CN109880778A - A kind of pair of capsicum has composite bacteria agent and its application of growth-promoting production-increasing function - Google Patents
A kind of pair of capsicum has composite bacteria agent and its application of growth-promoting production-increasing function Download PDFInfo
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- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
Abstract
The invention discloses composite bacteria agent and its applications that a kind of pair of capsicum has growth-promoting production-increasing function.Composite bacteria agent includes the fermentation liquid of Bacillus flexus HGD12, Pseudomonas putida HGD3 and Bacillus velezensis HP9 bacterial strain；The Bacillus flexus HGD12 is Bacillus flexus, and China typical culture collection center is preserved on April 1st, 2019, and deposit number is CCTCC NO:M 2019224；The Pseudomonas putida HGD3 is pseudomonas putida, and China typical culture collection center is preserved on April 1st, 2019, and deposit number is CCTCC NO:M 2019223；The Bacillus velezensis HP9 is Bei Laisi bacillus, and China typical culture collection center is preserved on April 8th, 2019, and deposit number is CCTCC NO:M 2019243.The present invention, which has, promotes chili growth, improves yield of hot pepper, reduces chemical fertilizer application, reaches the beneficial effect of Reducing amount of chemical fertilizer applied, capsicum volume increase.
Description
Technical field
The invention belongs to the microorganism of field of agricultural microbial technology more particularly to a kind of promotion chili growth and volume increase is multiple
Combined bacteria agent and its application.
Background technique
China is the big capsicum producing country of the first in the world and country of consumption, about 133.3 ten thousand hectares of sown area of year, but capsicum kind
Generally existing blindness fertilising and excessive the phenomenon that applying fertilizer in plant, the application of a large amount of chemical fertilizer result in soil hardening and fertility decline,
The decline of yield of hot pepper and quality can be finally caused, this phenomenon has influenced the sustainable development of agricultural.
Bacterial strain in microbial bacterial agent is due to the beneficial microbe in soil or vegetable material, to the nontoxic nothing of environment
Evil, can be improved after application soil, increase soil organic matter content, may additionally facilitate crop growth, play increase yield and
The effect of improving quality.Cooperate the excessive use that can also reduce chemical fertilizer with chemical fertilizer, preferably preserve the ecological environment.
The report of research and the application of capsicum complex bacterial agent special is less at present.Patent CN108130289 A is disclosed by 7
Plant the preparation of the capsicum complex bacterial agent special of Bacillus strain and a kind of pseudomonas strain composition and in big sharp green pepper field
Using upper effect.Patent CN107418906 A discloses the composite bacteria agent being made of 3 kinds of lactobacillus strains and is improving color green pepper
Application in yield and anti-root rot.But its effect of increasing production is carried out under common fertilizer amount of application, and is only compound bacteria
Field trial of the agent to 1 capsicum variety.
Summary of the invention
The object of the present invention is to provide composite bacteria agent and its applications that a kind of pair of capsicum has growth-promoting production-increasing function.This
Invention, which has, promotes chili growth, improves yield of hot pepper, reduces chemical fertilizer application, achievees the purpose that Reducing amount of chemical fertilizer applied, capsicum volume increase.
Technical solution of the present invention: a kind of pair of capsicum has composite bacteria agent and its application of growth-promoting production-increasing function, includes
The hair of Bacillus flexusHGD12, Pseudomonasputida HGD3 and Bacillus velezensis HP9 bacterial strain
Zymotic fluid；The Bacillus flexus HGD12 is Bacillus flexus, is preserved in Chinese Typical Representative culture on April 1st, 2019
Object collection, deposit number are CCTCC NO:M 2019224；The Pseudomonas putida HGD3 is Pseudomonas putida
Bacterium, China typical culture collection center is preserved on April 1st, 2019, and deposit number is CCTCC NO:M 2019223；It is described
Bacillus velezensis HP9 is Bei Laisi bacillus, and Chinese Typical Representative culture guarantor is preserved on April 8th, 2019
Hiding center, deposit number are CCTCC NO:M 2019243.
