CN108841761A - The method and its microbial inoculum used of promotion Growth of Red Clover and/or raising clover yield - Google Patents
The method and its microbial inoculum used of promotion Growth of Red Clover and/or raising clover yield Download PDFInfo
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Abstract
The invention discloses the method for promoting Growth of Red Clover and/or raising clover yield and its microbial inoculums used.The promotion Growth of Red Clover and/or the method for improving clover yield include the steps that being sown into soil again after mixing the seed of clover and microbial inoculum；The microbial inoculum contains the metabolin of clover rhizobia or/and the clover rhizobia；The bacterial strain number of the clover rhizobia is 92-29, is CGMCC No.15533 in the deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms center.Clover rhizobia 92-29 of the invention remarkably promotes Growth of Red Clover under the conditions of acid soil and significantly improves clover yield：Improve plant height, dross rate, root nodule numbers and/or the nodule weight of clover；The plant fresh weight and dry weight for increasing clover, increase green forage yield and the hay yield of clover.The present invention has broad application prospects in clover planting industry.
Description
Technical field
The present invention relates in field of agricultural microorganism promote Growth of Red Clover and/or improve clover yield method and its
Microbial inoculum used.
Background technique
Trifolium (Trifolium) is to be distributed a most wide category in pulse family, is short-term perennial leguminous herbaceous plant.Three
Leaf grass seeds class is all over the world, there are about more than 360 kinds, wherein higher about 25 kinds of agronomic value.China is meadow big country,
Clover is planted extensively in China, is mainly distributed on temperate zone to torrid areas, south to Yunnan, north have to Heilungkiang wild species and
The distribution of cultivar, China cultivate more for red clover, Trifolium repense and miscellaneous three leaf.Clover is have nitrogen fixing capacity excellent
Herbage contains protein abundant and minerals, and cold resistance is strong, and the green phase is very long, while also very heat-resisting, not tight to soil requirement,
It is adapted on acid and basic soil, resistance to trimming, resistance to trample, power of regeneration is strong, anti-harmful gas pollution and disease and insect resistance
Ability is strong, can also increase soil fertility, and the soil moisture is kept to control weed growth, increases covered ground and reduces soil and invades
Erosion.Clover can be used as domestic animal excellent food, the good preceding stubble of crops, green manure, be also good soil-and-water conservation effect and front yard
Garden green plants and nectar source and medicinal material.
Nitrogen is one of most important nutrient in plant growth, and rhizobium are that one kind can infect leguminous plant root
(minority is stem) forms the bacterium that root nodule carries out biological nitrogen fixation, and rhizobium with legume symbiosis system are made in biological nitrogen fixation
With strongest system, the nitrogen-fixing microorganism in soil converts the nitrogen in air in the ammonia of plant available, the nitrogen fixed
About the 65% of biological nitrogen fixation total amount.Currently, leguminous plant is planted in degenerated soil to have been to be concerned by more and more people, because
Nitrogen nutrition is poorer in degenerated soil, and soil fertilizer can be improved by the symbiotic azotification of leguminous plant and rhizobium
Power, Rhizobium Inoculation can be improved yield and the nitrogen reserves of leguminous plant.If the leguminous plant of plantation is without corresponding high
Imitate nitrogen of the Rhizobium strains therewith in symbiosis dross and fixed air, the reference state nitrogen that they will be completely dependent in soil, not only
The nitrogen that can be consumed in soil instead cannot be supplemented, soil fertility is caused to decline.Therefore, in order to give full play to rhizobium and pulse family
The symbiotic azotification of plant needs to screen high efficient strain and carries out artificial infection to maintain soil nitrogen balance.
Summary of the invention
The technical problem to be solved by the present invention is to how Growth of Red Clover and/or raising be remarkably promoted in acid soil
Clover yield.
In order to solve the above technical problems, the present invention provides promote Growth of Red Clover and/or improve clover yield
Microbial inoculum.
The microbial inoculum provided by the present invention for promoting Growth of Red Clover and/or improving clover yield contains clover rhizobia
The metabolin of (Rhizobium trifolii) or/and the clover rhizobia (Rhizobium trifolii)；
The bacterial strain number of the clover rhizobia (Rhizobium trifolii) is 92-29, in China Microbiological bacterium
The deposit number of kind preservation administration committee common micro-organisms center is CGMCC No.15533.
