CN113817625B - Flavobacterium acidophilus and application thereof in improvement of saline-alkali soil - Google Patents
Flavobacterium acidophilus and application thereof in improvement of saline-alkali soil Download PDFInfo
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Abstract
The invention discloses a strain of flavobacterium acidophilus and application thereof in improving saline-alkali soil. The acid flavobacterium is acid flavobacterium (Flavobacterium acidificum) 237T2, and the registration number of the acid flavobacterium in the China general microbiological culture Collection center is CGMCC No.19869. The acid flavobacterium disclosed by the invention has the advantages of reducing the pH value of the microenvironment of rhizosphere soil, being capable of symbiotic with plants, promoting plant growth and adapting to the unfavorable growth environment of saline-alkali soil.
Description
Surgical field
The invention relates to a strain of flavobacterium acidophilus in the field of microorganisms and application thereof in improving saline-alkali soil.
Background
Soil salinization is one of the most serious environmental problems in the world at present, and is also a direct influencing factor for limiting farmland efficient utilization and causing low agricultural productivity level. The salinized soil is widely distributed in more than 100 countries and regions of the world, and the area reaches 10 hundred million hm 2 . In addition 2.3X10 9 hm 2 4.5X10 of irrigated land 7 hm 2 Is endangered by salinization and occupies 19.5 percent of the irrigation area. The area of the salty soil in China is large, and the total area of various salty soil is about 9900 ten thousand hm 2 Mainly concentrated in northwest, northeast and coastal areas. Excessive salinity in the soil can cause changes in the physical and chemical properties of the soil, resulting in degradation of the growing environment of most crops, and soil salinization is also a major cause of agricultural yield degradation in many arid and semiarid regions of the world.
Saline-alkali soil is an important cultivated land reserve resource in China, and how to develop and utilize the saline-alkali soil resource is one of the important problems in agricultural production in China. For a long time, measures for improving the saline-alkali soil at home and abroad generally comprise four types: (1) water conservancy improvement measures, including irrigation, drainage, desilting, rice planting, seepage prevention and the like; (2) agricultural improvement measures: including leveling land, deep ploughing and sunning upturned soil, improving tillage, applying soil, fertilizing, sowing, rotation, interplanting and the like; (3) chemical improvement measures: comprising applying a modifying substance such as gypsum, phosphogypsum, calcium sulfite, calcium superphosphate, etc.; (4) biological improvement measures: including planting salt-tolerant plants and pasture, green manure, afforestation, etc. Because each measure has a certain application range and condition. Therefore, the treatment should be performed comprehensively according to the local conditions. At present, the microbial agent for improving the saline-alkali soil becomes a research hot spot, and the microbial agent has the main effects that microorganisms in the microbial agent can reproduce in soil, improve the soil nutrient condition, improve the soil enzyme activity, increase the microbial diversity and change the microbial flora, so that the effect of regulating the growth of crops is achieved.
Disclosure of Invention
The technical problem to be solved by the invention is how to improve the saline-alkali soil, and/or how to improve the photosynthetic performance of plants, and/or how to promote the growth of plants, and/or how to improve the biomass of plants, and/or how to improve the crop yield, and/or how to improve the salt and alkali tolerance of plants.
In order to solve the technical problems, the invention firstly provides a strain of flavobacterium acidophilus.
The Flavobacterium acidi provided by the invention is Flavobacterium acidi (Flavobacterium acidificum) 237T2, and the registration number of the Flavobacterium acidi in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No.19869. The strain is preserved in China general microbiological culture collection center (CGMCC) of China general microbiological culture Collection center (CGMCC) for 5 and 25 days in 2020. Hereinafter, flavobacterium acidophilus 237T2.
The acid flavobacterium 237T2 thallus is in a rod shape, gram-negative, circular in colony, irregular in edge, smooth in surface, moist, yellow, slightly protruding and opaque.
Flavobacterium acidophilus 237T2 has 16S rDNA shown in sequence 1 in the sequence table.
The following uses of Flavobacterium acidophilus 237T2 or/and a metabolite of Flavobacterium acidophilus 237T2 or/and a culture of Flavobacterium acidophilus 237T2 are also within the scope of the invention:
U1, application in improving saline-alkali soil or alkaline soil or in preparing products for improving saline-alkali soil or alkaline soil,
u2, the application in reducing the pH value of the saline-alkali soil or alkaline soil or the application in preparing a product for reducing the pH value of the saline-alkali soil or alkaline soil,
u3, in improving the salt alkali resistance or alkali resistance of plants or in preparing products for improving the salt alkali resistance or alkali resistance of plants,
u4, na in reducing the saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
u5, K in improving the saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of K for improving saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
u6, ca in improving the saline-alkali soil or alkaline soil of plants 2+ Application in absorption or preparation of Ca in improving saline-alkali soil or alkaline soil of plants 2+ The application in the field of absorption is that,
u7, na in reducing the saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
u8, na in reducing the saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil or alkaline soil by plants 2+ Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil by plants 2+ The use of the absorbent article in the manufacture of an absorbent article,
u9, the application in improving the photosynthetic performance of plants or the application in preparing products for improving the photosynthetic performance of plants,
u10, reduction of plant intercellular CO 2 Use in concentration or in the preparation of reduced plant intercellular CO 2 The application of the product with the concentration is that,
u11, the application in improving the net photosynthetic rate of plants or the application in preparing products for improving the net photosynthetic rate of plants,
u12, in the application of improving plant stomata conductivity or in the preparation of products for improving plant stomata conductivity,
u13, application in improving plant transpiration rate or in preparing product for improving plant transpiration rate,
u14, in the application of improving the water utilization efficiency of plant leaves or in the preparation of products for improving the water utilization efficiency of plant leaves,
u15, in the application of improving the chlorophyll content of plants or in the preparation of products for improving the chlorophyll content of plants,
U16, use in the production of auxin (IAA) or in the preparation of an auxin (IAA) producing product,
u17, application in promoting plant growth or application in preparing product for promoting plant growth,
u18, in the improvement of crop yield or in the preparation of products for improving crop yield,
u19, use in increasing plant biomass or use in preparing a product for increasing plant biomass.
In the above application, the product may be a microbial agent, a microbial ecological agent containing the microbial agent, or a biofertilizer containing the microbial agent.
In the above application, the culture of Flavobacterium acidophilus 237T2 may be a substance obtained by culturing Flavobacterium acidophilus 237T2 in a microorganism culture medium (i.e., a fermentation product such as a substance containing Flavobacterium acidophilus 237T2 and secreted into a liquid culture medium, i.e., a fermentation broth, or such as a substance containing Flavobacterium acidophilus 237T28 and secreted into a solid culture medium, i.e., a solid fermentation product).
