CN109929777A - A kind of Halomonas strain H6, composition and its application in salt tolerant growth-promoting - Google Patents
A kind of Halomonas strain H6, composition and its application in salt tolerant growth-promoting Download PDFInfo
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Abstract
The purpose of the present invention is by separation screening obtain one plant of Halomonas (Halomonas sp.) novel bacterial, which, which has, secretes heteroauxin, produces the abilities such as acc deaminase.Can low temperature or high temperature, it is saline and alkaline under the conditions of well grow.Composition using the strain and the microbial-bacterial fertilizer using liquid fermentation technology preparation, alleviate salt Saline Alkali Stress caused by crop, promote growth of the crop under salt stress.Novel bacterial or composition provided by the invention are the effects of being suitable for Special Geographic weather conditions and improving soil physico-chemical property and feeder capability, effective use environmental resource and reduce pollution by pesticides, and have the characteristics that effect is good, the period is short, environment compatibility is good, there is important practical value for solving the current soil salinization.
Description
Technical field:
The present invention relates to microorganism fields, in particular to a kind of new bacterium of the Halomonas with salt tolerant growth-promoting functions
Strain, composition, and its application in terms of improving crops Salt Stress Tolerance and promoting crops seedling stage growth.
Background technique:
The soil salinization and because irrigate caused by soil secondary salinization problem be that an important ecological environment is asked
Topic typically occurs in the area that drought, soil evaporation intensity is big, level of ground water is high and contains more solubility salt.Soil
Earth salination problem has become influence arid, semiarid zone agricultural, animal husbandry and sustainable economic development, has threatened ecology
Safe and stable key factor.
The soil salinization is a kind of main environmental risk as caused by nature or mankind's activity.According to joint state religion section
Text tissue and food and agricultural organization's incomplete statistics, the area in whole world salt-soda soil are 9.5 hundred million hm2.The distribution of China's salinized soil
Extensively, area is big, type is more, about 100,000,000 hm of the gross area2, wherein modern salinized soil accounts for about 37%, remain salinized soil and account for about 45%, dive
Account for about 18% in salinized soil, occurs mainly in arid, semiarid and subhumid regions.Harm master of the salinized soil to plant growth
In terms of showing the salt damage of plant and the salt-resistance of plant.Salinity is excessive in soil, can lead to plant root cell to soil
Interior moisture absorption is relatively difficult, or even cases of dehydration occurs, while plant also being made to generate mono-salt toxic action, causes normal life
Manage the destruction of metabolism.After salt damage occurs for plant, gently then growth and development is affected, heavy then lead to Plant death.
Xinjiang is the maximum oasis cultivation region in China, is increased recently as agricultural land mining inetesity, and water resource does not conform to
Reason such as utilizes to show the soil salinization and soil degradation problem gradually at the factors.They all have salination and desertification and deposit,
A series of salification and the features such as the poly- property of table is stronger and soil salt complicated component, to bring ecological environment problems and restrict
Agricultural production and socio-economic development.Xinjiang salinization soil mainly utilizes water-saving irrigation and physics, chemistry, biology etc.
A series of measures carries out comprehensive treatment improvement.By the improvement of many years, although Saline Ground in Xinjiang has different degrees of improvement, but
With the implementation of large-scale water-saving irrigation, Soil Salinization Development shows new problem and rule, such as under-film drip irrigation item again
Salification problem, dry basin inland salinity problem of outlet under part etc., and it is all very high to capital investment and technical requirements, to big
The reparation in area salt-soda soil implements relatively difficult.Blindly the reclamation of wasteland, soil out-of-flatness, long-term cropping are single etc. simultaneously causes
Soil with support imbalance, organic manure application rate reduce caused by crop anti-salt property reduce the problems such as also get worse.And conduct
The most economical effective measures that salinization land restores, use saline alkali tolerant plant kind and use biological agent to improve plant
Saline-alkali tolerant characteristic is that Xinjiang can Devoting Major Efforts To Developing and utilization to overcome the saline and alkaline biological modification technology to plant stress effect
One of technology.
But how to improve the Saline alkali tolerance of crops, saline-alkali tolerance has become the key subjects that researcher faces.Nothing
By the angle from improvement soil, or the angle of crop tolerance to salt alkalinity is improved, promotion plant is all solution in the growth of saline environment
The certainly key of problem.Plant growth-promoting rhizobacteria (Plantgrowth-promoting rhizobacteria, PGPR) is plant
It is a kind of in rhizosphere soil to promote plant growth, controlling disease, the microorganism for increasing crop yield.It will be helped using PGPR
In mitigating adverse effect of the salinity to plant, promote the utilization of decomposition of soil organic matter, Nutrient Cycling and plant nutrient, in turn
Phytomass or yield are improved, has become environmental-friendly, the cost-effective promotion plant of one kind at present in salinization soil
The development tactics of growth.Studies have shown that plant growth-promoting rhizobacteria is by symbiotic nitrogen fixation, dissolved metals hydrochlorate and improves nutrient
Several different mechanisms such as absorption, generation plant hormone promote plant growth in halomorphic soils, increase crop yield, pass through raising
The accumulation of plant osmosis Auto-regulator increases K+Concentration and the high K of maintenance+/Na+Than etc. mitigate salt stress on crop influence, root
Border bacterium generates acc deaminase, inducible system resistance, inducible system tolerance and also helps to the control of plant pathogenic microorganisms
Alleviate salt stress in plant.
