CN117229963B - Korean pseudomonas YBZ2 for preventing and treating walnut Jiao Shezheng and application thereof - Google Patents
Korean pseudomonas YBZ2 for preventing and treating walnut Jiao Shezheng and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of microorganisms, in particular to Korean pseudomonas YBZ2 for preventing and treating walnuts Jiao Shezheng and application thereof. Korean pseudomonas (Pseudomonas koreenis) YBZ2 for preventing and treating walnut Jiao Shezheng is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.27756. The Korean pseudomonas YBZ2 can effectively reduce the occurrence of walnut seedlings Jiao Shezheng.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to Korean pseudomonas YBZ2 for preventing and treating walnuts Jiao Shezheng and application thereof.
Background
Walnut (Juglans Regia l.) is also known as walnut, which is a stone fruit of Juglans (Juglans) belonging to the family Juglandaceae (Juglandaceae), belongs to a perennial temperate deciduous arbor, and has important edible, material, medicinal and ecological values. The walnut is used as one of the main cultivated tree species in the south Xinjiang forest fruit industry in the Xinjiang area, the cultivated area is up to 580 tens of thousands of mu, and the yield value are the first characteristic forest fruit in the Xinjiang.
The first report of walnut Jiao Shezheng in 2012 occurs in the main production area of juglans regia in south China, which has been 10 years ago. The walnut Jiao Shezheng is stressed by unfavorable environmental factors such as saline alkali in soil, hardening, high temperature, dry hot air, drought, saline alkali in irrigation well water and the like, and causes physiological diseases together with the weakening of tree vigor. After the walnut is damaged by salt, the leaf is dehydrated and wilted, and yellow brown scorched spots appear; salt stress with high concentration for a long time causes the walnut leaves to become black and fall off gradually, and the plant death can be caused when the salt stress is serious. The phenomenon of damage to walnuts caused by salt stress is found to be very similar to the phenotype of the plants when walnuts Jiao Shezheng occur through indoor replay, which indicates that salt stress is a factor for inducing the occurrence of walnuts Jiao Shezheng. The long-term high-concentration salt stress not only affects the quality and yield of the walnut fruits, but also causes huge economic loss. If effective measures are not taken in time, the healthy development of the walnut industry in Xinjiang areas is seriously threatened.
At present, the traditional method for improving and utilizing the saline soil in Xinjiang comprises the following steps: drainage (salt washing of rice), irrigation (reasonable irrigation and seepage prevention of channels), leveling (land leveling), fertilizer (grass rotation, green manure planting and farmyard manure, gypsum and the like) and forests (building of a forest belt). The implementation of the method needs to consume a great deal of manpower and resources, and has high control cost and slow effect. It is difficult to solve the current situation that the walnut grows in a high-salt environment for a long time. The orchard in which the walnuts Jiao Shezheng occur mainly adopts measures of reducing the groundwater level, balancing and applying N, P, K, B, zn, mn, fe and other fertilizers, adding potassium sulfate fertilizer, adding Shi Fushi acid organic fertilizer, spraying physiological regulator on the leaf surfaces, spraying anti-transpiration agent on the leaf surfaces and the like, and the occurrence of the walnuts Jiao Shezheng can be effectively prevented and controlled, but the implementation of the method still faces higher economic cost and large input of human resources.
Plant rhizosphere growth promoting bacteria (Plant Growth Promoting Rhizobacteria, PGPR) are a type of microorganism living in soil or attached to plant roots to promote plant growth and development in a direct or indirect manner and relieve environmental stress to plants. PGPR has been shown to enhance plant salt tolerance by modulating hormonal changes in plants, inducing plants to produce anti-reactive oxidases and osmoprotectant, modulating ionic balance in plants, improving photosynthetic rate changes in plants, and increasing plant water and fertilizer utilization by secreting extracellular polysaccharides. PGPR having a function of enhancing salt resistance of plants is mainly distributed in the genus Bacillus, pseudomonas, enterobacter, azospirillum and the like. The method has the advantages of low cost, no pollution and wide attention, and is used for excavating high-efficiency salt-tolerant PGPR strain, developing and applying salt-tolerant PGPR preparation to reduce the occurrence of walnut Jiao Shezheng caused by salty soil.