In order to solve the technical problems, the invention also provides a microbial agent, a microecological preparation containing the microbial agent or a biological fertilizer containing the microbial agent.
The microbial inoculum provided by the invention contains Flavobacterium acidophilus 237T2 or/and metabolites of Flavobacterium acidophilus 237T 2.
The microbial inoculum can be specifically any one of the following microbial inoculum:
a1, a microbial inoculum for improving saline-alkali soil or alkaline soil,
a2, a microbial inoculum for reducing the pH value of the saline-alkali soil or alkaline soil,
a3, a microbial agent for improving the salt-alkali resistance or alkali resistance of plants,
a4, reducing Na in the saline-alkali soil or alkaline soil of plants + The bacteria agent is absorbed by the bacteria agent,
a5, improving K in the saline-alkali soil or alkaline soil of plants + The bacteria agent is absorbed by the bacteria agent,
a6, improving Ca in the saline-alkali soil or alkaline soil of plants 2+ The bacteria agent is absorbed by the bacteria agent,
a7, reducing Na in the saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + The bacteria agent is absorbed by the bacteria agent,
a8, reducing Na in the saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil or alkaline soil by plants 2+ The bacteria agent is absorbed by the bacteria agent,
a9, a microbial inoculum for improving the photosynthetic performance of plants,
A10reducing plant intercellular CO 2 The concentration of the microbial inoculum is calculated,
a11, a microbial inoculum for improving the net photosynthetic rate of plants,
a12, a microbial inoculum for improving plant stomata conductivity,
a13, a microbial inoculum for improving the plant transpiration rate,
a14, a microbial inoculum for improving the water utilization efficiency of plant leaves,
a15, a microbial inoculum for improving the chlorophyll content of plants,
a16, a bacterial agent for producing auxin (IAA),
A17, a microbial inoculum for promoting plant growth,
a18, a microbial inoculum for improving crop yield,
a19, a microbial inoculum for improving plant biomass.
The active ingredients of the microbial inoculum can be Flavobacterium acidophilus 237T2 or/and metabolites of Flavobacterium acidophilus 237T2, and the active ingredients of the microbial inoculum can also contain other biological components or non-biological components, and the other active ingredients of the microbial inoculum can be determined by one skilled in the art according to the effects of the microbial inoculum.
The microbial agent may also include a carrier. The carrier may be a solid carrier or a liquid carrier. The solid carrier is a mineral material and a biological material; the mineral material may be at least one of turf, clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, and diatomaceous earth; the biological material is at least one of straws, pine shells, rice straws, peanut shells, corn flour, bean flour, starch, turf and animal excrement of various crops; the liquid carrier may be water; in the microbial inoculum, the Flavobacterium acidophilum 237T2 and/or the metabolite of Flavobacterium acidophilum 237T2 may exist in the form of living cells to be cultured, a fermentation broth of the living cells, a filtrate of a cell culture, or a mixture of cells and the filtrate. The dosage form of the microbial inoculum can be various dosage forms, such as liquid, emulsion, suspending agent, powder, granule, wettable powder or water dispersible granule.
Surfactants (such as Tween 20, tween 80, etc.), binders, stabilizers (such as antioxidants), pH regulators, etc. can also be added into the microbial inoculum according to the need.
In the above, the metabolite of Flavobacterium acidophilus 237T2 may be a fermentation broth of Flavobacterium acidophilus 237T 2. The fermentation broth of Flavobacterium acidophilus 237T2 can be prepared as follows: culturing Flavobacterium acidophilus 237T2 in a liquid fermentation medium, and collecting fermentation broth (containing Flavobacterium acidophilus 237T2 and substances secreted into the liquid fermentation medium), wherein the fermentation broth is metabolite of Flavobacterium acidophilus 237T 2.
Cultures of Flavobacterium acidophilus 237T2 as described above are also within the scope of the invention. The culture of Flavobacterium acidophilus 237T2 described above may have at least one of the following functions:
b1, improving the saline-alkali soil or alkaline soil,
b2, reducing the pH value of the saline-alkali soil or alkaline soil,
b3, a microbial agent for improving the salt-alkali resistance or alkali resistance of plants,
b4, reducing Na in the saline-alkali soil or alkaline soil of plants + The absorption of the liquid is carried out,
b5, improving K in the saline-alkali soil or alkaline soil of plants + The absorption of the liquid is carried out,
b6, improving Ca in the saline-alkali soil or alkaline soil of plants 2+ The absorption of the liquid is carried out,
b7, reducing Na in the saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil by plants + The absorption of the liquid is carried out,
b8, reducing Na in the saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil by plants 2+ The absorption of the liquid is carried out,
b9, improving the photosynthetic performance of the plants,
b10, decrease plant intercellular CO 2 The concentration of the water in the water is higher,
b11, improving the net photosynthetic rate of the plants,
b12, improving the air permeability of the plant air holes,
b13, improving the transpiration rate of the plants,
b14, improving the water utilization efficiency of the plant leaves,
b15, improving the chlorophyll content of the plants,
b16, producing auxin (IAA),
b17, promote the growth of plants,
b18, the yield of crops is improved,
b19, improving plant biomass.
Any of the following applications of the microbial inoculum also fall within the scope of the invention:
c1, application in improving saline-alkali soil or alkaline soil or in preparing products for improving the saline-alkali soil or alkaline soil,
c2, the application in reducing the pH value of the saline-alkali soil or alkaline soil or the application in preparing a product for reducing the pH value of the saline-alkali soil or alkaline soil,
c3, application in improving salt and alkali resistance of plants or in preparing products for improving salt and alkali resistance of plants,
c4, na in the saline-alkali soil or alkaline soil of the reduced plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
c5, K in improving saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of K for improving saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
c6, ca in improving saline-alkali soil or alkaline soil of plants 2+ Application in absorption or preparation of Ca in improving saline-alkali soil or alkaline soil of plants 2+ The application in the field of absorption is that,
c7, na in the saline-alkali soil or alkaline soil of the reduced plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
c8, na in saline-alkali soil or alkaline soil of plants is reduced + Absorbing and improving Ca in saline-alkali soil or alkaline soil by plants 2+ Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorption andimproving Ca in saline-alkali soil or alkaline soil of plants 2+ The use of the absorbent article in the manufacture of an absorbent article,
c9, application in improving photosynthetic performance of plants or in preparing products for improving photosynthetic performance of plants,
c10 and reduction of plant intercellular CO 2 Use in concentration or in the preparation of reduced plant intercellular CO 2 The application of the product with the concentration is that,
c11, in the application of improving the net photosynthetic rate of plants or in the preparation of products for improving the net photosynthetic rate of plants,
c12, application in improving plant stomata conductivity or in preparing products for improving plant stomata conductivity,
c13, application in improving plant transpiration rate or application in preparing product for improving plant transpiration rate,
c14, application in improving the water utilization efficiency of the plant leaves or application in preparing products for improving the water utilization efficiency of the plant leaves,
c15, application in improving plant chlorophyll content or in preparing product for improving plant chlorophyll content,
c16, use in the production of auxin (IAA) or in the preparation of an auxin (IAA) producing product,
c17, application in promoting plant growth or application in preparing product for promoting plant growth,
c18, use in increasing crop yield or use in preparing a product for increasing crop yield,
c19, use in increasing plant biomass or use in preparing a product for increasing plant biomass.
