CN115340968B - New application of pseudomonas microphylla and method thereof, pseudomonas microphylla 21.1.9.2-14 and products thereof - Google Patents

New application of pseudomonas microphylla and method thereof, pseudomonas microphylla 21.1.9.2-14 and products thereof Download PDF

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CN115340968B
CN115340968B CN202211038593.1A CN202211038593A CN115340968B CN 115340968 B CN115340968 B CN 115340968B CN 202211038593 A CN202211038593 A CN 202211038593A CN 115340968 B CN115340968 B CN 115340968B
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pseudomonas
microphylla
versuta
tobacco
strain
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CN115340968A (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 a new application of pseudomonas microphylla and a method thereof, and pseudomonas microphylla 21.1.9.2-14 and products thereof, belonging to the technical field of microorganisms. The invention provides the use of Pseudomonas microphylla (Pseudomonas versuta) for promoting plant growth and a plant growth promoting method based on Pseudomonas microphylla (Pseudomonas versuta). The invention also provides a pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with the preservation number of GDMCC NO:62683, and provides a biofertilizer 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 21.1.9.2-14 can obviously improve the plant height, the stem circumference, the leaf length and the leaf width of the plant, and has obvious promotion effect on plant growth.

Description

New application of pseudomonas microphylla and method thereof, pseudomonas microphylla 21.1.9.2-14 and products thereof
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a novel application of pseudomonas microphylla and a method thereof, pseudomonas microphylla 21.1.9.2-14, and products, applications and methods thereof.
Background
Tobacco is an important commercial crop. In 2020, the total amount of interest tax paid on Chinese tobacco accounts for 1.184% of the total proportion of GDP in China. With the development of related technologies in the fields of agriculture and biology, the future prospect of the development of the tobacco industry is also wider and wider. At the same time, however, tobacco planting is faced with some difficult problems, the continuous cropping obstacle of tobacco being the most troublesome of which.
Continuous cropping obstacle mainly refers to the phenomenon that under normal agricultural management measures, continuous cropping of the same crop or the same crop in the same soil causes various diseases and insect pests of the crop to occur frequently, the growth condition is poor, the quality is poor and the yield is reduced. The continuous cropping area of tobacco is enlarged year after year under the influence of factors such as cultivation conditions, economic benefits and the like, and the occurrence of continuous cropping obstacle is further aggravated. The continuous cropping of tobacco causes the soil environment to deteriorate, which results in the influence of tobacco plants on the absorption of soil nutrients, further causes the weakening of photosynthesis and the reduction of transpiration rate of tobacco, and finally leads to the reduction of tobacco yield and quality. For these reasons, tobacco continuous cropping obstacles have become a bottleneck that limits further development in the tobacco industry. Therefore, the method for overcoming the problem of continuous cropping obstacle of tobacco is found, and has important significance for sustainable development of tobacco industry in China.
The main factors causing the tobacco continuous cropping obstacle are the change of the physicochemical properties of the soil, the enrichment of the autotoxic substances secreted by the root system and the change of microbial communities in the soil. Wherein, the change of the microbial community in the soil is concentrated and reflected in the reduction of beneficial bacteria and the enrichment of harmful bacteria of the tobacco by bacillus and the like in the soil. Microorganisms are not only a reservoir of plant available nutrients in soil, but also the power for soil organic matter and nutrient conversion and circulation. The decomposition of beneficial microorganisms such as partial bacteria can decompose organic matters in soil into inorganic nutrients such as P, K which can be utilized by plants, so that the organic matters are supplied to the plants for normal growth and development. Harmful microorganisms can form competition with plants to obstruct the plants from absorbing nutrient substances, thereby causing the plants to be ill. In summary, the coordination of soil microorganisms with tobacco plants, and the maintenance of dominant status of beneficial microorganism flora in tobacco planting soil are of great importance in tobacco planting production. With the development of related technologies in the microorganism field, the preparation of the screened microorganism into a pollution-free fertilizer such as a biological bacterial fertilizer and the like has become 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 developed by microbiological technology according to the modern organic sustainable agriculture principle by relying on the related knowledge in the field of soil ecology and plant nutrition. The active microorganisms in the microbial fertilizer can improve the diversity of microorganisms in soil, keep the microbial flora beneficial to plant growth in a dominant position, promote the absorption of plants to nutrition, improve the plant yield and increase the economic benefit. In the production process of the microbial fertilizer, the most important step is to screen strains with the effect of promoting plant growth.
