CN107267414A - One plant of plant azotobacteria DJG211 and its application in soil and plant vigor is improved - Google Patents
One plant of plant azotobacteria DJG211 and its application in soil and plant vigor is improved Download PDFInfo
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Abstract
The invention discloses one plant of plant azotobacteria（Phytobacter diazotrophicus）DJG211 and its application in soil and plant vigor is improved, the bacterial strain were deposited in Guangdong Province's Culture Collection on June 5th, 2017（GDMCC）, deposit number is GDMCC No：60195.DJG211 bacterial strains can degrade soil pollutant, cleaning soil, diversity of soil microorganism is improved, so as to promote the nitrogen-fixing plants bacterium of plant growth function in activating soil and Nei Sheng crop plant tissues.It can be obviously promoted the plant growths such as paddy rice, Lettuce in the common wild-rice plant tissue of health.
Description
Technical field
The present invention relates to plant nitrogen fixation technology field, in particular it relates to fixed nitrogen microorganism resource, more particularly, to one plant
Plant azotobacteria DJG211 and its application in soil and plant vigor is improved.
Background technology
Soil pollution can substantially be summarized as organic pollution and the major class of inorganic pollution two.Inorganic matter pollution is mainly salt
Class, acid, alkali, heavy metal, radioactive element compound etc.；Organic Pollution mainly includes synthetic detergent, organic agricultural chemicals, cyaniding
Nuisance that thing, phenols, municipal sewage, oil, sludge and barnyard manure are brought etc..It is more when accumulating harmful substance in soil, exceed
Itself net energy power of soil, structure, composition and the function that may result in soil is changed, and beneficial microbe activity is suppressed,
Harmful substance or its catabolite are built up in soil.By " soil → plant → human body ", or by " soil → water →
Human body " enters absorption of human body indirectly, causes human health damage.Present peasant generally feels soil hardening, and crops is than difficult in the past
Kind.Resident generally feels fruit not as fragrant, melon is not as past sweet tea, dish tasteless in the past, although vegetables increasing number,
It is poorer more than taste as a child.In fact, because we covet increase grain yield in the past so that soil and its production
Environment receives serious destruction.The usage amount whole world first of Chinese chemical fertilizer, excessive fertilizer application causes the life of agricultural production
State key element quality decline, here it is the very crux.China's applications of pesticide amount, up to 1,300,000 tons, is 2.5 times of world average level.
In China, the practical efficiency of agricultural chemicals and chemical fertilizer is less than 30%, and remaining more than 70% all pollutes environment.Applying for pollutant increases
Plus, cause the beneficial bacterium in soil largely to reduce, soil quality declines, soil self purification activity weakens, so as to influence crops
Yield and quality, the harm mankind's is healthy, or even environment revenge risk occurs.
Nitrogen-fixing plants bacterium can within be born in crop plant tissue body, exist in natural environment, with faster breeding
Speed and metabolic process.The category bacterium separated for us and description a new category, with the bacterium affiliation of Enterobacter compared with
Closely.It is derived from common wild-rice tissue, can hydrolysis starch, decomposing protein, fine micro- element etc. of degrading.They exist in soil
In earth, water, plant rhizosphere and its tissue, material conversion and cycle, soil fertility conversion with nature, surrounding material metabolism are close
Cut is closed.While their decomposing force to organic matter are strong, while propagation, the decomposed substance of activity can be disengaged, promotes crop life
It is long.As nitrogen-fixing plants bacterial number increases in soil, he can provide various enzymes for soil, remove the pollutant of soil
Etc. harmful substance and increase diversity of soil microorganism.Using the nitrogen-fixing plants bacterium of the specific pollutant in soil that can degrade,
Bio-feritlizer is configured to, applied to soil restoration, promotes to have great importance in terms of plant growth, offer plant vigor.
The content of the invention
The invention aims to overcome the deficiencies in the prior art, there is provided one plant of plant azotobacteria DJG211.
It is a further object to provide one plant of plant azotobacteria DJG211 in soil and plant vigor is improved
Application.
