CN106399147A - Potassium-dissolved bacterium special for oilseed rape and potassium-dissolved inoculant prepared by adopting potassium-dissolved bacterium - Google Patents
Potassium-dissolved bacterium special for oilseed rape and potassium-dissolved inoculant prepared by adopting potassium-dissolved bacterium Download PDFInfo
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Abstract
The invention relates to a potassium-dissolved bacterium special for oilseed rape and a potassium-dissolved inoculant prepared by adopting the potassium-dissolved bacterium. A strain of the potassium-dissolved bacterium special for oilseed rape is named as YK309, the classification name is Arthrobacter defluvii, and the strain is preserved in the China Center for Type Culture Collection (CCTCC) (located in Wuhan University, Wuhan, China) on May 23, 2016, and is assigned with the accession number of CCTCC M 2016275. The potassium-dissolved bacterium special for oilseed rape is subjected to amplified culture, and thus the inoculant is obtained. The inoculant is used for seed dressing, film coating, root watering or root dipping, so that the effective inoculation is realized, and thus rapidly available potassium nutrition with high concentration is provided for oilseed rape. The strain provided by the invention can have agglutination reaction with lectin of oilseed rape, therefore, the strain has compatibility with the target crop and can ecize and survive for a long term at the root of the oilseed rape, and thus the efficacies of promoting the growth of the oilseed rape, improving the potassium content of the plant, and enhancing the stress resistance of oilseed rape are achieved. With the adoption of the potassium-dissolved bacterium and the potassium-dissolved inoculant, the use amount of the potassium fertilizer for oilseed rape culture can be reduced, the utilization ratio of the potassium fertilizer can be improved, the defects that the existing biofertilizer is poor in specificity and not obvious in use effect are overcome, and the development of the ecological agriculture is benefited.
Description
Technical field
The invention belongs to microbial technology field is and in particular to a kind of special molten B. mucilaginocus of rape and its microbial inoculum.
Background technology
Rape is a kind of important industrial crops, and potassium has influence on yield of rape and quality.Potassium promotes robust plant life
Long, the winter strengthens the spring and sends out, and increases effective branch amount, silique number and mass of 1000 kernel, improves yield；Potassium promotes photosynthesis, and accelerates light
Close product to shift to seed, the quality of rapeseed can be improved, increase rapeseed oil content；The resistant to lodging, anti-of rape can also be strengthened
Tremble with fear and drought-resistant ability, and can preferably resist multiple pest and disease damages of rape.Therefore, rape cultivation needs a large amount of chemical potassium applied
Fertile.Chemical fertilizer application promotes the production of rape, but crop only has 30% about to the average utilization of chemical fertilizer, there are about 30% chemical fertilizer
Lost by loss, or be fixed in soil.Therefore affect rapeseed quality using chemical fertilizer in a large number, lead to resource, the energy
Waste, and produce serious environmental pollution.
Full potassium content actually in agricultural land soil is very high, but the quick-acting potassium concns that in soil, crop can directly absorb
Typically between 40mg/kg～200mg/kg, only account for the 1%～5% of full potassium it is impossible to meet the needs of crop.Have one in soil
The bacterium of solvable potassium a bit, the potassium of not solvent can be converted into the water-soluble potassium nutrition that crop can absorb utilization by this kind of bacterium,
Promote plant growth.The existing molten B. mucilaginocus of some research and utilizations make bio-feritlizer and are applied to field, increase soil effective K dense
Degree, reduces the amount of application of potash fertilizer.But, due to use molten B. mucilaginocus and rape between lack compatibility it is impossible to effectively exist
Crop root is settled down, thus using effect produces fluctuation it is impossible to play expected growth-promoting yield increasing effect, and this predicament hinders molten potassium
The application of bacterium.
Can occur all possess certain compatibility between the biology interacting in ecological environment, so both can phase
Mutually identify, and then produce stable relations.It is intended to effectively utilize molten B. mucilaginocus and with rape, there is compatibility it is necessary to screen
Bacterial strain, just can make the special molten potassium microbial inoculum of rape using this compatibility bacterial strain.Up to the present still not with regard to oil
Dish has the research of special molten B. mucilaginocus and the application of compatibility.