Clover rhizobia 92-29 belongs to Gram-negative, short and small rod-shaped, no gemma, and the raw single flagellum in tool end can be transported
It is dynamic.Thallus size is 0.7 × 2.2 μm.It is grown on yeast juice mannite agar culture medium flat plate, the rounded microprotrusion of bacterium colony,
Neat in edge, surface is smooth relatively wet, light milky white, and lawn is sticky.Clover rhizobia 92-29 has sequence 1 in sequence table
16S rDNA sequence.
Clover rhizobia seed coat agent containing the microbial inoculum or the bio-feritlizer containing the microbial inoculum also belong to the present invention
Protection scope.
Obducens clover seed containing the microbial inoculum also belongs to protection scope of the present invention.
Above, the active constituent of above-mentioned microbial inoculum can be clover rhizobia 92-29 or/and clover rhizobia 92-29
Metabolin, the active constituent of above-mentioned microbial inoculum can also contain other biological ingredient or abiotic component, above-mentioned microbial inoculum other work
Property ingredient those skilled in the art clover Plant weight and/or plant height facilitation effect can be determined according to microbial inoculum.
Above, the microbial inoculum may also include carrier.The carrier can be solid carrier or liquid-carrier.The solid carries
Body is mineral material, biomaterial；The mineral material can for turf, clay, talcum, kaolin, montmorillonite, white carbon, zeolite,
At least one of silica and diatomite；The biomaterial is the stalk of all kinds of crops, loose shell, straw, peanut shell, corn
At least one of powder, bean powder, starch, turf and excrement of animal；The liquid-carrier can be water；In the microbial inoculum, three leaves
The metabolin of careless rhizobium 92-29 or/and clover rhizobia 92-29 can be the fermentation of the living cells, living cells that are cultured
The form of liquid, the filtrate of cell culture or cell and the mixture of filtrate exists.The dosage form of the microbial inoculum can be a variety of dosage forms,
Such as liquor, emulsion, suspending agent, pulvis, granule, wettable powder or water dispersible granules.
Above, as needed, surfactant (such as polysorbas20, Tween 80), bonding can be also added in the microbial inoculum
Agent, stabilizer (such as antioxidant), pH adjusting agent.
In order to solve the above technical problems, the present invention provides promote Growth of Red Clover and/or improve clover yield
Method.
The method provided by the present invention for promoting Growth of Red Clover and/or improving clover yield, including by clover
The step of seed and the microbial inoculum are sown into soil after mixing again.
In above-mentioned text, it is described promote Growth of Red Clover can for improve the plant height of clover, dross rate, root nodule numbers and/or
Nodule weight.
In above-mentioned text, the promotion Growth of Red Clover can be life of the promotion clover in neutral medium or acid medium
It is long.
It is above-mentioned text in, it is described improve clover yield can for improve clover single plant weight (fresh weight and/or dry weight) or
The yield of unit area.
In above-mentioned text, the clover yield that improves can be production of the raising clover in neutral medium or acid medium
Amount.
In above-mentioned text, the medium can be soil.
In above-mentioned text, the pH value of the acid medium may be greater than being equal to 4.2 less than 7, such as 4.8-6.2.
In above-mentioned text, the clover can be Trifolium repense.
Above-mentioned microbial inoculum, above-mentioned clover rhizobia seed coat agent and/or above-mentioned obducens clover seed are promoting clover raw
Application that is long and/or improving in clover yield also belongs to protection scope of the present invention.
It is demonstrated experimentally that clover rhizobia 92-29 of the invention is significant under acid (pH value 4.8-6.2) edaphic condition
Promote Growth of Red Clover and significantly improves clover yield：Improve plant height, dross rate, root nodule numbers and/or the root nodule of clover
Weight；The plant fresh weight and dry weight for increasing clover, increase green forage yield and the hay yield of clover.The present invention is in clover
It has broad application prospects in planting industry.
