CN107236689A - One fluorescent pseudomonads pf27 and its application in plant growth-promoting - Google Patents
One fluorescent pseudomonads pf27 and its application in plant growth-promoting Download PDFInfo
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Abstract
The invention discloses a fluorescent pseudomonads pf27 and its application in plant growth-promoting.The invention provides a fluorescent pseudomonads pf27, and prepare microbial bacterial agent pf27.It is experimentally confirmed to have plant height, root length of plant etc. after microbial bacterial agent pf27 processing and significantly improves, there are certain growth-promoting functions to plant, the crop handled through pf27 is compared with control group, growth-promoting effect surpasses 21.55%, reach as high as 829.14%, effect with wide spectrum, efficient promotion plant growth, enables in particular to promote solanaceous vegetable class crop growth, improves its quality.It is not good or the problems such as have residual that the present invention solves other method effects such as chemical fertilizer, and helps lend some impetus to agricultural sustainable development, with preferable application prospect.
Description
Technical field
The invention belongs to biological technical field, and in particular to a fluorescent pseudomonads pf27 and its in plant growth-promoting
Using.
Background technology
The Soil-sickness Problem that intensive manufacture is brought has badly influenced the yield and quality of crops, plant physiology
A series of problems, such as feature and rampant structure of soil microbial community and plant pest, badly influence the plantation effect of agricultural
Benefit and sound development.Large-scale excessive apply of chemical fertilizer causes great damage to environmental and human health impacts, it has also become
Influence environment, human health and the key factor of social development.In recent years, because chemical pesticide and chemical fertilizer are given birth on a large scale
Production is largely used with unconfined, and human habitat and Agriculture, forestry And Animal Husbandry product are polluted, human and livestock health and water and soil is compromised
Deng environmental resource, the sustainable development of agricultural production is constrained, oneself turns into one of social concern of current serious.Therefore, utilizing
Beneficial microbe in agroecosystem, preventing and treating insect pest of the plant, disease improve plant growth, increase crop yield and are increasingly becoming people
The focus studied.Growing for plant often takes with the microorganism close relation in surrounding environment, the growth potential of plant
Interaction result certainly between plant and microorganism.In plant -- in microflora, the aerial part of microorganism both with plant
(such as leaf encloses) and it is associated with ground portion, so as to affect the utilizability of nutrient and generation one in the structure of soil, soil
A little metabolites beneficial to plant or harmful.
Several chemical fertilizer for applying single mass partially too much are easily caused the trophic disturbance of crop, and chemical fertilizer is containing various mostly
The chemical substance such as different salts, wherein nitrogen, phosphorus, potassium is accumulated easily by soil consolidation in soil, and soil property salinization of soil is caused
Soil nutrient is unbalance, and the conversion of influence and obstruction crops to absorbed Nutrients absorbs, and makes fruits and vegetables growth traits inferior, makees
Thing mouthfeel and quality variation, Quality Down, such as vegetables not fragrant, melon and fruit of tasting are tasted sweetless, poor taste, and perishable, no
Preferably deposit.A quasi-microorganism such as Pseudomonas fluorescens, trichoderma, bacillus etc. can be by its metabolic process or metabolism in soil
Product improves plant growing condition, and such as increase nutrient, secreting hormone stimulate plant root system development etc., and induction plant is produced to planting
The defense reaction of thing disease root-rot disease as caused by late blight, sickle-like bacteria.In the secondary metabolite that Institute of Micro-biology produces, contain
There is the material to growth and development of plants with stimulation.Potato rhizosphere Pseudomonas fluorescens (Pseudomonas
Fluorescens group's dynamics and the quantity of dominant strain) and its influence to potato growth, are greenhouse test
The effect of acquired promotion plant growth provides the foundation and foundation applied to field.The polymer that microbial body is produced has
Drought-resistant, reduction water stress, improvement soil texture, supply organic nutrition of plant and the ability for adjusting ion activity, microorganism
The mineral element that plant is difficult by soil can be dissolved during its vital movement, and converts them into tool growth-promoting activity and is made
Material, so as to help the various mineral elements of plant absorption.Contain many B. mucilaginocus and phosphobacteria in soil, they can be by
The potassium and phosphorus of the invalid state of soil mineral are discharged, for growth and development of plants.Microorganism can promote the organic matter around root system
Humic acid is formed there is provided the resistance of plant, pollutant in soil of degrading reduces humic in the toxicity to plant, increase soil
Acid content, promotes growth and development of plants.Therefore, people are in the urgent need to safer and more effective replacement product.In recent years, microorganism system
Agent receives much concern the features such as with environment-friendly, growth-promoting volume increase, diseases prevention.Micro- life is developed into using beneficial microbe in soil
Thing fertilizer is research and development focus in recent years.
