CN115340968A - Novel application and method of pseudomonas spinosa, pseudomonas spinosa 21.1.9.2-14 and product thereof - Google Patents

Novel application and method of pseudomonas spinosa, pseudomonas spinosa 21.1.9.2-14 and product thereof Download PDF

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CN115340968A
CN115340968A CN202211038593.1A CN202211038593A CN115340968A CN 115340968 A CN115340968 A CN 115340968A CN 202211038593 A CN202211038593 A CN 202211038593A CN 115340968 A CN115340968 A CN 115340968A
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pseudomonas
spinosa
strain
veruta
tobacco
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CN115340968B (en
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陈雅琼
欧阳进
詹莜国
许东亚
孔德旬
李卓衡
周文瑾
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Kunming Company of Yunnan Tobacco Co
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Kunming Company of Yunnan Tobacco Co
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • C12N1/205Bacterial isolates
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/20Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00Botany in general
    • A01G7/06Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00Other organic fertilisers
    • C05F11/08Organic fertilisers containing added bacterial cultures, mycelia or the like
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05GMIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
    • C05G3/00Mixtures of one or more fertilisers with additives not having a specially fertilising activity
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/01Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/38Pseudomonas

Abstract

The invention relates to novel application of pseudomonas spinosa, a method thereof, pseudomonas spinosa 21.1.9.2-14 and a product thereof, belonging to the technical field of microorganisms. The invention provides application of Pseudomonas spinosa (Pseudomonas veruta) in promoting plant growth, and a plant growth promoting method based on the Pseudomonas spinosa (Pseudomonas veruta). The invention also provides a Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.9.2-14 with the preservation number of GDMCC NO:62683, a biological fertilizer based on the strain 21.1.9.2-14 and a method for promoting the growth of tobacco or ryegrass. A large number of experiments prove that the strain 21.1.2-14 can obviously improve the plant height, stem circumference, leaf length and leaf width of the plant and has obvious promotion effect on the growth of the plant.

Description

Novel application and method of pseudomonas spinosa, pseudomonas spinosa 21.4.1.9.2-14 and product thereof
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a novel application of pseudomonas spinosa and a method thereof, pseudomonas spinosa 21.1.2-14 and a product, application and method thereof.
Background
Tobacco is an important commercial crop. In 2020, the total amount of interest and tax paid by Chinese tobacco accounts for 1.184% of the total specific gravity of GDP in China. With the development of related technologies in the fields of agriculture and biology, the future development prospect of the tobacco industry is wider and wider. However, at the same time, tobacco planting faces some difficulties which are difficult to solve, and continuous cropping obstacles of tobacco are the most troublesome problems.
The continuous cropping obstacle mainly refers to the phenomenon that under normal agricultural management measures, the same crops or the same crops are planted in the same soil in multiple continuous cropping modes, so that various plant diseases and insect pests of the crops frequently occur, the breeding condition is poor, the quality is deteriorated, and the yield is reduced. Under the influence of factors such as cultivation and production conditions, economic benefits and the like, the area of continuous cropping of tobacco is enlarged year by year, and the continuous cropping obstacles are further aggravated. The continuous cropping of the tobacco deteriorates the soil environment, so that the absorption of the tobacco plants to soil nutrients is influenced, the photosynthesis of the tobacco is further weakened, the transpiration rate is reduced, and finally the yield and the quality of the tobacco are reduced. For the above reasons, the continuous cropping obstacle of tobacco has become a bottleneck problem restricting further development of tobacco industry. Therefore, a method for overcoming the problem of continuous cropping obstacle of tobacco is found, and the method has important significance for the sustainable development of the tobacco industry in China.
The main factors causing the continuous cropping obstacle of tobacco are the change of the physical and chemical properties of soil, the enrichment of autotoxic substances secreted by root systems and the change of microbial communities in the soil. Wherein, the change of the microbial community in the soil is reflected by the reduction of beneficial bacteria and the enrichment of harmful bacteria of the tobacco by bacillus and the like in the soil. The microorganisms are not only a reserve bank of available nutrients of plants in the soil, but also power for conversion and circulation of organic matters and nutrients of the soil. The decomposition of beneficial microbes such as partial bacteria and the like enables organic matters in the soil to be decomposed into inorganic nutrients such as P, K and the like which can be utilized by plants, so that the organic matters are required for the normal growth and development of the plants. Harmful microorganisms form a competitive relationship with plants, and the harmful microorganisms hinder the plants from absorbing nutrient substances and cause diseases of the plants. In conclusion, coordinating the relationship between soil microorganisms and tobacco plants, and keeping beneficial microbial flora in tobacco planting soil in an advantageous position has great significance in tobacco planting production. With the development of related technologies in the field of microorganisms, the preparation of screened microorganisms into pollution-free fertilizers such as biological bacterial manure and the like becomes an effective means for improving the diversity of soil microorganisms, promoting the growth of tobacco plants and further improving the quality of tobacco leaves.