Preservation explanation
Strain name：Clover rhizobia (Rhizobium trifolii)
Strain number：92-29
Preservation mechanism：China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is referred to as：CGMCC
Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date：On March 30th, 2018
Collection is registered on the books number：CGMCC No.15533
Specific embodiment
The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining
The bright present invention, the range being not intended to be limiting of the invention.Examples provided below can be used as the art ordinary skill
The guide that personnel are further improved, is not construed as limiting the invention in any way.
Experimental method in following embodiments is unless otherwise specified conventional method.Material as used in the following examples
Material, reagent etc., are commercially available unless otherwise specified.
The preparation method of solid medium in following embodiments is as follows：Glycerine 5ml, mannitol 5g, yeast extract 1g,
K2HPO40.5g, anhydrous MgSO40.2g, CaSO4·2H2O 0.2g, NaCl 0.1g, 1% (NH4)6Mo7O24·4H2O1ml、
1%H3BO31ml, agar 20g are settled to 1000ml with water, and tune pH value is 6.8-7.0,121 DEG C of sterilizing 30min.
The preparation method of fluid nutrient medium in following embodiments is as follows：Glycerine 5ml, mannitol 5g, yeast extract 1g,
K2HPO40.5g, anhydrous MgSO40.2g, CaSO4·2H2O 0.2g, NaCl 0.1g, 1% (NH4)6Mo7O24·4H2O1ml、
1%H3BO31ml is settled to 1000ml with water, and tune pH value is 6.8-7.0,121 DEG C of sterilizing 30min.
Clover rhizobia (Rhizobium trifolii) ACCC 18019 in following embodiments is in December, 1981
It is concealed within 19th China Committee for Culture Collection of Microorganisms agricultural microorganism center (abbreviation ACCC, address：Beijing Haidian
Area Zhong Guan-cun South Street 12, INST OF AGRICULTURAL RESOURCES, postcode 100081), from the receipts
The public can obtain the bacterial strain from China Committee for Culture Collection of Microorganisms agricultural microorganism center from hiding day.Clover root nodule
Bacterium (Rhizobium trifolii) ACCC 18019 hereinafter abbreviation clover rhizobia ACCC 18019.
Embodiment 1, the separation of clover rhizobia (Rhizobium trifolii) 92-29CGMCC No.15533 and mirror
It is fixed
1, the separation of bacterial strain
Wild Trifolium repens grass roots tumor is acquired from Qujing of Yunnan area, (takes mannitol 5g, glycerine 5g, yeast leaching with plate
Powder 1g, dipotassium hydrogen phosphate 0.5g, calcium sulfate 0.2g, magnesium sulfate 0.2g, sodium chloride 0.1g, the ammonium molybdate water that mass content is 1%
Solution 1ml and mass content are 1% boric acid aqueous solution 1ml, 0.5% Congo red 1ml, 20g agar, distilled water 1L, pH value are
6.8-7.0) the isolated bacterial strain 92-29 of scribing line.
2, the identification of bacterial strain
2.1, Morphological Identification
Logarithmic growth phase will be in, and bacterium colony size is stablized, above-mentioned steps 1 separate and the bacterial strain 92-29 that purifies into
Row single colonie state observation, main includes size, color, transparency, wettability, the bacterium colony surface state, colony edge of bacterium colony
State.On the other hand, to the bacterial strain 92-29 for being in logarithmic growth phase, optical microphotograph sem observation thallus is used after smear staining
Form.
The result shows that bacterial strain 92-29 belongs to Gram-negative, short and small rod-shaped, no gemma, the raw single flagellum in tool end can be transported
It is dynamic.Thallus size is 0.7 × 2.2 μm.It is grown on yeast juice mannite agar culture medium flat plate, the rounded microprotrusion of bacterium colony,
Neat in edge, surface is smooth relatively wet, and quality is uniform, light milky white, and lawn is sticky.
2.2,16S rDNA sequence homology analysis
Using the 16S rDNA segment of the resulting bacterial strain 92-29 of colony polymerase chain reaction (PCR) method amplification step 1, related reagent is by full formula
King Company provides.To 16S rDNA genetic fragment expand and cloning and sequencing the result shows that, the 16S rDNA of bacterial strain 92-29
Nucleotide sequence with sequence 1 in sequence table.The 16S rDNA and Rhizobium trifolii bacterial strain USDA of bacterial strain 92-29
16S rRNA gene (the Sequence ID of 2370 (T):MRDL01000029 similitude) is up to 99.93%.