To achieve these goals, the present invention is achieved by the following technical programs：
One plant of plant azotobacteria（Phytobacter diazotrophicus）DJG211, the bacterial strain was on June 5th, 2017
It is deposited in Guangdong Province's Culture Collection（GDMCC）, deposit number is GDMCC No：60195.Guangdong Province's microbial bacteria
The preservation address for planting collection is 5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100.
Nitrogen-fixing plants bacterium（Phytobacter diazotrophicus）Amylase, catalase, color can be produced
Propylhomoserin deaminase, lysine decarboxylase etc., thus energy hydrolysis starch, protein and fine micro- element etc., so that cleaning soil, increase soil
Microbial diversity and raising soil vitality in earth.The nitrogen-fixing plants bacterium in energy while be born in plant tissue, and generation is biological
Active material, with biological nitrogen fixation function, pollutant that can be in cleaning soil and the growth and breeding for suppressing pathogenic bacteria in soil,
Plant vigor is improved, delays crop aging.Soil vitality and edaphon can be improved using nitrogen-fixing plants bacterial preparation
Diversity, so as to improve quality of agricultural product, effectively improves the yield of crops.
The invention provides one plant from healthy common wild-rice（Oryza rufipogon）It is raw in plant tissue to consolidate
Nitrogen vegetative bacteria, growth potential is good on NFB and PDA culture medium, can carry out fixed nitrogen growth using nitrogen in air.Can be in plant
Survived in rhizosphere and plant tissue, improve plant to pathogen and plant vigor, delay crop aging.16S rRNA gene orders
Analyze bacterial strain DJG211 and nitrogen-fixing plants bacterium type strain LS8THomology is 99.6%.Described nitrogen-fixing plants bacterium
The cellulose and corrupt substance that can be degraded in soil, cleaning soil and improve soil vitality, are obviously promoted paddy rice, Lettuce
Growth and raising yield Deng crop, can be applied to various proportion of crop planting applications.Handled in the nitrogen-fixing plants bacterium through the present invention
Afterwards, remarkably promote the growth of the crops such as paddy rice, Lettuce and delay crop aging.
Plant azotobacteria of the present invention（Phytobacter diazotrophicus）DJG211 has following shape
State and physiology, biochemical characteristic：
A, thalli morphology characteristic：Nitrogen-fixing plants bacterium is Gram-negative bacteria, elongated rod shape, amphimicrobian；Bacterium colony is creamy white, no
Transparent, edge rounding, surface is smooth, cell size（It is long × wide）0.9~1.2 × 0.6~0.9 mm.
B, colonial morphology characteristic：Bacterium colony speed of growth on NFB culture medium flat plates is very fast, and bacterium colony is circular, opaque.30
DEG C, grow 24 hours, 1-3 millimeters of colony diameter.5 DEG C~40 DEG C of growth temperature range and pH growth scopes are 4.0~10.0, can
With the optimal pH growth scope 5.0~7.0 of growth, the NaCl higher than 5% suppresses growth.
