabolized the application of liquid or its bacteria suspension or its culture solution in the product of preparation prevention and treatment potato disease.
The present invention also provides Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its fermentation liquid or
It is metabolized liquid or its bacteria suspension or its culture solution antibacterial and/or it is antibacterial in application.
The present invention also provides Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its fermentation liquid or
It is metabolized the application of liquid or its bacteria suspension or its culture solution in preparation antibacterial and/or antibacterial product.
The present invention also provides Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its fermentation liquid or
It is metabolized the application of liquid or its bacteria suspension or its culture solution in prevention and treatment agricultural pests.
The present invention also provides Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its fermentation liquid or
It is metabolized the application of liquid or its bacteria suspension or its culture solution in the product of preparation prevention and treatment agricultural pests.
In above-mentioned application, the agricultural pests are Bemisia tabaci Bemisia tabaci.
The present invention also provides Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its fermentation liquid or
It is metabolized liquid or its bacteria suspension or its culture solution and is improving the application in resistivity of the plant to Bemisia tabaci.
The present invention also provides Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its fermentation liquid or
It is metabolized liquid or its bacteria suspension or its culture solution and improves the application in product of the plant to the resistivity of Bemisia tabaci in preparation.
It is a further object to provide a kind of products；The product has following 1) -4) in any function:
1) potato disease is prevented and treated；
2) antibacterial and/or antibacterial.
3) agricultural pests are prevented and treated；The agricultural pests are specially Bemisia tabaci；
4) plant is improved to the resistivity of Bemisia tabaci；
The active constituent of product provided by the invention is Pseudomonas fluorescens Pseudomonas fluorescens or its bacterium
Agent or its fermentation liquid or its metabolism liquid or its bacteria suspension or its culture solution；
In above-mentioned application or product, the Pseudomonas fluorescens Pseudomonas fluorescens is Pseudomonas
Bacterium Pseudomonas fluorescens pf27.
In above-mentioned application or product, the potato disease is potato epiphyte diseases, potato oomycetes disease and Ma Ling
Potato bacteriosis；
The potato epiphyte diseases are specially black scurf of potato；
The potato oomycetes disease is specially the late blight of potato；
The Potato Bacterial Diseases are specially potato scab.
In above-mentioned application or product, the bacterium is black scurf of potato bacterium and/or phytophthora infestans and/or potato
Streptomyces scabies.The black scurf of potato bacterium is specially black scurf of potato bacterium GS-3；The phytophthora infestans are specially
Phytophthora infestans 88069 and T30-4；The potato scab bacterium be specially potato scab bacterium 4.1789,
4.0076 with 4.1756.
It is a still further object of the present invention to provide a kind of methods for preventing and treating potato disease.
The method of prevention and treatment potato disease provided by the invention includes using Pseudomonas fluorescens Pseudomonas
Fluorescens pf27 or its microbial inoculum or its fermentation liquid or its metabolism liquid or its bacteria suspension or its culture solution processing potato children
The step of seedling.
The last one purpose of the invention is to provide a kind of method for preventing and treating agricultural pests Bemisia tabaci.
The method of prevention and treatment agricultural pests Bemisia tabaci provided by the invention includes using Pseudomonas fluorescens Pseudomonas
Fluorescens pf27 or its microbial inoculum or its fermentation liquid or its metabolism liquid or its bacteria suspension or its culture solution handle plant seedlings
The step of.
In the above method, the plant can be the plants such as Cruciferae, Curcurbitaceae, pulse family, Solanaceae and Malvaceae；The calabash
Lu Ke plant concretely cucumber, the plant of Solanaceae concretely tomato, potato and tobacco；The malvaceae plant is specific
It can be cotton.
In above-mentioned application or product or method,
The pf27 microbial inoculum the preparation method is as follows: Pseudomonas fluorescens pf27 is inoculated in KB culture medium, at 28 DEG C,
Shaken cultivation 12-16h under the conditions of 200rpm, then 4000rpm is centrifuged 20min, abandons supernatant, collects bacterial sediment, then with going out
The bacterial sediment is diluted to arrive pf27 microbial inoculum by bacterium water.
The pf27 fermentation liquid the preparation method is as follows: Pseudomonas fluorescens pf27 is inoculated in KB culture solution, 28
DEG C, shaken cultivation 12-16h under the conditions of 200rpm, then 4000rpm is centrifuged 20min, collects supernatant and ferments to get to pf27
Liquid.
Pf27 metabolism liquid the preparation method is as follows: by pf27 fermentation liquid by 0.22 μm of membrane filtration (step
Purpose is degerming), filtrate, which is collected, to get to pf27 is metabolized liquid.
The classification naming of Pseudomonas fluorescens Pseudomonas fluorescens pf27 of the invention is that fluorescence is false single
Born of the same parents bacterium Pseudomonas fluorescens, the bacterial strain are preserved in Chinese microorganism strain preservation pipe on May 8th, 2017
Reason committee common micro-organisms center (abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences
Institute of microbiology, postcode 100101), deposit number is CGMCC No.14105.