A kind of preparation method of the composite bacteria agent to capsicum as described in claim 1 with growth-promoting production-increasing function, according to
Following methods preparation；
A, slant activation: by Bacillus flexus HGD12, Pseudomonas putida HGD3 and Bacillus
Velezensis HP9 bacterial strain is inoculated in solid medium respectively, is activated overnight bacterial strain；
B, liquid activates: the lawn by step a activated spawn is picked them separately in fluid nutrient medium, in 28-30 DEG C of condition
Under, 150rpm shaking table shaken cultivation to logarithmic growth phase；
C, it obtains the fermentation liquid of each bacterial strain: each strain cultured solution for being in logarithmic growth phase in step b is pressed into 5% volume ratio
It is inoculated in fluid nutrient medium respectively, shaking table 150rpm shaken cultivation 18-36h, respectively obtains Bacillus at 28-30 DEG C
The fermentation liquid of flexus HGD12, Pseudomonas putida HGD3 and Bacillus velezensis HP9 bacterial strain；
D, the preparation of composite bacteria agent: by Bacillus flexus HGD12, Pseudomonas putida in step c
The fermentation liquid equal proportion of HGD3 and Bacillus velezensis HP9 bacterial strain is uniformly mixed, and obtains composite bacteria agent.
In the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function above-mentioned, in the step a, solid training
Supporting base is NA solid medium.
It is described based on following ratios in the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function above-mentioned
Contain beef extract 5.0g, peptone 10.0g, sodium chloride 5.0g, distilled water 1000mL and agar 15-20g in NA solid medium；
The NA solid medium is pH 7.0-7.2, uses the preceding and 20min that sterilizes at 121 DEG C.
In the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function above-mentioned, in step b, c, liquid
Culture medium is NA fluid nutrient medium.
In the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function above-mentioned, the NA fluid nutrient medium is every
It rises and contains: beef extract 5.0g, peptone 10.0g, sodium chloride 5.0g and distilled water 1000mL；The NA fluid nutrient medium is pH
7.0-7.2 uses the preceding and 20min that sterilizes at 121 DEG C.
In the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function above-mentioned, in the step d, compound bacteria
Agent is 3 × 10 containing total viable count8-3×109Between CFU/mL.
A kind of capsicum growth promoting bacteria agent product, the active constituent of the product include composite bacteria agent described in claim 1.
A kind of capsicum growth promoting bacteria agent product, the active constituent of the product are composite bacteria agents described in claim 1.
Capsicum growth promoting bacteria agent product above-mentioned further includes the conventional ingredient for growth promoting bacteria agent.
The present invention also provides the composite bacteria agents to promote chili growth, improve yield of hot pepper and reduce chemical fertilizer application side
The application in face, specific method are beared fruit the phase in chilli seedling (10d after transplanting), capsicum early flowering season (colonizing rear 30-40d), capsicum
(colonizing rear 60-70d) applies respectively, and each dosage is 2-4 liter/mu, and total viable count is 3 × 108-3×109Between CFU/mL,
Pouring root is in capsicum root after being watered dilution, and 3 times in total.
Compared with prior art, composite bacteria agent bacterium source of the invention is in the plant growth-promoting rhizobacteria of tea place rhizosphere soil, no
There is fixed nitrogen, phosphorus decomposing, potassium decomposing or the growth-promoting characteristic for generating acc deaminase with bacterial strain, and without Antagonism between each bacterial strain, mixing is applied
With can promote chili growth, yield of hot pepper is improved, and achieve the purpose that Reducing amount of chemical fertilizer applied.
In conclusion the present invention, which has, promotes chili growth, yield of hot pepper is improved, chemical fertilizer application is reduced, reaches chemical fertilizer and subtract
Amount, the beneficial effect of capsicum volume increase.
Detailed description of the invention
Influence of Fig. 1 composite bacteria agent to gorgeous peppery 425 plant height；
Influence of Fig. 2 composite bacteria agent to gorgeous 425 stem thickness of green pepper；
Influence of Fig. 3 composite bacteria agent to peppery No. 10 plant heights are abided by；
Influence of Fig. 4 composite bacteria agent to peppery No. 10 stem thicknesses are abided by；
Fig. 5 composite bacteria agent promotes to abide by peppery No. 10 root system development figures in Field information.
Specific embodiment
The present invention is further illustrated with reference to the accompanying drawings and examples, but be not intended as to the present invention limit according to
According to.