Content of the invention
For above-mentioned problem, the present invention provides a kind of special molten B. mucilaginocus of rape and its microbial inoculum.
A kind of bacterial strain of the special molten B. mucilaginocus of rape is named as YK309, and the Classification And Nomenclature of the special molten B. mucilaginocus of rape is
Dirt arthrobacterium（Arthrobacter defluvii）, it is deposited in China typical culture collection center, address：China. military
The Chinese. Wuhan University；Preservation date is on May 23rd, 2016, and deposit number is CCTCC M 2016275.
By special for described rape molten B. mucilaginocus in beef extract-peptone fluid nutrient medium Amplification Culture to obtain rape special
Molten potassium microbial inoculum；Special for rape molten potassium microbial inoculum is used for seed dressing or seed pelleting or seedling pours root or seedling root dipping, to realize effectively
Inoculation, increases soil effective K concentration.
The concrete preparation manipulation step of the special molten potassium microbial inoculum of rape is as follows：
1 environmental pollution thing arthrobacterium is inoculated in 50 mL～100 mL beef extract-peptone fluid nutrient mediums, 25～35 DEG C of temperature,
Under the conditions of rotating speed 100 r/min～180 r/min, concussion and cultivate 20h～60h, that is, obtain liquid culture；
Described liquid culture is the special molten potassium liquid bacterial agent of rape；
The application dosage of the special molten potassium liquid bacterial agent of rape is 1L/ mu～3L/ mu.
Add dry peat 50g～150g to mix in liquid culture described in 50 mL～100 mL, that is, rape is obtained
Special molten potassium solid fungicide；
The application dosage of the special molten potassium solid fungicide of rape is 1kg/ mu～3kg/ mu.
The Advantageous Effects of the present invention embody in the following areas：
1. the special molten B. mucilaginocus of the rape of the present invention can occur agglutinating reaction with rape agglutinin, therefore have affine with rape
Property, survival can be settled down for a long time in rapeseed root, play the effect promoting growth of rape, improving yield.
2. significantly molten potassium effect is had using the liquid bacterial agent or solid fungicide of the special molten B. mucilaginocus preparation of rape of the present invention
Really, using the metabolic capability of itself, the potassium of not solvent is discharged, increase substantially in bacterium solution or quick-acting potassium in soil
Content, ratio comparison increase by 50%～110%, for rape provide high concentration quick-acting potassium nutritions.
3. multinomial excellent life is had using the liquid bacterial agent or solid fungicide of the special molten B. mucilaginocus preparation of rape of the present invention
Reason activity, the siderophore of secretion high concentration, promote the absorption to ferrite rolling technology for the rape；The plant growth that debita spissitudo can be synthesized swashs
Plain IAA, promotes plant growth.
4. the liquid bacterial agent of the present invention or the application dosage of solid fungicide are low, liquid bacterial agent 1L/ mu～4L/ mu or solid
Microbial inoculum 1kg/ mu～5kg/ mu.Application process of the present invention is simple, can adopt or dress seed or be coated or pour the method such as root or root dipping
Inoculated.Compared with other use the heavy dose of biological fertilizer products of 30kg/ mu ~ 50kg/ mu, the present invention can significantly drop
Low production cost, energy-conserving and environment-protective.
In sum, the microbial inoculum of the present invention has monospecific to rape, can colonize simultaneously in rapeseed root long-term surviving
Play physiological function.The microbial inoculum of the application present invention can promote the absorption to potassium for the rape, improves the increment of rape；Reduce potash fertilizer
Usage amount, improves the content of soil available nitrogen, improves the potassium utilization rate of rape.The application present invention can change existing bio-fertilizer
Material selectivity is not strong, the unconspicuous disadvantage of using effect, the development of the favourable ecological agriculture.