The present invention screens by the exploitation again to existing bacterial strain and endangers Bemisia tabaci to more hosts and potato is mainly sick
Evil has the Pseudomonas fluorescens pf27 of control efficiency.It is experimentally confirmed: can be induced kind after pf27 microbial inoculum processing tomato seedling
The resistance of eggplant plant pair pest Bemisia tabaci, inhibitory effect reach about 50% in terms of laying eggs with larvae development, two-way choice
Experimental data shows that pf27 has apparent anthelminthic effect to Bemisia tabaci.In the fungistatic effect to the Major Diseases of potato,
Pf27 fermentation liquid is 82.56% to the fungistatic effect of black scurf of potato GS-3, and 88069 fungistatic effect of the late blight of potato is
47.33%, while pf27 metabolism liquid is respectively to the fungistatic effect that is averaged of potato scab 4.0076,4.1576 and 4.1789
49.74%, 39.39% and 45.47%, and E. coli is to the average bacteriostasis rate point of the bacteriosis of these three strains
It Zhi You 5.73%, 17.37% and 11.89%.The above results absolutely prove that pf27 fermentation liquid or metabolism liquid are main to potato
Disease has the fungistatic effect of wide spectrum.Pf27 microbial inoculum or pf27 fermentation liquid of the invention or pf27 metabolism liquid are as a kind of biological source
Pesticide has the advantages that wide spectrum, efficient and safe, not only avoids chemical pesticide and bring is largely used to remain and environmental pollution
The problems such as, and may advantageously facilitate environmental protection and agricultural sustainable development, with good application prospect.
Detailed description of the invention
Fig. 1 is the pf27 phylogenetic tree based on 16s rDNA sequence construct.
Fig. 2 is resistivity of the microbial bacterial agent pf27 to Bemisia tabaci.A be Bemisia tabaci lay eggs on tomato seedling plant with
Develop the schematic diagram of four-age larva；B is that Bemisia tabaci is planted in the tomato of control tomato plant (Control) and the processing of pf27 microbial inoculum
The two-way choice schematic diagram of strain；C is the number that Bemisia tabaci is laid eggs in the processing of pf27 microbial inoculum and the tomato plant for compareing (Control)
Mesh statistics.D is ratio of the Bemisia tabaci adults to two-way choice on the tomato plant of the processing of pf27 microbial inoculum and control (Control)
Statistics.E is that four-age larva is arrived in Bemisia tabaci development in the processing of pf27 microbial inoculum and the tomato plant single blade for compareing (Control)
Number statistical.F is that Bemisia tabaci adults are handled in pf27 microbial inoculum and compareed on the potato seedling of (Control) under natural conditions
Growth conditions.Note: * indicates that in 0.05 horizontal upper significant difference, * * is indicated in 0.01 horizontal upper significant difference (under).
Fig. 3 is the incidence statistics that the 4th, 5 cauline leaf of potato is inoculated with black scurf of potato GS-3, photograph taking inoculation 6
After it.A is that pf27 microbial inoculum processing group and control group potato leaf are inoculated with the hair after black scurf of potato GS-3 spore suspension
State of an illness condition, wherein upper figure is the incidence of control group potato leaf inoculation black scurf of potato GS-3 spore suspension；Under
Figure is pf27 microbial inoculum processing group potato leaf inoculation GS-3 spore suspension incidence.Each 4 biology of processing repeat,
Experiment is repeated 3 times.B is the scab that pf27 microbial inoculum processing group and control group potato leaf are inoculated with black scurf of potato GS-3 morbidity
Area statistics, n=12.
Fig. 4 is the incidence that the 4th, 5 cauline leaf of potato is inoculated with the late blight of potato 88069 and T30-4, and photograph taking connects
Kind is after 6 days.A is the incidence that the 4th, 5 cauline leaf of potato is inoculated with late blight of potato T30-4, wherein upper figure is pf27 microbial inoculum
The incidence of processing group potato leaf inoculation late blight of potato T30-4 spore suspension；The following figure is control group potato
Blade inoculation late blight of potato T30-4 spore suspension incidence, each 4 biology of processing repeat, and experiment repeats 3
It is secondary.B is the lesion area data that pf27 microbial inoculum processing group and control group potato leaf are inoculated with late blight of potato T30-4 morbidity
Statistics, n=12.C is the incidence that the 4th, 5 cauline leaf of potato is inoculated with the late blight of potato 88069, wherein upper figure is pf27
The incidence of microbial inoculum processing group potato leaf inoculation 88069 spore suspension of the late blight of potato；The following figure is control group horse
88069 spore suspension incidence of the bell potato blade inoculation late blight of potato, each 4 biology of processing repeat, experiment weight
It is 3 times multiple.D is pf27 microbial inoculum processing group and control group potato leaf is inoculated with the lesion area that the late blight of potato 88069 is fallen ill,
N=12.
Fig. 5 is suppression of the microorganism pf27 fermentation liquid to black scurf of potato GS-3, late blight of potato T30-4 and 88069
Bacterium schematic diagram.