The content of the invention
It is an object of the present invention to provide a fluorescent pseudomonads Pseudomonas fluorescens pf27.
The deposit number for the Pseudomonas fluorescens Pseudomonas fluorescens pf27 that the present invention is provided is CGMCC
No.14105。
The Pseudomonas fluorescens Pseudomonas fluorescens pf27 of present invention Classification And Nomenclature is false single for fluorescence
Born of the same parents bacterium Pseudomonas fluorescens, the bacterial strain is preserved in Chinese microorganism strain preservation pipe on May 8th, 2017
The 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), preserving number is CGMCC No.14105.
It is a further object to provide Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension
Or the new application of its nutrient solution or its tunning or the microbial inoculum containing it.
The invention provides Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution or
Its tunning or application of the microbial inoculum in plant strain growth is promoted containing it.
Present invention also offers Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution
Or its tunning or application of the microbial inoculum in the product for promoting plant strain growth is prepared containing it.
Present invention also offers Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution
Or its tunning or application of the microbial inoculum in increase plant fresh weight and/or fresh biomass containing it.
Present invention also offers Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution
Or its tunning or application of the microbial inoculum in the product of increase plant fresh weight and/or fresh biomass is prepared containing it.
Present invention also offers Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution
Or its tunning or microbial inoculum containing it improve plant plant height and/or stem is thick and/or radical amount and/or blade quantity and/
Or the application in blade area and/or maximum blade growing height.
Present invention also offers Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution
Or its tunning or microbial inoculum containing it improve plant plant height and/or stem is thick and/or radical amount and/or blade quantity preparing
And/or the application in the product of blade area and/or maximum blade growing height.
In above-mentioned application, the Pseudomonas fluorescens Pseudomonas fluorescens concretely Pseudomonas
Bacterium Pseudomonas fluorescens pf27.
It is a still further object of the present invention to provide a kind of product.
The active component for the product that the present invention is provided is above-mentioned Pseudomonas fluorescens Pseudomonas fluorescens
Pf27 or its bacteria suspension or its nutrient solution or its tunning or the microbial inoculum containing it；
The product has following 1) -3) in any function：
1) plant strain growth is promoted；
2) increase plant fresh weight and/or fresh biomass；
3) improve that plant plant height and/or stem be thick and/or radical amount and/or blade quantity and/or blade area and/or highest
Leaf growth height.
Final object of the present invention is to provide a kind of method of promotion plant growth.
The method for the promotion plant growth that the present invention is provided comprises the following steps：Use above-mentioned Pseudomonas fluorescens
Pseudomonas fluorescens pf27 or its microbial inoculum processing plant seedlings.
In the above method, the bacteria suspension concentration of the processing plant seedlings is 1 × 105CFU/mL。
In above-mentioned application or the said goods or the above method, the specific preparation method of the microbial inoculum is as follows：Fluorescence is false single
Born of the same parents bacterium pf27 is cultivated in KB nutrient solutions, obtains pf27 bacterium solutions, and pf27 bacterium solutions are centrifuged, and collects precipitation；Will with aqua sterilisa
It is 1 × 10 that the precipitation, which is diluted to concentration,9-1×1010CFU/mL, obtains the microbial inoculum.
In above-mentioned application or the said goods or the above method, the plant is plant of Solanaceae or crucifer or grass
Section plant.The plant of Solanaceae concretely tomato and potato；The crucifer concretely wild cabbage, fast dish and big
Chinese cabbage；The grass concretely wheat and corn.
The present invention screens Pseudomonas fluorescens pf27, and prepare micro- life by the exploitation again to existing bacterial strain
Thing microbial inoculum pf27.It is experimentally confirmed to have plant height, root length of plant etc. after microbial bacterial agent pf27 processing and significantly improves, it is right
Plant has certain growth-promoting functions, and the crop handled through pf27 is compared with control group, and growth-promoting effect surpasses 21.55%, and highest can
Up to 829.14%, with wide spectrum, efficiently promote the effect of plant growth, enable in particular to promote solanaceous vegetable class crop growth,
Improve its quality.It is not good or the problems such as have residual that the present invention solves other method effects such as chemical fertilizer, and contributes to
Promote agricultural sustainable development, with preferable application prospect.