Specific embodiment
Embodiment 1
The screening of the special molten B. mucilaginocus of rape and identification
1. the young root root system 4g of collection health canola plants of normal growth in soil, root system is placed in the aseptic life of 200 mL
In reason salt solution, add 100 μ L Tween 80s, 150 r/min concussion 1h, using 10 times of dilution methods, make 10-1、10-2、10-3、10-4、
10-5、10-5、10-7Gradient suspension, takes 10 respectively-4、10-5、10-5、10-7Dilution suspension 100 μ L coats molten B. mucilaginocus culture medium
Flat board, 30 DEG C of culture 60h；The larger bacterial strain of picking transparent circle, line on molten B. mucilaginocus culture medium flat plate purifies, and obtains molten potassium
21 plants of bacterium bacterial strain.
2. prepare rape agglutinin, compatibility is carried out using rape agglutinin to 21 plants of molten B. mucilaginocus of above-mentioned acquisition and coagulates
Collection test, obtains 16 plants of the molten potassium bacterial strain with rape compatibility.Compare the molten potassium of these bacterial strains using flame spectrophotometric determination
Activity, finds the molten potassium activity highest of YK309 bacterial strain, and this bacterial strain is the special molten B. mucilaginocus of rape.
3. according to universal bacterial authentication method, measure physiological and biochemical property and its 16S rDNA sequence of this bacterial strain, should
Strain Designation is YK309, and Classification And Nomenclature is dirt arthrobacterium（Arthrobacter defluvii）, it is deposited in Chinese Typical Representative training
Foster thing collection, address：China. Wuhan. Wuhan University；Preservation date is on May 23rd, 2016, and deposit number is CCTCC
M 2016275.
Embodiment 2
The special molten B. mucilaginocus dirt arthrobacterium YK309 bacterial strain of rape（Arthrobacter defluviiYK309）Molten potassium activity
Mensure.
1. bacterium solution preparation
Formation determination molten potassium active liquid culture medium, described culture medium prescription is Na2HPO42.0g；MgSO4·7H2O
0.2g；FeCl30.05g；Sucrose 5.0g；CaCO30.1g；Feldspar in powder 1.0g；Distilled water 1000mL, pH7.4.Molten potassium is taken to live
Property culture medium 100 mL be loaded in 250 mL triangular flasks, every bottle of access one ring YK309 bacterial strain lawn, control group do not connect bacterial classification, often
Group 3 is parallel；30 DEG C, 120 r/min shaking table cultures 45h, obtain the YK309 bacterial strain bacterium solution measuring molten potassium activity.
2. determination of activity
Take YK309 bacterial strain bacterium solution 80mL, 1200 r/min are centrifuged 10 min, remove the precipitation such as feldspar in powder of residual；Plus 6%
H2O2Disappear after boiling 1 h, using ultrasonic cell disruption instrument smudge cells 6 min, 6000 r/min are centrifuged 12 min, take supernatant
Liquid, measures potassium content using flame spectrophotometer method.
3. molten potassium determination of activity result
Under these conditions, the molten potassium activity measuring YK309 bacterial strain is 52.5 μ g/ml, and compares as 28.8 μ g/ml, YK309 bacterium
The molten potassium activity of strain considerably improves the concentration of solution effective K for 182%, the YK309 bacterial strain compareing.
Embodiment 3
The preparation of the special molten B. mucilaginocus liquid bacterial agent of rape and application
1. the preparation of the special molten B. mucilaginocus liquid bacterial agent of rape
Picking 1 ring bacterial classification from molten B. mucilaginocus bacterial classification YK309 inclined-plane, is inoculated in equipped with 150 mL aseptic beef extract-peptone liquid
In 500 mL triangular flasks of culture medium, 30 DEG C, 150 r/min concussion 38h, that is, rape special molten B. mucilaginocus YK309 liquid is obtained
Microbial inoculum.
2. rape inoculation cultivation
By rape seed as soaking at room temperature 4～6h in distilled water, it is placed in Light To Dark Ratio 14 in 28 DEG C of growth cabinets:10 vernalization are extremely
Rape seed shows money or valuables one carries unintentionally.Air-dried drab soil is taken to be placed in flowerpot, 2.5 kg/ basins, every basin is planted 10～15 vernalization and is showed money or valuables one carries unintentionally rape kind
Son；Control group A presses normal field cultivation fertilising, and control group B and experimental group K Amounts all halve；Every basin inoculation rape is special
Molten B. mucilaginocus YK309 liquid bacterial agent 3 mL, control group only taps into 3 mL blank cultures；3 plants of every basin final singling, day after seedling all sprouts
Often management liquid manure, grows to after 80d after rapeseed plants, harvests rapeseed plants, measures rapeseed plants dry weight and blade potassium content, examination
Test measurement result such as following table.