The microbial fertilizer is a novel biological fertilizer which is developed by the microbiological technology based on the relevant knowledge in the fields of soil ecology and plant nutrition and according to the modern organic sustainable development agricultural principle. The active microorganisms in the microbial bacterial manure can improve the diversity of microorganisms in soil, keep microbial floras beneficial to plant growth in an advantageous position, promote the absorption of plants to nutrition, improve the yield of plants and increase economic benefits. In the production process of the microbial fertilizer, the most important step is to screen strains which have a promoting effect on plant growth.
Few reports have been made in the prior art in the field of microorganisms for such strains of Pseudomonas spinosa (Pseudomonas veruta), and no report has been made on the use of Pseudomonas spinosa (Pseudomonas veruta) for promoting plant growth.
Disclosure of Invention
In view of the above-mentioned needs and gaps in the prior art, the present invention provides a strain for promoting tobacco growth, named Pseudomonas spinosa 21.1.9.2-14 (Pseudomonas veruta 21.1.9.2-14), and a corresponding method for applying the strain.
The technical scheme of the invention is as follows:
use of Pseudomonas spinosa (Pseudomonas veruta) for promoting plant growth.
A method for promoting plant growth, characterized in that Pseudomonas spinosa (Pseudomonas versuta) is applied in a plant growth environment.
The applying mode comprises the following steps: mixing the Pseudomonas spinosa (Pseudomonas veruta) with soil to be used as a plant growth substrate, and/or performing root irrigation treatment on the plant by using fermentation liquor of the Pseudomonas spinosa (Pseudomonas veruta), and/or preparing the Pseudomonas spinosa (Pseudomonas veruta) into bacterial powder and then applying the bacterial powder in a furrow or directly scattering the bacterial powder in the field.
A Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14, wherein the preservation number is GDMCC NO:62683.
A biological fertilizer comprises a fertilizer active ingredient, and is characterized in that the fertilizer active ingredient comprises a Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14 with the preservation number of GDMCC NO:62683.
The biological fertilizer further comprises: an auxiliary material;
preferably, the fertilizer is in the form of powder or liquid.
Use of Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.2-14 with the collection number of GDMCC NO:62683 for promoting plant growth.
Preferably, the plant is tobacco or ryegrass.
A method for promoting the growth of tobacco or ryegrass, characterized in that Pseudomonas micropteria (Pseudomonas versuta) strain 21.1.2-14 with the collection number GDMCC NO:62683 is applied in the environment of tobacco growth.
The mode of application is selected from: mixing the Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.9.2-14 with soil to be used as a tobacco growth substrate, and/or performing root irrigation treatment on a tobacco plant by using fermentation liquor of the Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.9.2-14, and/or preparing the Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.9.2-14 into bacterial powder and then performing furrow application on a tobacco field or directly scattering the bacterial powder in the field;
preferably, the bacterial powder is prepared by mixing sterilized wheat bran and fermentation liquor of Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14 and drying;
preferably, the mixing refers to mixing the wheat bran and fermentation liquor of Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.9.2-14 according to a ratio of 1kg to 1L;
preferably, the drying refers to mixing, putting into an electrothermal blowing drying oven, and drying for 3h at 55 ℃;
preferably, the application amount of the bacterial powder is 5kg per mu of land;
preferably, in the root irrigation treatment, the fermentation liquor of the Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.9.2-14 is diluted into a dilution liquid for use or the fermentation liquor of the Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.9.2-14 is directly used;
preferably, the dilution factor is 10-50;
preferably, during the root irrigation treatment, the consumption of the diluent of each tobacco is 10-100ml;
preferably, the amount of fermentation broth used per ryegrass is 50ml during root irrigation.