Although both at home and abroad it has been found that a large amount of plant growth-promoting bacteria resources, in microbial control industry, screening promotes plant
The PGPR bacterial strain that grows and gesture exists for the research and development that bacterial strain is really applied to crop field under the conditions of arid, Salt Strees Condition
It must go.Due to Xinjiang region environment arid, there is related microorganisms bad adaptability, ineffective in saline and alkaline multiplicity.Cause
This, screens and develops and be suitable for Xinjiang Special Geographic weather conditions and have plant good alleviating salt stress, promoting crop raw
The effects of long, improvement soil physico-chemical property and feeder capability, effective use environmental resource and reduction pollution by pesticides, and there is effect
The microorganism novel bacterial that fruit is good, the period is short, environment compatibility is good, for solve the current soil salinization have it is important practical
Value.
Summary of the invention:
The purpose of the present invention is obtain one plant of Halomonas (Halomonas sp.) novel bacterial, the bacterium by separation screening
Kind there is secretion heteroauxin, produce the abilities such as acc deaminase, using the strain composition and utilize liquid fermentation technology system
Standby microbial-bacterial fertilizer, can significantly improve the salt-tolerant trait of plant, and can promote plant growth.
The purpose of the present invention is what is be achieved through the following technical solutions:
Technical solution of the present invention: by obtaining one plant of salt list from Xinjiang Hesuo area halophytes rhizosphere separation screening
Born of the same parents bacterium novel bacterial (Halomonas sp.) H6 is prepared for salt tolerant growth-promoting microbial-bacterial fertilizer using liquid fermentation technology, obtains good
Good effect.Present invention synthesis solves microbial-bacterial fertilizer in the adaptability in salt-soda soil and the effect of growth-promoting, while making letter
It is single, it is convenient and practical.
Specifically, the present invention provides the novel bacterial salt unit cells that one plant is isolated from Xinjiang Hesuo area halophytes rhizosphere
Bacterium (Halomonas sp.) H6CGMCC No.15758.The bacterial strain has salt tolerant growth-promoting and secretion heteroauxin and production ACC de-
The abilities such as adnosine deaminase can be applied to improve the saline and alkaline resistance of plant and promote growth, have the applications well for being developed as bio-feritlizer
Prospect.
Novel bacterial Halomonas (Halomonas sp.) H6 CGMCC No.15758 provided by the present invention, from Xinjiang and
The large saline and alkaline Desert Area plant rhizosphere in county separates, screening, tames acquisition, and bacterium numbering H6 detects mirror through microbial taxonomy
It is fixed, it is determined as the potential novel species of bacterium Halomonas (Halomonas sp.) category.Currently, the bacterial strain was protected before the applying date
It is hidden in budapest treaty microorganism International Depository Authority: in China Committee for Culture Collection of Microorganisms's common micro-organisms
The heart (CGMCC).Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode:
100101, the deposit date is on May 14th, 2018, deposit number was CGMCC No.15758.
Novel bacterial salt tolerant Promoting bacteria Halomonas (Halomonas sp.) H6CGMCC that the present invention specifically provides
No.15758 is Gram-positive, rod-shaped, 0.3-0.8 × 1.4-2.5 μm, does not form endospore.It is grown in containing 2%
After cultivating 3-5d on the TSA culture medium of NaCl, bacterium colony is circle, neat in edge, slightly convex faint yellow bacterium colony.Thallus is raw
For long temperature range at 10-45 DEG C, optimum temperature growth is 28 DEG C；PH growth scope 5.0-10, optimal pH 7.0 are resistant to
By the NaCl of 1-20%.
Using the genome DNA of novel bacterial Halomonas (Halomonas sp.) H6CGMCC No.15758 bacterial strain as mould
Plate expands its 16S rDNA sequence, is sequenced, and obtains the 16S rDNA sequence that length is 1431bp, sees sequence table, with
The related strain that GenBank database is included carries out sequence analysis analysis, determines that the bacterium is subordinate to Halomonas Pseudomonas, with
Category mode standard bacterium Halomonas titanicae BH1TWith Halomonas neptunia Eplume1THomology difference
For 98.6% and 98.3%, formal microbial taxonomy not yet is carried out to the bacterial strain at present and is named, fixing tentatively strain name is salt
Monad (Halomonas sp.) H6.