Brief description of the drawings
Fig. 1 is the pf27 phylogenetic trees based on 16s rDNA sequence constructs.
Fig. 2 is growth-promoting effects of the microbial bacterial agent pf27 to solanaceous crops tomato.A. left figure control group solanaceous crops tomato
In miscellaneous No. 9 plant strain growths state, the growth conditions of the lower tomato plant of right figure pf27 microbial inoculums processing, picture is 21 after plant transplanting
It shoots；The plant plant height of the lower tomato of B.pf27 microbial inoculums processing and control group data statistics；The lower tomato of C.pf27 microbial inoculums processing
Plant leaf sum is counted with control group plant leaf total data；The lower tomato plant fresh weight of D.pf27 microbial inoculums processing is with compareing plant
Strain fresh weight data statistics；The lower tomato plant diameter (stem is thick) of E.pf27 microbial inoculums processing is united with control group plant diameter (stem is thick) data
Meter；The lower new leaf area in 2, tomato plant top of F.pf27 microbial inoculums processing and control group data statistics, all data statistics plant move
33 days after cultivation, n=9,3 repetitions, note：* the significant difference in 0.05 level is represented, * * represent that difference shows in 0.01 level
Write, * * * represent the significant difference in 0.001 level, with.
Fig. 3 is growth-promoting effects of the microbial bacterial agent pf27 to solanaceous crops potato.A. left figure control group solanaceous crops horse
The state of bell potato plant strain growth, right figure pf27 microbial inoculums handle the growth conditions of Potato plant, and picture was clapped for 4 weeks after transplanting
Take the photograph；The plant plant height of B.pf27 microbial inoculums processing Potato and control group plant data statistics；C.pf27 microbial inoculums handle bell
Potato plant leaf sum and control group data statistics；D.pf27 microbial inoculums processing Potato plant takes root number and control group plant
Number data of taking root is counted；The plant root length of E.pf27 microbial inoculums processing Potato is united with control group plant root length data
Meter；F.pf27 microbial inoculums handle Potato and control group plant production potato situation compares；G.pf27 handles Potato and control group
Plant produces potato gross weight data statistics, n=6,3 repetitions.
Fig. 4 is growth-promoting effects of the microbial bacterial agent pf27 to crop in cruciferae wild cabbage.A. left figure control group Cruciferae
Cabbage plant, the growth conditions of the lower cabbage plant of right figure pf27 microbial inoculums processing, picture shooting is 33 days after plant is transplanted；B.pf27
The lower new leaf area in 2, cabbage plant top of microbial inoculum processing is counted with control group plant area data, and every plant of plant counts 2 tops
Young leaves；The lower wild cabbage of C.pf27 microbial inoculums processing is counted with control group plant leaf total data；The lower cabbage plant of D.pf27 processing is fresh
Weight and control group plant fresh weight data statistics；The lower cabbage plant diameter (stem is thick) of E.pf27 microbial inoculums processing and control group plant diameter
(stem is thick) compares, n=9,3 repetitions.
Fig. 5 is growth-promoting effects of the microbial bacterial agent pf27 to the fast dish of crop in cruciferae Chinese holly.A. left figure control group four
Season blue or green fast dish plant strain growth state, the growth conditions of the fast dish plant of the lower Chinese holly of right figure pf27 microbial inoculums processing, picture shooting in
22 days after plant transplanting；B.pf27 microbial inoculums handle the fast dish of Chinese holly and control group plant leaf total number data statistics；C.pf27
The lower fast dish plant fresh weight of Chinese holly of microbial inoculum processing and control group plant fresh weight；The lower fast dish plant top 2 of Chinese holly of D.pf27 processing
The new leaf area of piece and control group plant area, 2 top young leaves of every plant of statistics, n=9,3 repetitions.
Fig. 6 is influences of the microbial bacterial agent pf27 to the growth-promoting effect of crop in cruciferae Chinese cabbage.A. left figure is control
Group new No. 3 Reducing sugars in Chinese cabbage Beijing, right figure is the growth conditions that pf27 microbial inoculums handle lower Chinese cabbage plant, and picture is clapped
It is photographed on after plant transplanting 22 days；The lower Chinese cabbage of B.pf27 microbial inoculums processing counts with control group plant leaf total data；C.pf27
The lower Chinese cabbage plant fresh weight of microbial inoculum processing and control group plant fresh weight；Lower 2, the Chinese cabbage plant top of D.pf27 microbial inoculums processing is new
Leaf area and control group plant area, 2 top young leaves of every plant of statistics, n=8,3 repetitions.