In the above application, the product may be a microbial ecological agent containing the microbial agent or a biological fertilizer containing the microbial agent.
In this application, the plant or crop may be a monocot or dicot. The dicotyledonous plant may be a plant of the order platycodon. The tubular flower plant may be a solanaceae plant. The Compositae plant may be a sunflower plant. The sunflower plant may be sunflower.
The method for culturing the flavobacterium acidophilus 237T2 also belongs to the protection scope of the invention.
The method for culturing the flavobacterium acidophilus 237T2 provided by the invention comprises the step of culturing the flavobacterium acidophilus 237T2 in a culture medium for culturing microorganisms.
The method for preparing the microbial inoculum also belongs to the protection scope of the invention.
The method for preparing the microbial inoculum provided by the invention comprises the step of taking the flavobacterium acidophilus 237T2 and/or the metabolite of the flavobacterium acidophilus 237T2 as components of the microbial inoculum to obtain the microbial inoculum.
In the above method, the microbial inoculum may be a liquid microbial inoculum. In the method, the flavobacterium acidophilus 237T2 can be cultured in a fermentation medium to obtain a fermentation broth, and the fermentation broth is mixed with a carrier to obtain the liquid microbial inoculum. The carrier may be turfy soil.
The microbial inoculum for improving the saline-alkali soil prepared by utilizing the flavobacterium acidicum 237T2 can be propagated in the soil of a root zone to form microbial dominant bacterial groups beneficial to the growth of crops because of a large amount of beneficial living bacterial substances and a plurality of natural fermentation active substances, so that the rhizosphere nutrition environment is regulated, the nutrition is manufactured and assisted by the crops, the microecological balance of the soil is improved and restored, the microbial inoculum can be used for improving the physicochemical property and the soil structure of the saline-alkali soil, thereby improving the permeability, accelerating salt leaching, cutting off capillaries to prevent the saline-alkali from returning and reducing the salt content of the soil. The flavobacterium acidizing 237T2 has the advantages of reducing the pH value of the microenvironment of rhizosphere soil, being capable of symbiotic with plants, promoting plant growth and adapting to the unfavorable growth environment of saline-alkali soil. The mechanism of the invention for improving the saline-alkali soil is as follows: (1) the activity of the flavobacterium acidophilus 237T2 can generate a large number of metabolic derivatives, and can effectively replace indissolvable elements solidified by saline-alkali soil for plant absorption and utilization; (2) the grass carbon contains a large amount of organic matters, has high organic matter content and rich humic acid, and also contains cellulose, hemicellulose, nitrogen, phosphorus, potassium and other substances; (3) flavobacterium acidophilus 237T2 can reduce the alkalinity of soil. (4) Flavobacterium acidophilus 237T2 is selected from plants, has symbiotic advantages with plants, can generate IAA and promote crop growth.
Biological material preservation instructions.
Classification naming of biological materials: flavobacterium acidicum.
Latin Wen Xueming of biological material: flavobacterium acidificum.
Strain number of biological material: 237T2.
The preservation units are fully named: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit is abbreviated as: CGMCC.
Address: no.1 and No. 3 of the north cinquefoil of the morning sun area of beijing city.
Preservation date: 25 days 5 and 25 months 2020.
Preservation number: CGMCC No.19869.
Drawings
FIG. 1 is a phylogenetic tree constructed based on the 16S rRNA gene sequence.
FIG. 2 shows the effect of Flavobacterium acidophilus 237T2 liquid inoculant on the pH of saline-alkali soil. The inter-treatment differences with different lower case letters were significant (P < 0.05).
FIG. 3 shows the results of qualitative detection of the secretion of auxin by Flavobacterium acidophilus 237T2. The left panel shows the treatment with Flavobacterium acidophilus 237T2 broth, and the right panel shows the negative Control (CK).
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The soil salinity and pH determination methods in the following examples are as follows: square for sampling at five pointsIn the method, 100g of soil (hereinafter referred to as initial soil) is taken at each point and brought back to a laboratory for natural air drying, ground, sieved by a 2mm sieve, supernatant of soil solution is extracted at a soil-to-water ratio of 1:5, and pH and conductivity of the soil are measured by a pH meter (FE 20) and a conductivity meter (DDS-307), respectively, and the conductivity is converted into soil salt content according to an empirical formula: soil salinity (g.kg) -1 ) =conductivity×0.064×5×10/1000.
Plant K in the following examples + 、Na + And Ca 2+ The content determination method comprises the following steps: weighing about 0.5g of plant leaf, adding 4ml of concentrated nitric acid (high-grade pure) into digestion tank, and adding H 2 O 2 (superior purity) 2ml. Standing for 2 hours, capping, screwing, slightly shaking, putting on a microwave digestion instrument for digestion by a preset program, opening the cover, putting on an acid-expelling instrument for expelling acid to the full extent at about 160 ℃, transferring to a 50ml volumetric flask with 5% nitric acid for constant volume, dry filtering, and detecting by an atomic absorption spectrophotometer.
Na in the following examples + /K + ＝Na + content/K + Content of Na + /Ca 2+ ＝Na + content/Ca 2+ The content is as follows.
Net photosynthetic rate, stomatal conductance, intercellular CO in the examples below 2 The concentration and transpiration rate were determined as follows: 8:00-11:30 measuring plant leaves by using LI-6400 portable photosynthetic apparatus (Beijing Ligaotai technology Co., ltd.) with intensity of light being set to 1900 umol/(m) uniformly 2 ·s)。
The method for measuring the Water Use Efficiency (WUE) in the following examples is as follows: wue=gy/(p+. DELTA.W).
GY is the yield (kg); p is precipitation (mm); Δw is the difference (mm) in water storage of soil at the beginning and end of the observation period.