The invention provides a bacterial fermentation method for promoting tobacco growth and application thereof, and tobacco seedlings are treated by using fermented bacterial liquid.
Furthermore, the bacterial strain is Pseudomonas spinosa (Pseudomonas veruta 21.1.2-14), is obtained by screening grass rhizosphere soil in Daguan county of Showtong city in Yunnan province, is preserved in Guangdong province microorganism strain preservation center, and has a preservation number of 62683, wherein the preservation number of the Pseudomonas veruta 21.1.2-14 (Pseudomonas veruta 21.1.2-14) is 62683.
Further, the preparation method of the bacterial fermentation liquor comprises the following steps:
(1) Preparing a culture medium: LB culture medium: adding 10g tryptone, 5g yeast powder and 10g sodium chloride into 1000ml deionized water, and uniformly stirring to obtain a 1LLB culture medium, wherein 15-20g agar powder is required to be added into a solid culture medium. The pH is natural, and the temperature is about 25-28 ℃.
(2) After the culture medium is cooled, the activated thalli are inoculated into a liquid culture medium and put into a 120r/min shaking table, and the culture temperature is 28 ℃. Culturing for 2-4 days;
further, the application mode of the inoculum is as follows:
(1) When crops are cultivated, a cultivation hole corresponding to the size of a seedling root is dug in soil, a seedling is placed in the cultivation hole, fermented bacterial liquid is poured on the root of the plant, and the using amount of the bacterial liquid is 10ml-100ml per plant according to the size of the root.
(2) Mixing the fermented bacteria liquid with water for root watering in any proportion, and then watering the mixture into a cultivation hole when crops are cultivated, or watering the mixture after soil is covered.
(3) Adding the fermented bacteria liquid into sterilized wheat bran, drying at low temperature to obtain bacteria powder, and applying the dried bacteria powder to field during plant cultivation.
(4) Adding the fermented bacteria liquid into sterilized wheat bran, drying at low temperature to obtain bacteria powder, activating the bacteria powder in nutrient solution, and pouring into plant root during cultivation.
(5) Freeze drying the fermented bacteria liquid to obtain pure bacteria powder, and directly applying the pure bacteria powder during plant cultivation.
(6) Freeze drying the fermented bacteria liquid to obtain pure bacteria powder, and mixing the pure bacteria powder with other matters for application.
(7) Freeze drying the fermented bacteria liquid to obtain pure bacteria powder, activating the bacteria powder and pouring the bacteria liquid containing activated bacteria onto the root of plant.
The invention has the following beneficial effects:
(1) The invention provides a new application of Pseudomonas spinosa (Pseudomonas veruta) in promoting plant growth for the first time, and fills the gap of the prior art in the field.
(2) The invention obtains a Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14 capable of promoting plant growth for the first time, and a large number of experiments prove that the Pseudomonas spinosa strain can obviously improve the plant height, stem circumference, leaf length and leaf width of the plant and has obvious promotion effect on plant growth.
(3) Compared with other fertilization modes, the fermentation liquid prepared by the invention has the characteristics of green, safety and environmental protection.
(4) The bacterial liquid prepared by the invention has good growth promoting effect on tobacco, can be applied to the production of tobacco in future, increases the yield and quality of tobacco leaves, and improves economic benefit.
The preservation information of the Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.2-14 of the invention is as follows:
the strain preservation name is as follows: 21 4.19.2-14
The preservation number is as follows: GDMCC NO:62683
And (3) classification and naming: micro-piercing pseudomonas
Latin name: pseudomonas versuta
The preservation unit: guangdong province microbial strain preservation center
The address of the depository: microbiological research institute of Chaireli Zhou 100 college No. 59 building 5 building Guangdong province academy of sciences
The preservation date is as follows: 8/4/2022.
Drawings
FIG. 1 is a front view and a back view showing the growth of the strain 21.4.1.2-14 of the present invention on a coated plate.
FIG. 2 is a phylogenetic tree of the strain 21.1.9.2-14 of the present invention.
FIG. 3 is a photograph showing the comparison between the growth forms of tobacco subjected to root irrigation with fermentation broth of strain 21.1.9.2-14 in Experimental example 1 of the present invention and control tobacco.
FIG. 4 is a photograph showing the comparison between the growth patterns of ryegrass root-irrigated with the fermentation broth of strain 21.1.9.2-14 and ryegrass of the control group in Experimental example 5 of the present invention.
