CN111778197A

CN111778197A - Methylobacterium strain with high growth promoting efficiency and application thereof

Info

Publication number: CN111778197A
Application number: CN202010783850.9A
Authority: CN
Inventors: 杨婷; 史发超; 安玉兴; 孙东磊; 卢颖林
Original assignee: Guangdong Institute of Bioengineering Guangzhou Cane Sugar Industry Research Institute
Current assignee: Guangdong Institute of Bioengineering Guangzhou Cane Sugar Industry Research Institute
Priority date: 2020-08-06
Filing date: 2020-08-06
Publication date: 2020-10-16

Abstract

The invention relates to a methylobacterium strain capable of promoting growth efficiently and application thereof. The methylobacterium has a deposit number of GDMCC No: 61052. the methylobacterium can secrete the plant auxin IAA, and can effectively promote the seed germination and the plant growth of plants.

Description

Methylobacterium strain with high growth promoting efficiency and application thereof

Technical Field

The invention relates to the field of microbiology, in particular to a methylobacterium strain capable of promoting growth efficiently and an application thereof.

Background

During the growth process, the plant interacts with various microorganisms, mainly including plant rhizosphere microorganisms, phyllospheric microorganisms and endophytes. These microorganisms can promote plant growth by direct or indirect means such as nitrogen fixation, phosphate solubilization, auxin secretion, increased plant resistance, and the like. In agricultural production, the beneficial microorganisms inoculated by external sources can promote the growth of plants and increase the crop yield, and meanwhile, the environmental friendliness and the sustainability of the action effect are advocated in modern agriculture with more and more importance on the ecological environment.

Methylobacterium (Methylobacterium) is a type of facultative aerobic bacteria that can utilize monocarbons including methanol, methylamine, etc. as carbon sources. Bacteria of the genus Methylobacterium are pink due to their ability to synthesize carotenoids, and are also called Pink-segmented Facultivative methylotrophics (PPFM). Methylobacterium is widely present in soil, air, water areas and other environments, and in plant phyllosphere and rhizosphere, has the functions of generating plant hormone, antagonizing plant pathogenic microorganisms, relieving abiotic stress and the like, and has great development and utilization values in agricultural production.

Disclosure of Invention

Based on the above, the invention aims to provide the methylobacterium YT2019B002, which can secrete plant auxin and can effectively promote the germination of plant seeds and the growth of plants.

The specific technical scheme is as follows:

a methylobacterium YT2019B002, wherein the deposited number of the methylobacterium YT2019B002 is GDMCC No. 61052.

The invention also aims to provide an application of the methylobacterium YT2019B002 in preparation of plant auxin.

The invention also aims to provide an application of the methylobacterium YT2019B002 in preparing a plant growth regulating preparation.

The invention also aims to provide application of the methylobacterium YT2019B002 in preparation of fertilizers for promoting plant growth.

The invention also aims to provide an application of the methylobacterium YT2019B002 in plant growth promotion.

In some of these embodiments, the promoting plant growth comprises promoting plant seed germination.

In some of these embodiments, the promoting plant growth comprises promoting plant growth.

Another object of the present invention is to provide a biological agent, the active ingredient of which comprises the above methylobacterium YT2019B 002.

Another objective of the present invention is to provide a method for culturing the methylobacterium YT2019B002, comprising the following steps: inoculating methylobacterium YT2019B002 into a culture medium, and culturing at 20-30 ℃; the culture medium is AMS culture medium.

Another object of the present invention is to provide a methylobacterium YT2019B002 culture prepared by culturing the methylobacterium YT2019B002 according to claim 1.

The Methylobacterium YT2019B002(Methylobacterium sp.) is preserved in the Guangdong province microbial strain preservation center (GDMCC, address: No. 59 building 5 of Michelia furiosu 100 of Guangzhou, Guangzhou province) specified by the national intellectual property office in 6.9.2020, and the preservation date is as follows: year 2020, 6/9, accession number: GDMCCNo: 61052.

Compared with the prior art, the invention has the following beneficial effects:

the invention discovers a new microorganism methylobacterium YT2019B002 belonging to the methylobacterium for the first time, which can produce auxin, has good promotion effect on seed germination and plant growth of plants, and has wide development and utilization values in agricultural production.