2.3, physiological and biochemical property is identified
With reference to《Common bacteria system identification handbook》(east show pearl, the wonderful Beijing English common bacteria system identification handbook of Cai:Section
Publishing house, 2011.) and《Microbiology Experiment》(Beijing Shen Ping, Fan Xiurong, Li Guang force Microbiology Experiment (third edition):
Higher Education Publishing House, 1999.) physiological and biochemical property of measurement bacterial strain 92-29.The result shows that bacterial strain 92-29 is chemoheterotrophy
Type, aerobic growth can utilize glucose, lactose, D-ribose, D- cellobiose, D-arabinose, mannitol, xylose, D- gala
Sugar, fructose, dulcitol and inositol growth；Using ammonium salt, nitrate and most amino acid as nitrogen source, Congo red suction colour response is micro-
Color is inhaled, B.T.B reaction produces acid, and gelatin liquefaction is positive, and cannot hydrolyze starch, cannot utilize cellulose, be unable to hydrolyzed casein, no
Using 3- ketone group lactose, citrate cannot be utilized, extracellular mucus abundant is being often accompanied by containing growth on sugar culture-medium, in beef
It is not grown on cream-peptone culture medium.
In view of above-mentioned form, analysis of physio biochemical characteristics and 16S rDNA sequence homology analysis as a result, step 1 is separated
It purifies obtained bacterial strain 92-29 and is accredited as clover rhizobia (Rhizobium trifolii).The clover rhizobia
(Rhizobium trifolii) 92-29 is preserved in China Committee for Culture Collection of Microorganisms on March 30th, 2018
(abbreviation CGMCC, address are common micro-organisms center：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), deposit number CGMCC
No.15533, hereinafter referred to as clover rhizobia 92-29.
The preparation of embodiment 2, clover rhizobia 92-29 microbial inoculum
1, the inclined-plane culture of clover rhizobia 92-29
The clover rhizobia 92-29 of picking embodiment 1 is inoculated in solid medium and carries out inclined-plane culture, trains at 28 DEG C
It supports 56 hours, obtains the clover rhizobia 92-29 of inclined-plane culture.
2, the activation of strain
The clover rhizobia 92-29 of picking step 1 inclined-plane culture, is inoculated in 200mL fluid nutrient medium, 28
DEG C, shaken cultivation 48 hours under 150rpm, obtain clover rhizobia 92-29 bacterium solution.
3, seed tank culture
The clover rhizobia 92-29 bacterium solution for taking 3L step 2, is accessed in 100L seeding tank (containing 60L Liquid Culture
Base) seed culture is carried out, air agitation culture 48 hours at 28 DEG C, 150rpm obtain clover rhizobia 92-29 seed
Liquid.
4, fermentation tank culture
It will be in the clover rhizobia 92-29 seed liquor access 1000L fermentor of step 3 in 10% ratio (volume ratio)
(containing 600L fluid nutrient medium) carries out fermented and cultured, and air agitation culture 48 hours, obtain clover at 28 DEG C, 150rpm
Rhizobium 92-29 fermentation liquid, in the clover rhizobia 92-29 fermentation liquid content of clover rhizobia 92-29 be 4 ×
109cfu/mL。
5, the preparation of clover rhizobia 92-29 microbial inoculum
It crushes, sieves with 100 mesh sieve after turf is spontaneously dried, be then 7.0 with limewash tune pH value, go out at 121 DEG C
Bacterium 60min obtains sterile turf.
The clover rhizobia 92-29 fermentation liquid of step 4 and the sterile turf are mixed, then are proliferated training under the conditions of 28 DEG C
50h is supported, clover rhizobia 92-29 microbial inoculum is obtained.It is qualified through detection finished product, three in clover rhizobia 92-29 microbial inoculum
The content of leaf grass rhizobium 92-29 is 3.0 × 108Cfu/ grams.