Embodiment 1: the Molecular Identification of 3 bacterial strains of composite bacteria agent is formed
Molecular Identification is carried out to 3 plants of bacterial strains: DNA of bacteria being extracted to bacterial strain HGD12, HGD3 and HP9, is with the DNA respectively
Template carries out PCR amplification using bacterial 16 S rDNA universal primer, amplified production is sequenced, the sequencing result of 3 plants of bacteriums
As shown in sequence table, the amplification length of 16S rDNA sequence is respectively 1417,1442,1454bp.Through sequence analysis, HGD12
Bacillus flexus NBRC15715 16S rDNA sequence (NCBI in bacterial strain 16S rDNA sequence and GenBank database
Accession number: BCVD01000224) homology is up to 99.93%；In HGD3 bacterial strain 16S rDNA sequence and GenBank database
Pseudomonas putida AA7 16S rDNA sequence (NCBI accession number: CP018846) homology is up to 99.86%；HP9 bacterium
Strain 16S rDNA sequence and GenBank database in Bacillus velezensis CR502 (Genbank accession number:
AY603658 homology) is 99.86%.
The above amplification 16S rDNA universal primer sequence are as follows:
27f:5'-AGAGTTTGATCMTGGCTCAG-3',
1492r:5'-TACGGYTACCTTGTTACGACTT-3'
The PCR system are as follows: 1 × premix rTaq, 0.4 μM of primer 2 7f, 0.4 μM of primer 1492r, 50ng DNA.
The PCR program are as follows: 95 DEG C of initial denaturation 10min, 95 DEG C of denaturation 45sec, 55 DEG C of annealing 45sec, 72 DEG C extend
90sec, 35 circulations；72 DEG C of extension 7min later.
According to Molecular Identification as a result, HGD12 bacterial strain is accredited as Bacillus flexus, HGD3 bacterial strain is accredited as
Pseudomonas putida, HP9 bacterial strain is accredited as Bacillus velezensis.
Embodiment 2: the growth-promoting characteristic measurement of 3 bacterial strains of composite bacteria agent is formed
Soluble phosphorus quantitative determination: the bacterium solution of activation is transferred in 100mL NBRIP fluid nutrient medium, shaking table under the conditions of 30 DEG C
Shaken cultivation 7d takes culture solution to measure pH value after 4 DEG C, 10000rpm centrifugation 10min, and molybdenum blue colorimetric method measures in supernatant
Titanium pigment content deducts titanium pigment content in blank control, is the amount of phosphorus dissolved of each bacterial strain.
Potassium decomposing quantitative determination: by activation bacterium solution switching in sieve husband's fluid nutrient medium of the Alexandria 50mL, under the conditions of 30 DEG C
Shaking table shaken cultivation 5d, the processing of culture solution use hydrogen peroxide ashing method, with water-soluble in aas determination supernatant
Property potassium content, deduct blank control in Content of soluble potassium, be the potassium decomposing amount of each bacterial strain.
Produce ACC deaminase activity measurement: bacterial strain through 60mL TSB fluid nutrient medium activation culture for 24 hours, culture solution 8000rpm
After being centrifuged 10min, being washed 2 times with DF culture solution, it is resuspended in 24mL ADF culture medium and cultivates for 24 hours；Thalline were collected by centrifugation uses
After 0.1mol/L Tris-HCl buffer (pH7.6) washing, it is resuspended in 600 μ L 0.1mol/L Tris-HCl buffers
(pH8.5) in；It adds 30 μ L toluene oscillation 30sec and obtains crude enzyme liquid with smudge cells；Thallus egg is measured with Bradford method
White concentration, with α -one butyric acid content in 2,4-dinitrophenylhydrazine method measurement crude enzyme liquid, ACC deaminase activity representation method are as follows: anti-
Under the conditions of answering, every milligram of mycoprotein is catalyzed ACC deamination per hour and forms α-batanone acid micromole's number, unit μm ol α-butanone
Acid/hmg albumen.
Siderophore assay: the strain inoculated shaking table shaken cultivation 2d in MKB culture medium, 3500rpm centrifugation 15min takes
Supernatant measures siderophore content using CAS method.
Above-mentioned culture medium prescription are as follows:
NBRIP culture medium: glucose 10g, tricalcium phosphate 5g, MgCl2·6H2O 5g, MgSO4·7H2O 0.25g, KCl
0.2g, (NH4)2SO40.1g, 1000mL, pH7.0,115 DEG C of sterilizing 30min of distilled water.
Alexandria sieve husband's culture medium: sucrose 5g, CaCO30.1g, Na2HPO45g, MgSO4·7H2O 0.5g, soil mine
Object 1g, FeCl30.005g, 1000mL, pH7.0-7.5,121 DEG C of sterilizing 20min of distilled water.