The chlorophyll content determination method in the following examples is as follows: the leaf parts of plants were measured using a portable chlorophyll meter (SPAD-502,KONICAMINOLTA SENSING,INC.) for a period of 8 a.m.: 00-8:30.SPAD value is a parameter that measures the relative content of chlorophyll in a plant.
The following examples were tested for the aroma index, the simpson index, the ACE index and the Chao1 index as follows: sunflower leaves were harvested and stored in a refrigerator at 4℃and then sent to the Shanghai Meiji biological medicine technologies Co.
All data in the examples below were analyzed for significance using IBM SPSS Statistics 23.
Example 1 isolation and identification of Flavobacterium acidophilus (Flavobacterium acidificum) 237T2
1. Isolation of Flavobacterium acidophilus (Flavobacterium acidificum)
Collecting rice seeds in a greenhouse of a national academy of agricultural sciences and agricultural division institute in the sea lake area of Beijing city, and carrying out surface sterilization after husking, wherein the method comprises the following steps: washing 1g of dehulled rice seeds with sterile water to remove surface dust and the like, sequentially soaking in 70% ethanol for 3min, soaking in 5% sodium hypochlorite for 5min, washing with 70% ethanol for 30s, washing with sterile water for 5-7 times, simultaneously taking sterilized rice seeds, slightly pressing and smearing the sterilized rice seeds on a TSA (Shanghai ai Yes Biotechnology Co., ltd.) plate, and culturing at 28 ℃ for 72h to check the surface sterilization effect. Grinding the surface sterilized seeds into powder in a sterile mortar, transferring the powder into 45ml of sterile water containing glass beads, oscillating at 28deg.C for 30min, and performing series from 10 respectively -3 、10 -4 10 -5 0.1ml of the diluted solution is spread on a flat plate, and after 3 times of culture at 28 ℃ for 3-5 days, single colony is picked up for separation, thus obtaining endophytic bacteria 237T2.
2. Identification of Flavobacterium acidophilus (Flavobacterium acidificum)
1. Morphological identification
The endophyte 237T2 in the logarithmic growth phase was observed for colony status by streaking, and the colony status mainly includes size, color, transparency, wettability, colony surface status (whether flat, protruding, wrinkled, depressed, etc.), colony edge status (whether regular, irregular, radial, etc.). On the other hand, the form of the cells was observed by an optical microscope after gram staining of endophyte 237T2 in the logarithmic growth phase. Biochemical identification was performed using the API20NE system.
The results show that endophytic bacteria 237T2 bacteria are rod-shaped, gram-negative, circular in colony, irregular in edge, smooth in surface, moist, yellow, slightly protruding and opaque. The physiological and biochemical characteristics of the acid flavobacterium 237T2 are shown in table 1, and the result is that the nitrate reduction and utilization test is negative, tryptophan can be decomposed to produce indole, glucose can be hydrolyzed to produce acid, escin hydrolysis is positive, gelatin hydrolysis is positive, arginine hydrolysis is negative, urea hydrolysis is negative, and PNPG test is positive. Glucose, arabinose, mannose, mannitol, N-acetyl-glucosamine, maltose, gluconate, malic acid and citric acid assimilation test positive, and capric acid, adipic acid and phenylacetic acid assimilation test negative.
TABLE 1 physiological and biochemical index of Flavobacterium acidophilus 237T2
| Test | Substrate(s) | Reaction | Results |
| NO 3 | Potassium nitrate | Nitrate reduction utilization | - |
| TRP | L-tryptophane | Indole production | + |
| GLU | D-glucose | Glucose acidification | + |
| ADH | L-arginine | Arginine bishydrolase | - |
| URE | Urea | Hydrolysis of urea | - |
| ESC | Esculin | Hydrolysis of esculin | + |
| GEL | Gelatin | Gelatin hydrolysis | + |
| PNPG | 4-nitroso-βD-methyl galactose | Galactosidase enzyme | + |
| GLU | D-glucose | Glucose assimilation reaction | + |
| ARA | L-arabinose | Arabinose assimilation reaction | + |
| MNE | D-mannose | Mannose assimilation reaction | + |
| MAN | D-mannitol | Mannitol assimilation reaction | + |
| NAG | N-acetyl glucosamine | N-acetyl-glucosamine assimilation reaction | + |
| MAL | D-maltose | Maltose assimilation reaction | + |
| GNT | Gluconate | Gluconate assimilation reaction | + |
| CAP | Decanoic acid | Capric acid assimilation reaction | - |
| ADI | Adipic acid | Adipic acid assimilation reaction | - |
| MLT | Malic acid | Malic acid assimilation reaction | + |
| CIT | Citric acid | Citric acid assimilation reaction | + |
| PAC | Phenylacetic acid | Phenylacetic acid assimilation reaction | - |
2. 16S rRNA Gene and genomic sequence homology analysis
The total genome DNA of endophytic bacteria 237T2 is extracted by adopting a root bacterium genome extraction kit. The 16S rRNA gene was amplified using the universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3'). The reaction system was 50. Mu.L: comprises 10 Xbuffer (5. Mu.L), dNTP (2.5 mmol/L) 4. Mu.L, primer 27F (10 mmol/L) 2. Mu.L, primer 1492R (10 mmol/L) 2. Mu. L, taq enzyme (5U/L) 0.25. Mu.L, 3. Mu.L DNA as template, ddH 2 O was made up to 50. Mu.L. PCR reaction procedure: 94 ℃ for 5min;94℃1min,52℃1min,72℃1min,30 cycles; and at 72℃for 10min. The PCR product was sent to the Shanghai worker for two-way sequencing and the sequence was analyzed by http:// eztax on-e.ezbiocloud.
The length of the obtained 16S rRNA fragment is 1286bp, and the sequence of the 16S rRNA fragment is shown in sequence 1 in a sequence table. The comparison result of the Eztax on website shows that the endophytic bacteria 237T2 has the highest similarity with Flavobacterium acidificum mode strains, is 99.45%, and is shown to be a type of 237T2 and Flavobacterium acidificum according to a phylogenetic tree constructed by the 16S rRNA gene sequence (figure 1), and the result shows that the endophytic bacteria 237T2 are Flavobacterium acidophilus (Flavobacterium acidificum).
Based on the above characteristics, endophytic bacterium 237T2 was identified as Flavobacterium acidophilus (Flavobacterium acidificum). Flavobacterium acidi (Flavobacterium acidificum) 237T2 was preserved in China general microbiological culture collection center (CGMCC) for 5 months and 25 days in 2020, and the registration number of the Flavobacterium acidi (Flavobacterium acidificum) in the China general microbiological culture collection center is CGMCC No.19869. Hereinafter, flavobacterium acidophilus 237T2.