Embodiment 3, clover rhizobia the 92-29 growing state of difference pH on acid medium compare (resistance test)
The pH of culture medium is adjusted to the 1%NaOH solution of sterilizing and 1%HCl solution by aforesaid liquid culture medium after sterilization
4.2,4.4,4.5,4.6,4.8,5.0,5.5,6.0,7.0, obtain the fluid nutrient medium of different pH value, 3 repetitions of each processing,
Clover rhizobia 92-29 and 18019 bacterial strain of clover rhizobia ACCC are inoculated in the fluid nutrient medium of different pH value respectively
In, under the conditions of 150rpm, 28 DEG C after constant-temperature shaking culture 48 hours, the fluid nutrient medium of bacterium is not inoculated with as empty using each handle
Its OD of white blank determination600Value.The results are shown in Table 1, shows that clover rhizobia 92-29 can be in the range that pH value is 4.2-7.0
Interior growth, clover rhizobia ACCC 18019 are only grown in the range of pH value is 4.8-7.0, and are 4.2- in pH value
In the range of 7.0, the upgrowth situation of clover rhizobia 92-29 is better than clover rhizobia ACCC 18019.Illustrate clover
The acid resistance of rhizobium 92-29 is better than clover rhizobia ACCC 18019, is one plant of acidproof rhizobium.
Table 1, clover rhizobia the fluid nutrient medium culture 48 hours of each pH value OD600Value
Embodiment 3, clover rhizobia 92-29 promote Growth of Red Clover and improve clover yield
One, the preparation of 18019 microbial inoculum of clover rhizobia ACCC
1, the inclined-plane culture of clover rhizobia ACCC 18019
Picking clover rhizobia ACCC 18019 is inoculated in solid medium and carries out inclined-plane culture, cultivates 56 at 28 DEG C
Hour, obtain the clover rhizobia ACCC 18019 of inclined-plane culture.
2, the activation of strain
The clover rhizobia ACCC 18019 of picking step 1 inclined-plane culture, is inoculated in 200mL fluid nutrient medium
In, 28 DEG C, shaken cultivation 48 hours under 150rpm, obtain 18019 bacterium solution of clover rhizobia ACCC.
3, seed tank culture
18019 bacterium solution of clover rhizobia ACCC for taking 3L step 2, is accessed in 100L seeding tank (containing 60L liquid
Body culture medium) seed culture is carried out, air agitation culture 48 hours at 28 DEG C, 150rpm obtain clover rhizobia ACCC
18019 seed liquors.
4, fermentation tank culture
18019 seed liquor of the clover rhizobia ACCC access 1000L of step 3 is fermented in 10% ratio (volume ratio)
(containing 600L fluid nutrient medium) carries out fermented and cultured in tank, and air agitation culture 48 hours, obtain three at 28 DEG C, 150rpm
18019 fermentation liquid of leaf grass rhizobium ACCC, clover rhizobia ACCC in 18019 fermentation liquid of clover rhizobia ACCC
18019 content is 4 × 109cfu/mL。
5, the preparation of 18019 microbial inoculum of clover rhizobia ACCC
It crushes, sieves with 100 mesh sieve after turf is spontaneously dried, be then 7.0 with limewash tune pH value, go out at 121 DEG C
Bacterium 60min obtains sterile turf.
18019 fermentation liquid of clover rhizobia ACCC of step 4 and the sterile turf are mixed, then under the conditions of 28 DEG C
Multiplying culture 50h obtains 18019 microbial inoculum of clover rhizobia ACCC.Qualified, the clover rhizobia ACCC through detection finished product
The content of clover rhizobia ACCC 18019 in 18019 microbial inoculums is 3.0 × 108Cfu/ grams.
Two, clover rhizobia 92-29 promotes Growth of Red Clover in the soil of pH6.15 and improves clover yield
Test site is in Yongzhou City, Hubei Province Qiyang County.This test uses RANDOMIZED BLOCK DESIGN, is randomly provided three processing
Area, each treatment region set three cells and repeat.Three treatment regions are respectively Bu Jiejun control treatment area, 92-29 microbial inoculum treatment region
With 18019 microbial inoculum treatment region of ACCC.Each cell size is 4 × 4 ㎡, soil pH value 6.15, Indigenous Rhizobia par
2.0×102Cfu/g dry ground.It is 2kg/hm according to clover application rate2Prepare seed.