The prior art in the field of microbiology has very few reports about strains of Pseudomonas microphylla (Pseudomonas versuta), and more never has reported about the use of Pseudomonas microphylla (Pseudomonas versuta) 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, which has a growth promoting effect on tobacco and is named as pseudomonas microphylla 21.1.9.2-14 (Pseudomonas versuta 21.4.1.9.2-14), and a corresponding application method thereof.
The technical scheme of the invention is as follows:
use of pseudomonas microphylla (Pseudomonas versuta) for promoting plant growth.
A method for promoting plant growth, characterized in that pseudomonas microphylla (Pseudomonas versuta) is applied in a plant growth environment.
The application mode comprises the following steps: mixing Pseudomonas microphylla (Pseudomonas versuta) with soil to be used as a plant growth matrix, and/or irrigating roots of plants with fermentation liquor of Pseudomonas microphylla (Pseudomonas versuta), and/or preparing Pseudomonas microphylla (Pseudomonas versuta) into bacterial powder and then applying the bacterial powder in furrows or directly scattering the bacterial powder in the field.
Pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14, characterized in that it has a deposit number GDMCC NO:62683.
A biofertilizer comprising a fertilizer active ingredient characterized in that said fertilizer active ingredient comprises pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC No. 62683.
The biological fertilizer also comprises: an auxiliary material;
preferably, the fertilizer is in the form of powder or liquid.
Use of pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC No. 62683 for promoting plant growth.
Preferably, the plant is tobacco or ryegrass.
A method for promoting the growth of tobacco or ryegrass, which is characterized in that pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with the preservation number of GDMCC NO:62683 is applied in the tobacco growth environment.
The manner of application is selected from: mixing the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with soil to be used as a tobacco growth matrix, and/or carrying out root irrigation treatment on tobacco plants by using fermentation liquor of the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14, and/or carrying out ditch application or direct sprinkling on tobacco fields after the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 is made into bacterial powder;
preferably, the bacterial powder is prepared by mixing sterilized wheat bran and fermentation liquor of pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 and drying;
preferably, the mixing means that the fermentation broth of wheat bran and pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 is mixed according to the ratio of 1kg to 1L;
preferably, the drying finger is put into an electrothermal blowing drying oven after being mixed, and dried for 3 hours at the temperature of 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 microphylla (Pseudomonas versuta) strain 21.1, 9.2-14 is diluted into a diluted liquor and then the diluted liquor is used or the fermentation liquor of the pseudomonas microphylla (Pseudomonas versuta) strain 21.1, 9.2-14 is directly used;
preferably, the dilution factor is 10-50;
preferably, the dilution liquid dosage of each plant of tobacco is 10-100ml during root irrigation treatment;
preferably, the amount of fermentation broth of each ryegrass is 50ml during the root irrigation treatment.
The invention provides a bacterial fermentation method for promoting tobacco growth and application thereof, and tobacco seedlings are treated by fermented bacterial liquid.
Further, the bacterial strain is Pseudomonas microphylla (Pseudomonas versuta, 4.1, 9.2-14), is obtained by screening grass rhizosphere soil in Shaogon county of Zhaotong, yunnan province, and is preserved in the microorganism strain preservation center of Guangdong province, and the preservation number of 21, 4.1, 9.2-14 (Pseudomonas versuta, 4.1, 9.2-14) is 62683.