Meanwhile the present invention provides the culture using above-mentioned Halomonas (Halomonas sp.) H6 bacterial strain.
The present invention further provides the combinations using above-mentioned Halomonas (Halomonas sp.) H6 strain culture
Object, the composition are liquid, freezing or dry powdered form.
It include auxiliary agent in above-mentioned composition in the present invention.
It further include solid-state carrier when the composition is freezing or dry powdered form in the present invention.
In the present invention, the solid-state carrier includes peat, turf, talcum, lignite, pyrophyllite, montmorillonite, alginates, pressure
Filter mud slurry, sawdust, perlite, mica, silica, silica flour, calcium-base bentonite, vermiculite, kaolin, precipitated calcium carbonate, diatom
One of soil, medical stone, calcite, zeolite, white carbon black, fine sand and clay are a variety of.
In the present invention, the auxiliary agent includes neopelex, sodium butylnaphthalenesulfonate, trehalose, glycerol, wooden
It is plain sodium sulfonate, Negel condensation polymer, niacin, alcohol, buffer salt, sodium chloride, amino acid, vitamins, protein, more
One of peptide, polysaccharide or monosaccharide, yeast extract, white carbon black, Tea Saponin, defatted milk are a variety of.
It also include a effective amount of chemical combination selected from the following of agricultural using the composition of above-mentioned offer series in the present invention
Object or composition: nutrient, fertilizer, microbial-bacterial fertilizer.
In the present invention, cotton is being improved using Halomonas (Halomonas sp.) H6 or composition of above-mentioned offer
Salt Stress Tolerance and the application for promoting crops seedling stage growth aspect.
Specifically, novel bacterial Halomonas (Halomonas sp.) H6 CGMCC No.15758 bacterial strain of the present invention
Feature be can low temperature or high temperature, it is saline and alkaline under the conditions of well grow, salt can be relieved, the economy based on cotton is made
Saline Alkali Stress caused by object can especially promote life of the cotton under salt stress by growth metabolism plant auxin secretion
It is long.
In the present invention, cotton is being improved using Halomonas (Halomonas sp.) H6 or composition of above-mentioned offer
Or wheat Salt Stress Tolerance and promote crops seedling stage growth aspect application.
The invention has the following beneficial technical effects:
(1) novel bacterial Halomonas (Halomonas sp.) H6CGMCC No.15758 bacterial strain provided by the invention, has
The characteristics of low temperature, high growth temperature, resistance to high salt and high pH, thus it can be relieved Saline Alkali Stress, promote plant growth.And have
Can synthesis of indole acetic acid, produce acc deaminase, play the role of promoting growth of plants.
(2) novel bacterial Halomonas (Halomonas sp.) H6CGMCC No.15758 bacterial strain provided by the invention, can make
After the cotton seed impregnated through bacterium solution dilution is grown 15 days, there is apparent facilitation.Compared with the control group, microbial inoculum has been used
Processing group root long and plant weight significantly increase 11.54% and 15.38% respectively；Wheat is through Halomonas H6 microbial inoculum dilution
After impregnating growth 15 days, the facilitation of root long and fresh weight is obvious, significantly increases 23.08% and 41.67% respectively.It proves
Cotton can be significantly improved under salt stress using microbial inoculum Halomonas (Halomonas sp.) H6 to grow, and there is obvious salt tolerant to promote
Raw effect.
(3) present invention is improving cotton and small using Halomonas (Halomonas sp.) H6 or composition provided
Wheat Salt Stress Tolerance and the application for promoting crops seedling stage growth aspect, have obvious salt tolerant growth-promoting functions, this can not only subtract
The dosage of few chemical fertilizer can also save peasant's spending, increase income, also have good ecological benefits and society
Benefit.
Detailed description of the invention:
Fig. 1 show Halomonas (Halomonas sp.) H6CGMCC No.15758 phylogenetic tree.
Fig. 2 show the colonial morphology of Halomonas (Halomonas sp.) H6CGMCC No.15758.
Fig. 3 show what different NaCl concentrations grew Halomonas (Halomonas sp.) H6 CGMCC No.15758
It influences.
Fig. 4 show the influence that different pH value grow Halomonas (Halomonas sp.) H6CGMCC No.15758.
Fig. 5 show the growth of Halomonas (Halomonas sp.) H6CGMCC No.15758 in the fermentation medium
Curve.
Fig. 6 show Halomonas (Halomonas sp.) H6CGMCC No.15758 and makees to the growth-promoting of cotton seedling
With.
Fig. 7 show Halomonas (Halomonas sp.) H6CGMCC No.15758 and makees to the growth-promoting of wheat seedling
With.