In view of this, the present invention has been made.
Disclosure of Invention
The invention aims to provide Korean pseudomonas YBZ for preventing and controlling walnut Jiao Shezheng and application thereof, wherein Korean pseudomonas (Pseudomonas koreenis) YBZ2 is obtained by separating and screening the rhizosphere of walnut trees, and walnut Jiao Shezheng can be effectively prevented and controlled.
In order to achieve the above object of the present invention, the following technical solutions are specifically adopted:
in one aspect, the present invention relates to a strain of Pseudomonas korea (Pseudomonas koreenis) YBZ deposited in China general microbiological culture Collection center, accession number address: the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms, no. 3, north Chen Xilu No.1, north Chaoyang, beijing, has a collection number of CGMCC No.27756 and a collection date of 2023, 6, 30 days.
The invention is used for effectively preventing and controlling the occurrence of the walnut Jiao Shezheng in Xinjiang from the perspective of improving the salt tolerance of the walnut. Screening salt-tolerant bacteria by combining a flat plate culture technology with living seedling culture, separating from the rhizosphere of walnut trees with the occurrence of the severe Jiao Shezheng to obtain salt-tolerant bacteria, inoculating the salt-tolerant bacteria to the root of the walnut under the greenhouse condition, and finally screening a bacterial strain-Pseudomonas koraiensis YBZ which is used for effectively preventing and controlling the walnut leaves Jiao Shezheng and promoting the growth of the root of the walnut by observing the reduction degree of the upper coke She Xianxiang of the walnut leaves.
According to the method, the experimental method of screening seedlings living bodies is combined under the greenhouse condition by utilizing the flat-plate culture technology, indexes such as the severity of damage to leaves, the content of malondialdehyde in the leaves and the activity of plant root systems are measured, and the effects of salt-tolerant bacteria on prevention and control of the occurrence of the walnut Jiao Shezheng and salt tolerance promotion of the walnut seedlings are reasonably evaluated, so that the method is an efficient and simple screening mode.
According to the invention, a greenhouse experiment is carried out by using walnut seedlings, when the seedlings grow to VI-phase seedlings, bacterial suspension of the strain YBZ is inoculated, when the seedlings grow to VIII-phase seedlings, salt treatment is started, and after the salt treatment is finished, obvious difference between the growth and development conditions of the walnut seedlings in the NaCl control group and the NaCl+ YBZ strain treatment group can be observed.
Another aspect of the present invention relates to a microbial preparation comprising the cells of Pseudomonas koraiensis (Pseudomonas koreenis) YBZ2 and a bacterial suspension thereof.
The microbial preparation provided by the invention belongs to an environment-friendly microbial preparation, and has the advantages of quick response, low cost, no pollution and sustainable use. The beneficial microorganism preparation is applied to plants per se or the surrounding environment, so that microorganisms can directly or indirectly participate in soil nutrient circulation, promote the plants to absorb nutrients and enhance the capability of the plants to resist abiotic stress.
Preferably, the microbial preparation comprises: solid or liquid formulations.
Preferably, when the microbial preparation is a solid preparation, the microbial preparation contains the living bacteria of Pseudomonas koraiensis (Pseudomonas koreenis) YBZ2 in an amount of 10 8 CFU/g or more.
Preferably, when the microbial preparation is a liquid preparation, the microbial preparation contains the living bacteria of Pseudomonas koraiensis (Pseudomonas koreenis) YBZ2 in an amount of 10 8 CFU/mL or more.
Another aspect of the present invention relates to a method for controlling walnuts Jiao Shezheng, which comprises applying the thalli of pseudomonas koraiensis (Pseudomonas koreenis) YBZ2 and/or the microbial preparation to walnuts.
Another aspect of the present invention relates to a method for promoting the growth of walnuts, which comprises administering the thalli of pseudomonas koraiensis (Pseudomonas koreenis) YBZ2 and/or the microbial preparation to walnuts.
Preferably, korean Pseudomonas (Pseudomonas koreenis) YBZ2 is applied to walnut by root irrigation.
Another aspect of the present invention relates to the use of Pseudomonas koraiensis (Pseudomonas koreenis) YBZ2 or the microbial preparation in controlling plant diseases and/or plant growth promotion.