TSB culture medium: soy peptone 3g, tryptone 17g, NaCl 5g, glucose 2.5g, K2HPO42.5g, distillation
1000mL, pH7.1-7.5,115 DEG C of sterilizing 30min of water.
DF culture medium: KH2PO44g, Na2HPO46g, MgSO4·7H2O 0.2g, glucose 2g, sodium gluconate 2g, lemon
Sour 2g, (NH4)2SO42g, component one, each 0.1mL of two solution of component, 1000mL, pH7.2,115 DEG C of sterilizing 30min of distilled water.
Wherein group is divided into H3BO310mg, MnSO4·H2O 11.19mg, ZnSO4·7H2O 124.6mg, CuSO4·5H2O 78.22mg,
MoO310mg is dissolved in 100mL sterile purified water；Component two is FeSO4·7H2O 100mg is dissolved in 10mL sterile purified water.
ADF culture medium: ACC (1- amino-cyclopropane -1- carboxylic acid) is dissolved in ultrapure water, filtration sterilization is added to without (NH4)2SO4Sterilizing DF culture medium in, final concentration of 3.0mmol/L.
MKB culture medium: casamino acid 5g, glycerol 15mL, K2HPO42.5g,MgSO4·7H2O 2.5g, distilled water
7.2,115 DEG C of sterilizing 20min of 1000mL, pH.
Growth-promoting characteristic measurement the result shows that, the amount of phosphorus dissolved of HGD12 bacterial strain is 218.88mg/L, and potassium decomposing amount is 2.63mg/L,
Its ACC deaminase activity secreted is 0.27 μm of ol α -one butyric acid/mgh；The amount of phosphorus dissolved of HGD3 bacterial strain is 334.90mg/L, solution
Potassium amount is 2.7mg/L, and the siderophore relative amount of generation is 21.20%；The amount of phosphorus dissolved of HP9 bacterial strain is 149.30mg/L, and ACC is de-
Adnosine deaminase living is 1.58 μm of ol α -one butyric acid/mgh, and siderophore content is 5.70%；3 bacterial strains all have a variety of growth-promoting characteristics.
Embodiment 3: the preparation to the composite bacteria agent of capsicum growth-promoting volume increase
To the composition of the composite bacteria agent of capsicum growth-promoting volume increase: Bacillus flexus HGD12, Pseudomonas
The fermentation liquid of putida HGD3 and Bacillus velezensis HP9 bacterial strain.
The preparation method of the above-mentioned composite bacteria agent to the volume increase of capsicum growth-promoting, includes the following steps:
A, slant activation: by the Bacillus flexus HGD12 of conservation, Pseudomonas putida HGD3 and
Bacillus velezensis HP9 bacterial strain is inoculated in solid medium respectively, is activated overnight bacterial strain；
B, liquid activates: the lawn by step a activated spawn is picked them separately in fluid nutrient medium, in 28-30 DEG C of condition
Under, 150rpm shaking table shaken cultivation to logarithmic growth phase；
C, it obtains the fermentation liquid of each bacterial strain: each strain cultured solution for being in logarithmic growth phase in step b is pressed into 5% volume ratio
It is inoculated in fluid nutrient medium respectively, shaking table 150rpm shaken cultivation 18-36h, respectively obtains Bacillus at 28-30 DEG C
The fermentation liquid of flexus HGD12, Pseudomonas putida HGD3 and Bacillus velezensis HP9 bacterial strain；
D, the preparation of composite bacteria agent: by Bacillus flexus HGD12, Pseudomonas putida in step c
The fermentation liquid equal proportion of HGD3 and Bacillus velezensis HP9 bacterial strain is uniformly mixed, and obtains composite bacteria agent, wherein compound bacteria
Agent is 3 × 10 containing total viable count8-3×109Between CFU/mL.
In above-mentioned preparation method, solid medium used in the inclined-plane culture in step a is NA solid medium；In step
In rapid b, c, fluid nutrient medium used is NA fluid nutrient medium.
In above-mentioned preparation method, based on following ratios, beef extract 5.0g, peptone are contained in NA solid medium used
10.0g, sodium chloride 5.0g, distilled water 1000mL and agar 15-20g；The NA solid medium be pH 7.0-7.2, using it is preceding simultaneously
Sterilize 20min at 121 DEG C.
In above-mentioned preparation method, every liter of NA fluid nutrient medium used contains: beef extract 5.0g, peptone 10.0g, sodium chloride
5.0g and distilled water 1000mL；The NA fluid nutrient medium is pH 7.0-7.2, uses the preceding and 20min that sterilizes at 121 DEG C.