Uniform Trifolium repense seed is selected, after being impregnated 1 minute with the concentrated sulfuric acid for several times with sterile water washing, with removal
Until acid, the seed that obtains that treated.Should treated seed carry out following three kinds of processing：
Clover rhizobia 92-29 microbial inoculum and Trifolium repense seed prepared by embodiment 2 is according to 1：10 mass ratio mixing,
It mixes thoroughly, obtained seed is known as 92-29- Trifolium repense seed.
By 18019 microbial inoculum of clover rhizobia ACCC of step 1 preparation and Trifolium repense seed according to 1：10 mass ratio
It mixes, mix thoroughly, obtained seed is known as ACCC 18019- Trifolium repense seed.
By sterile turf and Trifolium repense seed according to 1：10 mass ratio mixing is mixed thoroughly, and obtained seed is known as without Juncao
Charcoal-Trifolium repense seed.
Above-mentioned seed is uniformly seeded into soil according to Quarter Design, and type of seeding is drilling, line-spacing 10cm.It is above-mentioned each
For treatment region in addition to the seed sowed is different, other conditions are identical.Wherein, the seed of Bu Jiejun control treatment area sowing is
Sterile turf-Trifolium repense seed；The seed of 92-29 microbial inoculum treatment region sowing is 92-29- Trifolium repense seed；18019 bacterium of ACCC
The seed of agent treatment region sowing is ACCC 18019- Trifolium repense seed.
Sowing 100 days after under the same conditions, record each plant under ground portion root nodule numbers (abbreviation nodule number),
Lower part root nodule fresh weight (abbreviation nodule weight), under ground portion dross rate (abbreviation dross rate), aerial part plant height, ground
Plant part fresh weight, aerial part plant weights, and the average value of each group is calculated, respectively obtain average dross rate, average root
Tumor quantity, average root nodule fresh weight, aerial part plant average height (abbreviation plant average height), aerial part plant are average fresh
Weight (abbreviation plant mean fresh), aerial part fresh grass weight (abbreviation fresh grass weight), aerial part plant average dry weight are (referred to as
Hay weight).
Table 2, the test result respectively handled
The results are shown in Table 2, shows：
1, compared with the Trifolium repense in Bu Jiejun control treatment area, dross rate increases the Trifolium repense of 92-29 microbial inoculum treatment region
1125%, nodule number increases by 2800%, nodule weight and increases by 143.75%, and every plant of plant average height increases by 96.15%, is every
Strain plant mean fresh increases by 213.33%, fresh grass weight and increases by 108.91%, hay weight increase by 87.67%.
2, compared with the Trifolium repense of 18019 microbial inoculum treatment region of ACCC, dross rate increases the Trifolium repense of 92-29 microbial inoculum treatment region
Add 30.67%, nodule number to increase by 112.20%, nodule weight and increase by 85.71%, every plant of plant average height increases
37.84%, every plant of plant mean fresh increases by 51.61%, fresh grass weight and increases by 36.61%, hay weight increase by 48.91%.
The above results show clover rhizobia 92-29 of the invention to Trifolium repense than clover rhizobia ACCC 18019
There is more significant promotion growth and effect of increasing production in neutral Acid soil.
Three, clover rhizobia 92-29 promotes Growth of Red Clover in the soil of pH4.8 and improves clover yield
Test sets and does not connect bacterium control treatment, 92-29 microbial inoculum is handled and 18019 microbial inoculum of ACCC handles these three processing.
Uniform Trifolium repense seed is selected, after being impregnated 1 minute with the concentrated sulfuric acid for several times with sterile water washing, with removal
Until acid, the seed that obtains that treated.Should treated seed carry out following three kinds of processing：
Clover rhizobia 92-29 microbial inoculum and Trifolium repense seed prepared by embodiment 2 is according to 1：10 mass ratio mixing,
It mixes thoroughly, obtained seed is known as 92-29- Trifolium repense seed.
By 18019 microbial inoculum of clover rhizobia ACCC of step 1 preparation and Trifolium repense seed according to 1：10 mass ratio
It mixes, mix thoroughly, obtained seed is known as ACCC 18019- Trifolium repense seed.
By sterile turf and Trifolium repense seed according to 1：10 mass ratio mixing is mixed thoroughly, and obtained seed is known as without Juncao
Charcoal-Trifolium repense seed.