Specific embodiment:
In order to better explain the present invention, below in conjunction with the specific embodiment main contents that the present invention is furture elucidated, but
The contents of the present invention are not limited solely to following embodiment.Unless otherwise specified, technological means used in embodiment is ability
Conventional means known to field technique personnel, raw materials used is commercial goods, can be bought by public channel, in technique
Used equipment and instrument are the common equipment in this field.
2%NaCl TSA culture medium: pancreas casein peptone 15g, soy peptone 5g, NaCl 20g, agar 15g, H2O
1000mL, pH 7.3.
Fermentation medium component (g/L): glucose 3.0, peptone 5.0, yeast extract 2.0, K2HPO43.0, NaCl
20.0 pH7.0-7.2.
DF culture medium (g/L): KH2PO44.0, Na2HPO46.0, MgSO4·7H2O 0.2, glucose 2, gluconic acid
Sodium 2, citric acid 2, (NH4)2SO42.0, FeSO4·7H2O 0.001, each 0.1mL of the trace element solution of component one and two, sample
It is added, sufficiently dissolves, 121 DEG C of high pressure sterilization 20min one by one.Trace element solution: component one: H3BO3 10mg、MnSO4·
H2O 11.19mg、ZnSO4·7H2O 124.6mg、CuSO4·5H2O 78.22mg、MoO3 10mg、 H2O 100mL, is dissolved in
In 100mL sterile distilled water.Component two: by 100mg FeSO4·7H2O is dissolved in 100mL sterile distilled water.
Embodiment one: separation, the screening and identification of Halomonas (Halomonas sp.) H6
(1) it separates, screen:
The saline and alkaline Desert Area plant rhizosphere soil sample in Xinjiang Heshuo County is acquired, microorganism is carried out by gradient dilution method
Separation and purifying.Soil sample 10g is weighed, is put into 100mL sterile water, the bead of sterilizing is added, 150rpm, shakes by 30 DEG C
20min keeps sample fully dispersed.100 μ L of sterile working Aspirate supernatant is added in 900 μ L sterile waters, is diluted and is made with sterile water
At 10-2-10-5The dilution of concentration.Each concentration dilution liquid for taking 100 μ L, using customary coating methods in the TSA containing 2%NaCl
It is coated with, is placed in 30 DEG C of constant incubators after culture 48 hours in solid medium tablets, after growing bacterium colony on plate,
Single colonie on picking plate carries out purifying culture, up to no miscellaneous bacteria falls.Therefrom preferably go out the bacterial strain that one plant of number is H6, turns
It is connected in the medium slant of TSA containing 2%NaCl and saves backup.
(2) classify, identify:
1, the sequence and analysis of the 16S rDNA of Halomonas (Halomonas sp.) H6:
(1) extraction of pcr template DNA:
Strain H6 after purification is inoculated into the TSA culture medium of 2%NaCl, 30 DEG C shaking table culture 2 days, collect thallus,
Genome DNA is extracted using DNA extraction agent box.
(2) PCR amplification
Using specific primer:
27F:5'-AGAGTTTGATCCTGGCTCAG-3',
1492R:5'-GGTTACCTTGTTACGACTT-3'；
25 μ L of PCR reaction system total volume, PCR amplification condition are 94 DEG C of 5min；94 DEG C of 45s, 56 DEG C of 45s, 72 DEG C
45s, 30 circulations；72℃10min.
(3) sequencing
Pcr amplification product is sequenced through electrophoresis detection and after purification, obtains the 16S rDNA sequence that length is 1431bp, sequence
Shown in SEQ ID NO:1 referring to attached offer, gained sequence is compared through the common website NCBI, and constructs and be
System development tree, the phylogenetic tree of the bacterium are compared analysis and are found referring to attached drawing 1, bacterial strain H6 and mode standard bacterium
Halomonas titanicae BH1TWith Halomonas neptunia Eplume1TAffiliation is nearest, 16S rDNA sequence
Column are respectively 98.6% and 98.3% with the two homology, are passed through using 5.0 software of MEGA of the common use in this field
Neighbor-Joining method establishes systematic evolution tree, as a result finds through comparing analysis, bacterial strain H6 and Halomonas
titanicae BH1TFor cluster on a branch, the value of the confidence is respectively 86%, shows a possibility that strain is as novel bacterial
It is high, fabulous stability and high supporting rate are embodied in chadogram, it can should by the identification of system fungus molecular level
Bacterial strain H6 is determined as Halomonas (Halomonas sp.) novel species, has the characteristics that the typicalness that novel bacterial has, from taxology
Angle is temporarily named as Halomonas (Halomonas sp.) H6.