Detailed Description
The following detailed description of the present invention is further described in detail with reference to specific examples and experimental examples, but the scope of the present invention is not limited thereto.
Sources of biological material
The tobacco variety Honghuadajinyuan used in the experimental examples 1 to 4 of the present invention is a publicly known and used tobacco variety, and is stored in the laboratory of the applicant unit or is commercially available.
The ryegrass used in Experimental example 5 of the present invention was a commonly used grass plant known to the public, and was stored in the applicant's unit laboratory, or was obtained commercially.
Group 1 example, novel use of Pseudomonas spinosa (Pseudomonas veruta)
The present group of embodiments provides the use of Pseudomonas spinosa (Pseudomonas versuta) for promoting plant growth.
The use of Pseudomonas microscopia (Pseudomonas versuta) for promoting plant growth is the first suggestion of the present invention that any application of Pseudomonas microscopia (Pseudomonas versuta) for the growth of any plant and/or the application of Pseudomonas microscopia (Pseudomonas versuta) or to the growth substrate of any plant and/or the action of Pseudomonas microscopia (Pseudomonas versuta) or to any link of the growth process of any plant, the application of Pseudomonas microscopia (Pseudomonas versuta) or to any form of growth, cultivation or planting environment of any plant falls within the scope of protection of the present invention.
Group 2 examples, methods of the present invention for promoting plant growth
The present group of embodiments provides a method of promoting plant growth. All embodiments of this group share the following common features: pseudomonas spinosa (Pseudomonas versuta) is applied in the plant growth environment.
In some specific embodiments, the applying comprises: mixing the micro-piercing Pseudomonas (Pseudomonas verussa 21.1.9.2-14) with soil to be used as a plant growth substrate, and/or performing root irrigation treatment on the plant by using fermentation liquor of the micro-piercing Pseudomonas (Pseudomonas verussa 21.1.9.2-14), and/or preparing the micro-piercing Pseudomonas (Pseudomonas verussa 21.1.9.2-14) into bacterial powder and then applying the bacterial powder in a furrow or directly scattering the bacterial powder in a field.
Example 3, pseudomonas spinosa Strain 21.1.2-14 of the invention
The embodiment of the group provides a Pseudomonas spinosa (Pseudomonas versita) strain 21.1.9.2-14, and the preservation number is GDMCC NO:62683.
Any use, replication, culture, propagation, inoculation, enrichment, marketing, promissory sale, production, preparation, addition, application, administration of the Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.9.2-14 with the accession number GDMCC NO:62683 falls within the scope of the present invention.
In some embodiments, the present invention provides a strain selected from a soil sample, wherein the growth morphology of the strain coated with a plate is shown in FIG. 1, and the molecular identification result is shown in FIG. 2, and the strain belongs to Pseudomonas spinosa (Pseudomonas veruta) in taxonomy. The invention names the strain as 21.1.2-14 and sends the preservation, and the preservation information is as follows:
the strain preservation name is as follows: 21 4.19.2-14
The preservation number is as follows: GDMCC NO 62683
And (3) classification and naming: micro-piercing pseudomonas
Latin name: pseudomonas versuta
The preservation unit is as follows: guangdong province microbial strain preservation center
The address of the depository: microbiological research institute of Chaireli Zhou 100 college No. 59 building 5 building Guangdong province academy of sciences
The preservation date is as follows: 8/month/4/2022.
Group 4 example, biofertilizer of the present invention
The present group of embodiments provides a biofertilizer. All embodiments of this group share the following common features: the biological fertilizer comprises a fertilizer active ingredient which comprises a Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.9.2-14 with the preservation number of GDMCC NO:62683.
In a further embodiment, the biofertilizer further comprises: an auxiliary material;
preferably, the fertilizer is in the form of powder or liquid.
The preparation of the microbial inoculum or bacterial manure or microecological preparation can be carried out on the strains 21.1.2-14 of the invention by adopting auxiliary materials commonly used in the field of microbial inoculum preparation, and the strains can be prepared into various possible dosage forms, such as powder, liquid, tablets, capsules, sprays and the like, based on technical common knowledge in the field or conventional technical means well known by the technical knowledge in the field, such as technical means recorded in tool books such as Microecological preparations, agricultural microbial concentrated preparations, pharmaceutical auxiliary material manuals, fertilizer use technical manuals and the like.