EXAMPLE 2 Protoxalbumin secretion by Flavobacterium acidophilus (Flavobacterium acidificum) 237T2
According to the reference (Eric G lickm ann, yves Dessaux.A critical exam ination of the specificity of the salkowski reagent for indolic compounds produced by phytopathogentic bacteria [ J)]Applied and Environ mental Microbiology,1995 (2): 793-796) Salkowski colorimetry was used to determine the plant growth hormone (IAA) secreting properties of Flavobacterium acidophilus 237T2. Inoculating Flavobacterium acidophilus 237T2 into R 2 A liquid medium (Shanghai research industries Co., ltd.) was shake-cultured at 28℃with 180rpm for 4d to obtain 237T2 fermentation broth.
Qualitative detection was performed as follows: mu.L of 237T2 fermentation broth was taken with 50. Mu.L of Salkowski broth (Salkowski broth from 50mL of 35% HClO) 4 Solution and 1mL of 0.5M FeCl 3 The solution mixed liquid) was dropped on a white ceramic plate and left at room temperature in a dark place for 0.5 hours, and then observed to turn red, indicating that IAA was secreted. mu.L of a colorimetric solution of 50mg/L IAA (the colorimetric solution of 50mg/L IAA is a solution obtained by adding IAA to a Salkowski colorimetric solution to an IAA content of 50 mg/L) was added to 50. Mu.L of 237T2 fermentation broth as a positive control, and 50. Mu. L R was used as a positive control 2 A liquid medium was treated with 50. Mu.L of a colorimetric solution of 50mg/L IAA as a negative Control (CK). The experiment was repeated three times.
Qualitative detection showed that after the 237T2 fermentation dropwell was added with Salkowski colorimetric solution, the bacterial solution became red, indicating that 237T2 was able to secrete plant growth hormone (IAA) (fig. 3).
Quantitative detection: centrifuging 237T2 fermentation broth on a centrifuge at 12000rpm for 1 min, collecting 400ul of the supernatant, adding Salkowski colorimetric solution with equal volume, standing in dark for 30min, and measuring OD 530 Values. Standard curves were drawn using analytically pure IAA gradient dilutions. Simultaneously, the OD is measured on an ELISA reader in 800ul to 48 holes of 237T2 fermentation liquor 600 . The experiment was repeated three times.
Drawing IAA standard curve, mixing IAA solutions with different concentrations and Salkowski colorimetric solution in equal volume, standing for 30min at room temperature under dark condition, and measuring OD 530 Values, IAA at different concentrations were configured as follows:
IAA concentration quantification: 250mg/L:0.025g IAA+100ml absolute ethanol
200mg/L:8.0ml 250mg/L IAA+2.0ml absolute ethanol
150mg/L:6.0ml 250mg/L IAA+4.0ml absolute ethanol
100mg/L:4.0ml 250mg/L IAA+6.0ml absolute ethanol
50mg/L:2.0ml 250mg/L IAA+8.0ml absolute ethanol
35mg/L:1.4ml 250mg/L IAA+8.6ml absolute ethanol
30mg/L:1.2ml 250mg/L IAA+8.8ml absolute ethanol
25mg/L:1.0ml 250mg/L IAA+9.0ml absolute ethanol
0.0mg/L:10.0ml of absolute ethanol.
OD of 237T2 supernatant and Salkowski colourimetric liquid was measured 530 Value sum 237T2 fermentation broth OD 600 The quantitative result (Table 2) was obtained by conversion from the standard curve equation, and the IAA content in the 237T2 fermentation broth was 276.3978mg/L.
TABLE 2 quantitative determination of IAA secretion by Flavobacterium acidophilus 237T2
| OD530 | OD600 | OD530/OD600 | IAA concentration (mg/L) |
| 0.7105 | 0.2618 | 2.7139 | 276.3978 |
Example 3 preparation of microbial agent for improving saline-alkali soil
The activated Flavobacterium acidophilus 237T2 is inoculated into a 500mL triangular flask filled with 100mL of fermentation medium, and is subjected to shaking culture at 37 ℃ and 180rpm for 48 hours to obtain a fermentation broth of Flavobacterium acidophilus 237T2, wherein the content of the Flavobacterium acidophilus 237T2 in the fermentation broth of Flavobacterium acidophilus 237T2 is 10 9 cfu/ml. Wherein, the fermentation medium is a neutral TSB medium, and the preparation method comprises the following steps: according to the instruction of commercial culture medium, 30g of TSB (BD company) dry powder is taken, distilled water is added to a volume of 1000ml, then NaOH is used for adjusting the pH value of the dry powder to 7.0, and the liquid obtained by sterilizing at 121 ℃ for 20 minutes is the neutral TSB culture medium.
Uniformly mixing turfy soil (carrier) and the Flavobacterium acidophilus 237T2 fermentation liquor to obtain the microbial agent for improving the saline-alkali soil. The bacterial content of Flavobacterium acidophilum 237T2 in the bacterial agent for improving saline-alkali soil is 5 multiplied by 10 8 cfu/g。
EXAMPLE 4 laboratory test for reducing the pH of saline-alkaline soil Using Flavobacterium acidophilum 237T2
Inoculating Flavobacterium acidophilus 237T2 in neutral TSB culture medium (same as in example 2), shake culturing at 30deg.C and 180rpm (rotation radius 20 mm) for 48 hr to obtain Flavobacterium acidophilus 237T2 fermentation broth, wherein the Flavobacterium acidophilus 237T2 content in the Flavobacterium acidophilus 237T2 fermentation broth is 10 9 cfu/ml。
The fermentation broth of Flavobacterium acidophilus 237T2 was applied to a moderately saline-alkali soil having a pH of 8.45 (saline-alkali soil was taken from corn field in the artificial long irrigation area administration test station in Hechen of Hevea, inner Mongolia) on day 3, and the fermentation broth of Flavobacterium acidophilus 237T2 was applied to the soil at an amount of 2kg per mu, during which the pH changes were measured on days 12, 4, 6, 9, and 18, respectively, while the moderately saline-alkali soil having a pH of 8.45, to which the fermentation broth of Flavobacterium acidophilus 237T2 was not applied, was used as a Control (CK). The acid flavobacterium 237T2 has a remarkable effect on lowering the pH of soil, and the lowering effect is an instant effect, wherein the pH of the acid flavobacterium 237T2 fermentation broth is remarkably lowered by 0.34 unit (P < 0.05) compared with CK in the first day, and the pH lowering amplitude of the acid flavobacterium 237T2 fermentation broth is similar to that of the first day (12 months 4 days) in days 3 to 15 (i.e., 12 months 6 to 18 days) (fig. 2).