Further, the preparation method of the bacterial fermentation broth comprises the following steps:
(1) Preparing a culture medium: LB medium: 10g of tryptone, 5g of yeast powder and 10g of sodium chloride are added into 1000ml of deionized water, and then uniformly stirred to obtain a 1LLB culture medium, and 15-20g of agar powder is added into the solid culture medium. The pH is natural, and the temperature is about 25-28 ℃.
(2) After the medium cooled, the activated cells were inoculated into a liquid medium and placed in a shaking table at 120r/min at a culture temperature of 28 ℃. Culturing for 2-4d;
further, the application mode of the strain is as follows:
(1) During crop cultivation, a cultivation hole corresponding to the seedling root size is dug in soil, a small seedling is placed in the cultivation hole, fermented bacterial liquid is poured on the root of a plant, and the bacterial liquid consumption is 10ml-100 ml/plant according to the root size.
(2) Mixing the fermented bacterial liquid with water for root irrigation in any proportion, and irrigating into cultivation holes or irrigating after earthing during crop cultivation.
(3) And adding the fermented bacterial liquid into the sterilized wheat bran, drying at low temperature to prepare bacterial powder, and applying the dried bacterial powder to the field when cultivating plants.
(4) And adding the fermented bacterial liquid into sterilized wheat bran, drying at low temperature to prepare bacterial powder, activating the bacterial powder in the nutrient solution, and pouring the bacterial powder into the roots of plants during cultivation.
(5) And freeze-drying the fermented bacterial liquid to prepare pure bacterial powder, and directly applying the dried pure bacterial powder when cultivating plants.
(6) And freeze-drying the fermented bacterial liquid to prepare pure bacterial powder, and mixing the pure bacterial powder with other substances for application.
(7) Freeze-drying the fermented bacterial liquid to prepare pure bacterial powder, activating the bacterial powder, and pouring the bacterial liquid containing activated bacteria on the root of the plant.
The invention has the following beneficial effects:
(1) The invention provides a new application of pseudomonas microphylla (Pseudomonas versuta) for promoting plant growth for the first time, and fills the blank of the prior art in the field.
(2) The invention obtains the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 capable of promoting plant growth for the first time, and a large number of experiments prove that the pseudomonas microphylla strain can obviously promote plant height, stem circumference, leaf length and leaf width of plants, and has obvious promotion effect on plant growth.
(3) Compared with other fertilization modes, the fermentation broth 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, and improves the economic benefit.
The preservation information of the Pseudomonas microphylla (Pseudomonas versuta) strain 21.4.1.9.2-14 of the invention is as follows:
name of strain preservation: 21 4.1 9.2-14
Deposit number: GDMCC NO 62683
Classification naming: pseudomonas micropunctata
Latin name: pseudomonas versuta
Preservation unit: microorganism strain collection center of Guangdong province
Deposit unit address: guangzhou city first middle road 100 # college 59 # building 5 Guangdong national academy of sciences of microbiology
Preservation date: 2022, 8 and 4.
Drawings
FIG. 1 shows the front and back side views of the growth of the coated plates of strains 21.4.1.9.2-14 according to the invention.
FIG. 2 is a phylogenetic tree of strains 21.4.1.9.2-14 according to the present invention.
FIG. 3 is a photograph showing the comparison of the growth patterns of tobacco root-irrigated with the fermentation broth of strain 21.1.9.2-14 according to experimental example 1 of the present invention and tobacco of a control group.
FIG. 4 is a comparison of growth patterns of ryegrass rooted with strain 21.1.9.2-14 in experimental example 5 of the present invention versus control ryegrass.
Detailed Description
The following describes the present invention in detail with reference to specific examples and experimental examples, but is not intended to limit the scope of the present invention.
Sources of biological materials
The tobacco variety Honghuadajinyuan used in experimental examples 1-4 of the present invention is a well-known and public tobacco variety, which is stored in the laboratory of applicant's unit, and also can be obtained by a commercial route.
The ryegrass used in experimental example 5 of the present invention is a publicly known and commonly used pasture plant, which is stored in the applicant's unit laboratory, and is also commercially available.