Fig. 6 is bacteriostatic experiment of the pf27 microbial inoculum to black scurf of potato GS-3, the late blight of potato 88069 and T30-4.A
For the growing state of pf27 microbial inoculum processing group and the control group plating late blight of potato 88069 and T30-4 bacterial plaque, wherein
Left figure is the growing state of 88069 bacterial plaque of pf27 microbial inoculum processing group and the control group plating late blight of potato, and right figure is
Pf27 microbial inoculum processing group and control group plating late blight of potato T30-4 bacterial plaque growing state.B is pf27 microbial inoculum processing group
With the growing state of control group plating black scurf of potato GS-3 bacterial plaque.C is pf27 microbial inoculum to the late blight of potato
88069, the column diagram statistics of the inhibiting rate of T30-4 and black scurf of potato GS-3.Each 4 biology of processing repeat, experiment
It is repeated 3 times.
Fig. 7 is that pf27 is metabolized liquid to the bacteriostatic experiment of potato scab 4.1789,4.0076 and 4.1756.A is inoculation
The experimental group of potato scab 4.1789,4.0076 and 4.1756, control group, the bacterial plaque in blank group culture medium grow feelings
Condition.B is to be inoculated with the experimental group of potato scab 4.1789,4.0076 and 4.1756, control group, the suppression in blank group culture medium
The column diagram of rate processed counts.Each 4 biology of processing repeat, and experiment is repeated 3 times.
Preservation explanation
Strain name: Pseudomonas fluorescens
Latin name: Pseudomonas fluorescens
Strain number: pf27
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism abbreviation: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on May 8th, 2017
Collection is registered on the books number: CGMCC No.14105
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Quantitative test in following embodiments, is respectively provided with three repeated experiments, and results are averaged.
(fluid nutrient medium only removes the agar powder in solid medium used medium formula as follows in following embodiments
Remove, and keep other components and concentration constant):
MEA culture medium (solid): Malt extract 30g, Mycological peptone 5g, agar powder 15g,
pH5.4；
Beef-protein medium (solid): peptone 10g, beef extract 3g, sodium chloride 5g, agar powder 12g, water
1000mL, pH 7.2-7.4；
LB culture medium (solid): sodium chloride 10g, peptone 10g, yeast extract 5g, agar powder 12g, water 1000mL, pH
7.0；
KB culture medium (solid): peptone 20g, K2HPO41.5g, glycerol 8ml, MgSO40.74g, H2O supplies 1L,
pH 7.0；Solid medium adds agar powder 12g.
PDA culture medium (solid): peeled potatoes 200g, glucose 20g, agar powder 15g, water 1000mL.
10%V8 culture medium (solid): the V8 juice 4000rpm bought in supermarket (is centrifuged 10min, collected by V8 juice supernatant
Supernatant) 100mL, CaCO31g, agar powder 15g, water 1000mL.
Black scurf of potato bacterium GS-3 in following embodiments is recorded in following document: Zhao Wei congruence, 2006, Chinese Ma Ling
The identification of potato Streptomyces scabies, the public can obtain from applicant, which only attaches most importance to used in the related experiment of duplicate invention,
It not can be used as other purposes to use.
Phytophthora infestans T30-4 in following embodiments is recorded in following document: Sansome E etc., Disploidy
and chromosomal structural hybridity in Phytophthora infestans.Nature)；Potato
Late disease bacteria 88069 is recorded in following document: Liu Wei, the clone of late blight of potato resistance related gene StBL-SLGPK and function
It can analysis；The public can obtain from applicant, which only attaches most importance to used in the related experiment of duplicate invention, not can be used as it
Its purposes uses.
Potato scab bacterium 4.0076 in following embodiments is purchased from Chinese agriculture Microbiological Culture Collection administrative center,
Strain catalog number is ACCC40076；Potato scab bacterium 4.1756 and 4.1789 is purchased from Chinese microorganism strain preservation pipe
Reason committee common micro-organisms center, deposit number are respectively CGMCC No.4.1756 and CGMCC No.4.1789.
The separation and identification of embodiment 1, pf27 bacterial strain
One, the separation of pf27 bacterial strain
Pf27 is isolated from Yunnan Province, oryza meyeriana area, Puer City rhizosphere soil (north latitude 22。33 ' 56 ", 100。35 ' 12 ", sea
Pull out 737 meters).Specific separation method is as follows: will pick up from the rhizosphere soil in oryza meyeriana area, picking root system scrapes Rhizosphere Soil, claims
5g is measured, is put into the sterilizing triangular flask being ready under aseptic condition, with aqua sterilisa gradient dilution to 10-6Times, 3 weights are set
It is multiple.By the pedotheque of gradient dilution take 200 μ L be coated onto respectively MEA culture medium, beef-protein medium, LB culture medium and
It on KB culture medium flat plate, is cultivated in 28 DEG C, 37 DEG C of incubators respectively, observes the growth conditions of bacterium colony daily, it is dilute as the result is shown
It is interpreted as 10-5、10-6Times when plate on there is no that bacterium colony is grown, and be diluted to 10-4Times when, single bacterium can be grown on plate
It falls, and picking grows single bacterium colony on KB culture medium, is named as pf27 bacterial strain, then expand in KB fluid nutrient medium
Further bacterial strain identification is done in big culture.