Drawings

FIG. 1 shows the shape and gram stain profile of the strain YT2019B002 on the medium (A: shape of the culture on AMS medium; B: gram stain);

FIG. 2 is a gel electrophoresis chart of the PCR amplification result of the specific primer (mxaF gene), (M: DL2000 Marker; 1, 2: YT2019B002 strain; 3, 4: negative control);

FIG. 3 is a phylogenetic tree constructed by the Neighbor-Joining (NJ) method based on the 16S rRNA sequence; the strain Escherichia coli 562(J01859) is an outlier,^T: a model strain;

FIG. 4 shows the effect of YT2019B002 on seed germination (A: CK treatment for 3D; B, C: CK treatment for 5D; D: YT2019B002 treatment for 3D; E, F: YT2019B002 treatment for 5D);

FIG. 5 shows the effect of different treatments on pepper seedling growth after transplantation 23D (A: CK 1; B: CK 2; C: YT2019B002 (G); D: YT2019B002 (P));

FIG. 6 is a graph showing the capability of strain YT2019B002 in producing IAA (a: IAA standard curve; B: fermentation supernatant color reaction after 7d culture, A: CK; B: YT2019B 002).

Detailed Description

Experimental procedures according to the invention, in which no particular conditions are specified in the following examples, are generally carried out under conventional conditions, or under conditions recommended by the manufacturer. The various chemicals used in the examples are commercially available.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

The terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusions. For example, a process, method, apparatus, article, or device that comprises a list of steps is not limited to only those steps or modules listed, but may alternatively include other steps not listed or inherent to such process, method, article, or device.

The "plurality" referred to in the present invention means two or more. "and/or" describes the association relationship of the associated objects, meaning that there may be three relationships, e.g., a and/or B, which may mean: a exists alone, A and B exist simultaneously, and B exists alone. The character "/" generally indicates that the former and latter associated objects are in an "or" relationship.

The present invention will be described in further detail with reference to specific examples.

The material and the method are as follows:

separating the material source: the separation material, namely, the sugarcane seedlings (Saccharum officinarum 'Badila') is collected from the east Yong town of south sand area of Guangzhou, Guangdong province, and after collection, the ice box is preserved at low temperature and brought back to a laboratory, and the ice box is preserved at low temperature of-20 ℃ for functional microorganism separation.

Solid AMS medium: according to the Methylobacillus bacteria obligate culture Medium in Whittenbury et al 1970, the main component comprises K₂HPO₄0.7g，KH₂PO₄0.54g，MgSO₄·7H₂O 1.0g，CaCI₂·2H₂O 0.2g，FeSO₄·7H₂O 4mg，NH₄Cl 0.5g，ZnSO₄·7H₂O 100.0mcg，MnCl₂·4H₂O 30.0mcg，H₃BO₃300.0mcg，CoCl₂·6H₂O 200.0mcg，CuCl₂·2H₂O 10.0mcg，NiCl₂·6H₂O 20.0mcg，Na₂MoO₄·2H₂O60.0 mcg, Agar 15.0g, distilled water 1L, pH 6.8, sterilizing at 121 deg.C for 30min, cooling to 50-55 deg.C, adding 1mL of filter sterilized methanol per 100mL of culture medium, mixing, and pouring onto a flat plate.

AMS liquid medium: agar was not added to the medium, and the other formulation was the same as solid AMS medium.

The separation method comprises the following steps: the sugarcane rhizosphere soil is prepared into 10 by a gradient dilution method^-6-10^-8g/mL soil suspension, uniformly coating the suspension on a solid AMS culture medium, carrying out inversion culture at a constant temperature of 25 ℃, observing, separating and purifying a strain producing pink pigment, and naming the strain as YT2019B002, inoculating the strain into a nutrient broth liquid culture medium, shaking at a speed of 160rpm of 25 ℃ for 48 hours, adding 30% glycerol aqueous solution (v/v) according to a volume ratio of 1:1, and storing the strain at a temperature of-80 ℃ for later use.

Identification of the strain YT2019B 002:

1. determination of physicochemical Properties of Strain YT2019B002

Streak-culturing strain YT2019B002 on soybean casein agar culture medium TSA (OXOID) solid culture medium at 25 deg.C for 5 days, picking single colony, gram-staining, and culturing

2Systems gram negative bacteria identification card (GN card) for carbon source utilization, enzyme activity and drug resistance detection.