New use of Pseudomonas microphylla (Pseudomonas versuta) in group 1 example
The present set of examples provides the use of pseudomonas microphylla (Pseudomonas versuta) for promoting plant growth.
The use of pseudomonas microphylla (Pseudomonas versuta) in promoting plant growth is the first time proposed by the present invention, any application of pseudomonas microphylla (Pseudomonas versuta) to the growth of any plant and/or application of pseudomonas microphylla (Pseudomonas versuta) to the growth substrate of any plant and/or application of pseudomonas microphylla (Pseudomonas versuta) to any part of the growth process of any plant, application of pseudomonas microphylla (Pseudomonas versuta) to any form of growth, cultivation or planting environment of any plant, falls within the scope of the present invention.
Group 2 examples, methods of the invention for promoting plant growth
The present set of embodiments provides a method of promoting plant growth. All embodiments of this group share the following common features: pseudomonas microphylla (Pseudomonas versuta) is applied in a plant growth environment.
In some specific embodiments, the means of applying comprises: mixing Pseudomonas microphylla (Pseudomonas versus, 21, 4.1, 9.2-14) with soil to be used as a plant growth matrix, and/or irrigating roots of plants with fermentation liquor of Pseudomonas microphylla (Pseudomonas versuta, 21, 4.1, 9.2-14), and/or preparing Pseudomonas microphylla (Pseudomonas versuta, 21, 4.1, 9.2-14) into bacterial powder and then furrowing or directly scattering in the field.
Group 3 example, pseudomonas micropipes (Pseudomonas versuta) Strain 21 4.1.9.2-14 of the invention
This example provides a Pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC NO:62683.
Any use, replication, culture, propagation, inoculation, enrichment, sales, offers, production, preparation, addition, application, and administration of Pseudomonas microphylla (Pseudomonas versuta) strain 21.4.1.9.2-14 with deposit number GDMCC NO. 62683 falls within the scope of the present invention.
In some examples, the invention screens soil samples for a strain whose growth morphology of coated plates is shown in FIG. 1, and whose molecular characterization is shown in FIG. 2, and which belongs to Pseudomonas microphylla (Pseudomonas versuta) in taxonomic fashion. The invention names the strain 21.1.9.2-14 and sends the strain to be preserved, and the preservation information is as follows:
name of strain preservation: 21 4.1 9.2-14
Deposit number: GDMCC NO 62683
Classification naming: pseudomonas micropunctata
Latin name: pseudomonas versuta
Preservation unit: microorganism strain collection center of Guangdong province
Deposit unit address: guangzhou city first middle road 100 # college 59 # building 5 Guangdong national academy of sciences of microbiology
Preservation date: 2022, 8 and 4.
Group 4 example, biofertilizer of the invention
The present set of embodiments provides a biofertilizer. All embodiments of this group share the following common features: the biological fertilizer comprises a fertilizer active ingredient, wherein the fertilizer active ingredient comprises pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with the preservation number of GDMCC NO:62683.
In a further embodiment, the biological fertilizer further comprises: an auxiliary material;
preferably, the fertilizer is in the form of powder or liquid.
The bacterial strain 21.1.2-14 can be prepared into various possible formulations, such as powder, liquid, tablets, capsules and sprays, by adopting auxiliary materials commonly used in the field of microbial preparation and preparing microbial agents based on technical common knowledge in the field or conventional technical means known in the field, for example, technical means recorded in the technical books of microecological preparation, agricultural microbial concentrated preparation, pharmaceutical auxiliary materials handbook, new-compiled fertilizer application technical handbook and the like.
The auxiliary materials commonly used in the field of microbial preparation can be selected from medicinal auxiliary materials or common culture medium components for microorganisms; the pharmaceutical excipients include, but are not limited to: solvents, propellants, solubilizing agents, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure modifiers, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-binding agents, integration agents, permeation promoters, pH modifiers, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, release retarders.