Fig. 7 is influences of the microbial bacterial agent pf27 to the growth-promoting effect of crop in cruciferae.A. right figure pf27 microbial inoculums are handled
Lower crop in cruciferae cabbage plant total amount, left figure is control group cabbage plant total amount, n=9；B. under the processing of right figure pf27 microbial inoculums
The fast dish plant total amount of Cruciferae Chinese holly, left figure control cabbage plant total amount, n=9；C. right figure pf27 microbial inoculums handle lower cross
The flower new No. 3 plant total amounts in section Chinese cabbage Beijing, left figure control cabbage plant total amount, n=8；D. wild cabbage, the fast dish of Chinese holly and great Bai
The new No. 3 biomass varieties statistics in dish Beijing.
Fig. 8 is influences of the microbial bacterial agent pf27 to the growth-promoting effect of gramineous crop wheat.A. left figure control group grass
Section's wheat JN17 Reducing sugars compare, the processing of right figure pf27 microbial inoculums lower wheat plant growth conditions, n=9；B.pf27 microbial inoculums
Lower and control group grass family plant wheat JN17 plant leafs minimum altitude (Low) and maximum height (High) difference the system of processing
Meter, n=9, picture shooting is in after after planting 27 days.
Fig. 9 is influences of the microbial bacterial agent pf27 to the growth-promoting effect of gramineous crop corn.A. left figure control group grass
Section's plant Zea mays Mo17 plant strain growth tendency charts, the lower grass corn Mo17 plant of right figure pf27 microbial inoculums processing, picture is clapped
Take the photograph 22 days, n=9,3 repetitions after transplanting；The processing of B.pf27 microbial inoculums is lower and control group grass family plant corn Mo17 plants stems are thick
Comparison；Lower and control group grass family plant corn Mo17 plants stems the comparison of C.pf27 microbial inoculums processing；The processing of D.pf27 microbial inoculums
The comparison of lower grass corn Mo17 and control group plant plant height；The lower grass corn of E.pf27 microbial inoculums processing
The comparison of Mo17 and control group plant stem slightly.
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 is referred to as：CGMCC
Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date：On May 8th, 2017
Collection is registered on the books numbering：CGMCC No.14105
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material, reagent used etc., unless otherwise specified, are commercially obtained in following embodiments.
Quantitative test in following embodiments, is respectively provided with three repetition experiments, results averaged.
The bacteria distribution culture medium used in following embodiments is as follows：
MEA culture mediums：Malt extract 30g, Mycological peptone 5g, agar powder 15g, pH5.4；
Beef-protein medium：Peptone 10g, beef extract 3g, sodium chloride 5g, agar powder 12g, water 1000ml, pH
7.2-7.4；
LB culture mediums：Sodium chloride 10g, peptone 10g, yeast extract 5g, agar powder 12g, water 1000ml, pH 7.0；
KB culture mediums：Peptone 20g, K2HPO41.5g, glycerine 8ml, MgSO40.74g, H2O supplies 1L, pH 7.0；
Solid medium adds agar powder 12g.
The separation and identification of embodiment 1, pf27 bacterial strains
First, the separation of pf27 bacterial strains
Pf27 is isolated from Yunnan Province Puer City oryza meyeriana area rhizosphere soil (north latitude 22。33 ' 56 ", 100。35 ' 12 ", sea
Pull out 737 meters).Specific separation method is as follows：The rhizosphere soil in oryza meyeriana area will be picked up from, picking root system scrapes Rhizosphere Soil, claimed
Measure and put under 5g, aseptic condition in the sterilizing triangular flask being ready for, 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 mediums, beef-protein medium, LB culture mediums and
On KB culture medium flat plates, respectively at 28 DEG C, cultivated in 37 DEG C of incubators, the growth conditions of observation bacterium colony, as a result show dilute daily
It is interpreted as 10-5、10-6Times when flat board on there is no that bacterium colony is grown, and be diluted to 10-4Times when, single bacterium can be grown on flat board
Fall, and picking grows single bacterium colony on KB culture mediums, is named as pf27 bacterial strains, then expands in KB fluid nutrient mediums
Big culture, does further bacterial strain identification.
2nd, the identification of pf27 bacterial strains
1st, the Morphological Identification of pf27 bacterial strains
Pf27 bacterial strains are cultivated on KB culture mediums can form orange colonies after 24h~48h, bacterium colony state is rounded, surface
It is smooth to have raised brim neat, it is more sticky, easily provoke.