2. Molecular biological identification of strain YT2019B002

YT2019B002 single colony DNA was extracted by a thermal cracking method, and the type of the strain was confirmed by mxaF gene amplification using the specific primers mxa f1003(5-GCGGCACCAACTGGGGCTGGT-3 ', SEQ ID NO:2) and mxa r1561 (5-GGGCAGCATGAAGGGCTCCC-3', SEQ ID NO: 3). Meanwhile, the 16S rRNA fragment is amplified by 27F (5'-AGAGTTTGATCMTGGCTCAG-3', SEQ ID NO:4) and 1492R (5'-TACGGYTACCTTGTTACGACTT-3', SEQ ID NO:5), the PCR product is sent to Shanghai biological engineering (Shanghai) GmbH for sequencing, and the sequence is used for identifying the accurate type of YT2019B002 by constructing a phylogenetic tree with the sequence of a similar species.

3. And (3) measuring the growth promoting capability:

(1) seed germination experiment: the method comprises the steps of selecting cabbage heart seeds (four-nine cabbage hearts) as experimental materials, sterilizing the surface of a 2% sodium hypochlorite solution for 15 minutes, washing the sodium hypochlorite solution on the surface of the seeds with sterile water, uniformly placing the seeds in culture dishes paved with sterile gauze, adding 20mL of YT2019B002 bacterial liquid cultured by AMS liquid culture medium shaking at 28 ℃ for 5 days into each dish, taking the AMS liquid culture medium with the same volume as a control, and culturing the seeds in the dark at the constant temperature of 25 ℃ for 5 days to observe and count the germination rate.

(2) Growth-promoting pot experiment: mixing imported Denmark Torpu peat soil and vermiculite at a ratio of 5:1 (V/V), and sterilizing at 121 deg.C for 3 times at 20 min. A500 ml plastic flowerpot is soaked in a 2 wt% sodium hypochlorite solution for 30-60min, and then the residual sodium hypochlorite solution is fully washed away by sterile water for later use. Subpackaging sterilized soil into sterilized flowerpots, selecting pepper seedlings with the same growth vigor for 3-4 weeks, transplanting the pepper seedlings into the flowerpots, and repeating the treatment for 4 plants in each flowerpot for 4 treatments, wherein the treatments comprise (1) YT2019B002 (G): inoculating 30ml of bacterial liquid into each pot in a root irrigation mode 1 day after seedling transplantation; ② YT2019B002 (P): inoculating 10ml of bacterial liquid in each pot in a leaf surface spraying mode on

days

1, 8 and 15 after seedling transplantation; ③ CK 1: replacing the bacterial liquid in the first step with AMS liquid culture medium with the same volume for root irrigation treatment; fourthly, CK 2: and (4) replacing the bacterial liquid in the treatment step II with the same volume of AMS liquid culture medium for carrying out foliar spraying treatment. All treatments are placed in a light incubator with 28 ℃ and 16h/8h photoperiod for culture, water is poured once every two days, and after 23 days of transplantation, measurement statistics and single-factor analysis of variance (Fisher's LSD test) are carried out on the plant height, the fresh weight/dry weight of the overground part/underground part, the chlorophyll content of leaves and the number of true leaves of the pepper seedlings subjected to different treatments.

Determination of enzyme activity of strain YT2019B002

Determination of dextranase Activity: the strain was streaked on KM medium and contained 0.4% (NH) as a main component₄)₂SO₄，0.01％NaCI，0.01％MgSO₄，0.01％CaCI₂0.05% yeast extract, 33mg Fe (III) -EDTA, 0.05M potassium phosphate buffer (pH 7.0), 0.04% sodium carboxymethylcellulose (CMC), Agar 15.0g, distilled water 1L, sterilized at 121 ℃ for 30 min. After 3 days of inverted culture at constant temperature of 30 ℃, 0.1% Congo red solution is poured into the flat plate to dye for 30min, 1M NaCI solution fades for 10min, and whether a transparent ring is generated or not is observed.