Above-mentioned sterile turf-Trifolium repense seed, 92-29- Trifolium repense seed and ACCC 18019- Trifolium repense seed are distinguished
It is seeded into the basin equipped with identical soil and (1.5kg soil is housed, the pH value of the soil is 4.80, is taken in the basin of 15cm × 18cm
From Yongzhou City, Hubei Province Qiyang County), every basin sows 30 seeds.In each processing in addition to the seed sowed is different, other conditions
It is identical.Wherein, the seed for not connecing the sowing of bacterium control treatment is sterile turf-Trifolium repense seed；The processing sowing of 92-29 microbial inoculum
Seed be 92-29- Trifolium repense seed；The seed of 18019 microbial inoculum of ACCC processing sowing is ACCC 18019- Trifolium repense seed.
Experiment is repeated four times, and repeats 4 basins of each processing every time.It after planting puts greenhouse to be cultivated under the same conditions to florescence, record is each
The under ground portion dross rate (abbreviation dross rate) of a processing, above-ground plant parts height, aerial part fresh weight (plant fresh weight) and
Aerial part dry weight (plant weights), and the average value of each group is calculated, respectively obtain dross rate, aerial part plant mean height
It is average to spend (abbreviation plant average height), aerial part plant mean fresh (abbreviation plant mean fresh) and aerial part plant
Dry weight (abbreviation plant average dry weight).
Table 3, the test result respectively handled
The results are shown in Table 3, shows：
1,92-29 microbial inoculum processing Trifolium repense compared with the Trifolium repense for not connecing bacterium control treatment, dross rate increase by 1550%,
Every plant of plant average height increases by 66.67%, every plant of plant mean fresh and increases by 121.43%, every plant of plant average dry weight increase
107.14%.
2, compared with the Trifolium repense that 18019 microbial inoculum of ACCC is handled, dross rate increases the Trifolium repense of 92-29 microbial inoculum processing
88.57%, every plant of plant average height increases by 40.00%, every plant of plant mean fresh increase by 55.00%, every plant of plant is averaged
Dry weight increases by 51.63%.
The above results show clover rhizobia 92-29 of the invention to Trifolium repense than clover rhizobia ACCC18019
There is more significant promotion growth and effect of increasing production in acid soil.
The present invention is had been described in detail above.To those skilled in the art, do not depart from spirit of the invention and
Range, and without carrying out under unnecessary experimental conditions, can synchronization parameters, concentration and under the conditions of, it is real in a wider range
Apply the present invention.Although The present invention gives particular embodiments, it is understood that, the present invention can be improved further.
In short, pressing the principle of the present invention, the application is intended to include any change, purposes or improvement of the present invention, including departing from this Shen
Please in the open scope, and the change carried out with routine techniques known in the art.By the range of following attached claims,
It can carry out the application of some essential characteristics.
<110>INST OF AGRICULTURAL RESOURCES
<120>The method and its microbial inoculum used of promotion Growth of Red Clover and/or raising clover yield
<130> GNCFH181205
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1341
<212> DNA
<213>Clover rhizobia（Rhizobium trifolii）
<400> 1
tgcctccttg cggttagcgc actaccttcg ggtaaaacca actcccatgg tgtgacgggc 60
ggtgtgtaca aggcccggga acgtattcac cgcggcatgc tgatccgcga ttactagcga 120
ttccaacttc atgcactcga gttgcagagt gcaatccgaa ctgagatggc ttttggagat 180
tagctcacac tcgcgtgctc gctgcccact gtcaccacca ttgtagcacg tgtgtagccc 240
agcccgtaag ggccatgagg acttgacgtc atccccacct tcctctcggc ttatcaccgg 300
cagtcccctt agagtgccca actgaatgct ggcaactaag ggcgagggtt gcgctcgttg 360
cgggacttaa cccaacatct cacgacacga gctgacgaca gccatgcagc acctgtgtcc 420
cggtccccga agggaacctt gcatctctgc aagtagccgg gcatgtcaag ggctggtaag 480
