CN106399147B - A kind of dedicated molten B. mucilaginocus of rape and its microbial inoculum - Google Patents
A kind of dedicated molten B. mucilaginocus of rape and its microbial inoculum Download PDFInfo
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- CN106399147B CN106399147B CN201610425774.8A CN201610425774A CN106399147B CN 106399147 B CN106399147 B CN 106399147B CN 201610425774 A CN201610425774 A CN 201610425774A CN 106399147 B CN106399147 B CN 106399147B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/06—Arthrobacter
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The present invention relates to a kind of dedicated molten B. mucilaginocus of rape and its microbial inoculums.A kind of bacterial strain of the dedicated molten B. mucilaginocus of rape is named as YK309, classification naming be dirt arthrobacterium (Arthrobacter defluvii), on May 23rd, 2016 is deposited in China typical culture collection center, address: the Chinese Wuhan Wuhan University；Deposit number is CCTCC M 2016275；The dedicated molten B. mucilaginocus of the rape is expanded into culture and obtains microbial inoculum；Microbial inoculum is used to dress seed or be coated or be poured root or root dipping to realize effective inoculation, provides the quick-acting potassium nutritions of high concentration for rape.With rape agglutinin agglutinating reaction can occur for bacterial strain of the invention, therefore affinity with target crop, can settle down for a long time survival in rapeseed root, play and promote growth of rape, improve Potassium Quantity in Plant, increase rape resistance.The present invention can reduce rape cultivation Potassium Fertilizer, improve the utilization rate of potash fertilizer, change that existing bio-feritlizer specificity is strong, the unconspicuous disadvantage of using effect, the development of the advantageous ecological agriculture.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to a kind of dedicated molten B. mucilaginocus of rape and its microbial inoculum.
Background technique
Rape is a kind of important industrial crops, and potassium influences yield of rape and quality.Potassium promotes robust plant raw
Long, the winter strengthens spring hair, increases effective branch amount, silique number and mass of 1000 kernel, improves yield；Potassium promotes photosynthesis, and accelerates light
It closes 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 enhanced
Cold and drought-resistant ability, and can preferably resist a variety of pest and disease damages of rape.Therefore, the chemical potassium that rape cultivation needs largely to apply
Fertilizer.Chemical fertilizer application promotes the production of rape, but crop only has 30% or so to the average utilization of chemical fertilizer, and there are about 30% chemical fertilizer
It is lost by loss, or is fixed in soil.Therefore rapeseed quality largely is influenced using chemical fertilizer, leads to resource, the energy
Waste, and generate serious environmental pollution.
Full potassium content actually in agricultural land soil is very high, but the quick-acting potassium concns that crop can directly be absorbed and utilized in soil
Generally between 40mg/kg~200mg/kg, the 1%~5% of full potassium is only accounted for, is not able to satisfy the needs of crop.There are one in soil
The bacterium of a little solvable potassium, this kind of bacterium can convert the potassium of not solvent to the absorbable water-soluble potassium nutrition utilized of crop,
Promote plant growth.Have the molten B. mucilaginocus production bio-feritlizer of some research and utilizations and be applied to field, it is dense to increase soil effective K
Degree, reduces the amount of application of potash fertilizer.But due to lacking compatibility between the molten B. mucilaginocus used and rape, cannot effectively it exist
Crop root is settled down, thus using effect generates fluctuation, can not play expected growth-promoting yield increasing effect, this predicament hinders molten potassium
The application of bacterium.
All has certain compatibility between the biology that can be interacted in ecological environment, the two can phase in this way
Mutually identification, and then generate stable relations.It is intended to effectively utilize molten B. mucilaginocus, it is necessary to screen affinity with rape
Bacterial strain can just make the dedicated molten potassium microbial inoculum of rape using this compatibility bacterial strain.Up to the present still not about to oil
The research and application of the affinity dedicated molten B. mucilaginocus of dish.
Summary of the invention
In view of the above problems, the present invention provides a kind of dedicated molten B. mucilaginocus of rape and its microbial inoculum.
A kind of bacterial strain of the dedicated molten B. mucilaginocus of rape is named as YK309, and the classification naming of the dedicated molten B. mucilaginocus of rape is
Dirt arthrobacterium (Arthrobacter defluvii), be deposited in China typical culture collection center, address: Chinese is military
Chinese Wuhan University；The deposit date is on May 23rd, 2016, deposit number was CCTCC M 2016275.
The dedicated molten B. mucilaginocus of the rape is expanded culture in beef extract-peptone fluid nutrient medium, and to obtain rape dedicated
Molten potassium microbial inoculum；The dedicated molten potassium microbial inoculum of rape is used for seed dressing or seed pelleting or seedling pours root or seedling root dipping, it is effective to realize
Inoculation increases soil effective K concentration.