Two, the identification of pf27 bacterial strain
1, the Morphological Identification of pf27 bacterial strain
Pf27 bacterial strain can form orange colonies after cultivating for 24 hours~48h on KB culture medium, and bacterium colony state is rounded, surface
Smooth have that raised brim is neat, more sticky, easily provokes.
2, the Molecular Identification of pf27 bacterial strain
The DNA for extracting pf27 bacterial strain carries out PCR amplification using universal primer Eubac27F/Eubac1492R, is contained
The PCR product of the conserved region pf27 bacterial strain 16S rDNA.Primer sequence is as follows:
Eubac27F:5 '-AGAGTTTGATCCTGGCTCAG-3 '；
Eubac1492R:5 '-GGTTACCTTGTTACGACTT-3 '.
PCR product is connected to pEASY-T3 (buying from the complete biological Co., Ltd of formula gold in Beijing) through TA clone, with general
Primer is sequenced in Invitrogen Corp..
Sequencing result shows: the nucleotide sequence of PCR product is as shown in sequence 1.It will sequencing gained sequence and NCBI data
After library carries out blast comparison, by MEGA5.1 software to pf27 and other represent 22 kinds of different pseudomonad kinds fluorescence it is false single
Born of the same parents' type strain carries out Phylogenetic Analysis, and phylogenetic tree is as shown in Figure 1.
In summary qualification result determines that the classification naming of pf27 bacterial strain is Pseudomonas fluorescens Pseudomonas
Fluorescens, the bacterial strain are preserved in China Committee for Culture Collection of Microorganisms's commonly micro- life on May 8th, 2017
Object center (abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postal
It compiles 100101), deposit number is CGMCC No.14105.
The preparation of embodiment 2, microorganism pf27 microbial inoculum, microorganism pf27 fermentation liquid and microorganism pf27 metabolism liquid
1, the preparation of microorganism pf27 microbial inoculum
The Pseudomonas fluorescens pf27 obtained in embodiment 1 is inoculated in KB culture medium, at 28 DEG C, under the conditions of 200rpm
Shaken cultivation 12-16h, then 4000rpm is centrifuged 20min, abandons supernatant, collects bacterial sediment, then thallus is sunk with aqua sterilisa
Shallow lake is diluted, and microorganism pf27 microbial inoculum (microbial inoculum pf27) is made, and pf27 viable bacteria total concentration is 1 in microorganism pf27 microbial inoculum finished product
×109-1×1010CFU/mL。
2, the preparation of microorganism pf27 fermentation liquid
The Pseudomonas fluorescens pf27 obtained in embodiment 1 is inoculated in KB culture solution, at 28 DEG C, under the conditions of 200rpm
Shaken cultivation 12-16h, then 4000rpm is centrifuged 20min, collects supernatant, as microorganism pf27 fermentation liquid, microorganism
Pf27 viable bacteria total concentration is 1 × 10 in pf27 fermentation liquid finished product2-1×103CFU/mL。
3, microorganism pf27 is metabolized the preparation of liquid
Microorganism pf27 fermentation liquid in step 2 is passed through into 0.22 μm of membrane filtration (purpose of the step is degerming), is collected
Filtrate, as microorganism pf27 are metabolized liquid.
Embodiment 3, microorganism pf27 microbial inoculum are to the resistivity of agricultural pests Bemisia tabaci
One, experimental method
1, tomato plant
(1) miscellaneous No. 9 tomato seeds (scientific and technological (Beijing) the Co., Ltd production of middle vegetable kind industry) are disappeared with 3%NaClO solution by
After malicious 10min, ddH is used2O is cleaned 6 times, seed after being sterilized.Then seed after disinfection is uniformly layered on to the vermiculite mud of sterilizing
In charcoal soil matrix matter (vermiculite and peat soil are mixed according to the ratio of 1:1,121 DEG C of high pressure sterilization 20min), at 26 DEG C, illumination: dark
Under the conditions of (12h:12h) cultivate seed sprout, when seed germination and growth arrive two panels true leaf when, be transplanted to respectively uniformly add pf27
(the pf27 microbial inoculum that every kg vermiculite peat soil matrix addition 50mL OD value is 1, pf27 microbial inoculum make the vermiculite peat soil matrix of microbial inoculum
With final concentration of 1 × 105CFU/mL in the nutrition cave for) neutralizing control (not add any microbial inoculum as control, Control).
Tomato seedling is transplanted in seedling culture hole plate (hole tray specification: 9 centimetres of back cut diameter, 6 centimetres of lower port diameter, 8 centimetres of height), is placed on
Grown in greenhouse (growth conditions: light application time 12h, 26 DEG C of temperature, relative humidity 50%).
(2) single blade for the tomato plant for growing into 5 weeks or so is placed in the plastic cup of ounce (bottom, height, bore
Size is 32mm × 34mm × 40mm, the opening that diameter is 5mm or so is opened in plastic cup side, for Bemisia tabaci to be placed in cup
In；The opening of 2mm or so is opened in the other side from bore position, for placing petiole), the Bemisia tabaci adults (3 that then will just hatch
Head female adult pest and 3 male imagos) from opening it is inoculated into monolithic tomato leaves, then be open envelope with the medical adhesive tape breathed freely
Firmly.The egg laying amount on each tomato leaf is counted after 10 days.Each 8 biology of processing repeat, and experiment is repeated 3 times.