Example 5 field test for improving saline-alkali soil Using Flavobacterium acidicum 237T2 for crop growth
1. Test site
The field plot test is arranged in the field of the inner Mongolia Wuyuan county, and the field soil is silt loam. Before fertilization, a five-point sampling method is adopted, 100g of soil (hereinafter referred to as initial soil) is taken at each point and brought back to a laboratory, and each greenhouse soil is subjected to salinity and pH detection and nutrient content detection (the nutrient content detection method is that 0-40cm layers of soil samples are taken after the sunflower harvest and brought back to the laboratory for natural air drying, crushing and sieving, and then soil organic matters, quick-acting nitrogen (alkaline hydrolysis method) and quick-acting potassium (ammonium acetate extraction) are measured, and specific measurement methods are shown in soil agrochemicals analysis (Bao Shidan, 2005). A soil salinity and pH measurement method is that the sampled soil is brought back to the laboratory for natural air drying, ground and sieved by a 2mm sieve, a soil solution supernatant is extracted with a soil water ratio of 1:5, and the soil conductivity is measured by a conductivity meter (DDS-307), and the conductivity is converted into the soil salinity content (g.kg-1) =soil salinity (g.kg-1) ×0.064X10/1000 according to an empirical formula, and the detection result is shown in Table 3.
TABLE 3 physicochemical Properties of the initial soil
2. Experimental design and implementation
A field micro-area test was performed in the county of pentayuan, inner mongolia, 5-10 months in 2019. And 2 treatment areas are randomly arranged by adopting a random granule design, and three repetitions are arranged in each treatment area. The 2 treatment areas are respectively a non-microbial agent treatment area (CK) and a microbial agent treatment area (JJ).
The study area was divided into 6 cells, each cell having an area of 1.8m x 1.8m. And distributes the processes in a random drawing manner. The above 2 treatment zones were the same except for whether or not to inoculate Flavobacterium acidophilus 237T 2.
Edible sunflower JK601 is sown in 2019, 5 and 26 days, and the base fertilizer is applied by referring to a local method when the sunflower is sown in both treatments, and the specific fertilization method is as follows: 0.067kg of organic silicon water-soluble slow-release fertilizer compound fertilizer is applied per square meter to serve as base fertilizer. After 18 days of 6 months 2019, seedling-stage topdressing is carried out on the two treatment areas, and the microbial inoculum treatment area is 600kg/hm 2 The microbial agent for improving a saline-alkali soil of example 3 was applied in an amount such that the bacterial cell content of Flavobacterium acidophilus 237T2 in the soil was 3X 10 10 cfu/m 2 The area treated without microbial inoculum was treated in accordance with 600kg/hm 2 Grass carbon (carrier in microbial agent for saline-alkali soil improvement of example 3); the application method is that ditching is carried out in a row close to the root of the plant by 5-10cm, the ditch depth is 10-15cm, and soil is used for covering after the application. Leaf photosynthesis (table 4), plant height and SPAD values (table 5) were measured during the test period for sunflower at the bud stage, soil salinity, pH and plant ion content (table 6), soil bacterial diversity (table 7).
(1) Effect of Flavobacterium acidophilus 237T2 on photosynthetic characteristics of saline-alkali soil
TABLE 4 influence of microbial inoculant for saline-alkali soil improvement on photosynthetic property of saline-alkali soil
Note that: the different lower case letters following the same column of data represent significant differences (P < 0.05).
As can be seen from table 4, the net photosynthetic rate, stomatal conductance, transpiration rate, and leaf moisture utilization efficiency and SPAD of the microbial inoculant treated areas were increased by 52.90%, 46.15%, 33.33%, 14.51% and 12.99%, respectively, as compared to the non-inoculant treated areas; while the microbial agent treatment area pair reduces the intercellular CO 2 The concentration has a remarkable effect, and the concentration is reduced by 20.29% compared with that of a treatment area without applying the microbial inoculum. It is demonstrated that Flavobacterium acidophilus 237T2 can enhance photosynthesis performance of plants in saline-alkali soil.
The planting area of the inner mongolian sunflower accounts for about 30% of the total area of the national sunflower, and the total yield accounts for about 39% of the total yield of the national sunflower. In recent years, sunflower planting has become the mainstay industry for local agriculture and is one of the main economic sources for local farmers to expand reproduction. Photosynthesis is a very complex biological redox process, and can convert the surrounding simple inorganic substances into organic substances for self-use, which are important influencing factors in the process of dry matter accumulation and yield formation of crops. The organic matters formed by photosynthesis account for about 95% of the total dry matters of plants. Net photosynthetic rate, stomatal conductance and intercellular CO 2 The concentration and other indexes can measure the photosynthetic performance of plants. Soil salinization can influence respiration, photosynthesis and the like of plants, so that the soil salinization is one of important factors restricting the sustainable development of crops and agriculture in river irrigation areas. The research on the photo-biological characteristics of sunflower in the saline-alkali soil has great significance for improving the productivity.
(2) Influence of Flavobacterium acidicum 237T2 on plant height and aboveground biological quality and yield of saline-alkali soil
TABLE 5 influence of microbial inoculant for saline-alkali soil improvement on plant height, aboveground biomass and yield of saline-alkali soil
| Treatment of | Height of plant (cm) | Dry weight of overground biomass (g) | Yield (kg. Hm) -2 ) |
| CK | 93.05a | 125.85a | 2050a |
| JJ | 107.33b | 186.20b | 2400b |
Note that: the different lower case letters following the same column of data represent significant differences (P < 0.05).
As shown in Table 5, the plant height of the microbial inoculum treatment area is obviously increased by 15.35% compared with CK, the biological quality of the overground part is obviously increased by 47.95% compared with CK, and the yield is obviously increased by 17.07% compared with CK. It is demonstrated that Flavobacterium acidophilus 237T2 can promote plant growth and increase plant yield in saline-alkali soil.
(3) Influence of Flavobacterium acidophilus 237T2 on pH of saline-alkali soil and ion content of plants
TABLE 6 microbial inoculant for saline-alkali soil improvement for pH and plant K of saline-alkali soil + 、Na + And Ca 2+ Influence of ion content
| Treatment of | pH | K + (g·kg -1 ) | Na + (g·kg -1 ) | Ca 2+ (g·kg -1 ) | Na + /K + | Na + /Ca 2+ |
| CK | 8.45a | 29.55a | 12.84a | 3.43a | 0.43a | 3.74a |
| JJ | 8.21b | 36.33b | 10.45b | 4.76b | 0.29b | 2.19b |
Note that: the different lower case letters following the same column of data represent significant differences (P < 0.05).