2nd, the Molecular Identification of pf27 bacterial strains
The DNA of pf27 bacterial strains is extracted, performing PCR amplification is entered using universal primer Eubac27F/Eubac1492R, contained
The PCR primer of pf27 bacterial strain 16S rDNA conserved regions.Primer sequence is as follows：
Eubac27F：5’-AGAGTTTGATCCTGGCTCAG-3’；
Eubac1492R：5’-GGTTACCTTGTTACGACTT-3’.
PCR primer is connected to pEASY-T3 (buying from the complete biological Co., Ltd of formula gold in Beijing) through TA clones, with general
Primer is sequenced in Invitrogen Corp..
Sequencing result shows：The nucleotide sequence of PCR primer is as shown in sequence 1.Will sequencing gained sequence and NCBI data
Storehouse is carried out after blast comparisons, and the fluorescence for representing 22 kinds of different pseudomonad kinds with other to pf27 by MEGA5.1 softwares is false single
Born of the same parents' type strain carries out Phylogenetic Analysis, and phylogenetic tree is as shown in Figure 1.
Summary qualification result, it is Pseudomonas fluorescens to determine pf27 strain names, and its Classification And Nomenclature is that fluorescence is false single
Born of the same parents bacterium Pseudomonas fluorescens, the bacterial strain is preserved in Chinese microorganism strain preservation pipe on May 8th, 2017
The 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), preserving number is CGMCC No.14105.
The preparation of embodiment 2, microbial bacterial agent pf27
By the Pseudomonas fluorescens pf27 obtained in embodiment 1 in KB culture mediums shaken cultivation, in 28 DEG C, 200rpm bars
Shaken cultivation 12-16h under part, then 4000rpm centrifugations 20min, abandons supernatant, collects thalline bacterial sediment, then use aqua sterilisa
Bacterial sediment is diluted, pf27 viable bacterias in microbial bacterial agent pf27 (microbial inoculum pf27), microbial bacterial agent pf27 finished products are made
Total concentration is 1 × 109-1×1010CFU/mL。
The growth-promoting effect of embodiment 3, microbial bacterial agent pf27 to solanaceous crops tomato seedling
First, experimental method
Miscellaneous No. 9 tomato seeds (scientific and technological (Beijing) Co., Ltd production of middle vegetable kind industry) use 3%NaClO solution disinfections by
Used after 10min, ddH2O is cleaned 6 times, seed after being sterilized.Then seed after sterilization is uniformly layered on to the vermiculite peat of sterilizing
(vermiculite and peat soil are according to 1 for soil matrix matter:1 ratio is mixed, 121 DEG C of autoclavings, 20min) hair seedling, in 26 DEG C, 12h:12h
Illumination：Seed is cultivated under dark condition to sprout, and when seed germination and growth is to two panels true leaf, uniform addition microbial inoculum is transplanted to respectively
(concentration is that every kg vermiculites peat soil substrate mixture is to add the bacterium solution that 50ml OD values are 1 to pf27, and bacterium solution uses dense eventually
Degree is 1 × 105CFU/mL) and in the nutrition cave of control, tomato seedling is transplanted in seedling culture hole plate (9 lis of hole tray specification back cut diameter
Rice, 6 centimetres of lower port diameter, 8 centimetres of height), it is placed on growth (condition in greenhouse：Light application time 12h, 26 DEG C of temperature is relatively wet
Degree is 50%).Continue to place under above-mentioned condition and cultivate, 1 plant of seedling is transplanted in each Culture basin, each 27 repetitions of processing.With not
Any microbial inoculum is added as control (Control).The plant height, the number of blade, tomato plant of each plant of plant are counted after transplanting 33 days
Fresh weight, tomato plant stem are thick, 2, tomato top young leaves leaf area, and calculate biomass increase.Biomass increase (%)=(place
Reason group plant fresh weight-control group plant fresh weight)/control group plant fresh weight × 100%.
2nd, growth-promoting effect is counted
As a result it is as shown in Figure 2.Microbial bacterial agent pf27 can produce obvious growth-promoting effect to tomato seedling, with control group
Compare, the average plant height of tomato plant adds 18%, number of blade increase by 23%, plant diameter after microbial bacterial agent pf27 processing
(stem is thick) is 2.5 times of control group, and the blade area of pf27 processing increases up to as many as 5 times, and the biomass of tomato plant increases
Plus reach 771.16%.