Embodiment 4: to the composite bacteria agent Field information effect of capsicum growth-promoting volume increase
Test site: the sweetgum town village Feng Yuan, the Zunyi City, Guizhou Province area Bo Zhou (106 ° 34 ' 23 " of east longitude, 27 ° 35 ' 39 " of north latitude,
Height above sea level 938m).
Test period: 2018.02-2018.10
Trial crops: capsicum
Test variety: gorgeous green pepper 425 abides by peppery No. 10
Test soil: cultivation soil
Experimental design uses RANDOMIZED BLOCK DESIGN, totally 2 district's groups, and each district's groups set 3 replicated plots, minizone away from
60cm；Every cell is arranged 2 ridges altogether, every ridge ridge face width 60cm, is spaced 40cm between two ridges, and when plantation uses a ridge duplicate rows, and spacing in the rows is
30-40cm, each plot area are 64m2.Capsicum uses floating seedlings mode, is transplanted when growing 6-8 piece true leaf, plants
Training density is 3800-4000 nest/667m2.Each cell water management, the prevention and control of plant diseases, pest control etc. are all the same, by local pepper cultivation side
Formula is managed.
This test sets two processing: (1) as control group, amount of application is compound fertilizer (N-P for common fertilizer application2O5-K2O, always
Nutrient >=45%) 60kg, organic fertilizer (total nutrient N+P2O5+K2O >=5%, organic matter >=45%) 100kg, Diammonium phosphate (DAP)
20kg is applied as base manure.(2)+80% chemical fertilizer application of composite bacteria agent is processing group, and 80% applying quantity of chemical fertilizer is compound fertilizer
48kg/ mus, 16kg/ mus of Diammonium phosphate (DAP), 80kg/ mus of organic fertilizer, 80% chemical fertilizer is applied as base manure；Composite bacteria agent is given birth in capsicum
Application in phase.
The method of administration of composite bacteria agent: in chilli seedling (10d after transplanting), the capsicum early flowering season (colonizing rear 30-40d), peppery
Green pepper bears fruit the phase and (colonizes rear 60-70d) applies respectively, and each dosage is 2-4 liter/mu, and total viable count is 3 × 108-3×
109Between CFU/mL, to carry out pouring root after 25 times of tap water dilutions, 3 times in total.
Test result:
Influence to gorgeous green pepper 425 plant height and stem thickness
As shown in Figure 1, measuring its plant height respectively in seedling stage, early flowering season and the phase of bearing fruit of the growth of gorgeous green pepper 425, compound bacteria is found
Agent processing group has been respectively increased 7.33%, 11.08% and 12.63% compared with the plant height of control group, makes after composite bacteria agent pouring root gorgeous
The plant height of green pepper 425 significantly improves (P < 0.05).
As shown in Fig. 2, measuring its stem thickness respectively in seedling stage, early flowering season and the phase of bearing fruit of the growth of gorgeous green pepper 425, compound bacteria is found
Agent processing group has increased separately 11.86%, 22.14% and 22.14% compared with the stem thickness of control group, makes after composite bacteria agent pouring root gorgeous
The stem thickness of green pepper 425 dramatically increases (P < 0.05).
As shown in table 1, it to the influence of gorgeous green pepper 425 single plant yield, single plant fruiting number and per mu yield, is used reducing by 20% chemical fertilizer
On the basis of amount, inoculating compound bacterium agent group is compared compared with control, has been respectively increased 38.76%, 52.89% and 38.76%, compound bacteria
The inoculation of agent promotes the single plant yield, single plant fruiting number and per mu yield (P < 0.05) of gorgeous green pepper 425 significantly.
Influence of 1 composite bacteria agent of table to gorgeous 425 yield of green pepper
Note: numerical value by the way of " mean+SD ", in same row different lowercases represent significant difference (P <
0.05)。
Influence to peppery No. 10 plant heights, stem thickness and root system development is abided by
As shown in figure 3, measuring its plant height respectively in seedling stage, early flowering season and the phase of bearing fruit for abiding by peppery No. 10 growths, find compound
Microbial inoculum processing group has been respectively increased 9.83%, 17.00% and 16.34% compared with the plant height of control group, makes after composite bacteria agent pouring root
It abides by peppery No. 10 plant heights and significantly improves (P < 0.05).