(3) single blade for the tomato plant for growing into 5 weeks or so is placed in the plastic cup of ounce, by 16 it is female at
Worm is inoculated on single tomato leaf, by 2 days egg-laying times, whole adults is removed, counts each tomato after 22 days
The quantity to 4 instar larvaes is developed on blade.Each 8 biology of processing repeat, and experiment is repeated 3 times.
(4) the pf27 microbial inoculum processing group and the tomato seedling of control group that grow into 5 weeks or so are individually positioned in 120 purposes
The two sides of cage (40cm × 40mm × 40mm), then freeze Bemisia tabaci adults 2 minutes on ice, 100 tobacco powder of every number
Lice adult is placed between two plants of tomatoes, and the number of the Bemisia tabaci on two plants of tomatoes is counted after 15 minutes, calculates Bemisia tabaci at two plants
Selection rate on tomato.The calculation method of selection rate: respectively in pf27 processing group and control group plant in 100 Bemisia tabacis of statistics
On Bemisia tabaci number, and be added selection Bemisia tabaci number summation, calculate cigarette on pf27 processing group and control group plant
The shared total Bemisia tabaci percentage selected of aleyrodid.Each 6 biology of processing repeat, and experiment is repeated 3 times.
2, potato plant
By purchase from the environment that the potato in the maiden voyage supermarket of Beijing is placed on moist dark, by after a week, picking out
The bud of the consistent healthy potato of bud, with being cut after 70% alcohol sterilization blade.Then young shoot is transplanted to respectively
Uniformly (every kg vermiculite peat soil matrix addition 50mL OD value is 1 to the vermiculite peat soil matrix of the sterilizing of addition pf27 microbial inoculum
Pf27 microbial inoculum, pf27 microbial inoculum use final concentration of 1 × 105CFU/mL) and control (using do not add any microbial inoculum as compare,
Control in nutrition cave), being placed in plastic flowerpot, (specification: back cut diameter is 14 centimetres, and lower port diameter is 9.5 centimetres, high
Degree be 12.5 centimetres) in culture, surface cover one layer of preservative film moisturizing, preservative film is removed after it grows cotyledon, place from
It is grown under the conditions of so, obtains potato seedling.
(2) growth of the Bemisia tabaci adults under the processing of pf27 microbial inoculum and on the potato seedling of control is observed under natural conditions
State and quantity.Each 6 biology of processing repeat, and experiment is repeated 3 times.
Two, insect resistant effect counts
The quantity and tobacco powder of egg laying amount of the Bemisia tabaci on the blade of selection, development to 4 instar larvaes are counted under anatomical lens
Selection rate of the lice in pf27 microbial inoculum processing group and the tomato plant of control group.
As a result as shown in Figure 2.It is compared with control group, Bemisia tabaci adults are laid eggs on the tomato of microorganism pf27 microbial inoculum processing group
Amount and the quantity developed to 4 instar larvaes significantly reduce, and egg laying amount reduces about 50.11%, and the worm amount developed to 4 instar larvaes is about reduced
33.51%.Bemisia tabaci is shown in the two-way choice experimental data of pf27 microbial inoculum processing group and the tomato plant of control group: pf27 bacterium
The tomato plant of agent processing group has apparent anthelmintic action, the selection rate point of pf27 microbial inoculum processing group and control group to Bemisia tabaci
It is not 35.28% and 64.72%, anthelminthic effect 29.44%.Observe the horse of pf27 microbial inoculum processing group in its natural state simultaneously
Influence of the bell potato seedling plants to Bemisia tabaci, the Bemisia tabaci quantity on potato seedling that discovery microorganism pf27 microbial inoculum is handled are aobvious
Write the Bemisia tabaci quantity for being less than control group.
The influence of embodiment 4, microorganism pf27 microbial inoculum to the pathogenicity of black scurf of potato GS-3
One, experimental method
1, the plantation of potato seedling
By purchase from the environment that the potato in the maiden voyage supermarket of Beijing is placed on moist dark, by after a week, picking out
The bud of the consistent healthy potato of bud, with being cut after 70% alcohol sterilization blade.Then young shoot is transplanted to respectively
Uniformly (every kg vermiculite peat soil matrix addition 50mL OD value is 1 to the vermiculite peat soil matrix of the sterilizing of addition microbial inoculum pf27
Pf27 microbial inoculum, pf27 microbial inoculum use final concentration of 1 × 105CFU/mL) and control (using do not add any microbial inoculum as compare,
Control in nutrition cave), being placed in plastic flowerpot, (specification: back cut diameter is 14 centimetres, and lower port diameter is 9.5 centimetres, high
Degree be 12.5 centimetres) in culture, surface cover one layer of preservative film moisturizing, preservative film is removed after it grows cotyledon, place from
It is grown under the conditions of so, obtains potato seedling.