The growth-promoting effect of embodiment 4, microbial bacterial agent pf27 to solanaceous crops potato seedling
First, experimental method
It will buy and be placed on after being selected from the super incity potato of Beijing maiden voyage in the environment of moist dark, after one week,
The bud of the consistent healthy potato of budding is selected, is cut with 70% alcohol sterilization blade.Then young shoot is transplanted to
(50ml/Kg, bacterium solution final concentration is 10 by uniform addition microbial inoculum pf275CFU/mL plastic flowerpot (specification back cut diameter):14 lis
Rice, 9.5 centimetres of lower port diameter, height 12.5 centimetres) and sterilize vermiculite matrix (according to 1:1 ratio is mixed, and 121 DEG C of high pressures are gone out
Bacterium, 20min) in, surface covers one layer of preservative film moisturizing, is removed after it grows cotyledon, is placed on growth (bar in greenhouse
Part：Light application time 12h, 26 DEG C of temperature, relative humidity 50%).Using without any microbial inoculum as control (Control), each
Handle 16 young plants, 3 repetitions.Plant height, the number of blade, the length of root of potato, radical mesh and Ma Ling are counted after transplanting 28 days
Potato knot potato situation, and calculate biomass increase.Biomass increase (%)=(treatment group plant fresh weight-control group plant fresh weight)/
Control group plant fresh weight × 100%.
2nd, growth-promoting effect is counted
As a result as shown in figure 3, microbial bacterial agent pf27 can remarkably promote the growth of potato seedling, potato is shown
Up to 4 times of plant height increase, compared with the control, the number of blade and radical mesh increase by 114% and 50% respectively.Without chemical fertilizer
Under service condition, the potato production potato amount of microbial bacterial agent pf27 processing is dramatically increased, and statistical result shows microbial bacterial agent
Pf27 increases up to 1115.15% to the biomass of potato.
The growth-promoting effect of embodiment 5, microbial bacterial agent pf27 to crop in cruciferae wild cabbage
First, experimental method
According to No. one rich to head cabbage varieties capital (Chinese Academy of Agricultural Sciences's vegetable or flower research of the experimental method in embodiment 3
Institute) seed carry out vernalization, until grow 2 true leaves by seedling replanting (9 lis of the hole tray specification back cut diameter into seedling culture hole plate
Rice, 6 centimetres of lower port diameter, 8 centimetres of height), continue to place under above-mentioned condition and cultivate, 1 plant of seedling is transplanted in each Culture basin, often
9 repetitions of individual processing, are repeated 3 times, to be used as control (Control) without any microbial inoculum.Each strain is counted after transplanting 33 days
The number of blade of plant, plant fresh weight, cabbage plant stem are thick, 2, wild cabbage top young leaves leaf area, and calculate biomass increase.It is raw
Object amount increases (%)=(treatment group plant fresh weight-control group plant fresh weight)/control group plant fresh weight × 100%.
2nd, growth-promoting effect is counted
As a result as shown in figure 4, microbial bacterial agent pf27 can remarkably promote the growth of cabbage seedling, it is mainly reflected in wild cabbage
The number of blade, stem is thick and fresh weight compared with the control, have a significant increase, the increase of leaf area pole conspicuousness compared with the control.
Under the service condition without chemical fertilizer, the statistical result showed of the fresh biomass of the wild cabbage of microbial bacterial agent pf27 processing is micro-
Bacteria agent pf27 increases up to 400.00% to the biomass of wild cabbage.
Growth-promoting effects of the microbial bacterial agent pf27 of embodiment 6 to the fast dish of crop in cruciferae
First, experimental method
According to the experimental method in embodiment 3 to fast dish of the fast vegetable kind four seasons 1 (in national vegetables engineering technology research and development
The heart) seed carry out vernalization, until grow 2 true leaves by seedling replanting (9 lis of the hole tray specification back cut diameter into seedling culture hole plate
Rice, 6 centimetres of lower port diameter, 8 centimetres of height), continue to place under above-mentioned condition and cultivate, 1 plant of seedling is transplanted in each Culture basin, often
9 repetitions of individual processing, are repeated 3 times, control are used as to be not added with any bacterium solution.The blade of each plant of plant is counted after transplanting 22 days
Number, plant fresh weight, 2, fast dish top young leaves leaf area, and calculate biomass increase.Biomass increase (%)=(treatment group is planted
Strain fresh weight-control group plant fresh weight)/control group plant fresh weight × 100%.