Preferably, the plant disease comprises: diseases caused by salt stress.
Preferably, the disease caused by salt stress comprises: and Jiao Shezheng of walnuts.
Preferably, the plant growth promoting object comprises: walnut.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention screens 397 strain separated from the root of walnut tree with the weight of Jiao Shezheng by flat streak, and obtains Korean pseudomonas (Pseudomonas koreenis) YBZ2 which is walnut rhizosphere symbiotic bacteria after screening.
(2) The area of the scorched leaves of walnut seedlings in a high-salt environment can be obviously reduced after the strain YBZ is inoculated, the occurrence of walnut Jiao Shezheng is slowed down, and the severity of damage to the leaves of the walnut seedlings is reduced; after the strain YBZ is inoculated, the fresh weight of the main root and the root system activity of the walnut seedlings are obviously increased, and the growth promotion effect on the root of the seedlings is realized; after the strain YBZ is inoculated, the content of malondialdehyde in walnut leaves is obviously reduced, and the strain YBZ is verified from the plant physiological point of view to be capable of preventing and controlling the occurrence of walnut seedlings Jiao Shezheng.
(3) The microbial preparation provided by the invention belongs to an environment-friendly microbial preparation, has the effect of promoting the growth of plants, and can effectively improve the control effect of the plants on abiotic stress, especially Jiao Shezheng, so as to improve the adversity viability of the plants.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the morphology of walnut seedling plants in different growth periods;
FIG. 2 is a leaf symptom classification of greenhouse walnut seedlings Jiao Shezheng;
FIG. 3 is a graph showing relief of symptoms of walnut seedlings Jiao Shezheng by strain YBZ 2;
FIG. 4 is a graph of strain YBZ2 for mitigation of leaf damage in focal She Zheng walnut seedlings;
FIG. 5 is a promotion of growth of the root system of the focal She Zheng walnut seedlings by strain YBZ;
FIG. 6 shows the reduction of malondialdehyde content of the focused She Zheng walnut seedlings by strain YBZ;
FIG. 7 shows the improvement of the root system activity of the walnut seedlings of the focusing She Zheng by the strain YBZ;
FIG. 8 shows colony morphology of Pseudomonas aeruginosa YBZ2 (scale bar: 2 mm).
Detailed Description
The technical solution of the present invention will be clearly and completely described below with reference to the accompanying drawings and detailed description, but it will be understood by those skilled in the art that the examples described below are some, but not all, examples of the present invention, and are intended to be illustrative of the present invention only and should not be construed as limiting the scope of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1 sample treatment and Strain isolation
The test strain was isolated from the walnut root system and surrounding soil of severe Jiao Shezheng occurring in the acksu region of Uygur autonomous region in Xinjiang, 4 months in 2021. During collection, firstly removing fallen leaf layers and surface soil, taking out walnut roots along the growth direction of the walnut roots, respectively collecting soil around the walnut roots, soil on the root system surface and root system samples, taking the soil and the root system samples back to a laboratory, and separating and screening symbiotic bacteria of the walnut roots.
(1) Transferring 0.25g of walnut root peripheral soil into a 15mL centrifuge tube, adding 10mL of 1 XPBS solution to prepare a sample stock solution, and performing 10-time gradient dilution to 10 -9 The method comprises the steps of carrying out a first treatment on the surface of the And (3) carrying out vortex vibration on the diluted sample solution, uniformly mixing, sucking 20 mu L of bacterial liquid, uniformly dripping the bacterial liquid into a flat-plate culture medium, and uniformly coating by using a coater. The sample solutions of each gradient were applied to a lysis broth (LB medium: 10g/L of sodium chloride, 5g/L of yeast extract, 10g/L of tryptone, pH 7.0), an R2A medium (yeast extract 0.5g/L, peptone 0.5g/L, casein hydrolysate 0.5g/L, glucose 0.5g/L, soluble starch 0.5g/L, potassium dihydrogen phosphate 0.5g/L, sodium pyruvate 0.3g/L, anhydrous magnesium sulfate 0.025g/L, pH 7.0-7.4), a 1/10 tryptone agar medium (1/10 TSA medium: tryptone 1.5g/L, pH 7.1-7.5), a tryptone glucose medium (TYG medium: 3.0g/L, yeast extract 3.0g/L, potassium dihydrogen phosphate 0.03g, pH 0.5g, sodium dihydrogen phosphate 0.03g, pH 0.5g, pH 0.03g/L, sodium dihydrogen phosphate 0.5g, pH 0.5 g/L). Culturing at 30deg.C overnight; the colony morphology was observed the next day, and the monoclonal streaks with different shapes and morphologies were picked and purified until no bacteria grew out. The monoclonals were picked up into 5mL LB liquid medium and cultured at 28℃and 160rpm for 12-14h. When the strain is in logarithmic growth phase, 1mL is sucked into a bacteria-preserving tube, 200 mu L of 87% glycerol is added, and the mixture is uniformly mixed by vortex and stored in a refrigerator at-80 ℃ for a long time.