As shown in figure 4, measuring its plant height respectively in seedling stage, early flowering season and the phase of bearing fruit for abiding by peppery No. 10 growths, find compound
Microbial inoculum processing group has increased separately 16.07%, 18.46% and 18.32% compared with the stem thickness of control group, makes after composite bacteria agent pouring root
The stem thickness of gorgeous green pepper 425 dramatically increases (P < 0.05).
As shown in figure 5, after different disposal group is abided by peppery No. 10 plant taking-up, observing root system development after picking time
Situation is shown in that the plant root development of composite bacteria agent processing group is more vigorous, and root is coarseer, and root hair is more.
As shown in table 2, the influence to peppery No. 10 single plant yields, single plant fruiting number and per mu yield is abided by is reducing by 20% chemical fertilizer
On the basis of dosage, inoculating compound bacterium agent group is compared compared with control, has been respectively increased 22.10%, 7.24% and 22.10%, compound
The inoculation of microbial inoculum promotes significantly abides by peppery No. 10 single plant yields and per mu yield (P < 0.05).
Influence of 2 composite bacteria agent of table to peppery No. 10 yield is abided by
Note: numerical value by the way of " mean+SD ", in same row different lowercases represent significant difference (P <
0.05)。
Conclusion: showing gorgeous green pepper 425 and the Field information for abiding by peppery No. 10 two capsicum varieties, is reducing fertilizer amount 20%
Under the premise of, application composite bacteria agent increases the plant height and stem thickness of pepper plant significantly, improves single plant yield, single plant fruiting
Several and per mu yield has achieved the effect that Reducing amount of chemical fertilizer applied, capsicum volume increase.
Sequence table
<110>Guizhou University
<120>a kind of pair of capsicum has composite bacteria agent and its application of growth-promoting production-increasing function
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1417
<212> DNA
<213>Bacillus flexus HGD12 (Bacillus flexus)
<400> 1
ggctggctcc ataaaggtta ccccaccgac ttcgggtgtt acaaactctc gtggtgtgac 60
gggcggtgtg tacaaggccc gggaacgtat tcaccgcggc atgctgatcc gcgattacta 120
gcgattccag cttcatgtag gcgagttgca gcctacaatc cgaactgaga atggttttat 180
gggattggct tgacctcgcg gtcttgcagc cctttgtacc atccattgta gcacgtgtgt 240
agcccaggtc ataaggggca tgatgatttg acgtcatccc caccttcctc cggtttgtca 300
ccggcagtca ccttagagtg cccaactaaa tgctggcaac taagatcaag ggttgcgctc 360
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cactctgtcc cccgaagggg aacgctctat ctctagagtt gtcagaggat gtcaagacct 480
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cgtcaattcc tttgagtttc agtcttgcga ccgtactccc caggcggagt gcttaatgcg 600
ttagctgcag cactaaaggg cggaaaccct ctaacactta gcactcatcg tttacggcgt 660
ggactaccag ggtatctaat cctgtttgct ccccacgctt tcgcgcctca gcgtcagtta 720
cagaccaaaa agccgccttc gccactggtg ttcctccaca tctctacgca tttcaccgct 780
acacgtggaa ttccgctttt ctcttctgca ctcaagttcc ccagtttcca atgaccctcc 840
acggttgagc cgtgggcttt cacatcagac ttaagaaacc gcctgcgcgc gctttacgcc 900
caataattcc ggataacgct tgccacctac gtattaccgc ggctgctggc acgtagttag 960
ccgtggcttt ctggttaggt accgtcaagg tacaagcagt tactcttgta cttgttcttc 1020
cctaacaaca gagttttacg acccgaaagc cttcatcact cacgcggcgt tgctccgtca 1080
gactttcgtc cattgcggaa gattccctac tgctgcctcc cgtaggagtc tgggccgtgt 1140
ctcagtccca gtatggccga tcccctctca ggtcggctat gcatcgttgc cttggtgagc 1200
cgttacctca ccaactagct aatgcaccgc gggcccatct gtaagtgata gccgaaacca 1260
tctttcaatt ttctcttatg caagagaaaa tgttatccgg tattagctcc ggtttcccgg 1320
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catagaagca agcttctaat cagttcgctc gactgca 1417