2, bacterium solution configures
Black scurf of potato bacterium GS-3 is inoculated into PDA culture medium, culture to can observe maturation under the microscope
The black scurf of potato GS-3 of sporangium collects spore, is made using the distilled water flushing culture dish containing 0.01% polysorbas20
Spore concentration is 1.0 × 105The GS-3 spore suspension of CFU/ml, for being inoculated with after then standing 2~3 hours at room temperature.
3, artificial infection and Disease investigation
The single leaf in 10, leaf top that 4 and 5 leaf positions are taken when inoculation, is inserted in the solid agar medium that concentration is 1% for petiole
In upper flat plate, two panels list leaf is placed in each plate, with moisturizing on the distilled water to plate for adding 2mL to sterilize, in each single blade
The heart drips 20 μ L GS-3 spore suspensions in the upper surface of blade, finally covers culture ware lid, is sealed with ventilative medical adhesive tape,
Being placed on temperature is 20 DEG C, illumination: dark (12h:12h), humidity are cultivated under conditions of being 60%.Simultaneously any bacterium is not added dropwise
0.01% polysorbas20 of liquid is as control.Handle the 6th day investigation lesion area, lesion area calculation formula: S=(scab length
× scab width) × π/4.Each 4 biology of processing repeat, and experiment is repeated 3 times.
Two, disease resisting effect counts
As a result as shown in Figure 3.It is compared with control group, the black scurf of potato GS-3 morbidity of microorganism pf27 microbial inoculum processing group
Reduce, lesion area be averaged conspicuousness reduction about 46.61%.It is aobvious to illustrate that microorganism pf27 microbial inoculum has black scurf of potato GS-3
The disease resisting effect of work.
The influence of embodiment 5, microorganism pf27 microbial inoculum to late blight of potato T30-4 and 88,069 two kinds of bacterium pathogenicities
One, experimental method
1, the plantation of potato seedling
With 1 in 4 step 1 of embodiment.
2, bacterium solution configures
Phytophthora infestans T30-4 and 88069 is inoculated into respectively on 10%V8 solid medium, 20 DEG C of dark surrounds
Culture cut mycelia block of uniform size after 2~3 days, was placed in the 10%V8 fluid nutrient medium of 20mL, in 20 DEG C of dark standings
Mycelia clump is moved into sterile culture dish after culture 2~3 days, suitable sterilizing pure water is added, it is left to stand 12h in 26 DEG C of illumination
The right side can produce a large amount of zoosporangiums；Using the distilled water flushing culture dish containing 0.01% polysorbas20, spore, system are collected
It is 1.0 × 10 at spore concentration5Then the T30-4 suspension and 88069 suspension of CFU/ml stands 2~3 hours at room temperature
Afterwards for being inoculated with.
3, artificial infection and Disease investigation
Artificial infection and Disease investigation are the same as 3 in 4 step 1 of embodiment.Each 4 biology of processing repeat, and experiment repeats
3 times.
Two, disease resisting effect counts
As a result as shown in Figure 4.It is compared with control group, the potato leaf on piece of microorganism pf27 microbial inoculum processing group is inoculated with Ma Ling
After potato late blight T30-4 and 88069 pathogens, lesion area conspicuousness is reduced, and the lesion area for being inoculated with T30-4 reduces
53.85%, being inoculated with 88069 lesion area then reduces 79.80%.Illustrate microorganism pf27 microbial inoculum to the late blight of potato
T30-4 and 88069 has significant disease resisting effect.
Embodiment 6, microorganism pf27 fermentation liquid are to black scurf of potato GS-3, late blight of potato T30-4 and 88069
The influence of bacteriostasis rate
One, experimental method
1, pf27 fermentation liquid is added to according to the ratio of 1:50 and has melted and be cooled to 50 DEG C or so of PDA culture medium
In, rear paved plate is mixed well, the PDA culture medium containing pf27 microbial inoculum is obtained.The culture of any microbial inoculum is not added in setting simultaneously
Base is control.
Pf27 fermentation liquid is added to according to the ratio of 1:50 and has melted and be cooled to 50 DEG C or so of 10%V8 culture
In base, rear paved plate is mixed well, obtains the V8 culture medium containing pf27 microbial inoculum.The culture of any microbial inoculum is not added in setting simultaneously
Base is control.
2, bacteriostatic experiment
The black scurf of potato GS-3 disease fungus colony edge cultivated in PDA culture medium 5 days is beaten take diameter be 6mm,
Uniform bacterium dish is grown, mycelia downwards, is inoculated in the center of the PDA culture medium containing pf27 microbial inoculum, in 20 DEG C of constant temperature incubations.
Each 4 biology of processing repeat, and experiment is repeated 3 times.
5 days late blight of potato T30-4 will be cultivated on V8 culture medium respectively and 88069 disease fungus colony edges are beaten and taken
Diameter is 6mm, grows uniform bacterium dish, and mycelia downwards, is inoculated in the center of the V8 culture medium containing pf27 microbial inoculum, in 20 DEG C
Constant temperature incubation.Each 4 biology of processing repeat, and experiment is repeated 3 times.