Data from table, experimental group rapeseed plants dry weight increases by 42% than control group B, exceedes control group A；Experimental group
Rape root dry weight is than control group B increase by 34.4%, suitable with control group A；The control group of blade potassium content same horizontal potash fertilizer than applying
B increase by 10.1%, suitable with control group A.The liquid bacterial agent applying the special molten B. mucilaginocus YK309 of rape can be greatly lowered potash fertilizer
Consumption, improves the absorption to potassium for the rape, promotes growth of rape.
Embodiment 4
The preparation of the special molten B. mucilaginocus solid fungicide of rape and application
1. the preparation of the special molten B. mucilaginocus solid fungicide of rape
Picking 1 ring bacterial classification from molten B. mucilaginocus bacterial classification YK309 inclined-plane, is inoculated in equipped with 50 mL aseptic beef extract-peptone liquid
In 250 mL triangular flasks of culture medium, 32 DEG C, 130 r/min concussion 30h, add dry peat 80 g to mix i.e. and rape is obtained
Special molten B. mucilaginocus YK309 solid fungicide.
2. rape inoculation cultivation
Method is the same, and Fertilization Level improves（See table）, every basin inoculates YK309 solid fungicide 4.5g, comparison connect through 100 DEG C,
The solid fungicide 4.5g of 20min inactivation treatment；Normal management, harvests rapeseed plants after 90d, measures rapeseed plants dry weight and blade
Potassium content；The content of soil effective K before and after determination experiment, calculates the balance of potassium；Measurement result such as following table.
*：In experiment, control group A applies the KCl of 502mg/ basin, and containing K in KCl is 52.4%, calculates amount of potassium applied 263mg, comparison
Group B and experimental group are 131.5mg.
Data from table, the experimental group rape dry weight of inoculation YK309 solid fungicide increases by 42.4% than control group B, slightly above
Control group A；The experimental group Potassium Quantity in Plant of inoculation YK309 solid fungicide is than control group B increase by 9.9%, lower by 5.3% than control group A.
Apply the special molten B. mucilaginocus YK309 solid fungicide of rape and Potassium Fertilizer can be greatly lowered, improve Potassium Quantity in Plant, promote rape
Growth.
Calculate the balance of potassium according to the data measuring：Potassium amount after potassium amount-test before Potassium balance=test, on the occasion of
The potassium representing Shi Jin is fixed, and the potassium that negative value represents original stationary state in soil is released.
The average quick-acting potassium content of soil × every basin soil weight+amount of potassium applied before potassium uptake=test before test
The average quick-acting potassium content of soil × every basin soil weight+rape potassium accumulation after potassium uptake=test after test
Knowable to the data calculating, due to application of more potash fertilizer in control group A, lead to apply in a large number the potassium quilt into soil
Fixing, cause to waste；Control group B potassium is fixed 33.9mg；In control group C, due to not applying into potash fertilizer, original in soil
The acquisition of some microorganisms existing releases a small amount of available potassium, but total amount is not, and plant strain growth is very weak.In inoculation YK309
In the experimental group soil of solid fungicide, in soil, the potassium of original stationary state is also released, and plant strain growth is good.
Utilization rate of fertilizer computing formula：Utilization rate of fertilizer=（A-A0）/B×100%
In formula
A, potassium application processes plant total amount containing potassium, mg；
A0, not potassium application plant total amount containing potassium, mg；
B, potassium total amount of applying fertilizer, mg.