C, physio-biochemical characteristics：Gram-negative, bacterial strain growth production acid in improvement PDA liquid medium.Starch Hydrolysis,
Urase, methyl red test, VP (Voges Proskauer) experiment, indole test, casein hydrolysis, catalase are sun
Property, nitrate reductase, gelatin liquefaction it is negative.Can with PEARLITOL 25C, lactulose, maltose, mannosamine, apricot glycosides, ursin, α-
Cyclodextrin, D- melibioses, N- acyl group-D-Glucose amine, N- acyl-beta-D-MANNOSE amine, PEARLITOL 25C, water glycosides poplar,
β-cyclodextrin, i- erythritols, L- trehaloses, D- galactolipins, D- galacturonic acids, alpha-D-glucose, Cori ester
Salt, G-6-P salt, Pfansteihl, vinegar milk acid, D, L- α-glycerine phosphate, Beta-methyl-D-Glucose, m- tartaric acid,
L- urocanic acids, Alpha-Methyl-D-Glucose glycosides, m- inositols, maltotriose, 3- methyl-D-glucoses, Beta-methyl-D-Glucose
Glycosides, 6-o-D- glucopyranose acyl-D- fructofuranoses, D- melitrioses, D-glucitol, stachyose, sucrose, alpha-hydroxybutyric acid,
Beta-hydroxy-butanoic acid, α-ketoglutaric acid, L MALIC ACID, formicester pyruvic acid, D- glucosaccharic acids, succinamic acid, α-butanedioic acid, amber
Sour formicester, L- phenylalanines, Serine, altheine acid, glycyl-L-glutamic acid, L-threonine, thymidine, uridine,
The ￠ of thymidine-5-monophosphate, the ￠ of uridine-5-monophosphate are growth on the culture medium of carbon source.Glycerine, D- sweet dews can not be utilized
It is sugar, gentiobiose, pyruvonitrile acid, Alpha-Methyl-D- galactosides, propylamine acid amide, the amino-histidines of L- third, Pidolidone salt, sweet
Glycyl proline, aminoacyl-L- amine, METHIONINE are growth on the culture medium of sole carbon source.PH growth scopes pH4.0~
pH10.0；There is slight tolerance to NaCl, can be grown under less than 5.0%NaCl concentration.
The 16S rDNA sequences such as SEQ ID NO of the DJG211 bacterial strains：Shown in 1.
Claimed described plant azotobacteria（Phytobacter diazotrophicus）DJG211 exists
Application in degraded parathion.
Claimed described plant azotobacteria（Phytobacter diazotrophicus）DJG211 exists
It is ethene by Acetylene Reduction, the nitrogen in air is converted into ammonia, the application in being utilized for plant.
Claimed described plant azotobacteria（Phytobacter diazotrophicus）DJG211 exists
Promote the application in paddy rice or/and Lettuce growth.
A kind of method for promoting paddy rice or Lettuce to grow, by plant azotobacteria of the present invention（Phytobacter diazotrophicus）DJG211 is prepared into liquid or solid bacterial manure, and liquid or solid bacterial manure is applied into paddy rice or Lettuce.
A kind of microbial inoculum for promoting paddy rice or Lettuce to grow, its preparation method is, plant fixed nitrogen of the present invention is thin
Bacterium DJG211 is based on 28 DEG C using Liquid Culture and cultivated 48 hours, obtains bacterium number up to 4 ' 109CFU.mL-1Zymotic fluid, use vermiculite
Make carrier, zymotic fluid and vermiculite are pressed 2：1 mass ratio mixing, is prepared into bacterium number up to 1.1~2.8 ' 109 CFU.g-1Microbial inoculum.
Compared with prior art, the present invention has the advantages that：
The present invention is one plant of soil pollutant that can degrade, cleaning soil, raising diversity of soil microorganism, so that activating soil
With the nitrogen-fixing plants bacterium for promoting plant growth function in interior raw crop plant tissue.It is from healthy common wild-rice plant group
In knitting, the plant growths such as paddy rice, Lettuce can be obviously promoted.
Brief description of the drawings
Fig. 1 is degradation rate of the nitrogen-fixing plants bacterium DJG211 bacterial strains to 20 mg/L parathion.
Fig. 2 influences for inoculation nitrogen-fixing plants bacterium DJG211 bacterial strains on the aerobic bacteria of soil containing parathion, inoculum concentration 0.2g/
Kg soil.
Fig. 3 influences for inoculation nitrogen-fixing plants bacterium DJG211 bacterial strains on soil actinomycete containing parathion, inoculum concentration 0.2g/kg
Soil.
Fig. 4 is that nitrogen-fixing plants bacterium promotes Lettuce growth.
Embodiment
The present invention is made with reference to Figure of description and specific embodiment and further being elaborated, the embodiment
It is served only for explaining the present invention, is not intended to limit the scope of the present invention.Test method used in following embodiments is such as without spy
Different explanation, is conventional method；Used material, reagent etc., are the reagent commercially obtained unless otherwise specified
And material.