Steps are as follows for the specific preparation manipulation of the dedicated molten potassium microbial inoculum of rape:
1 environmental pollution object arthrobacterium is inoculated in the mL beef extract-peptone fluid nutrient medium of 50 mL~100, temperature 25~35
DEG C, under the conditions of revolving speed 100 r/min~180 r/min, shake culture 20h~60h is to get arriving liquid culture；
The liquid culture is the dedicated molten potassium liquid bacterial agent of rape；
The administration dosage of the dedicated molten potassium liquid bacterial agent of rape is 1L/ mus~3L/ mus.
Dry peat 50g~150g is added in the liquid culture described in the mL of 50 mL~100 to mix, obtains rape
Dedicated molten potassium solid fungicide；
The administration dosage of the dedicated molten potassium solid fungicide of rape is 1kg/ mus~3kg/ mus.
Advantageous effects of the invention embody in the following areas:
1. agglutinating reaction can occur with rape agglutinin for the dedicated molten B. mucilaginocus of rape of the invention, therefore have parent with rape
And property, survival can be settled down for a long time in rapeseed root, play the effect for promoting growth of rape, improving yield.
2. the liquid bacterial agent or solid fungicide using the dedicated molten B. mucilaginocus preparation of rape of the present invention have significant molten potassium effect
Fruit is released the potassium of not solvent using the metabolic capability of itself, is increased substantially in bacterium solution or quick-acting potassium in soil
Content, than control increase by 50%~110%, provide the quick-acting potassium nutritions of high concentration for rape.
3. the liquid bacterial agent or solid fungicide using the dedicated molten B. mucilaginocus preparation of rape of the present invention have multinomial excellent life
Reason activity, secretes the siderophore of high concentration, promotes absorption of the rape to ferrite rolling technology；The plant growth that debita spissitudo can be synthesized swashs
Plain IAA promotes plant growth.
4. the administration dosage of liquid bacterial agent or solid fungicide of the invention is low, 1L/ mus~4L/ mus of liquid bacterial agent or solid
1kg/ mus~5kg/ mus of microbial inoculum.Method of administration of the present invention is simple, can be used or dresses seed or be coated or pours the methods of root or root dipping
It is inoculated with.Compared with other biological fertilizer products using 30kg/ mus ~ 50kg/ mus large dosages, the present invention can significantly drop
Low production cost, energy conservation and environmental protection.
In conclusion microbial inoculum of the invention has monospecific to rape, can be colonized simultaneously in rapeseed root long-term surviving
Play physiological function.It can promote absorption of the rape to potassium using microbial inoculum of the invention, improve the increment of rape；Reduce potash fertilizer
Usage amount improves the content of soil available nitrogen, improves the potassium utilization rate of rape.It can change existing bio-fertilizer using the present invention
Expect that specificity is strong, the unconspicuous disadvantage of using effect, the development of the advantageous ecological agriculture.
Specific embodiment
Embodiment 1
The screening and identification of the dedicated molten B. mucilaginocus of rape
1. the young root root system 4g of acquisition health canola plants of normal growth in the soil, by root system be placed in 200 mL without
In bacterium physiological saline, 100 μ L Tween 80s are added, 150 r/min shake 1h and are made 10 using 10 times of dilution methods-1、10-2、10-3、
10-4、10-5、10-5、10-7Gradient suspension, takes 10 respectively-4、10-5、10-5、10-7Dilution 100 μ L of suspension is coated on molten B. mucilaginocus training
Support base plate, 30 DEG C of culture 60h；The biggish bacterial strain of picking transparent circle, purifying of crossing on molten B. mucilaginocus culture medium flat plate, obtains
21 plants of molten B. mucilaginocus bacterial strain.
2. preparing rape agglutinin, it is solidifying to carry out compatibility using 21 plant molten B. mucilaginocus of the rape agglutinin to above-mentioned acquisition
Collection test, obtains 16 plants of 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, which is the dedicated molten B. mucilaginocus of rape.
3. the physiological and biochemical property and its 16S rDNA sequence of the bacterial strain are measured, by this according to universal bacterial identification method
Strain Designation is YK309, classification naming be dirt arthrobacterium (Arthrobacter defluvii), it is deposited in Chinese Typical Representative training
Support object collection, address: the Chinese Wuhan Wuhan University；The deposit date is on May 23rd, 2016, deposit number CCTCC
M 2016275。
Embodiment 2
The dedicated molten B. mucilaginocus dirt arthrobacterium YK309 bacterial strain of rape (Arthrobacter defluvii YK309) molten potassium
Active measurement.