The potted plant rape of inoculation YK309 solid fungicide is all remarkably higher than to the utilization rate of potash fertilizer applies the right of phase same level fertilizer
Apply 100% level control group A according to group B, illustrate that inoculating YK309 solid fungicide can increase substantially utilization rate of fertilizer, promote rape
Absorption to nutrient.The utilization rate of fertilizer of YK309 is 117.5%, illustrates that molten potassium bacterium YK309 can improve the utilization rate of potash fertilizer,
Slowly available potassium in soil can be converted into available potassium, increase Soil Available potassium content, for Absorption of Rape and utilization, promote rape life
Long.
Claims (4)
1. a kind of special molten B. mucilaginocus of rape it is characterised in that：A kind of bacterial strain of the special molten B. mucilaginocus of described rape is named as
YK309, the Classification And Nomenclature of the special molten B. mucilaginocus of rape is dirt arthrobacterium（Arthrobacter defluvii）, in being deposited in
State's Type Tissue Collection, preservation date is on May 23rd, 2016, and deposit number is CCTCC M 2016275.
2. by the special molten potassium microbial inoculum of rape of rape a kind of described in claim 1 special molten B. mucilaginocus preparation it is characterised in that：
By special for described rape molten B. mucilaginocus, in beef extract-peptone fluid nutrient medium, Amplification Culture obtains the special molten potassium of rape
Microbial inoculum；
Special for rape molten potassium microbial inoculum is used for seed dressing or seed pelleting or seedling pours root or seedling root dipping, to realize effectively inoculating,
Increase soil effective K concentration.
3. the method for the special molten potassium microbial inoculum of rape described in preparation claim 2 is it is characterised in that concrete operation step is as follows：
1 environmental pollution thing arthrobacterium is inoculated in 50 mL～100 mL beef extract-peptone fluid nutrient mediums, 25～35 DEG C of temperature,
Under the conditions of rotating speed 100 r/min～180 r/min, concussion and cultivate 20h～60h, that is, obtain liquid culture；
Described liquid culture is the special molten potassium liquid bacterial agent of rape；
The application dosage of the special molten potassium liquid bacterial agent of rape is 1L/ mu～3L/ mu.
4. the special molten potassium microbial inoculum of rape according to claim 3 preparation method it is characterised in that：
Add dry peat 50g～150g to mix in liquid culture described in 50 mL～100 mL, that is, rape is obtained special
Molten potassium solid fungicide；
The application dosage of the special molten potassium solid fungicide of rape is 1kg/ mu～3kg/ mu.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107828691A (en) * | 2017-11-28 | 2018-03-23 | 安徽农业大学 | One grows tobacco molten potassium microbial inoculum of specific complex and its preparation method and application |
| CN110317744A (en) * | 2019-05-08 | 2019-10-11 | 安徽农业大学 | A method of producing Marseille bacterium and its production basket purpurin of Indigo pigment |
| CN115197878A (en) * | 2022-06-16 | 2022-10-18 | 安徽农业大学 | Potassium-increasing and quality-improving microbial agent for tobacco and preparation method and application thereof |
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| CN101013135A (en) * | 2007-02-12 | 2007-08-08 | 安徽农业大学 | Method for sifting functional fungoid having specific affinity with non-leguminous plant by using agglutinin |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828691A (en) * | 2017-11-28 | 2018-03-23 | 安徽农业大学 | One grows tobacco molten potassium microbial inoculum of specific complex and its preparation method and application |
| CN107828691B (en) * | 2017-11-28 | 2019-10-08 | 安徽农业大学 | A kind of tobacco compound molten potassium microbial inoculum and its preparation method and application |
| CN110317744A (en) * | 2019-05-08 | 2019-10-11 | 安徽农业大学 | A method of producing Marseille bacterium and its production basket purpurin of Indigo pigment |
| CN110317744B (en) * | 2019-05-08 | 2021-11-05 | 安徽农业大学 | Marseillea fungus for producing blue-violet pigment and method for producing blue-violet pigment by using Marseillea fungus |
| CN115197878A (en) * | 2022-06-16 | 2022-10-18 | 安徽农业大学 | Potassium-increasing and quality-improving microbial agent for tobacco and preparation method and application thereof |
| CN115197878B (en) * | 2022-06-16 | 2023-11-10 | 安徽农业大学 | Potassium-increasing quality-improving microbial agent for tobacco and preparation method and application thereof |
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