Observed day by day after inoculation, and with right-angled intersection method measure it is each for examination disease fungus it is straight on culture medium flat plate
Diameter, and bacteriostasis rate is calculated according to the following formula: bacteriostasis rate (%)=(control colony diameter-processing colony diameter)/(control bacterium colony
Diameter-bacteria cake diameter) × 100%, bacteria cake diameter 6mm.Antibacterial schematic diagram is as shown in Figure 5.
Two, fungistatic effect counts
As a result as shown in table 1 and Fig. 7.In the Major Diseases black scurf of potato GS-3 to potato, the late blight of potato
In the fungistatic effect of 88069 and T30-4, pf27 microbial inoculum is 82.56% to the bacteriostasis rate of black scurf of potato GS-3, potato evening
The bacteriostasis rate of epidemic disease 88069 is 47.33%, and the bacteriostasis rate of late blight of potato T30-4 is about 10%.
Table 1, pf27 count the bacteriostasis rate of black scurf of potato, the late blight of potato and potato scab
Embodiment 7, microorganism pf27 metabolism liquid are to the bacteriostasis rate of potato scab 4.0076,4.1756 and 4.1789
It influences
One, experimental method
1, pf27 metabolism liquid is added in the LB solid medium for being cooled to 40 DEG C or so according to the ratio of 1:50, is mixed
Solid culture plate is prepared in the super-clean bench of sterilizing afterwards, making a call to a diameter with punch in plate center after plate solidification is
The hole of 6mm or so obtains experimental group culture medium.
By E. coli DH5 α in LB liquid medium, 37 DEG C, 200rpm shaken cultivation 12-16h, then
4000rpm is centrifuged 20min, collects supernatant, and the membrane filtration for being 0.22 μm with aperture by it, collects filtrate, obtain DH5 α
It is metabolized liquid；DH5 α metabolism liquid is added in the LB solid medium for being cooled to 40 DEG C or so according to the ratio of 1:50, after mixing
Solid culture plate is prepared in the super-clean bench of sterilizing, making a call to a diameter with punch in plate center after plate solidification is
The hole of 6mm or so obtains control group culture medium.
The hole that a diameter is 6mm or so is made a call to punch in LB solid medium center simultaneously, obtains blank group culture
Base.
2, potato scab germ 4.0076,4.1756 and 4.1789 is inoculated into LB liquid medium respectively, In
28 DEG C, under the conditions of 200rpm after shaken cultivation 12-16h, 4.0076,4.1756 He of potato scab cause of disease bacterium solution is obtained respectively
4.1789.The potato scab cause of disease bacterium solution 4.0076,4.1756 and 4.1789 of 20 μ L is taken to be added separately to step 1 preparation
In the hole 6mm of experimental group culture medium, control group culture medium and blank group culture medium, observe day by day, and count bacteriostasis rate.Bacteriostasis rate
(%)=(blank group colony diameter-processing group colony diameter)/(blank group colony diameter-bacteria cake diameter) × 100%.Processing group
Respectively experimental group and control group.Bacteria cake diameter is 6mm.
Two, fungistatic effect counts
As a result as shown in table 1 and Fig. 7.Pf27 be metabolized liquid to the 4.0076 of the Major Diseases potato scab of potato,
4.1576 and 4.1789 average bacteriostasis rates have respectively reached 49.74%, 39.39% and 45.47%, and E. coli
DH5 α only has 5.73%, 17.37% and to 4.0076,4.1576 and 4.1789 average bacteriostasis rate of potato scab respectively
11.89%.Illustrate that pf27 metabolism liquid has good fungistatic effect to three bacteriosises of potato.
In conclusion pf27 microbial inoculum or pf27 fermentation liquid or pf27 metabolism liquid have the suppression of wide spectrum to potato Major Diseases
Bacterium effect.