2, physiological and biochemical test
(1) by growth conditions result of study show Halomonas (Halomonas sp.) H6, Gram-positive,
It is rod-shaped, 0.3-0.8 × 1.4-2.5 μm, do not form endospore.It is grown in containing cultivating 3-5d on 2%NaCl TSA culture medium
Afterwards, bacterium colony is circle, neat in edge, slightly convex faint yellow bacterium colony, and colonial morphology is referring to shown in attached drawing 2.Thalli growth temperature
Range is spent at 10-45 DEG C, and optimum temperature growth is 28 DEG C；PH growth scope 5.0-10, best pH value are 7.0, can tolerate 1-
20% NaCl.
(2) hydrogen oxide enzyme and oxidase active are the positive, hydrolyzable junket otoginsenoside.
(3) it is identified using Biolog strain idenfication instrument GNIII test board, determines Halomonas (Halomonas sp.) H6
The carbon source that can be utilized be a-D- glucose, D-glucitol, pectin, ρ-Hydroxy-phenyl-acetic acid, PEARLITOL 25C, D- galacturonic acid,
Gamma-amino-butyric acid, D-Fructose, D-arabitol, l-Alanine, L- galacturonic acid lactone, D-ALPHA-Hydroxypropionic acid methyl esters, D- seaweed
Sugar, D- galactolipin, inositol, L-arginine, maltonic acid, L- lactose, beta-hydroxy-d, L butyric acid, D- cellobiose, 3- first
Acyl glucose, glycerol, L-Aspartic acid, D-Glucose aldehydic acid, citric acid, D-Fructose, D-Glucose -6- phosphoric acid, L- paddy ammonia
Acid, α-ketoglutaric acid, acetoacetate, sucrose, N- acetyl-β-D-MANNOSE amine, L- fructose, D-malic acid, propionic acid, D- pine two
Sugar, quininic acid, L MALIC ACID, acetic acid, inosine, Serine.
Comprehensive 16S rDNA sequence analysis, Phylogenetic Analysis, Microbiological Characteristics analysis are as a result, show that the present invention mentions
The Halomonas H6 of confession is really one plant of typical bacterium novel species of Halomonas, it is temporarily named as Halomonas
(Halomonas sp.)H6.It is common that the bacterium on May 14th, 2018 was preserved in China Committee for Culture Collection of Microorganisms
Microorganism center (CGMCC), address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica,
Culture presevation number is CGMCC No.15758.
Embodiment two: the growth-promoting ability detection of Halomonas (Halomonas sp.) H6 bacterial strain
(1) heteroauxin (IAA) ability is secreted
Halomonas H6 is inoculated in the TSA fluid nutrient medium of the 2%NaCl of the L-Trp containing 100mg/L, 180r/
After 30 DEG C of culture 1d of min shaking table, take 50 μ L bacteria suspensions drop on whiteware plate, while isometric Salkowski ratio is added
Color liquid, Salkowski color solution are 50mL 35%HClO4With 1mL 0.5mol/L FeCl3Mixing, and do not connect so that 50 μ L are added
The TSA fluid nutrient medium of the 2% of bacterium is to compare with the mixed solution of isometric color solution.Whiteware plate is protected from light in room temperature
After placing 30min, color in the whiteware plate of 50 μ L Halomonas H6 bacteria suspensions is added and reddens.
(2) acc deaminase test is produced
Halomonas H6 is inoculated in the DF solid medium of the ACC containing 3mM, after passing on 3 times, Halomonas H6 is in ACC
For the well-grown on only nitrogen source culture medium, the bacterial strain of growth is to produce deaminase positive strain.
Research shows that: Halomonas (Halomonas sp.) H6 has the ability of secretion heteroauxin and acc deaminase.
Embodiment three: Halomonas (Halomonas sp.) H6 bacterial strain salt resistance ability detection
Halomonas Halomonas sp.H6CGMCC No.15758 is inoculated in TSA fluid nutrient medium, 30 DEG C,
After being cultivated for 24 hours under the conditions of 180rpm, bacteria suspension is respectively connected to containing 1%, 5%, 10%, 15%, the TSA of 20%NaCl concentration
In fluid nutrient medium, 30 DEG C, 180rpm shaken cultivation for 24 hours, surveys the OD value at 600nm, observes Halomonas Halomonas
The growing state of sp.H6, to determine its salt tolerant degree.
Result of study is as shown in Fig. 3, in the range of NaCl concentration is 1-20%, Halomonas Halomonas
Sp.H6 can be grown, but with the raising of salinity, the increment of bacterium is in significant downward trend,
Example IV: the resistance to pH ability detection of Halomonas (Halomonas sp.) H6 bacterial strain
Halomonas Halomonas sp.H6CGMCC No.15758 is inoculated in TSA fluid nutrient medium, 30 DEG C,
After being cultivated for 24 hours under the conditions of 180rpm, by bacteria suspension be respectively connected to pH value be respectively 3,5,7,9 TSA fluid nutrient medium in, 30
DEG C, 180rpm shaken cultivation for 24 hours, surveys the OD value at 600nm, the growing state of Halomonas Halomonas sp.H6 is observed,
To determine the ability of its resistance to pH.