(2) About 20 sections of root system with the length of 2cm and the diameter of about 2mm are cut by a sterile scalpel and transferred into a 1.5mL centrifuge tube. 1mL of 1 XPBS solution and 3-5 sterile glass beads are added into a centrifuge tube, and vortex vibration is carried out to enable soil samples attached to a root system to fall down until the surface of the root system is in a clean state. By using nothingThe root and glass beads were removed from the forceps, the solution was centrifuged at 13000rpm for 15min at 4℃and the supernatant was discarded to give a precipitate. 1mL of 1 XPBS solution was added to the centrifuge tube to prepare a stock sample solution, which was diluted 10-fold to 10 -9 . The step after dilution is the same as (1).
(3) Transferring the root system after shaking and cleaning in the step (2) into a sterile culture dish, and adding 20mL of 1% sodium hypochlorite for sterilization; after complete soaking for 1min, the root system was transferred to a petri dish containing sterile water and washed with shaking (this step was repeated at least 3 times, ensuring removal of residual sodium hypochlorite from the root system surface). Naturally air-drying the cleaned root system under aseptic condition, transferring into an aseptic mortar, adding a proper amount of 1 XPBS solution, and fully grinding to a homogenate state. Transferring the homogenate into a 15mL centrifuge tube, fixing the volume of 1 XPBS solution to 10mL to prepare a sample stock solution, and performing 10-time gradient dilution to 10 -9 . The step after dilution is the same as (1).
EXAMPLE 2 salt tolerant bacterial screening
Bacteria were streaked onto LB plates and the monoclonal was picked into 5mL LB liquid medium. Culturing at 30deg.C and 200rpm overnight, and regulating bacterial liquid concentration OD 600 1.0. A small amount of bacterial liquid is dipped and cultivated on an LB plate containing NaCl (the content of NaCl added into each liter of culture medium is 30g, hereinafter referred to as 3% NaCl-LB plate), and after 24 hours of cultivation at 30 ℃, whether normal growth can be observed.
EXAMPLE 3 preparation of bacterial suspension
The strain of example 2 capable of growing normally on a 3% NaCl-LB plate was picked up and cultured overnight at 30℃and 200rpm in 10mL of LB liquid medium; the next day, 10mL of bacterial liquid is transferred into a triangular flask containing 90mL of LB liquid medium, and is cultured at 30 ℃ and 200rpm overnight; on the third day, 100mL of the bacterial liquid was centrifuged at 8000rpm for 3min, and the supernatant was discarded to retain bacterial pellet. Resuspension of the bacterial pellet with sterile water and adjustment of OD 600 The value was calculated to give a final concentration of 10 at 100mL 9 CFU/mL of the bacterial suspension was diluted with 900mL of sterile distilled water to a final concentration of 1L of 10 8 CFU/mL of bacterial suspension.
Example 4 salt tolerance and growth promotion experiment of walnut seedling greenhouse
Walnut seedlings began to germinate to stage III seedlings (see fig. 1) for about 20 days. 200mL of clear water is poured every three days in the period, and the humidity in the basin is kept proper. When seedlings grow to stage IV seedlings, 200mL of 25% Hoagland's nutrient solution is poured once every five days. When the nutrient solution is treated until the seedlings grow to the VI-stage seedlings, about 30 days later, the nutrient solution can be stopped from being poured, salt-tolerant bacteria can be inoculated, 200mL of the bacterial suspension of the embodiment 3 is inoculated in each pot in a root-pouring mode every 5 days, and bacteria are inoculated for 3 times; during this period, the NaCl treated group and the fresh water control group were required to be poured with an equal amount of fresh water. After the inoculation of the strain YBZ2 bacterial suspension is finished, when seedlings grow to VIII-stage seedlings, the NaCl treatment group and the NaCl+strain YBZ2 treatment group begin to carry out salt treatment, 200mL of 125mM NaCl solution is poured every 5 days, 8 times of salt solution is required to be poured, and the clear water control group still is poured with equal amount of clear water.