Embodiment 1
The isolation and purification method of bacterial strain of the present invention is as follows：The fresh common wild-rice root sample of collection is net with distilled water flushing,
Then cut root samples to be placed in sterilized culture dish, with 3% hydrogen peroxide (H2O2) handle 4 minutes（To remove root surface
Decaying material）, sterilized water washed once, then with 0.1% mercuric chloride (HgCl2) processing 5 minutes, in sterilized water vibration washing 5~
7 times, 5~10 minutes every time, and last time cleaning solution is coated on NFB the and PDA solid mediums of nitrogen-free or few nitrogen, with
Whether detection sterilization is thorough.The complete common wild-rice root sample of surface sterilization is shredded with the scalpel that sterilized, 10 are made-3,
10-4, 10-5Bacteria suspension is respectively coated NFB, PDA solid culture primary surface, is cultivated in 30 DEG C of artificial incubator, whole
Individual operating process is aseptically completed.After bacterium colony is grown, picking single bacterium colony is coated flat board, until the face of bacterium colony
Color, shape, transparency, quality are consistent.Finally by simple microscopy and dyeing, its form is further looked at, it is consistent, long with width
It is bacterial strain purification standard to spend homogeneous, staining conditions homogeneous.
Preserve：Bacterial strain after isolating and purifying is cultivated after 2~3 d on NFB culture medium flat plates and inclined-plane, collected on flat board
Thalline.Part is stored in 15% sterile glycerol (- 20 DEG C of daily preservations) and (- 80 DEG C long-term to preserve).
The nitrogen-fixing plants bacterium of the present invention（Phytobacter diazotrophicus）DJG211 have following form and
Physiology, biochemical characteristic：
A, thalli morphology characteristic：Nitrogen-fixing plants bacterium is Gram-negative bacteria, elongated rod shape, amphimicrobian；Bacterium colony is creamy white, no
Transparent, edge rounding, surface is smooth, cell size（It is long × wide）0.9~1.2 × 0.6~0.9 mm.
B, colonial morphology characteristic：Bacterium colony speed of growth on NFB culture medium flat plates is very fast, and bacterium colony is circular, opaque.30
DEG C, grow 24 hours, 1-3 millimeters of colony diameter.5 DEG C~40 DEG C of growth temperature range and pH growth scopes are 4.0~10.0, can
With the optimal pH growth scope 5.0~7.0 of growth, the NaCl higher than 5% suppresses growth.
C, physio-biochemical characteristics：Gram-negative, bacterial strain growth production acid in improvement PDA liquid medium.Starch Hydrolysis,
Urase, methyl red test, VP (Voges Proskauer) experiment, indole test, casein hydrolysis, catalase are sun
Property, nitrate reductase, gelatin liquefaction it is negative.Can with PEARLITOL 25C, lactulose, maltose, mannosamine, apricot glycosides, ursin, α-
Cyclodextrin, D- melibioses, N- acyl group-D-Glucose amine, N- acyl-beta-D-MANNOSE amine, PEARLITOL 25C, water glycosides poplar,
β-cyclodextrin, i- erythritols, L- trehaloses, D- galactolipins, D- galacturonic acids, alpha-D-glucose, Cori ester
Salt, G-6-P salt, Pfansteihl, vinegar milk acid, D, L- α-glycerine phosphate, Beta-methyl-D-Glucose, m- tartaric acid,
L- urocanic acids, Alpha-Methyl-D-Glucose glycosides, m- inositols, maltotriose, 3- methyl-D-glucoses, Beta-methyl-D-Glucose
Glycosides, 6-o-D- glucopyranose acyl-D- fructofuranoses, D- melitrioses, D-glucitol, stachyose, sucrose, alpha-hydroxybutyric acid,
Beta-hydroxy-butanoic acid, α-ketoglutaric acid, L MALIC ACID, formicester pyruvic acid, D- glucosaccharic acids, succinamic acid, α-butanedioic acid, amber
Sour formicester, L- phenylalanines, Serine, altheine acid, glycyl-L-glutamic acid, L-threonine, thymidine, uridine,
The ￠ of thymidine-5-monophosphate, the ￠ of uridine-5-monophosphate are growth on the culture medium of carbon source.Glycerine, D- sweet dews can not be utilized
It is sugar, gentiobiose, pyruvonitrile acid, Alpha-Methyl-D- galactosides, propylamine acid amide, the amino-histidines of L- third, Pidolidone salt, sweet
Glycyl proline, aminoacyl-L- amine, METHIONINE are growth on the culture medium of sole carbon source.PH growth scopes pH4.0~
pH10.0；There is slight tolerance to NaCl, can be grown under less than 5.0%NaCl concentration.