2nd, growth-promoting effect is counted
As a result as shown in figure 5, microbial bacterial agent pf27 can remarkably promote the growth of fast dish seedling, it is mainly reflected in fast dish
The number of blade, fresh weight and leaf area, compared with not doing the control group of any processing, there is significant increase.Without chemical fertilizer
Service condition under, microbial bacterial agent pf27 processing the fast dish of Chinese holly fresh biomass statistical result showed microbial bacteria
Agent pf27 increases up to 137.27% to the biomass of fast dish.
The growth-promoting influential effect of embodiment 7, microbial bacterial agent pf27 to crop in cruciferae Chinese cabbage
First, experimental method
According to No. 3 new to Chinese cabbage cultivar Beijing of the experimental method in embodiment 3 (in national vegetables engineering technology research and development
The heart) seed carry out vernalization, until grow 2 true leaves by seedling replanting (9 lis of the hole tray specification back cut diameter into seedling culture hole plate
Rice, 6 centimetres of lower port diameter, 8 centimetres of height), continue to place under above-mentioned condition and cultivate, 1 plant of seedling is transplanted in each Culture basin, often
9 repetitions of individual processing, are repeated 3 times, control are used as to be not added with any bacterium solution.The blade of each plant of plant is counted after transplanting 22 days
Number, plant fresh weight, 2, Chinese cabbage top young leaves leaf area, and calculate biomass increase.Biomass increases (%)=(treatment group
Plant fresh weight-control group plant fresh weight)/control group plant fresh weight × 100%.
2nd, growth-promoting effect is counted
As a result as shown in fig. 6, microbial bacterial agent pf27 can promote the growth of Seedling of Chinese Cabbage, it is mainly reflected in Chinese cabbage
The number of blade compared with not doing the control group of any processing conspicuousness increase, fresh weight and leaf area are in a short time temporarily without conspicuousness
Difference.Under the service condition without chemical fertilizer, the statistics of the fresh biomass of the Chinese cabbage of microbial bacterial agent pf27 processing
As a result show that microbial bacterial agent pf27 increases up to 9.72% to the biomass of Chinese cabbage.
The fast dish of the wild cabbage of Cruciferae, Chinese holly and the growth-promoting effect comparative result of Chinese cabbage are as shown in fig. 7, microbial bacteria
Agent pf27 has the effect of significant growth-promoting to wild cabbage, the fast dish of Chinese holly of Cruciferae, but to Chinese cabbage Beijing new 3
Growth-promoting effect is not really obvious, illustrates that this pf27 microbial inoculum is only suitable for partial cross flower section crop.
The effect of embodiment 8, microbial bacterial agent pf27 to gramineous crop wheat growth-promoting
First, experimental method
First by wheat seed Jinan 17 (JN17) with being used after 3%NaClO solution disinfections 10min, ddH2O is cleaned 6 times, is obtained
Seed after sterilization, is then uniformly layered on and is placed with the massive plate of one layer of sterile filtering layer by seed after to sterilization, the ddH of sterilizing2O
Filter paper keeps moistening, 37 DEG C of incubator vernalization overnight is placed on, after after the embryo germination of wheat seed, according to the reality in embodiment 3
Proved recipe method, is transplanted in seedling culture hole plate (9 centimetres of hole tray specification back cut diameter, 6 centimetres of lower port diameter, 8 centimetres of height), after
Continuous placement is cultivated at ambient temperature, and several plants of seeds are uniformly transplanted in each Culture basin, and each 9 repetitions of processing, are repeated 3 times,
Control is used as to be not added with any bacterium solution.Wheat growth trend is observed after transplanting 27 days, the minimum blade and most of wheat seedling is counted
The height Long-term change trend of high blade.
2nd, growth-promoting effect is counted
As a result as shown in figure 8, microbial bacterial agent pf27 can promote the growth of wheat seedling early stage, be mainly reflected in not
The control group for doing any processing is compared, and the minimum growing height of wheat seedlings blade is not present significant difference, but highest leaf
The growing height of piece is apparently higher than control group.
The growth-promoting effect of embodiment 9, microbial bacterial agent pf27 to gramineous crop corn
First, experimental method
First by corn seed Mo17 (inbred corn system is given by Institute of Zoology, Academia Sinica Zhang Xiaoming researcher)
With being used after 3%NaClO solution disinfections 10min, ddH2O is cleaned 6 times, seed after being sterilized, and seed is uniform after then sterilizing
It is layered on and is placed with the massive plate of one layer of sterile filtering layer, the ddH of sterilizing2O filter paper keeps moistening, is placed on 37 DEG C of incubators and stays overnight and urges
Bud, after the embryo and root of corn seed are sprouted, according to the experimental method in embodiment 3, is transplanted in seedling culture hole plate (hole tray
9 centimetres of specification back cut diameter, 6 centimetres of lower port diameter, 8 centimetres of height), continue to place and cultivate at ambient temperature, each culture
The uniform corn seed for transplanting 1 plant of development indifference in basin, each 9 repetitions of processing, are repeated 3 times, and are made with being not added with any bacterium solution
For control.