Result of study is as shown in Fig. 4, and Halomonas Halomonas sp.H6 is more biased towards in raw under weak basic condition
Long, pH value is equal well-grown in the range of 7-9, and the most suitable growth pH value is 7.When pH value is equal to or less than 5, increment
It is substantially reduced.
Embodiment five: growth curve and the microbial inoculum preparation of Halomonas (Halomonas sp.) H6
Halomonas Halomonas sp.H6CGMCC No.15758 is seeded in the TSA solid culture containing 2% NaCl
It after being activated on base, transfers in TSA fluid nutrient medium (the bottled 100mL of 500mL triangle), 180rpm is cultivated at 30 DEG C
18h obtains seed liquor.Seed liquor is inoculated in fermentation medium in 1:100 ratio, 30 DEG C, 200rpm is cultivated for 24 hours, bacterium
Agent maximum concentration is up to 6 × 109To get microbial bacterial agent.In actual use can be as needed, fermenting agent is diluted one
It is used after determining multiple.It is the growth curve (10 times of dilution) of the bacterium in the fermentation medium referring to attached drawing 5.
Embodiment six: growth-promoting functions of Halomonas (Halomonas sp.) H6 to cotton seedling
Full grains and the cotton seed without obvious breakage are selected, sterilizes 5min using 0.1% mercuric chloride, after sterile water is cleaned,
Seed 4h is impregnated with 1/100 concentration bacterium solution；The control treatment seed for not connecing bacterium is dipped in sterile purified water.
Basin is planted using the cave in 4 × 5 caves, plants soil after yellow sand drying to wash for cave.The appropriate 5%NaCl solution of soil
After infiltration, addition is placed tap water overnight and is mixed, and so that soil moisture content is reached 20%, salt content reaches 0.75%.It is real
Design 10 caves of each processing are tested, each processing cotton seed point implantation cave are planted in basin, 4, every cave, depth about 1.5cm, and use plastic film
Covering, is placed in phjytotron and cultivates.20 DEG C of condition of culture, no light 8h；22 DEG C, 30% illumination 2h；25 DEG C, 100% light
According to 12h；30% illumination 2h.To germination, film is thrown off, poured primary (every cave about 10mL) every 5 days.15d to be germinateed
The root long and plant weight of test group and control group cotton plants are measured afterwards.
As shown in Fig. 6, it the experimental results showed that, compared with control treatment, is soaked through 1/100 concentration microbial inoculum of Halomonas H6
After cotton seeds after bubble, grows 15 days root longs and plant weight significantly increases 11.54% and 15.38% respectively, illustrate salt unit cell
The immersion of bacteria agent dilution has apparent facilitation to cotton seed.
Embodiment seven: growth-promoting functions of Halomonas (Halomonas sp.) H6 to wheat
Full grains and the wheat seed without obvious breakage are selected, sterilizes 5min, sterile washing using 0.1% mercuric chloride
After net, seed 4h is impregnated with 1/100 concentration bacterium solution；The control treatment seed for not connecing bacterium is dipped in sterile purified water.
Basin is planted using the cave in 4 × 5 caves, plants soil after yellow sand drying to wash for cave.The appropriate 5%NaCl solution of soil
After infiltration, addition is placed tap water overnight and is mixed, and so that soil moisture content is reached 20%, salt content reaches 0.75%.It is real
Design 10 caves of each processing are tested, each processing wheat seed point implantation cave are planted in basin, 4, every cave, depth about 1.5cm, and use plastics
Film covering, is placed in phjytotron and cultivates.20 DEG C of condition of culture, no light 8h；22 DEG C, 30% illumination 2h；25℃,
100% illumination 12h；30% illumination 2h.To germination, film is thrown off, poured primary (every cave about 10mL) every 5 days.To
The various forms parameter of test group and control group wheat plant, including plant height, root long and fresh weight are measured after germination 15d.
As shown in Fig. 7, it the experimental results showed that, compared with control treatment, is soaked through 1/100 concentration microbial inoculum of Halomonas H6
It after wheat seed after bubble, grows 15 days, the facilitation of plant height is relatively weak, increases only 3.06%, but to root long and fresh
The facilitation of weight is obvious, significantly increases 23.08% and 41.67% respectively, illustrates the immersion pair of Halomonas microbial inoculum dilution
Wheat plays the role of significantly promoting growth.
Embodiment eight: one kind includes the composition of Halomonas (Halomonas sp.) H6 bacterial strain
On the basis of the above embodiments, Halomonas (Halomonas sp.) H6 provided using above-described embodiment one
The culture of bacterial strain.