Sequence table
<110>Institute of Microorganism, Academia Sinica
<120>application of the Pseudomonas fluorescens pf27 in anti-Bemisia tabaci and prevention and treatment potato disease
<160>1
<210>1
<211>1508bp
<212>DNA
<213>artificial sequence
<220>
<223>
<400>1
agagtttgat cctggctcag attgaacgct ggcggcaggc ctaacacatg caagtcgagc 60
ggtagagaga agcttgcttc tcttgagagc ggcggacggg tgagtaatgc ctaggaatct 120
gcctggtagt gggggataac gttcggaaac gaacgctaat accgcatacg tcctacggga 180
gaaagcaggg gaccttcggg ccttgcgcta tcagatgagc ctaggtcgga ttagctagtt 240
ggtgaggtaa tggctcacca aggcgacgat ccgtaactgg tctgagagga tgatcagtca 300
cactggaact gagacacggt ccagactcct acgggaggca gcagtgggga aaattggaca 360
atgggcgaaa gcctgatcca gccatgccgc gtgtgtgaag aaggtcttcg gattgtaaag 420
cactttaagt tgggaggaag ggcagttacc taatacgtga ttgttttgac gttaccgaca 480
gaataagcac cggctaactc tgtgccagca gccgcggtaa tacagagggt gcaagcgtta 540
atcggaatta ctgggcgtaa agcgcgcgta ggtggtttgt taagtaggat gtgaaatccc 600
cgggctcaac ctgggaactg cattcaaaac tgactgacta gagtatggta gagggtggtg 660
gaatttcctg tgtagcggtg aaatgcgtag atataggaag gaacaccagt ggcgaaggcg 720
accacctgga ctaatactga cactgaggtg cgaaagcgtg gggagcaaac aggattgata 780
ccctggtagt ccacgccgta aacgatgtca actagccgtt ggaagccttg agcttttagt 840
ggcgcagcta acgcattaag ttgaccgcct ggggagtacg gccgcaaggt taaaactcaa 900
atgaattgac gggggcccgc acaagcggtg gagcatgtgg tttaattcga agcaacgcga 960
agaaccttac caggccttga catccaatga actttctaga gatagattgg tgccttcggg 1020
aacattgaga caggtgctgc atggctgtcg tcagctcgtg tcgtgagatg ttgggttaag 1080
tcccgtaacg agcgcaaccc ttgtccttag ttaccagcac gtaatggtgg gcatctaagg 1140
agactgccgg tgacaaaccg gaggaaggtg gggatgacgt caagtcatca tggcccttac 1200
ggcctgggct acacacgtgc tacaatggtc ggtacagagg gttgccaagc cgcgaggtgg 1260
agctaatccc ataaaaccga tcgtagtccg gatcgcagtc tgcaactcga ctgcgtgaag 1320
tcggaatcgc tagtaatcgc gaatcagaat gtcgcggtga atacgttccc gggccttgta 1380
cacaccgccc gtcacaccat gggagtgggt tgcaccagaa gtagctagtc taaccttcgg 1440
gaggacggtt accacggtgt gattcatgac tggggtgaag tcgtaacaag gtaaccaagg 1500
gcgatccc 1508
Claims (3)
1. Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its bacteria suspension are in prevention and treatment agricultural pests
Using；
Or, Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its bacteria suspension prevent and treat agricultural evil in preparation
Application in the product of worm；
The agricultural pests are Bemisia tabaci；
The Pseudomonas fluorescens Pseudomonas fluorescens is Pseudomonas fluorescens Pseudomonas
Fluorescens pf27, deposit number are CGMCC No. 14105.
2. Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its bacteria suspension are improving plant to Bemisia tabaci
Resistivity in application；
Or, Pseudomonas fluorescens Pseudomonas fluorescens or its microbial inoculum or its bacteria suspension improve plant pair in preparation
Application in the product of the resistivity of Bemisia tabaci；
The Pseudomonas fluorescens Pseudomonas fluorescens is Pseudomonas fluorescens Pseudomonas
Fluorescens pf27, deposit number are CGMCC No. 14105.
3. a kind of method for preventing and treating agricultural pests Bemisia tabaci, including with Pseudomonas fluorescens Pseudomonas fluorescens
The step of pf27 or its microbial inoculum or its bacteria suspension handle plant seedlings；
The Pseudomonas fluorescens Pseudomonas fluorescens pf27, deposit number are CGMCC No. 14105.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101619292A (en) * | 2009-06-04 | 2010-01-06 | 新疆农业科学院微生物应用研究所 | Endophyte with insecticidal active substances and application in biological control |
| CN104152380A (en) * | 2014-08-07 | 2014-11-19 | 领先生物农业股份有限公司 | Ultraviolet mutagenesis type pseudomonas florescens and application thereof |
| CN105367287A (en) * | 2015-12-09 | 2016-03-02 | 天津北洋百川生物技术有限公司 | Microbial fertilizer for controlling potato scab and preparation and application method thereof |
| US9528115B1 (en) * | 2013-03-15 | 2016-12-27 | The United States Of America, As Represented By The Secretary Of Agriculture | Pseudomonas fluorescens2-79 with genes for biosynthesis of pyrrolnitrin improves biocontrol activity |
-
2017
- 2017-07-21 CN CN201710599261.3A patent/CN107251909B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101619292A (en) * | 2009-06-04 | 2010-01-06 | 新疆农业科学院微生物应用研究所 | Endophyte with insecticidal active substances and application in biological control |
| US9528115B1 (en) * | 2013-03-15 | 2016-12-27 | The United States Of America, As Represented By The Secretary Of Agriculture | Pseudomonas fluorescens2-79 with genes for biosynthesis of pyrrolnitrin improves biocontrol activity |
| CN104152380A (en) * | 2014-08-07 | 2014-11-19 | 领先生物农业股份有限公司 | Ultraviolet mutagenesis type pseudomonas florescens and application thereof |
| CN105367287A (en) * | 2015-12-09 | 2016-03-02 | 天津北洋百川生物技术有限公司 | Microbial fertilizer for controlling potato scab and preparation and application method thereof |
Non-Patent Citations (1)
| Title |
|---|
| Biocontrol of Rhizoctonia solani in native patato (Solanum phureja) plants using native Pseudomonas fluorescens;Gloria Bautista et al.;《Acta boil. Colomb.》;20071231;第12卷(第1期);第19-32页 * |
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