1. prepared by bacterium solution
Preparation measures molten potassium active liquid culture medium, and the culture medium prescription is Na2HPO4 2.0g；MgSO4·7H2O
0.2g；FeCl3 0.05g；Sucrose 5.0g；CaCO3 0.1g；Feldspar in powder 1.0g；Distilled water 1000mL, pH7.4.Take molten potassium living
Property 100 mL of culture medium loaded in 250 mL triangular flasks, every bottle of one ring YK309 bacterial strain lawn of access, control group does not connect strain, often
Group 3 parallel；It 30 DEG C, 120 r/min shaking table culture 45h, obtains and measures the molten active YK309 bacterial strain bacterium solution of potassium.
2. determination of activity
It takes YK309 bacterial strain bacterium solution 80mL, 1200 r/min to be centrifuged 10 min, removes the precipitating such as remaining feldspar in powder；Add
6% H2O2Disappear after boiling 1 h, is centrifuged 12 min using 6 min of ultrasonic cell disruption instrument smudge cells, 6000 r/min, takes
Clear liquid measures potassium content using flame spectrophotometer method.
3. molten potassium determination of activity result
Under the above conditions, the molten potassium activity for measuring YK309 bacterial strain is 52.5 μ g/ml, and compareing is 28.8 μ g/ml,
The molten potassium activity of YK309 bacterial strain is that 182%, the YK309 bacterial strain of control improves the concentration of solution effective K significantly.
Embodiment 3
The preparation and application of the dedicated molten B. mucilaginocus liquid bacterial agent of rape
1. the preparation of the dedicated molten B. mucilaginocus liquid bacterial agent of rape
From 1 ring strain of picking on the molten inclined-plane B. mucilaginocus strain YK309, it is inoculated in equipped with the sterile beef extract-peptone of 150 mL
In 500 mL triangular flasks of fluid nutrient medium, 30 DEG C, 150 r/min shake 38h, obtain the dedicated molten B. mucilaginocus YK309 of rape
Liquid bacterial agent.
2. rape inoculation cultivation
By rape seed as 4~6h of soaking at room temperature in distilled water, it is placed in Light To Dark Ratio 14:10 in 28 DEG C of growth cabinets and urges
Bud to 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 plants 10~15 vernalization and shows money or valuables one carries unintentionally oil
Colza；Control group A is applied fertilizer by normal field cultivation, and control group B and experimental group K Amounts halve；Every basin is inoculated with rape
Dedicated molten 3 mL of B. mucilaginocus YK309 liquid bacterial agent, control group only tap into 3 mL blank cultures；Every basin final singling 3 after seedling all sprouts
Strain, daily management liquid manure harvest rapeseed plants after rapeseed plants grow to 80d, measure rapeseed plants dry weight and blade contains potassium
Amount tests measurement result such as following table.
The data from table are as it can be seen that experimental group rapeseed plants dry weight increases by 42% than control group B, more than control group A；Experimental group
Rape root dry weight is suitable with control group A than control group B increase by 34.4%；Control group of the blade potassium content than applying same horizontal potash fertilizer
B increase by 10.1%, it is suitable with control group A.Potash fertilizer can be greatly lowered in the liquid bacterial agent of the application dedicated molten B. mucilaginocus YK309 of rape
Dosage improves absorption of the rape to potassium, promotes growth of rape.
Embodiment 4
The preparation and application of the dedicated molten B. mucilaginocus solid fungicide of rape
1. the preparation of the dedicated molten B. mucilaginocus solid fungicide of rape
From 1 ring strain of picking on the molten inclined-plane B. mucilaginocus strain YK309, it is inoculated in equipped with the sterile beef extract-peptone of 50 mL
In 250 mL triangular flasks of fluid nutrient medium, 32 DEG C, 130 r/min shake 30h, and dry 80 g of peat mixing is added and obtains
The dedicated molten B. mucilaginocus YK309 solid fungicide of rape.
2. rape inoculation cultivation
Method is the same, and Fertilization Level improves (see the table below), and every basin is inoculated with YK309 solid fungicide 4.5g, and control connects through 100
DEG C, the solid fungicide 4.5g of 20min inactivation treatment；Normal management, harvests rapeseed plants after 90d, measurement rapeseed plants dry weight and
Blade potassium content；The content of soil effective K, calculates the balance of potassium before and after measurement experiment；Measurement result such as following table.
*: containing K in the KCl of control group A application 502mg/ basin in experiment, KCl is 52.4%, calculates amount of potassium applied 263mg, control
Group B and experimental group are 131.5mg.