Group 5 example, new use of the inventive Strain 21.1.9.2-14
This group of examples provides the use of Pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 deposited under accession number GDMCC NO:62683 for promoting plant growth.
In specific embodiments, the plant is tobacco or ryegrass.
The person skilled in the art can, in accordance with the teachings of the present invention, use the Pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC NO:62683 for the growth-promoting aspect of other plants including, but not limited to: rice, wheat, corn, soybean, cotton and other field crops; fruits such as strawberry, apple, blueberry, watermelon, etc.; flowers such as geranium, poinsettia, goldens, carnation and roses; pasture such as ryegrass; etc.
Group 6 examples, methods of the invention for promoting tobacco growth
The present set of embodiments provides a method of promoting tobacco growth. All embodiments of this group share the following common features: pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC NO:62683 was applied in a tobacco growing environment.
In a specific embodiment, the means of application are selected from: mixing the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with soil to be used as a tobacco growth matrix, and/or carrying out root irrigation treatment on tobacco plants by using fermentation liquor of the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14, and/or carrying out ditch application or direct sprinkling on tobacco fields after the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 is made into bacterial powder;
preferably, the bacterial powder is prepared by mixing sterilized wheat bran and fermentation liquor of pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 and drying;
preferably, the mixing means that the fermentation broth of wheat bran and pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 is mixed according to the ratio of 1kg to 1L;
preferably, the drying finger is put into an electrothermal blowing drying oven after being mixed, and dried for 3 hours at the temperature of 55 ℃;
preferably, the application amount of the bacterial powder is 5kg per mu of land;
preferably, during root irrigation treatment, the fermentation liquor of the pseudomonas micropunching (Pseudomonas versuta) strain 21.1, 9.2-14 is diluted and then used;
preferably, the dilution factor is 10-50;
preferably, the dilution of each tobacco plant is 10-100ml when the root irrigation treatment is performed.
In other embodiments, the promoting tobacco growth refers to: the plant height, and/or the stem circumference, and/or the leaf length and/or the leaf width of the tobacco plants are improved.
Experimental example 1, application of Pseudomonas microphylla 21.1.9.2-14 (Pseudomonas versuta 21.4.1.9.2-14) fermentation liquor in tobacco production.
Fermenting the strain to a flocculent turbid bacterial liquid in a culture medium according to fermentation conditions, adding the bacterial liquid into a conventional nutrient soil matrix according to the dosage of 50ml per basin to serve as a treatment group, and simultaneously taking a bacterial liquid-free group as a control group; firstly, raising seedlings of tobacco, and using a tobacco variety of safflower Dajinyuan as a test material. Firstly, uniformly broadcasting tobacco seeds in a seedling raising tray, putting the seedling raising tray in a greenhouse for raising seedlings, transplanting the tobacco seedlings into plastic pot bowls after the tobacco seedlings grow to a five-leaf period, putting one tobacco seedling per pot in the greenhouse for culturing, and treating 7 pots per pot. Sampling in the tobacco agglomeration period, and measuring various relevant indexes, wherein the specific measurement indexes and methods are as follows:
data acquisition is carried out in the period of the growing tobacco plants,
plant height: the height from the basal part of the plant ground stem to the growing point is measured by a flexible ruler.
The stem circumference: the circumference of the stem from the 5 th leaf position to the 6 th leaf position from the ground.
Maximum leaf length: and measuring the blade base to the blade tip by using a flexible rule to obtain the maximum blade length.
Maximum leaf width: and measuring the widest part of the blade by using a flexible rule to obtain the maximum blade width.
Specific data of various indexes of tobacco plants of a root irrigation group and a control group of bacterial liquid treated by the fermentation liquid of the strain 21.1.9.2-14 are shown in the following table 1. The growth forms of tobacco plants in the root irrigation group and the control group of the bacterial liquid are shown in figure 3.