gttctgcgcg ttgcttcgaa ttaaaccaca tgctccaccg cttgtgcggg cccccgtcaa 540
ttcctttgag ttttaatctt gcgaccgtac tccccaggcg gaatgtttaa tgcgttagct 600
gcgccaccga acagtatact gcccgacggc taacattcat cgtttacggc gtggactacc 660
agggtatcta atcctgtttg ctccccacgc tttcgcacct cagcgtcagt aatggaccag 720
tgagccgcct tcgccactgg tgttcctccg aatatctacg aatttcacct ctacactcgg 780
aattccactc acctcttcca tactccagat cgacagtatc aaaggcagtt ccagggttga 840
gccctgggat ttcacccctg actgatcgat ccgcctacgt gcgctttacg cccagtaatt 900
ccgaacaacg ctagccccct tcgtattacc gcggctgctg gcacgaagtt agccggggct 960
tcttctccgg ataccgtcat tatcttctcc ggtgaaagag ctttacaacc ctagggcctt 1020
catcactcac gcggcatggc tggatcaggc ttgcgcccat tgtccaatat tccccactgc 1080
tgcctcccgt aggagtttgg gccgtgtctc agtcccaatg tggctgatca tcctctcaga 1140
ccagctatgg atcgtcgcct tggtaggcct ttaccccacc aactagctaa tccaacgcgg 1200
gctcatcctt gaccgataaa tctttctccc gaaggacaca tacggtatta gcacaagttt 1260
ccctgcgtta ttccgtagtc aagggtagat tcccacgcgt tactcacccg tctgccgctc 1320
cccttgcggg gcgctcgact g 1341
Claims (10)
1. promoting Growth of Red Clover and/or improving the microbial inoculum of clover yield, it is characterised in that：The microbial inoculum contains clover root
The metabolin of tumor bacterium (Rhizobium trifolii) or/and the clover rhizobia (Rhizobium trifolii)；
The bacterial strain number of the clover rhizobia (Rhizobium trifolii) is 92-29, is protected in Chinese microorganism strain
The deposit number for hiding administration committee's common micro-organisms center is CGMCC No.15533.
2. microbial inoculum according to claim 1, it is characterised in that：The promotion Growth of Red Clover is the strain for improving clover
Height, dross rate, root nodule numbers and/or nodule weight, the clover yield that improves is to improve the single plant weight or list of clover
The yield of plane product.
3. microbial inoculum according to claim 1 or 2, it is characterised in that：The active constituent of the microbial inoculum is the clover root
The metabolin of tumor bacterium (Rhizobium trifolii) or/and the clover rhizobia (Rhizobium trifolii).
4. the clover rhizobia seed coat agent containing microbial inoculum as claimed in any one of claims 1-3 contains in claim 1-3
The bio-feritlizer of any microbial inoculum.
5. containing the obducens clover seed of microbial inoculum as claimed in any one of claims 1-3.
6. the method for promoting Growth of Red Clover and/or improving clover yield, including by the seed of clover and claim 1-3
In any microbial inoculum mix after the step of being sown into soil again.
7. microbial inoculum as claimed in any one of claims 1-3 is promoting Growth of Red Clover and/or is improving the application in clover yield.
8. clover rhizobia seed coat agent as claimed in claim 4 or bio-feritlizer are promoting Growth of Red Clover and/or raising three
Application in leaf Herbage harvest.
9. obducens clover seed described in claim 5 is promoting Growth of Red Clover and/or is improving answering in clover yield
With.
10. according to the method for claim 6, any application in claim 7-9, it is characterised in that：The rush
It is plant height, dross rate, root nodule numbers and/or the nodule weight for improving clover into Growth of Red Clover, the raising clover produces
Amount is the single plant weight of raising clover or the yield of unit area.
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| WO2020161352A1 (en) * | 2019-02-08 | 2020-08-13 | Aphea.Bio Nv | Means and methods for improving plant growth and yield |
| CN116267480A (en) * | 2023-03-16 | 2023-06-23 | 北京市农林科学院 | Method for improving yield and quality of red clover under forest |
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| WO2020161352A1 (en) * | 2019-02-08 | 2020-08-13 | Aphea.Bio Nv | Means and methods for improving plant growth and yield |
| CN113825400A (en) * | 2019-02-08 | 2021-12-21 | 阿法埃生物公司 | Means and methods for improving plant growth and yield |
| CN116267480A (en) * | 2023-03-16 | 2023-06-23 | 北京市农林科学院 | Method for improving yield and quality of red clover under forest |
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