The auxiliary materials commonly used in the field of microbial inoculum preparation can be selected from pharmaceutical auxiliary materials or culture medium components commonly used by microorganisms; such pharmaceutical excipients include, but are not limited to: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integration agents, permeation enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, antifoaming agents, thickeners, encapsulation agents, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, release retardants.
Group 5 examples, novel use of the strains 21.1.9.2-14 of the invention
The present group of examples provides the use of Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.2-14 with the accession number GDMCC NO:62683 for promoting plant growth.
In specific embodiments, the plant is tobacco or ryegrass.
One skilled in the art can use Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.9.2-14, deposited with accession number GDMCC NO:62683, for growth promoting applications in other plants, including but not limited to: field crops such as rice, wheat, corn, soybean, cotton and the like; fruits such as strawberry, apple, blueberry, watermelon, etc.; flowers such as geranium, poinsettia, goldenseal, carnation, rose, etc.; forage grass such as ryegrass; and the like.
Group 6 example, method of promoting tobacco growth of the present invention
The present set of embodiments provides a method of promoting tobacco growth. All embodiments of this group share the following common features: pseudomonas spinosa (Pseudomonas versuta) strain 21.1.9.2-14 with accession number GDMCC NO:62683 was applied in the tobacco growing environment.
In particular embodiments, the mode of application is selected from: mixing the Pseudomonas spinosa (Pseudomonas veruta) strain 21.4.1.2-14 with soil to be used as a tobacco growth substrate, and/or performing root irrigation treatment on a tobacco plant by using fermentation liquor of the Pseudomonas spinosa (Pseudomonas veruta) strain 21.4.1.2-14, and/or preparing the Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14 into bacterial powder and then performing furrow application or directly scattering the bacterial powder in a tobacco field;
preferably, the bacterial powder is prepared by mixing and drying sterilized wheat bran and fermentation liquor of Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.2-14;
preferably, the mixing refers to mixing the wheat bran and fermentation liquor of Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.9.2-14 according to a ratio of 1kg to 1L;
preferably, the drying refers to mixing, putting into an electrothermal blowing drying oven, and drying for 3h at 55 ℃;
preferably, the application amount of the bacterial powder is 5kg per mu of land;
preferably, during the root irrigation treatment, the fermentation liquor of the Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.2-14 is diluted for use;
preferably, the dilution factor is 10-50;
preferably, the diluted solution of each tobacco is 10-100ml when the root is irrigated.
In other embodiments, the tobacco growth promoting means: the plant height, and/or the stem circumference, and/or the leaf length, and/or the leaf width of the tobacco plant are/is improved.
Experimental example 1, application of fermentation liquor of Pseudomonas spinosa 21.1.2-14 (Pseudomonas versuta 21.1.2-14) in tobacco production.
Fermenting the strains according to fermentation conditions until flocculent turbid bacterial liquid appears in a culture medium, adding the flocculent turbid bacterial liquid into a conventional nutrient soil matrix according to the dosage of 50ml per pot to serve as a treatment group, and simultaneously taking a bacterial liquid group without the bacterial liquid as a control group; firstly, tobacco seedlings are grown, and tobacco variety Honghuadajinyuan is used as a test material. Firstly, uniformly scattering tobacco seeds in a seedling tray, placing the seedling tray in a greenhouse for seedling cultivation, transplanting the tobacco seedlings into plastic pot bowls after the tobacco seedlings grow to five leaves, placing one tobacco seedling in each pot, and culturing in the greenhouse, wherein 7 pots are treated for each pot. Sampling in the tobacco agglomeration stage, and determining various relevant indexes, wherein the specific measurement indexes and the method are as follows:
collecting data in the growing agglomeration stage of the tobacco plant,
plant height: the height from the base of the stem to the growing point on the surface of the plant is measured by a flexible ruler.
Stem circumference: the circumference of the stem between the 5 th and 6 th leaf positions from the ground.
Maximum leaf length: and measuring the distance from the base part to the tip of the blade by using a flexible rule to obtain the maximum blade length.
Maximum leaf width: and measuring the widest position of the blade by using a flexible rule to obtain the maximum blade width.
The concrete data of each index of tobacco plants of the root irrigation group and the control group treated by the bacterial liquid fermentation liquid of the strain 21.1.9.2-14 of the invention are shown in the following table 1. The growth forms of the tobacco plants in the bacteria-liquid root-irrigation group and the control group are shown in figure 3.