<210> 2
<211> 1442
<212> DNA
<213>pseudomonas putida HGD3 (Pseudomonas putida)
<400> 2
cattggaagc gcagctacca tgcaagtcga gcggatgacg ggagcttgct ccttgattca 60
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aggaacgcta ataccgcata cgtcctacgg gagaaagcag gggaccttcg ggccttgcgc 180
tatcagatga gcctaggtcg gattagctag ttggtgaggt aatggctcac caaggcgacg 240
atccgtaact ggtctgagag gatgatcagt cacactggaa ctgagacacg gtccagactc 300
ctacgggagg cagcagtggg gaatattgga caatgggcga aagcctgatc cagccatgcc 360
gcgtgtgtga agaaggtctt cggattgtaa agcactttaa gttgggagga agggcagtaa 420
gttaatacct tgctgttttg acgttaccga cagaataagc accggctaac tctgtgccag 480
cagccgcggt aatacagagg gtgcaagcgt taatcggaat tactgggcgt aaagcgcgcg 540
taggtggttt gttaagttgg atgtgaaagc cccgggctca acctgggaac tgcatccaaa 600
actggcaagc tagagtacgg tagagggtgg tggaatttcc tgtgtagcgg tgaaatgcgt 660
agatatagga aggaacacca gtggcgaagg cgaccacctg gactgatact gacactgagg 720
tgcgaaagcg tggggagcaa acaggattag ataccctggt agtccacgcc gtaaacgatg 780
tcaactagcc gttggaatcc ttgagatttt agtggcgcag ctaacgcatt aagttgaccg 840
cctggggagt acggccgcaa ggttaaaact caaatgaatt gacgggggcc cgcacaagcg 900
gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaggcct tgacatgcag 960
agaactttcc agagatggat tggtgccttc gggaactttg acacaggtgc tgcatggctg 1020
tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgta acgagcgcaa cccttgtcct 1080
tagttaccag cacgtaatgg tgggcactct aaggagactg ccggtgacaa accggaggaa 1140
ggtggggatg acgtcaagtc atcatggccc ttacggcctg ggctacacac gtgctacaat 1200
ggtcggtaca gagggttgcc aagccgcgag gtggagctaa tctcacaaaa ccgatcgtag 1260
tccggatcgc agtctgcaac tcgactgcgt gaagtcggaa tcgctagtaa tcgcgaatca 1320
gaatgtcgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca ccatgggagt 1380
gggttgcacc agaagtagct agtctaacct tcgggaggac ggtaccacgg ttatactgcc 1440
gc 1442
<210> 3
<211> 1454
<212> DNA
<213>Bei Laisi bacillus HP9 (Bacillus velezensis)
<400> 3
aaatttgtca cttcggcggc tggctcctaa aggttacctc accgacttcg ggtgttacaa 60
actctcgtgg tgtgacgggc ggtgtgtaca aggcccggga acgtattcac cgcggcatgc 120
tgatccgcga ttactagcga ttccagcttc acgcagtcga gttgcagact gcgatccgaa 180
ctgagaacag atttgtggga ttggcttaac ctcgcggttt cgctgccctt tgttctgccc 240
attgtagcac gtgtgtagcc caggtcataa ggggcatgat gatttgacgt catccccacc 300
ttcctccggt ttgtcaccgg cagtcacctt agagtgccca actgaatgct ggcaactaag 360
atcaagggtt gcgctcgttg cgggacttaa cccaacatct cacgacacga gctgacgaca 420
accatgcacc acctgtcact ctgcccccga aggggacgtc ctatctctag gattgtcaga 480
ggatgtcaag acctggtaag gttcttcgcg ttgcttcgaa ttaaaccaca tgctccaccg 540
cttgtgcggg cccccgtcaa ttcctttgag tttcagtctt gcgaccgtac tccccaggcg 600
gagtgcttaa tgcgttagct gcagcactaa ggggcggaaa ccccctaaca cttagcactc 660
atcgtttacg gcgtggacta ccagggtatc taatcctgtt cgctccccac gctttcgctc 720
ctcagcgtca gttacagacc agagagtcgc cttcgccact ggtgttcctc cacatctcta 780
cgcatttcac cgctacacgt ggaattccac tctcctcttc tgcactcaag ttccccagtt 840
tccaatgacc ctccccggtt gagccggggg ctttcacatc agacttaaga aaccgcctgc 900
gagcccttta cgcccaataa ttccggacaa cgcttgccac ctacgtatta ccgcggctgc 960
tggcacgtag ttagccgtgg ctttctggtt aggtaccgtc aaggtgccgc cctatttgaa 1020
cggcacttgt tcttccctaa caacagagct ttacgatccg aaaaccttca tcactcacgc 1080
ggcgttgctc cgtcagactt tcgtccattg cggaagattc cctactgctg cctcccgtag 1140
gagtctgggc cgtgtctcag tcccagtgtg gccgatcacc ctctcaggtc ggctacgcat 1200
cgtcgccttg gtgagccgtt acctcaccaa ctagctaatg cgccgcgggt ccatctgtaa 1260
gtggtagccg aagccacctt ttatgtctga accatgcggt tcaaacaacc atccggtatt 1320
agccccggtt tcccggagtt atcccagtct tacaggcagg ttacccacgt gttactcacc 1380
cgtccgccgc taacatcagg gagcaagctc ccatctgtcc gctcgacttg catgtatagc 1440
tgccccaatt ccac 1454
Claims (10)
1. the composite bacteria agent that a kind of pair of capsicum has growth-promoting production-increasing function, it is characterised in that: include Bacillus flexus