The composition of described Halomonas (Halomonas sp.) the H6 strain culture, the composition be liquid,
Freezing or dry powdered form.
It include auxiliary agent in above-mentioned composition.
It further include solid-state carrier when the composition is freezing or dry powdered form.
The solid-state carrier includes peat, turf, talcum, lignite, pyrophyllite, montmorillonite, alginates, press mud, saw
Bits, perlite, mica, silica, silica flour, calcium-base bentonite, vermiculite, kaolin, precipitated calcium carbonate, diatomite, medical stone,
One of calcite, zeolite, white carbon black, fine sand and clay are a variety of.
The auxiliary agent include neopelex, sodium butylnaphthalenesulfonate, trehalose, glycerol, sodium lignin sulfonate,
Negel condensation polymer, niacin, alcohol, buffer salt, sodium chloride, amino acid, vitamins, protein, polypeptide, polysaccharide or
One of monosaccharide, yeast extract, white carbon black, Tea Saponin, defatted milk are a variety of.
It also include a effective amount of compound selected from the following of agricultural or combination using the composition of above-mentioned offer series
Object: nutrient, fertilizer, microbial-bacterial fertilizer.
As described above, the present invention can be realized preferably, the above embodiments are only to preferred implementation side of the invention
Formula is described, and is not intended to limit the scope of the present invention, without departing from the spirit of the design of the present invention, this field
The various changes and improvements that those of ordinary skill makes technical solution of the present invention, should all fall into present invention determine that protection
In range.
Sequence table
<110>Microorgan Application Inst., Xinjiang Agricultural Academy is (in Xinjiang, China-Armenia's bio-engineering research exploitation
The heart)
<120>a kind of Halomonas strain H6, composition and its application in salt tolerant growth-promoting
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1431
<212> DNA
<213>Halomonas (Halomonas sp)
<400> 1
cggaaccacc gttggtgatc gccctcttgc gttaggctaa ccacttctgg tgcagtccac 60
tcccatggtg tgacgggcgg tgtgtacaag gcccgggaac gtattcaccg tgacattctg 120
attcacgatt actagcgatt ccgacttcac ggagtcgagt tgcagactcc gatccggact 180
gagaccggct ttaagggatt cgctgactct cgcgagctcg cagccctttg taccggccat 240
tgtagcacgt gtgtagccct acccgtaagg gccatgatga cttgacgtcg tccccacctt 300
cctccggttt gtcaccggca gtctccttag agttcccgac attactcgct ggcaaataag 360
gacaagggtt gcgctcgtta cgggacttaa cccaacattt cacaacacga gctgacgaca 420
gccatgcagc acctgtctta cagttcccga aggcacacca gaatctcttc cggcttctgt 480
agatgtcaag ggtaggtaag gttcttcgcg ttgcatcgaa ttaaaccaca tgctccaccg 540
cttgtgcggg cccccgtcaa ttcatttgag ttttaacctt gcggccgtac tccccaggcg 600
gtcgacttat cgcgttaact tcgccacaaa gtgctctagg cacccaacgg ctggtcgaca 660
tcgtttacgg cgtggactac cagggtatct aatcctgttt gctacccacg ctttcgcacc 720
tcagtgtcag tgtcagtcca gaaggccgcc ttcgccactg gtattcctcc cgatctctac 780
gcatttcacc gctacaccgg gaattctacc ttcctctcct gcactctagc ctgacagttc 840
cggatgccgt tcccaggttg agcccggggc tttcacaacc ggcttatcaa gccacctacg 900
cgcgctttac gcccagtaat tccgattaac gcttgcaccc tccgtattac cgcggctgct 960
ggcacggagt tagccggtgc ttcttctgcg agtgatgtct ttcctaccgg gtattaaccg 1020
ataggcgttc ttcctcgctg aaagtgcttt acaacccgag ggccttcttc acacacgcgg 1080
catggctgga tcagggttgc ccccattgtc caatattccc cactgctgcc tcccgtagga 1140
gttcgggccg tgtctcagtc ccgatgtggc tgatcatcct ctcagaccag ctacggatcg 1200
ttgccttggt aagccattac cttaccaact agctaatccg acataggctc atccaatagc 1260
gggagccgga gccccctttc tcccgtagga cgtatgcggt attagcctgg gtttccccag 1320
gttatccccc actatcgggc agattcctat gcattactca cccgtccgcc gctcgtcagc 1380
atgtagcaag ctagccctgt taccgctcga cttgcatgtg gttaggcccc g 1431
Claims (10)
1. a kind of Halomonas (Halomonas sp.) H6, which is characterized in that the Halomonas (Halomonas sp.)
The culture presevation number of H6 be CGMCC No.15758, Halomonas (Halomonas sp.) H6 16S rDNA sequence, gene sequence
Column are as shown in SEQ ID NO:1.