TABLE 1 indexes of fermentation liquor of strain 21.4.1.2-14 for treating tobacco
Figure BDA0003816434840000081
Experimental example 2
The application of the diluted fermentation liquor of Pseudomonas micro-piercing 21.1.9.2-14 (Pseudomonas versuta 21.1.2-14) in tobacco production: fermenting the strain according to fermentation conditionsFermenting until flocculent turbid bacterial liquid appears, diluting the bacterial liquid by 10, 20, 30, 40 and 50 times respectively (the concentration of the bacterial strain diluted by the bacterial liquid is 6.4 multiplied by 10 respectively 6 cfu/ml、 3.2×10 6 cfu/ml、2.1×10 6 cfu/ml、1.6×10 6 cfu/ml、1.28×10 6 cfu/ml) is used for root irrigation of the tobacco, and the other treatment methods are applied to the fermentation liquor.
Experimental example 3
The application of the bacterial powder prepared from the Pseudomonas spinosa 21.1.9.2-14 (Pseudomonas versuta 21.1.2-14) in the tobacco production: fermenting the strains according to fermentation conditions until flocculent turbid bacteria liquid appears, adding the fermented bacteria liquid into the wheat bran sterilized at high temperature and high pressure according to the proportion of 1kg wheat bran and 1L strain fermentation liquid, fully and uniformly mixing the mixture, putting the mixture into an electrothermal blowing drying box under the condition of 55 ℃, drying for 3h, storing the treated bacteria powder in a room-temperature drying place, applying the dried bacteria powder to fields when crops are cultivated, wherein the application amount is 5kg per mu of land, and the dried bacteria powder can be applied in ditches or directly scattered in the fields.
Experimental example 4 application of the bacterial strain 21.4.1.2-14 powder in tobacco production
The application of the activated bacterial powder prepared from the Pseudomonas spinosa 21.1.9.2-14 (Pseudomonas versuta 21.1.2-14) in tobacco production is as follows: the method for preparing the bacterial powder is the same as the experimental example 3, the activation of the strains is carried out before the use, firstly, the PDA culture medium is prepared, 5g of the bacterial powder is added into every 1L of the liquid culture medium, the mixture is put into a shaking table, the rotating speed is set to be 120r/min, the culture temperature is 28 ℃, and the bacterial liquid which obviously generates flocculent turbidity is cultured for 2-4 days and then can be used. The activated bacterial suspension was diluted 10, 20, 30, 40 and 50 times (the concentrations of the bacterial suspension diluted by these times were 6X 10 times, respectively 6 cfu/ml、3×10 6 cfu/ml、 2×10 6 cfu/ml、1.5×10 6 cfu/ml、1.2×10 6 cfu/ml) can be directly irrigated into roots according to the dosage of 10-100ml of diluent for each plant.
Data acquisition is carried out in the growing and agglomeration stage of the tobacco plant, and specific measurement indexes and methods are as follows:
plant height: the height from the base of the stem to the growing point on the surface of the plant is measured by a flexible ruler.
Stem circumference: the circumference of the stem between the 5 th leaf position and the 6 th leaf position from the ground.
Maximum leaf length: and measuring the distance from the base part to the tip of the blade by using a flexible rule to obtain the maximum blade length.
Maximum leaf width: and measuring the widest position of the blade by using a flexible rule to obtain the maximum blade width.
The concrete data of each index of tobacco plants of the root irrigation group and the control group by using the bacterial liquid activated by the bacterial strain 21.1.9.2-14 bacterial powder of the invention are shown in the following table 2:
TABLE 2
Figure BDA0003816434840000091
Experimental examples 5, 21.1.2-14 (Pseudomonas versuta 21.1.2-14) application in pasture grass production
Fermenting the strains according to fermentation conditions until flocculent turbid bacterial liquid appears in a culture medium, adding the flocculent turbid bacterial liquid into a conventional nutrient soil matrix according to the dosage of 50ml per pot to serve as a treatment group, and simultaneously taking a bacterial liquid group without the bacterial liquid as a control group; first, raising seedlings of pasture, and using a pasture variety ryegrass as a test material. Firstly, uniformly scattering pasture seeds in a seedling raising tray, placing the tray in a greenhouse for raising seedlings, transplanting the seedlings into plastic pot bowls after the pasture seedlings grow to five-leaf stage, placing one pasture seedling in each pot, and culturing in the greenhouse, wherein each pot is treated by 7 pots. Sampling when the pasture grows to about 20cm, and determining various related indexes, wherein the specific measurement indexes and the method are as follows:
plant height: the pasture is straightened, and the height from the base of the stem to the longest leaf on the surface of the pasture is measured by a flexible rule.