The 16S rDNA sequences such as SEQ ID NO of the DJG211 bacterial strains：Shown in 1.
The determination of the molecular classification status of DJG211 bacterial strains of the present invention：Using bacterial 16 S rRNA gene universal primers 25f
DNA amplification in vitro is carried out with 1492r, PCR primer is directly subjected to sequencing.The 16S rDNA sequences that acquisition is sequenced is defeated
Enter GenBank and carry out sequence B last comparisons, primarily determine that the nitrogen-fixing plants bacterium of the present invention（Phytobacter diazotrophicus）The position of genus and species of the bacterial strain in taxology.As a result the nitrogen-fixing plants bacterium of the present invention is found
（Phytobacter diazotrophicus）Belong to vegetative bacteria category（Phytobacter）, with nitrogen-fixing plants bacterium
（Phytobacter diazotrophicus）Type strain LS8THomology is 99.6%, but the sequence incomplete one of the two
Cause, have 0.4% difference, it is interior different bacterial strain of the same race to show the two.With reference to above-mentioned physio-biochemical characteristics, 16S rDNA sequences
Row analysis, nitrogen-fixing plants bacterium of the invention（Phytobacter diazotrophicus）Vegetative bacteria category should be belonged to
（Phytobacter）.
Therefore, the DJG211 bacterial strains of the present invention were deposited in Guangdong Province's Culture Collection on June 5th, 2017
（GDMCC）, deposit number is GDMCC No：60195.The preservation address of Guangdong Province's Culture Collection is that Guangzhou is first
5 building, the strong compound the 59th of Road 100 building.
Embodiment 2
PDA culture medium：200g potatos are weighed, peeling chopping is cleaned, the 1000ml that adds water boils half an hour, filtered through gauze, then
Plus 10-20g glucose, fully dissolving, pH is naturally, 121 °C, steam sterilizing 30min.
Using parathion as the culture medium of sole carbon source：K2HPO4×3H2O 1.30g, MgSO4 × 7H2O 0.50g, FeSO4
×7H2O 0.02g, NaNO33.0g, KCl 0.45g, parathion 20 mg/L, H2O 1000mL, pH value is natural（About 6.5-
7.2）, 121 °C, steam sterilizing 30min.
The application of DJG211 strains for degrading parathion：Nitrogen-fixing plants bacterium DJG211 bacterial strains preculture in PDA is arrived
Exponential phase, then be inoculated into 2% inoculum concentration in 5 mg/L parathion culture medium and adapt to culture 24h, then will adapt to cultivate
Liquid is inoculated into 20 mg/L parathion nutrient solution with 2% inoculum concentration, after cultivating 8 hours, and the degradation rate for determining parathion is
18.2%, a degradation rate was determined every 6 hours later, until after being measured to culture 50 hours, degradation rate has reached 80.5%, survey
Determine result such as Fig. 1.
Embodiment 3
DJG211 bacterial strains improve diversity of soil microorganism and soil activation：By nitrogen-fixing plants bacterium DJG211 bacterial strains in PDA
Thalline is collected by centrifugation to exponential phase in middle preculture, using glucose as carrier, prepares bacteria containing amount about 2x109/ g microbial inoculum.