Counted after transplanting 22 days each plant stem is thick and plant height.
2nd, growth-promoting effect is counted
As a result as shown in figure 9, microbial bacterial agent pf27 can remarkably promote the growth of corn seedling, it is mainly reflected in corn
The height and diameter (stem is thick) of plant, compared with not doing the control group of any processing, the stem of microbial bacterial agent pf27 processing slightly has
Pole significant difference.The stem of the plant of microbial bacterial agent pf27 processing is slightly 1.6 times of control group.
Sequence table
<110>Institute of Microorganism, Academia Sinica
<120>One fluorescent pseudomonads pf27 and its application in plant growth-promoting
<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 (10)
1. a fluorescent pseudomonads Pseudomonas fluorescens pf27, its deposit number is CGMCC
No.14105。
2. Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution or its tunning contain
There is application of its microbial inoculum in plant strain growth is promoted.
3. Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution or its tunning contain
There is application of its microbial inoculum in the product for promoting plant strain growth is prepared.
4. Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution or its tunning contain
There is application of its microbial inoculum in increase plant fresh weight and/or fresh biomass.
5. Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution or its tunning contain
There is application of its microbial inoculum in the product of increase plant fresh weight and/or fresh biomass is prepared.
6. Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution or its tunning contain
There is its microbial inoculum improving thick plant plant height and/or stem and/or radical amount and/or blade quantity and/or blade area and/or most
Application in high leaf growth height.
7. Pseudomonas fluorescens Pseudomonas fluorescens or its bacteria suspension or its nutrient solution or its tunning contain
Have its microbial inoculum prepare improve thick plant plant height and/or stem and/or radical amount and/or blade quantity and/or blade area and/
Or the application in the product of maximum blade growing height.
8. according to any described method in claim 2-7, it is characterised in that：The Pseudomonas fluorescens Pseudomonas
Fluorescens is the Pseudomonas fluorescens Pseudomonas fluorescens pf27 described in claim 1.
9. a kind of product, its active component is the Pseudomonas fluorescens Pseudomonasfluorescens described in claim 1
Pf27 or its bacteria suspension or its nutrient solution or its tunning or the microbial inoculum containing it；
The product has following 1) -3) in any function：
1) plant strain growth is promoted；
2) increase plant fresh weight and/or fresh biomass；
3) improve that plant plant height and/or stem be thick and/or radical amount and/or blade quantity and/or blade area and/or maximum blade
Growing height.
10. a kind of method of promotion plant growth, comprises the following steps：With the Pseudomonas fluorescens described in claim 1
Pseudomonas fluorescens pf27 or its bacteria suspension processing plant seedlings.
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| CN116042468A (en) * | 2022-12-28 | 2023-05-02 | 云南大学 | Multifunctional fluorescent pseudomonas with functions of promoting pasture growth and low-temperature composting fermentation |
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| CN102113521A (en) * | 2010-12-06 | 2011-07-06 | 山东中烟工业有限责任公司 | Plant virus-resisting preparation and applications thereof |
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| CN103146610A (en) * | 2013-03-12 | 2013-06-12 | 南京农业大学 | Plant growth-promoting rhizobacteria and application thereof |
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| CN102113521A (en) * | 2010-12-06 | 2011-07-06 | 山东中烟工业有限责任公司 | Plant virus-resisting preparation and applications thereof |
| CN102787090A (en) * | 2012-08-07 | 2012-11-21 | 哈尔滨师范大学 | Alfalfa root growth-promoting rhizobacteria MJM-5 and application of alfalfa root growth-promoting rhizobacteria MJM-5 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116042468A (en) * | 2022-12-28 | 2023-05-02 | 云南大学 | Multifunctional fluorescent pseudomonas with functions of promoting pasture growth and low-temperature composting fermentation |
| CN116042468B (en) * | 2022-12-28 | 2024-04-26 | 云南大学 | Multifunctional fluorescent pseudomonas with functions of promoting pasture growth and low-temperature composting fermentation |
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