Tiller number: the leaves were divided, and the branching occurred at the base of the grass stalk was counted.
Fresh weight: the root of the pasture is cut off, and the weight of the overground part of the pasture is weighed by an electronic scale.
Dry weight: the root of the pasture is cut off, and the pasture is dried and weighed by an electronic scale.
The specific data of various indexes of pasture plants of a root irrigation group and a control group treated by the bacterial liquid fermented liquid of the strain 21.1.9.2-14 are shown in the following table 3. The growth forms of the grass plants of the bacteria liquid root-irrigation group and the control group are shown in figure 4.
TABLE 3
Figure BDA0003816434840000101

Claims (10)

1. Use of Pseudomonas spinosa (Pseudomonas veruta) for promoting plant growth.
2. A method for promoting plant growth, characterized in that Pseudomonas spinosa (Pseudomonas versuta) is applied in a plant growth environment.
3. A method of promoting plant growth as claimed in claim 2, wherein said applying comprises: mixing the Pseudomonas spinosa (Pseudomonas veruta) with soil to be used as a plant growth substrate, and/or performing root irrigation treatment on the plant by using fermentation liquor of the Pseudomonas spinosa (Pseudomonas veruta), and/or preparing the Pseudomonas spinosa (Pseudomonas veruta) into bacterial powder and then applying the bacterial powder in a furrow or directly scattering the bacterial powder in the field.
4. A Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14, wherein the preservation number is GDMCC NO:62683.
5. A biological fertilizer comprises a fertilizer active ingredient, and is characterized in that the fertilizer active ingredient comprises a Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14 with the preservation number of GDMCC NO:62683.
6. A biofertilizer according to claim 5, further comprising: an auxiliary material;
and/or the fertilizer is in a powder or liquid form.
7. Use of Pseudomonas spinosa (Pseudomonas versuta) strain 21.19.2-14 with the collection number of GDMCC NO:62683 in promoting plant growth is provided.
8. Use of a strain of Pseudomonas spinosa (Pseudomonas versuta) 21.1.2-14 according to claim 5 for promoting the growth of a plant, wherein said plant is tobacco or ryegrass.
9. A method for promoting the growth of tobacco or ryegrass, characterized in that Pseudomonas micropteria (Pseudomonas versuta) strain 21.1.2-14 with the collection number GDMCC NO:62683 is applied in the environment of tobacco growth.
10. The method of claim 9, wherein the applying is performed by a method selected from the group consisting of: mixing the Pseudomonas spinosa (Pseudomonas veruta) strain 21.4.1.2-14 with soil to be used as a tobacco growth substrate, and/or performing root irrigation treatment on a tobacco plant by using fermentation liquor of the Pseudomonas spinosa (Pseudomonas veruta) strain 21.4.1.2-14, and/or preparing the Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14 into bacterial powder and then performing furrow application or directly scattering the bacterial powder in a tobacco field;
and/or the bacterial powder is prepared by mixing and drying sterilized wheat bran and fermentation liquor of Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14;
and/or, the mixing refers to mixing of the wheat bran and fermentation liquor of Pseudomonas spinosa (Pseudomonas versuta) strain 21.1.9.2-14 according to a ratio of 1kg to 1L;
and/or, the drying means mixing, putting into an electric heating air blast drying oven, and drying for 3h at 55 ℃;
and/or the application amount of the bacterial powder is 5kg per mu of land;
and/or, during the root irrigation treatment, diluting the fermentation liquor of the Pseudomonas spinosa (Pseudomonas veruta) strain 21.1.2-14 into a diluent for use or directly using the fermentation liquor of the Pseudomonas spinosa (Pseudomonas veruta) strain 21.19.2-14;
and/or the dilution multiple is 10-50 times;
and/or, during the root irrigation treatment, the consumption of the diluent of each tobacco plant is 10-100ml;
and/or the amount of fermentation liquor of each ryegrass is 50ml during root irrigation treatment.
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