3L is watered using the mL of 50% parathion 120, takes wherein 1L parathion solution to spray application in the plastic tub of the soil containing 10kg, is placed in
Under natural conditions.Do not connect nitrogen-fixing plants bacterium microbial inoculum for control, be inoculated with the nitrogen-fixing plants bacterium microbial inoculum 0.2g/kg of above-mentioned preparation
Soil for microbial inoculum processing.Three repetitions are often handled, the aerobic bacteria being measured by sampling every 15 days in soil and actinomyces sum,
As a result Fig. 2 and Fig. 3 are seen.After inoculation 15 days, control and processing soil aerobic bacteria quantity and Population of Actinomycetes have different degrees of
Improve.Because nitrogen-fixing plants bacterium DJG211 bacterial strains have degradation to parathion, parathion is relieved to edaphon
Inhibitory action, promotes the breeding of soil aerobic bacteria and actinomyces, improves the diversity and soil activation of edaphon.
Embodiment 4
DJG211 bacterial strains promote paddy growth effect：Nitrogen-fixing plants bacterium is tested using pot-culture method and connects good early effect in bacterium paddy rice.
After nitrogen-fixing plants bacterium bacterial strain is activated, in exponential phase, bacteria suspension 10 is made8Cell .mL-1Concentration processing has 10 days seedlings
Sterile water process is used in the rice seedlings root in age 40 hours, control.Then it is transferred in the plastic cup equipped with 500 grams of rice soil, often
The few nitrogen nutrition liquid of 50mL plants is added in individual plastic cup, 3 reprocessings are cultivated at room temperature.Observe, add water in time daily.
Culture was observed to after 25 days, and it is 10 that test group, which adds corresponding concentration,8Cell .mL-1The few nitrogen battalion of bacterium solution 40mL and 50mL plant
Nutrient solution, control group adds equivalent sterilized water and 50mL plant nitrogen-free nutrient solutions.Continue to cultivate observation to taking pictures after 20 days and record,
Determine root length, leaf length, fresh weight, tiller number, root weight and the dry weight of paddy rice.
The few nitrogen nutrition liquid composition of plant：Calcium sulfate 0.046%；Calcium nitrate 0.003%；Potassium chloride 0.0075%；Phosphoric acid hydrogen
Dipotassium 0.0136%；Ironic citrate 0.0075%；Magnesium sulfate 0.0046%；Liquid microelement 1ml.Liquid microelement is matched：
ZnSO40.22 g；H3BO32.86 g；MnSO41.81 g；H2MoO40.02 g；CuSO40.8 g；H2O 1000 ml〗。
Nitrogen-fixing plants bacterium DJG211 Inoculated Rices after potted plant culture 45 days, take pictures and measure the root length of paddy rice, leaf length,
Fresh weight, tiller number, root weight and dry weight.It the results are shown in Table 1.As can be seen from Table 1, nitrogen-fixing plants bacterium DJG211 Inoculated Rices it
Afterwards, the root length of paddy rice, leaf length, root weight, fresh weight have all reached significant difference compared with the control without inoculating strain, have to paddy rice
It is obvious to promote raw effect.Wherein root increases 52.7%, and leaf length adds 44.9%, and fresh weight adds 73.2%, and root is added again
52.3%, dry weight adds 53.1%.
Growth-promoting effect after the nitrogen-fixing plants bacterium DJG211 Inoculated Rices of table 1
| Bacterial strain | Leaf length (cm) | Root length (cm) | Fresh weight (g) | Dry weight (g) | Root weight (g) |
| CK | 35.07±1.02e | 9.45±0.73c | 1.31±0.09d | 0.52±0.02c | 0.1±0.02d |
| DJG211 | 50.82±0.81abc | 14.43±0.42a | 4.89±0.26bc | 1.11±0.05ab | 0.21±0.02abc |
Note：Significant difference between same column difference lowercase letter processing（p<0.05）
Embodiment 5
Nitrogen-fixing plants bacterium DJG211 promotees Lettuce growth result：After nitrogen-fixing plants bacterium bacterial strain is activated, in exponential phase
It is interior, thalline is collected by centrifugation, bacteria suspension 2x10 is configured to sterile distilled water8Cell .mL-1Concentration sprays the naked oats of 7 days seedling ages
Dish, control sprays processing with sterilized water.Every 10 square metres of vegetable plots go bacteria suspension 10mL to be diluted to 100mL to spray, and 100mL is used in control
Sterilized water is sprayed, after 15 days, effect such as Fig. 4.The Lettuce chlorophyll content for spraying nitrogen-fixing plants bacterium is compareed than spray clear water
Increase by 15.2%, Vc contents than spray clear water control increase by 11.2%, control of the soluble sugar than spray clear water increases by 8.2%, harvest
Fresh weight is than control increase by 20.2% afterwards.