As shown in table 6, the microbial agent treatment area can significantly reduce the pH value of soil by 0.24 units compared with CK; the micro-scale for improving the saline-alkali soil of example 3 was appliedThe biological bacterial agents can increase the K of plants to different degrees + And Ca 2+ Content of Na is reduced + Content of K in + The content is increased by 22.94% compared with CK; ca (Ca) 2+ The content is increased by 38.78 percent compared with CK; while Na is + The content is reduced by 22.87% compared with CK.
The method shows that the flavobacterium acidophilus 237T2 can obviously reduce the pH value of the saline-alkali soil and can obviously increase the K of plants + And Ca + Content of Na is obviously reduced + The content is as follows. Flavobacterium acidophilus 237T2 reduces plant pair Na + And the saline-alkali resistance of crops is improved.
(4) Effect of Flavobacterium acidicum 237T2 on bacterial diversity in saline-alkali soil
TABLE 7 influence of microbial inoculant for saline-alkali soil improvement on bacterial diversity of saline-alkali soil
| Treatment of | Shannon index | Simpson index | Chao1 index | ACE index |
| CK | 5.078±0.306a | 0.021±0.006a | 2749.026±144.236a | 2745.531±134.382a |
| JJ | 5.544±0.497a | 0.070±0.046a | 2888.423±186.830a | 2828.206±159.916a |
Note that: the different lower case letters following the same column of data represent significant differences (P < 0.05).
As shown in Table 7, the bacterial diversity under 237T2 treatment was improved as compared with CK, but the effect was not remarkable, but the aromatic concentration index, the Simpson index, the Chao1 index and the ACE index were improved by 9.18%, 233.33%, 5.07% and 3.01% as compared with CK, respectively.
The diversity of soil microorganisms is the most important control factor for the stability of the soil ecosystem and is also an important index for measuring the balance of the soil ecosystem. The aromatic index and the simpson index are indices used to estimate the microbial diversity in a sample, wherein a greater aromatic index value indicates a greater microbial community diversity in the sample, and the simpson index is the opposite; ACE index and Chao1 index are indices used to estimate the abundance of a bacterial population, and the larger the value, the higher the abundance.
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
Sequence listing
<110> institute of agricultural resource and agricultural division of national academy of agricultural sciences
<120> A Flavobacterium acidophilum and application thereof in improving saline-alkali soil
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1286
<212> DNA
<213> Flavobacterium acidophilus (Flavobacterium acidificum)
<400> 1
cggtgtgtac aaggcccggg aacgtattca ccgtggcatt ctgatccacg attactagcg 60
attccgactt cacggagtcg agttgcagac tccgatccgg actacgacgc actttatgag 120
gtccgcttgc tctcgcgagg tcgcttctct ttgtatgcgc cattgtagca cgtgtgtagc 180
cctactcgta agggccatga tgacttgacg tcatccccac cttcctccgg tttatcaccg 240
gcagtctcct ttgagttccc gaccgaatcg ctggcaacaa aggataaggg ttgcgctcgt 300
tgcgggactt aacccaacat ttcacaacac gagctgacga cagccatgca gcacctgtct 360
cacggttccc gaaggcacta aggcatctct gccgaattcc gtggatgtca agagtaggta 420
aggttcttcg cgttgcatcg aattaaacca catgctccac cgcttgtgcg ggcccccgtc 480
aattcatttg agttttaacc ttgcggccgt actccccagg cggtcgactt aacgcgttag 540
ctccggaagc cactcctcaa gggaacaacc ctccaagtcg acatcgttta cggcgtggac 600
taccagggta tctaatcctg tttgctcccc acgctttcgc accctgagcg tcagtcttcg 660
tccagggggc cgccttcgcc accggtattc ctccagatct ctacgcattt caccgctaca 720
cctggaattc tacccccctc tacgagactc aagcctgcca gtttcaaatg cagttcccag 780
gttaagcccg gggatttcac atctgactta acagaccgcc tgcgtgcgct ttacgcccag 840
taattccgat taacgcttgc accctccgta ttaccgcggc tgctggcacg gagttagccg 900
gtgcttcttc tgcgggtaac gtcaatcagt aaggttatta acctcattgc cttcctcccc 960
gctgaaagta ctttacaacc cgaaggcctt cttcatacac gcggcatggc tgcatcaggc 1020
ttgcgcccat tgtgcaatat tccccactgc tgcctcccgt aggagtctgg accgtgtctc 1080
agttccagtg tggctggtca tcctctcaga ccagctaggg atcgtcgcct aggtgggccg 1140
ttaccccgcc tactagctaa tcccatctgg gttcatccga tagtgagagg cccgaaggtc 1200
cccctctttg gtcttgcgac gttatgcggt attagccacc gtttccagtg gttatccccc 1260
tctatcgggc agatccccag acatta 1286
Claims (10)
1. The flavobacterium acidophilus is characterized in that: the Flavobacterium acidi is Flavobacterium acidi (Flavobacterium acidificum) 237T2, and the registration number of the Flavobacterium acidi in the China general microbiological culture Collection center is CGMCC No.19869.
2. The microbial inoculum is characterized in that: the microbial inoculum contains the flavobacterium acidophilus according to claim 1.
3. A probiotic comprising the microbial agent of claim 2.
4. A biofertilizer comprising the microbial agent of claim 2.
5. The culture of Flavobacterium acidicum of claim 1, which is obtained by culturing the Flavobacterium acidicum of claim 1 in a microorganism culture medium.