The fermentation liquid of HGD12, Pseudomonas putida HGD3 and Bacillus velezensis HP9 bacterial strain；It is described
Bacillus flexus HGD12 is Bacillus flexus, is preserved in China typical culture collection on April 1st, 2019
The heart, deposit number are CCTCC NO:M 2019224；The Pseudomonas putida HGD3 is pseudomonas putida, in
It is preserved in China typical culture collection center on April 1st, 2019, deposit number is CCTCC NO:M 2019223；It is described
Bacillus velezensis HP9 is Bei Laisi bacillus, and Chinese Typical Representative culture guarantor is preserved on April 8th, 2019
Hiding center, deposit number are CCTCC NO:M 2019243.
2. a kind of preparation method of the composite bacteria agent to capsicum as described in claim 1 with growth-promoting production-increasing function, feature
It is: prepares by the following method；
A, slant activation: by Bacillus flexus HGD12, Pseudomonas putida HGD3 and Bacillus
Velezensis HP9 bacterial strain is inoculated in solid medium respectively, is activated overnight bacterial strain；
B, liquid activates: the lawn by step a activated spawn is picked them separately in fluid nutrient medium, under the conditions of 28-30 DEG C,
150rpm shaking table shaken cultivation is to logarithmic growth phase；
C, it obtains the fermentation liquid of each bacterial strain: each strain cultured solution for being in logarithmic growth phase in step b is distinguished by 5% volume ratio
It is inoculated in fluid nutrient medium, shaking table 150rpm shaken cultivation 18-36h at 28-30 DEG C obtains Bacillus flexus
The fermentation liquid of HGD12, Pseudomonas putida HGD3 and Bacillus velezensis HP9 bacterial strain；
D, the preparation of composite bacteria agent: by Bacillus flexus HGD12, Pseudomonas putida HGD3 in step c and
The fermentation liquid equal proportion of Bacillus velezensis HP9 bacterial strain is uniformly mixed, and obtains composite bacteria agent.
3. the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function according to claim 2, feature exist
In: in the step a, solid medium is NA solid medium.
4. the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function according to claim 3, feature exist
In: based on following ratios, beef extract 5.0g, peptone 10.0g, sodium chloride 5.0g, distillation are contained in the NA solid medium
Water 1000mL and agar 15-20g；The NA solid medium is pH7.0-7.2, uses the preceding and 20min that sterilizes at 121 DEG C.
5. the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function according to claim 2, feature exist
In: in step b, c, fluid nutrient medium is NA fluid nutrient medium.
6. the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function according to claim 5, feature exist
In: described every liter of NA fluid nutrient medium contains: beef extract 5.0g, peptone 10.0g, sodium chloride 5.0g and distilled water 1000mL；
The NA fluid nutrient medium is pH7.0-7.2, uses the preceding and 20min that sterilizes at 121 DEG C.
7. the preparation method of the composite bacteria agent to capsicum with growth-promoting production-increasing function according to claim 2, feature exist
In: in the step d, composite bacteria agent is 3 × 10 containing total viable count8-3×109Between CFU/mL.
8. a kind of capsicum growth promoting bacteria agent product, it is characterised in that: the active constituent of the product includes described in claim 1 multiple
Combined bacteria agent.
9. a kind of capsicum growth promoting bacteria agent product, it is characterised in that: the active constituent of the product is described in claim 1 compound
Microbial inoculum.
10. capsicum growth promoting bacteria agent product as claimed in claim 8 or 9, it is characterised in that: further include for the normal of growth promoting bacteria agent
Advise ingredient.
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