SEQUENCE LISTING
<110>Guangdong Jie Shi agricultural science and technologys Co., Ltd
<120>One plant of plant azotobacteria DJG211 and its application in soil and plant vigor is improved
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1345
<212> DNA
<213>16S rDNA sequences
<400> 1
ggtagcacag ggagcttgct ctcgggtgac gagtggggga cgggtgagta atgtctggga 60
aactgcccga tggaggggga taactactgg aaacggtagc taataccgca taatgtggca 120
agaccaaaga gggggacctt cgggcctctt gccatcggat gtgcccagat gggattagct 180
tgttggtgag gtaatggctc accaaggcga caatccctag ctggtctgag aggatgacca 240
gccacactgg aactgagaca cggtccagac tcctacggga ggcagcagtg gggaatattg 300
cacaatgggc gcaagcctga tgcagccatg ccgcgtgtgt gaagaaggcc ttcgggttgt 360
aaagcacttt cagcggggag gaaggtgtta aggttaataa ccttgtcgat tgacgttacc 420
cgcagaagaa gcaccggcta actccgtgcc agcagccgcg gtaatacgga gggtgcaagc 480
gttaatcgga attactgggc gtaaagcgca cgcaggcggt ctgtcaagtc ggatgtgaaa 540
tccccgggct caacctggga actgcattcg aaactggcag gcttgagtct tgtagagggg 600
ggtagaattc caggtgtagc ggtgaaatgc gtaaagatct ggaggaatac cggtggcgaa 660
ggcggccccc tggacaaaga ctgacgctca ggtgcgaaag cgtggggagc aaacaggatt 720
agataccctg gtagtccacg ccgtaaacga tgtcgacttg gaggttgtgc ccttgaggcg 780
tggcttccgg agctaacgcg ttaagtcgac cgcctgggga gtacggccgc aaggttaaaa 840
ctcaaatgaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgatgcaa 900
cgcgaagaac cttacctggt cttgacatcc acggaatttg gcagagatgc cttagtgcct 960
tcgggaaccg tgagacaggt gctgcatggc tgtcgtcagc tcgtgttgtg aaatgttggg 1020
ttaagtcccg caacgagcgc aacccttatc ctttgttgcc agcggtccgg ccgggaactc 1080
aaaggasact gccagtgata aactggagga aggtggggat gacgtcaagt catcatggcc 1140
cttacgacta gggctacaca cgtgctacaa tggcatatac aaagagaagc gacctcgcga 1200
gagcaagcgg acctcataaa gtatgtcgta gtccggattg gagtctgcaa ctcgactcca 1260
tgaagtcgga atcgctagta atcgtggatc agaatgccac ggtgaatacg ttcccgggcc 1320
ttgtacacac cgcccgtcac accat 1345
Claims (6)
1. one plant of plant azotobacteria（Phytobacter diazotrophicus）DJG211, it is characterised in that the bacterial strain
Guangdong Province's Culture Collection is deposited on June 5th, 2017（GDMCC）, deposit number is GDMCC No：60195.
2. the plant azotobacteria described in claim 1（Phytobacter diazotrophicus）DJG211 is in degraded to sulphur
Application in phosphorus.
3. the plant azotobacteria described in claim 1（Phytobacter diazotrophicus）DJG211 by acetylene also
Originally it was ethene, the nitrogen in air is converted into ammonia, the application in being utilized for plant.
4. the plant azotobacteria described in claim 1（Phytobacter diazotrophicus）DJG211 is promoting paddy rice
Or/and the application in Lettuce growth.
5. a kind of method for promoting paddy rice or Lettuce to grow, it is characterised in that by the plant azotobacteria described in claim 1
（Phytobacter diazotrophicus）DJG211 is prepared into liquid or solid bacterial manure, and liquid or solid bacterial manure is applied to
Paddy rice or Lettuce.