6. Use of flavobacterium acidophilus according to claim 1, in any of the following:
u1, application in improving saline-alkali soil or alkaline soil or in preparing products for improving saline-alkali soil or alkaline soil,
u2, the application in reducing the pH value of the saline-alkali soil or alkaline soil or the application in preparing a product for reducing the pH value of the saline-alkali soil or alkaline soil,
U3, in improving the salt alkali resistance or alkali resistance of plants or in preparing products for improving the salt alkali resistance or alkali resistance of plants,
u4, na in reducing the saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
u5, K in improving the saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of K for improving saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
u6, ca in improving the saline-alkali soil or alkaline soil of plants 2+ Application in absorption or preparation of Ca in improving saline-alkali soil or alkaline soil of plants 2+ The application in the field of absorption is that,
u7, na in reducing the saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
u8, na in reducing the saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil or alkaline soil by plants 2+ Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil by plants 2+ The use of the absorbent article in the manufacture of an absorbent article,
u9, the application in improving the photosynthetic performance of plants or the application in preparing products for improving the photosynthetic performance of plants,
u10, reduction of plant intercellular CO 2 Use in concentration or in the preparation of reduced plant intercellular CO 2 The application of the product with the concentration is that,
u11, the application in improving the net photosynthetic rate of plants or the application in preparing products for improving the net photosynthetic rate of plants,
u12, in the application of improving plant stomata conductivity or in the preparation of products for improving plant stomata conductivity,
u13, application in improving plant transpiration rate or in preparing product for improving plant transpiration rate,
u14, in the application of improving the water utilization efficiency of plant leaves or in the preparation of products for improving the water utilization efficiency of plant leaves,
u15, in the application of improving the chlorophyll content of plants or in the preparation of products for improving the chlorophyll content of plants,
u16, use in the production of auxins or in the preparation of auxin-producing products,
u17, application in promoting plant growth or application in preparing product for promoting plant growth,
u18, in the improvement of crop yield or in the preparation of products for improving crop yield,
U19, use in increasing plant biomass or use in preparing a product for increasing plant biomass.
7. Use of the culture of claim 5 for any of the following:
u1, application in improving saline-alkali soil or alkaline soil or in preparing products for improving saline-alkali soil or alkaline soil,
u2, the application in reducing the pH value of the saline-alkali soil or alkaline soil or the application in preparing a product for reducing the pH value of the saline-alkali soil or alkaline soil,
u3, in improving the salt alkali resistance or alkali resistance of plants or in preparing products for improving the salt alkali resistance or alkali resistance of plants,
u4, na in reducing the saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
u5, K in improving the saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of K for improving saline-alkali soil or alkaline soil of plants + In absorbent productsThe application of the method is that,
u6, ca in improving the saline-alkali soil or alkaline soil of plants 2+ Application in absorption or preparation of Ca in improving saline-alkali soil or alkaline soil of plants 2+ The application in the field of absorption is that,
u7, na in reducing the saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
u8, na in reducing the saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil or alkaline soil by plants 2+ Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil by plants 2+ The use of the absorbent article in the manufacture of an absorbent article,
u9, the application in improving the photosynthetic performance of plants or the application in preparing products for improving the photosynthetic performance of plants,
u10, reduction of plant intercellular CO 2 Use in concentration or in the preparation of reduced plant intercellular CO 2 The application of the product with the concentration is that,
u11, the application in improving the net photosynthetic rate of plants or the application in preparing products for improving the net photosynthetic rate of plants,
u12, in the application of improving plant stomata conductivity or in the preparation of products for improving plant stomata conductivity,
u13, application in improving plant transpiration rate or in preparing product for improving plant transpiration rate,
u14, in the application of improving the water utilization efficiency of plant leaves or in the preparation of products for improving the water utilization efficiency of plant leaves,
U15, in the application of improving the chlorophyll content of plants or in the preparation of products for improving the chlorophyll content of plants,
u16, use in the production of auxins or in the preparation of auxin-producing products,
u17, application in promoting plant growth or application in preparing product for promoting plant growth,
u18, in the improvement of crop yield or in the preparation of products for improving crop yield,
u19, use in increasing plant biomass or use in preparing a product for increasing plant biomass.
8. The use of the microbial agent of claim 2 for any of the following:
c1, application in improving saline-alkali soil or alkaline soil or in preparing products for improving the saline-alkali soil or alkaline soil,
c2, the application in reducing the pH value of the saline-alkali soil or alkaline soil or the application in preparing a product for reducing the pH value of the saline-alkali soil or alkaline soil,
c3, application in improving salt and alkali resistance of plants or in preparing products for improving salt and alkali resistance of plants,
c4, na in the saline-alkali soil or alkaline soil of the reduced plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
c5, K in improving saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of K for improving saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
c6, ca in improving saline-alkali soil or alkaline soil of plants 2+ Application in absorption or preparation of Ca in improving saline-alkali soil or alkaline soil of plants 2+ The application in the field of absorption is that,
c7, na in the saline-alkali soil or alkaline soil of the reduced plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving K in saline-alkali soil or alkaline soil of plants + The use of the absorbent article in the manufacture of an absorbent article,
c8, in reducing the salt-alkali soil of plantsNa in soil or alkaline soil + Absorbing and improving Ca in saline-alkali soil or alkaline soil by plants 2+ Application in absorption or preparation of Na for reducing saline-alkali soil or alkaline soil of plants + Absorbing and improving Ca in saline-alkali soil or alkaline soil by plants 2+ The use of the absorbent article in the manufacture of an absorbent article,
c9, application in improving photosynthetic performance of plants or in preparing products for improving photosynthetic performance of plants,
c10 and reduction of plant intercellular CO 2 Use in concentration or in the preparation of reduced plant intercellular CO 2 The application of the product with the concentration is that,
c11, in the application of improving the net photosynthetic rate of plants or in the preparation of products for improving the net photosynthetic rate of plants,
c12, application in improving plant stomata conductivity or in preparing products for improving plant stomata conductivity,
c13, application in improving plant transpiration rate or application in preparing product for improving plant transpiration rate,
c14, application in improving the water utilization efficiency of the plant leaves or application in preparing products for improving the water utilization efficiency of the plant leaves,
c15, application in improving plant chlorophyll content or in preparing product for improving plant chlorophyll content,
c16, use in the production of auxins or in the preparation of auxin-producing products,
c17, application in promoting plant growth or application in preparing product for promoting plant growth,
c18, use in increasing crop yield or use in preparing a product for increasing crop yield,
c19, use in increasing plant biomass or use in preparing a product for increasing plant biomass.
9. The method of culturing the flavobacterium acidophilus of claim 1, comprising the step of culturing the flavobacterium acidophilus in a medium for culturing the microorganism.
10. A method for producing the microbial inoculum according to claim 2, comprising the step of obtaining the microbial inoculum by using the flavobacterium acidophilus according to claim 1 as a component of the microbial inoculum.
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| Application Number | Priority Date | Filing Date | Title |
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| CN107709275A (en) * | 2015-06-02 | 2018-02-16 | 科氏农艺服务有限责任公司 | Microbial bacterial agent composition and its purposes in agricultural |
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| CN107709275A (en) * | 2015-06-02 | 2018-02-16 | 科氏农艺服务有限责任公司 | Microbial bacterial agent composition and its purposes in agricultural |
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| Sahu,K.P.等.Pantoea ananatis strain OsEp_A& N_30A19 16S ribosomal RNA gene, partial sequence.Genbank：MT367861.1.2020,全文. * |
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