EXAMPLE 5 statistics of blade Damage
The main harm of NaCl to walnut seedlings is reflected in the occurrence of walnut scorched leaves, namely the leaves are obviously damaged in a yellow brown scorched state. When setting up the damage classification, the damage area appearing on the seedling leaves is used as the basis of the classification. The score was rated 0-4 according to the lesion area as shown in fig. 2 (a). And counting the number of the seedling leaves with different damage levels by taking each basin as a unit, and calculating the damage severity of the seedling leaves. Blade classification criteria are shown in table 1.
Table 1 criteria for classifying leaf symptoms of walnut seedlings Jiao Shezheng
The severity of damage to leaf symptoms of walnut seedlings Jiao Shezheng after NaCl treatment was calculated according to the following formula.
Severity of injury = 100×Σ (number of damaged leaves at each stage x representative value at each stage)/(total leaves investigated x representative value at highest stage).
EXAMPLE 6 determination of malondialdehyde content
1. And taking walnut seedling leaves of different concentration treatment groups, quickly flushing the leaves by sterile water, and quickly wiping the leaves by filter paper. The same parts of each group of blades were taken, and 0.3g of each group was repeatedly weighed.
2. Extracting solution: placing 0.3g of sample into a centrifugal tube cooled in advance, adding 3mL of 1% trichloroacetic acid, and quickly crushing to homogenate; the homogenate was transferred to a 50mL centrifuge tube, the centrifuge tube was rinsed with 2mL of 1% trichloroacetic acid, and the resulting liquid was transferred to a new centrifuge tube.
3. Determination of the samples: adding 5mL of 0.5% thiobarbituric acid into a centrifuge tube, fully and uniformly mixing by a vortex oscillator, and carrying out boiling water bath for 10min; centrifuging at 5000rpm for 10min at 4 ℃ after rapid cooling; OD values at wavelengths of 450nm, 532nm and 600nm are measured respectively, and are substituted into a formula to calculate the content of malondialdehyde.
Malondialdehyde concentration (μmol/L) =c=6.45× (a 532 -A 600 )-0.56×A 450 。
Malondialdehyde content (μmol/g) =y= (c×v)/W.
V was 5mL and W was 0.3g.
Example 7 determination of walnut root vigor
Plant root system activity was detected using the plant root system activity detection kit-TTC method (Solarbio).
Example 8 control Effect of Strain YBZ2 on walnut seedlings Jiao Shezheng
According to the invention, a bacterial strain YBZ2 for preventing and controlling walnut seedlings Jiao Shezheng is obtained by screening 397 bacteria separated from the rhizosphere of a severe Jiao Shezheng walnut tree by adopting salt flat-plate culture and walnut seedling measurement.
After the bacterial suspension of the strain YBZ is root-irrigated and inoculated to the root of the walnut seedling for three times, the walnut seedling is irrigated with NaCl solution (125 mM). The results showed that leaf scorch was significantly reduced in the salt-treated group inoculated with the strain YBZ bacterial suspension compared to the NaCl-treated group (see fig. 3); blade damage severity was reduced by 45% (see fig. 4); the fresh weight of the main root is remarkably increased by 5.57g under the condition of a greenhouse (see FIG. 5); the content of malondialdehyde in leaves was significantly reduced by 69.24% (see fig. 6); the root activity of seedlings was significantly increased to 1704.39. Mu.g/g.h, slightly decreased compared to the fresh water control group, but significantly increased to 609.72. Mu.g/g.h compared to the NaCl control group (see FIG. 7). The strain YBZ has good application prospect in prevention and control of walnut seedlings Jiao Shezheng.