TABLE 1 various indices of tobacco treated with Strain 21 4.1.9.2-14 fermentation broth
Experimental example 2
Application of Pseudomonas microphylla 21.1.9.2-14 (Pseudomonas versuta 21.4.1.9.2-14) fermentation liquor after dilution in tobacco production: fermenting the strain according to fermentation conditions until flocculent turbidity appears in the strain, respectively diluting the strain 10, 20, 30, 40 and 50 times (the concentration of strain diluted solution after dilution of the strain is 6.4X10) 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) root irrigation treatment is carried out on tobacco, and the rest treatment methods are applied with fermentation liquor.
Experimental example 3
Application of Pseudomonas microphylla 21.1.9.2-14 (Pseudomonas versuta 21.4.1.9.2-14) prepared into bacterial powder in tobacco production: fermenting the strain according to the fermentation condition until flocculent turbid bacterial liquid appears, adding the fermented bacterial liquid into wheat bran subjected to high-temperature high-pressure sterilization, adding 1L of strain fermentation liquid into 1kg of wheat bran, fully and uniformly mixing the materials, putting the mixture into an electrothermal blowing drying oven under the condition of 55 ℃, drying the mixture for 3 hours, storing the treated bacterial powder at a normal-temperature drying place, applying the dried bacterial powder to a field in an application amount of 5kg per mu of land during crop cultivation, and applying the bacterial powder in furrows or directly scattering the bacterial powder in the field.
Experimental example 4, application of the bacterial strain 21 4.1.9.2-14 bacterial powder in tobacco production
Application of pseudomonas microphylla 21.1.9.2-14 (Pseudomonas versuta 21.4.1.9.2-14) prepared into bacterial powder after activation in tobacco production: the method for preparing the bacterial powder is the same as that of experimental example 3, bacterial activation is carried out before use, firstly PDA culture medium is prepared, 5g of bacterial powder is added into each 1L of liquid culture medium, the liquid culture medium is put into a shaking table, the rotation speed is set to be 120r/min, the culture temperature is 28 ℃, and bacterial liquid with obvious flocculent turbidity appears in 2-4 days of culture can be used. Diluting the activated bacterial solutions 10, 20, 30, 40, and 50 times (the concentration of the diluted bacterial solutions after dilution of the bacterial solutions is 6×10 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), tobacco can be directly root-irrigated according to the dosage of 10-100ml of diluent for each plant.
The data acquisition is carried out in the period of the growing tobacco plant, and the specific measurement indexes and methods are as follows:
plant height: the height from the basal part of the plant ground stem to the growing point is measured by a flexible ruler.
The stem circumference: the circumference of the stem from the 5 th leaf position to the 6 th leaf position from the ground.
Maximum leaf length: and measuring the blade base to the blade tip by using a flexible rule to obtain the maximum blade length.
Maximum leaf width: and measuring the widest part of the blade by using a flexible rule to obtain the maximum blade width.
Specific data of various indexes of tobacco plants of a root irrigation group and a control group of bacterial liquid after bacterial powder activation of the bacterial strain 21.1.9.2-14 are shown in the following table 2:
TABLE 2
Experimental examples 5, 21.1.9.2-14 (Pseudomonas versuta 21.4.1.9.2-14) application in pasture production
Fermenting the strain to a flocculent turbid bacterial liquid in a culture medium according to fermentation conditions, adding the bacterial liquid into a conventional nutrient soil matrix according to the dosage of 50ml per basin to serve as a treatment group, and simultaneously taking a bacterial liquid-free group as a control group; firstly, raising seedlings of pasture, and using a pasture variety ryegrass as a test material. Firstly, uniformly broadcasting pasture seeds in a seedling raising tray, putting the pasture seeds in a greenhouse for raising seedlings, transplanting the pasture seedlings into plastic pot bowls after the pasture seedlings grow to a five-leaf period, putting one pasture seedling in the greenhouse for culturing, and treating 7 pots each. Sampling when the pasture grows to about 20cm, and measuring various related indexes, wherein the specific measuring indexes and the method are as follows:
plant height: straightening the pasture, and measuring the height from the basal part of the surface stem of the pasture to the longest leaf by using a flexible rule.