2. Halomonas as described in claim 1 (Halomonas sp.) H6, which is characterized in that the bacterial strain can
It is resistant to the NaCl of 1-20%.
3. composition, containing the described in any item Halomonas of claim 1 to 2 (Halomonas sp.) H6 bacterial strain training
Support object.
4. composition according to claim 3, which is characterized in that the composition is liquid, freezing or dried powder shape
Formula.
5. composition according to claim 4, which is characterized in that include auxiliary agent in the composition.
6. composition according to claim 4, which is characterized in that when the composition is freezing or dry powdered form,
It further includes solid-state carrier.
7. composition according to claim 6, which is characterized in that the solid-state carrier includes peat, turf, talcum, brown
Coal, pyrophyllite, montmorillonite, alginates, press mud, sawdust, perlite, mica, silica, silica flour, calcium-base bentonite, leech
One of stone, kaolin, precipitated calcium carbonate, diatomite, medical stone, calcite, zeolite, white carbon black, fine sand and clay or
It is a variety of.
8. composition according to claim 5, which is characterized in that the auxiliary agent includes neopelex, butyl
Sodium naphthalene sulfonate, trehalose, glycerol, sodium lignin sulfonate, Negel condensation polymer, niacin, alcohol, buffer salt, sodium chloride, ammonia
One of base acid, vitamins, protein, polypeptide, polysaccharide or monosaccharide, yeast extract, white carbon black, Tea Saponin, defatted milk are more
Kind.
9. according to composition described in claim 3 ~ 8 any one, which is characterized in that it, which also includes that agricultural is a effective amount of, is selected from
Compound or composition below: nutrient, fertilizer, microbial-bacterial fertilizer.
10. as described in claim 3 or 8 any one Halomonas (Halomonas sp.) H6 improving cotton or wheat
Salt Stress Tolerance and the application for promoting crops seedling stage growth aspect.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111100813A (en) * | 2019-12-30 | 2020-05-05 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Salt-tolerant growth-promoting bacterium JP-JH for relieving salt damage of plants and application thereof |
| CN113061550A (en) * | 2021-04-01 | 2021-07-02 | 新疆农业科学院微生物应用研究所(中国新疆—亚美尼亚生物工程研究开发中心) | Lactobacillus new strain Z6 and application thereof in food |
| CN113943684A (en) * | 2021-11-24 | 2022-01-18 | 青岛理工大学 | Tender haloplankton and application thereof in production of salinization farmland |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008061624A (en) * | 2006-09-11 | 2008-03-21 | Iib:Kk | New microorganism and liquid-waste disposal method using the same |
| CN102120973A (en) * | 2010-12-08 | 2011-07-13 | 清华大学 | Halomonas strain and application thereof |
-
2019
- 2019-02-26 CN CN201910142806.7A patent/CN109929777B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008061624A (en) * | 2006-09-11 | 2008-03-21 | Iib:Kk | New microorganism and liquid-waste disposal method using the same |
| CN102120973A (en) * | 2010-12-08 | 2011-07-13 | 清华大学 | Halomonas strain and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| FURKAN ORHAN: "Alleviation of salt stress by halotolerant and halophilic plant growth-promoting bacteria in wheat (Triticum aestivum)", 《BRAZILIAN JOURNAL OF MICROBIOLOGY》 * |
| PRITHVIRAJ DESALE ET AL.: "Plant growth promoting properties of Halobacillus sp. and Halomonas sp. in presence of salinity and heavy metals", 《J. BASIC MICROBIOL.》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111100813A (en) * | 2019-12-30 | 2020-05-05 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Salt-tolerant growth-promoting bacterium JP-JH for relieving salt damage of plants and application thereof |
| CN111100813B (en) * | 2019-12-30 | 2021-03-05 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Salt-tolerant growth-promoting bacterium JP-JH for relieving salt damage of plants and application thereof |
| CN113061550A (en) * | 2021-04-01 | 2021-07-02 | 新疆农业科学院微生物应用研究所(中国新疆—亚美尼亚生物工程研究开发中心) | Lactobacillus new strain Z6 and application thereof in food |
| CN113061550B (en) * | 2021-04-01 | 2023-03-24 | 新疆农业科学院微生物应用研究所(中国新疆—亚美尼亚生物工程研究开发中心) | Lactobacillus new strain Z6 and application thereof in food |
| CN113943684A (en) * | 2021-11-24 | 2022-01-18 | 青岛理工大学 | Tender haloplankton and application thereof in production of salinization farmland |
| CN113943684B (en) * | 2021-11-24 | 2024-02-13 | 青岛理工大学 | Salmonella pinnata and application thereof in salinized farmland production |
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| Publication number | Publication date |
|---|---|
| CN109929777B (en) | 2021-03-09 |
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