6. a kind of microbial inoculum for promoting paddy rice or Lettuce to grow, it is characterised in that its preparation method is, by described in claim 1
Plant azotobacteria DJG211 using Liquid Culture be based on 28 DEG C cultivate 48 hours, obtain bacterium number up to 4 ' 109CFU.mL-1Hair
Zymotic fluid, carrier is made with vermiculite, and zymotic fluid and vermiculite are pressed into 2：1 mass ratio mixing, is prepared into bacterium number up to 1.1~2.8 ' 109
CFU.g-1Microbial inoculum.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111763650A (en) * | 2020-08-06 | 2020-10-13 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Nitrogen-fixing methylobacterium strain and application thereof |
| CN112746049A (en) * | 2021-03-12 | 2021-05-04 | 浙江大学 | Diazotroph phytobacterium LM-H separated from smelting plant soil and application thereof |
| CN112980730A (en) * | 2021-03-12 | 2021-06-18 | 浙江大学 | Mixed microbial inoculum capable of degrading mixed wastewater of n-butyl alcohol and propylene glycol monobutyl ether and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101255403A (en) * | 2007-12-26 | 2008-09-03 | 北京大学 | Pyridine degradation bacterium strain and uses thereof |
| WO2012009641A2 (en) * | 2010-07-15 | 2012-01-19 | Fbsciences Holdings, Inc. | Microorganism compositions and methods |
| CN103789224A (en) * | 2013-11-14 | 2014-05-14 | 西北大学 | Agrobacterium with free living nitrogen fixation, phosphate dissolution and potassium dissolution capacity and application of agrobacterium |
| CN103789223A (en) * | 2013-11-14 | 2014-05-14 | 西北大学 | Acinetobacter calcoaceticus with free living nitrogen fixation and phosphorus and potassium dissolving capability and application of acinetobacter calcoaceticus |
| CN104805045A (en) * | 2015-05-05 | 2015-07-29 | 中国科学院烟台海岸带研究所 | Plant growth-promoting rhizobacteria (PGPR) and application thereof |
-
2017
- 2017-06-21 CN CN201710476749.7A patent/CN107267414B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101255403A (en) * | 2007-12-26 | 2008-09-03 | 北京大学 | Pyridine degradation bacterium strain and uses thereof |
| WO2012009641A2 (en) * | 2010-07-15 | 2012-01-19 | Fbsciences Holdings, Inc. | Microorganism compositions and methods |
| CN103789224A (en) * | 2013-11-14 | 2014-05-14 | 西北大学 | Agrobacterium with free living nitrogen fixation, phosphate dissolution and potassium dissolution capacity and application of agrobacterium |
| CN103789223A (en) * | 2013-11-14 | 2014-05-14 | 西北大学 | Acinetobacter calcoaceticus with free living nitrogen fixation and phosphorus and potassium dissolving capability and application of acinetobacter calcoaceticus |
| CN104805045A (en) * | 2015-05-05 | 2015-07-29 | 中国科学院烟台海岸带研究所 | Plant growth-promoting rhizobacteria (PGPR) and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 蒋宝贵等: "解磷解钾自生固氮菌的分离筛选及鉴定", 《华中农业大学学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111763650A (en) * | 2020-08-06 | 2020-10-13 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Nitrogen-fixing methylobacterium strain and application thereof |
| CN112746049A (en) * | 2021-03-12 | 2021-05-04 | 浙江大学 | Diazotroph phytobacterium LM-H separated from smelting plant soil and application thereof |
| CN112980730A (en) * | 2021-03-12 | 2021-06-18 | 浙江大学 | Mixed microbial inoculum capable of degrading mixed wastewater of n-butyl alcohol and propylene glycol monobutyl ether and application thereof |
| CN112746049B (en) * | 2021-03-12 | 2022-03-11 | 浙江大学 | Diazotroph phytobacterium LM-H separated from smelting plant soil and application thereof |
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| CN107267414B (en) | 2018-09-11 |
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