Tillering number: leaves were separated and branches were counted from the basal part of the grass stems.
Fresh weight: the roots of the pasture were cut off and the weight of the upper part of the pasture was weighed with an electronic scale.
Dry weight: cutting the roots of the pasture, drying the pasture, and weighing the pasture by an electronic scale.
Specific data of various indexes of pasture plants of a root irrigation group and a control group of bacteria liquid treated by the fermentation broth of the strain 21.1.9.2-14 of the present invention are shown in the following table 3. The growth forms of pasture plants in the root irrigation group and the control group of the bacterial liquid are shown in figure 4.
TABLE 3 Table 3

Claims (18)

1. Use of pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC No. 62683 for promoting plant growth.
2. A method for promoting plant growth, characterized in that pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC No. 62683 is applied in a plant growth environment.
3. A method of promoting plant growth according to claim 2, wherein the means of application comprises: mixing Pseudomonas microphylla (Pseudomonas versuta) with soil to be used as a plant growth matrix, and/or irrigating roots of plants with fermentation liquor of Pseudomonas microphylla (Pseudomonas versuta), and/or preparing Pseudomonas microphylla (Pseudomonas versuta) into bacterial powder and then applying the bacterial powder in furrows or directly scattering the bacterial powder in the field.
4. Pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14, characterized in that it has a deposit number GDMCC NO:62683.
5. A biofertilizer comprising a fertilizer active ingredient characterized in that said fertilizer active ingredient comprises pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC No. 62683.
6. The biofertilizer of claim 5, further comprising: an auxiliary material;
and/or the fertilizer is in the form of powder or liquid.
7. Use of pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with deposit number GDMCC No. 62683 for promoting plant growth.
8. Use of a strain 21.1.9.2-14 of pseudomonas microphylla (Pseudomonas versuta) according to claim 5 for promoting plant growth, wherein the plant is tobacco or ryegrass.
9. A method for promoting the growth of tobacco or ryegrass, which is characterized in that pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with the preservation number of GDMCC NO:62683 is applied in the tobacco growth environment.
10. The method of claim 9, wherein the applying is performed in a manner selected from the group consisting of: mixing the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 with soil to be used as a tobacco growth matrix, and/or carrying out root irrigation treatment on tobacco plants by using bacterial liquid of the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14, and/or carrying out ditch application or direct sprinkling on tobacco fields after the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 is made into bacterial powder.
11. The method for promoting the growth of tobacco or ryegrass according to claim 10, wherein the bacterial powder is prepared by mixing and drying sterilized wheat bran and bacterial liquid of pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14.
12. The method for promoting the growth of tobacco or ryegrass according to claim 11, wherein the mixed liquid of wheat bran and pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 is mixed according to the ratio of 1kg to 1L.
13. The method for promoting the growth of tobacco or ryegrass according to claim 11, wherein the drying finger is placed in an electrothermal blowing drying oven after mixing, and dried for 3 hours at 55 ℃.
14. The method for promoting the growth of tobacco or ryegrass according to claim 10 or 11, wherein the application amount of the bacterial powder is 5kg per mu of land.
15. The method for promoting the growth of tobacco or ryegrass according to claim 10, wherein the root-irrigation treatment is carried out by diluting the bacterial liquid of the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14 into a diluted liquid and then using or directly using the bacterial liquid of the pseudomonas microphylla (Pseudomonas versuta) strain 21.1.9.2-14.
16. The method of claim 15, wherein the dilution factor is 10-50.
17. The method for promoting the growth of tobacco or ryegrass according to claim 10 or 15, wherein the dilution of each plant of tobacco is 10-100ml when the root-irrigation treatment is performed.
18. The method for promoting the growth of tobacco or ryegrass according to claim 17, wherein the amount of bacterial liquid per ryegrass is 50ml during the root irrigation treatment.
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