The data from table are as it can be seen that the experimental group rape dry weight of inoculation YK309 solid fungicide increases by 42.4% than control group B, slightly
Higher than control group A；The experimental group Potassium Quantity in Plant of inoculation YK309 solid fungicide is lower than control group A than control group B increase by 9.9%
5.3%.Potassium Fertilizer can be greatly lowered in the application dedicated molten B. mucilaginocus YK309 solid fungicide of rape, improve Potassium Quantity in Plant, promote
Into growth of rape.
The balance of potassium is calculated according to the data of measurement: potassium amount, positive value after potassium amount-test before Potassium balance=test
Represent apply into potassium be fixed, the potassium that negative value represents original stationary state in soil is released.
Soil is averaged quick-acting potassium content × every basin soil weight+amount of potassium applied before potassium uptake=test before testing
Soil is averaged quick-acting potassium content × every basin soil weight+rape potassium accumulation after potassium uptake=test after test
From the data of calculating it is found that due to applied more potash fertilizer in control group A, cause largely to apply the potassium into soil
Element is fixed, and causes to waste；Control group B potassium is fixed 33.9mg；In control group C, due to not applying into potash fertilizer, in soil
The acquisition of unborn some microorganisms releases a small amount of available potassium, but total amount is inadequate, and plant strain growth is very weak.It is being inoculated with
In the experimental group soil of YK309 solid fungicide, the potassium of original stationary state is also released in soil, and plant strain growth is good.
Utilization rate of fertilizer calculation formula: utilization rate of fertilizer=(A-A0)/B × 100%
In formula
A, potassium application handle 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 potting rape of inoculation YK309 solid fungicide is all remarkably higher than pair for applying phase same level fertilizer to the utilization rate of potash fertilizer
100% level control group A is applied according to a group B, illustrates that utilization rate of fertilizer can be increased substantially by being inoculated with YK309 solid fungicide, promote rape
Absorption to nutrient.The utilization rate of fertilizer of YK309 is 117.5%, illustrates that the utilization rate of potash fertilizer can be improved in molten potassium bacterium YK309,
Slowly available potassium in soil can be converted to available potassium, increase Soil Available potassium content, for Absorption of Rape and utilization, promote rape raw
It is long.
Claims (3)
1. a kind of dedicated molten B. mucilaginocus of rape, it is characterised in that: a kind of bacterial strain of the dedicated molten B. mucilaginocus of rape is named as
YK309, the classification naming of the dedicated molten B. mucilaginocus of rape be dirt arthrobacterium (Arthrobacter defluvii), it is deposited in
State's Type Tissue Collection, the deposit date is on May 23rd, 2016, deposit number was CCTCC M 2016275.
2. a kind of dedicated molten potassium microbial inoculum of rape of the dedicated molten B. mucilaginocus preparation of rape as described in claim 1, it is characterised in that:
1 environmental pollution object arthrobacterium is inoculated in the mL beef extract-peptone fluid nutrient medium of 50 mL~100,25~35 DEG C of temperature,
Under the conditions of revolving speed 100 r/min~180 r/min, shake culture 20h~60h to get arrive liquid culture；
The liquid culture is the dedicated molten potassium liquid bacterial agent of rape；
The administration dosage of the dedicated molten potassium liquid bacterial agent of rape is 1L/ mus~3L/ mus；
The dedicated molten potassium liquid bacterial agent of rape is used for seed pelleting or seedling pours root or seedling root dipping, to realize effectively inoculation, is increased
Add soil effective K concentration.
3. the dedicated molten potassium microbial inoculum of rape of the dedicated molten B. mucilaginocus preparation of a kind of rape according to claim 2, it is characterised in that:
Dry peat 50g~150g is added in the liquid culture described in the mL of 50 mL~100 to mix, it is dedicated to obtain rape
Molten potassium solid fungicide；
The administration dosage of the dedicated molten potassium solid fungicide of rape is 1kg/ mus~3kg/ mus；
The dedicated molten potassium solid fungicide of rape is used to dress seed to realize effective inoculation, increases soil effective K concentration.
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| 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 |
| 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 |
| 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 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Non-Patent Citations (2)
| Title |
|---|
| Arthrobacter defluvii sp. nov., 4-chlorophenol-degrading bacteria isolated from sewage;Kwang Kyu Kim等;《International Journal of Systematic and Evolutionary Microbiology》;20081231;第58卷;1916-1921 * |
| 油菜亲和性根际菌株的筛选及其促生效果;崔松松等;《西北农林科技大学学报（自然科学版）》;20160131;第44卷(第1期);91-96 * |
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