Example 9 molecular biological identification and colony morphology of Strain YBZ2
(1) Amplification and identification of 16S rDNA of Strain YBZ2
Genomic DNA of strain YBZ was extracted as a template using universal primer 27F:5'-AGAGTTTGATCMTGGCTCAG-3' and 1492R:5'-TACGGYTACCTTGTTACGACTT-3', PCR amplified 16S rRNA gene fragment. The reaction system is shown in Table 2.
TABLE 2PCR reaction System
The PCR product was detected by 1% agarose gel electrophoresis and sent to Rui Bo XingKe Biotechnology Co., ltd for sequencing.
The 16S rDNA sequencing results are shown in SEQ ID No.1 below:
SEQ ID No.1：
AGCTTGCTCCTGGATTCAGCGGCGGACGGGTGAGTAATGCCTAGGAATCTGCCTGGTAGTGGGGGACAACGTTTCGAAAGGAACGCTAATACCGCATACGTCCTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGAAGGGTTGTAGATTAATACTCTGCAATTTTGACGTTACCGACAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGTAATACAGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTTGTTAAGTTGGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGGCAAGCTAGAGTATGGTAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGACTGATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGGAGCCTTGAGCTCTTAGTGGCGCAGCTAACGCATTAAGTTGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATCCAATGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACATTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACGTAATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCACCAGAAGTAGTAG。
the obtained sequence is subjected to BLAST search in the National Center for Biological Information (NCBI), and the sequence similarity with the 16S rRNA gene segment of the strain YBZ is Pseudomonas koreenis, wherein the similarity reaches 99.78%. Thus, the strain YBZ was identified as Pseudomonas korea, designated as Pseudomonas korea YBZ (Pseudomonas koreenis YBZ 2), and deposited with China general microbiological culture Collection center with the accession number CGMCC No.27756 for 2023, 6 months and 30 days.
(2) Colony morphology of strain YBZ2
After strain YBZ was inoculated onto LB solid medium and incubated at 30℃for 24 hours, the colonies were observed to be nearly circular, milky white, central microprotrusions, and the colony surfaces were moist and smooth and the edges were smooth (see FIG. 8).
While the invention has been illustrated and described with reference to specific embodiments, it is to be understood that the above embodiments are merely illustrative of the technical aspects of the invention and not restrictive thereof; those of ordinary skill in the art will appreciate that: modifications may be made to the technical solutions described in the foregoing embodiments, or equivalents may be substituted for some or all of the technical features thereof, without departing from the spirit and scope of the present invention; such modifications and substitutions do not depart from the spirit of the corresponding technical solutions; it is therefore intended to cover in the appended claims all such alternatives and modifications as fall within the scope of the invention.
Claims (8)
1. Korean pseudomonas strain for preventing and controlling walnut Jiao ShezhengPseudomonas koreenis) YBZ2 and is preserved in China general microbiological culture Collection center (CGMCC) No.27756.
2. A microbial preparation comprising the cells of pseudomonas koraiensis YBZ and a bacterial suspension thereof according to claim 1.
3. The microbial preparation according to claim 2, wherein the microbial preparation comprises: solid or liquid formulations.
4. The microbial preparation according to claim 3, wherein when the microbial preparation is a solid preparation, the number of viable bacteria of the microbial preparation containing the Pseudomonas aeruginosa YBZ2 is 10 8 CFU/g or more.
5. The microbial preparation according to claim 3, wherein when the microbial preparation is a liquid preparation, the number of viable bacteria of the microbial preparation containing the Pseudomonas aeruginosa YBZ2 is 10 8 CFU/mL or more.
6. A method for controlling walnuts Jiao Shezheng, characterized in that the thalli of pseudomonas koraiensis YBZ2 according to claim 1 and/or the microbial preparation according to any one of claims 2 to 5 are applied to walnuts.
7. A method for promoting the growth of walnuts, characterized in that the thalli of pseudomonas koraiensis YBZ according to claim 1 and/or the microbial preparation according to any one of claims 2 to 5 are applied to walnuts.
8. Use of the microorganism preparation of pseudomonas YBZ of claim 1 or any one of claims 2 to 5 for controlling plant diseases and/or promoting walnut growth;
the plant disease includes: diseases caused by salt stress;
the diseases caused by the salt stress comprise: and Jiao Shezheng of walnuts.
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