Assay of Hemophilus Activity reference is made to the method of Shin et al (2001.) A ① 60.5.5 mg Chromel S in 50ml deionised water ② 10ml ferric iron solution (1mM FeCl)₃·6H₂O,10mM hydrochloric acid as a solvent), ③ 72.9.9 mg of CTAB is dissolved in 40ml of deionized water, the three solutions are mixed and metered to 100ml, the pH is adjusted to be neutral, sterilization is carried out at 121 ℃ for 20min, 30.24g of pegs are added into 900ml of water agar culture medium, sterilization is carried out at 6.8,121 ℃ for 20min, A and B solutions are mixed and poured into a flat plate, inoculation is carried out at 30 ℃ and inverted for 3d, and then observation and recording are carried out.

Determination of auxin IAA Activity: the strain was inoculated with 5mL of L-tryptophan (2 mgL) using a Salkowski colorimetric assay^-1) The nutrient broth liquid medium of (1), cultured at 26 ℃ and 160rpm for 7 daysThe culture solution was centrifuged at 6500rpm for 15min, the supernatant was filtered through a 0.22 μm microfiltration membrane to obtain a sterile filtrate, and 1mL of Salkowski color developing solution (2mL of 0.5 MFeCl) was added to each mL of the filtrate₃，49mL 35％[v/v]HClO₄) Incubating at room temperature for 30-60min, and measuring the absorbance of the filtrate at 530nm with spectrophotometer with culture solution without bacteria as control.

ACC activity assay: referring to the Penrose and Glick (2003) method, the strain was inoculated into a 50mL Erlenmeyer flask containing 10mL AMS liquid medium, shaken at 28 ℃ for 4-5 days at 120rpm, centrifuged at 3000g for 5min to collect the cells, washed twice with filter-sterilized 0.1M Tris-HCI (pH 7.5), resuspended in 1mL0.1M Tris-HCI (pH 7.5), 200. mu.L of the resulting suspension was spread on DF microelement medium containing 3mM ACC as the only nitrogen source to add (NH)₄)₂SO₄The DF culture medium (0.2% w/v) is used as a positive control, and the growth condition of the strain is observed after the coated culture dish is placed at the constant temperature of 28 ℃ for 3d of inverted culture.

As a result:

1. identification

The strain YT2019B002 is gram-stained red (as shown in figure 1) and is a gram-negative bacterium. Colony morphology: culturing on AMS solid culture medium to obtain opaque light pink colony with diameter of 1-3mm, upward protrusion, and neat edge, and drying.

Physical and chemical Property characterization As shown in Table 1, using

2System and

2 gram negative bacteria identification card (GN), through the determination of 47 kinds of carbon source utilization, enzyme activity and drug resistance and a negative control reaction in total, the primary identification YT2019B002 is Methylobacterium sp bacteria.

TABLE 1 physicochemical Properties of Strain YT2019B002

Note: 95% to 100% positive; v-6% to 94% positive; positive-0% to 5%.

Using the DNA of the strain YT2019B002 as a template and amplification with a specific primer of Methylobacterium, a specific band with a fragment size of 560bp was obtained (fig. 2), and it was confirmed that YT2019B002 was a bacterium of the genus Methylobacterium. The 16S rRNA sequence (shown as SEQ ID NO: 1) of the strain YT2019B002 is compared with the sequence in the NCBI database, and the similarity of the sequence and the M.populi model strain BJ001(Access No.: AY251818) is the highest and reaches 99.49 percent. A phylogenetic tree is constructed by using 16S rRNA sequences of other species of Methylobacterium in NCBI as reference and Escherichia coli (Access No.: J01859) as outer population and adopting a Neighbor-Joining method (NJ), and strains YT2019B002 and M.populi (AY251818) in the obtained optimal tree form an independent branch (as shown in figure 3) with high support rate (96).

The 16S rRNA gene sequence of the methylobacterium YT2019B002 is shown as SEQ ID NO: 1, and the following components:

SEQ ID NO:1：

CAGCTTACACATGCAGTCGAACGGGCTTCTTCGGAAGTCAGTGGCAGACGGGTGAGTAACACGTGGGAACGTGCCCTTCGGTTCGGAATAACTCAGGGAAACTTGAGCTAATACCGGATACGCCCTTATGGGGAAAGGTTTACTGCCGAAGGATCGGCCCGCGTCTGATTAGCTTGTTGGTGGGGTAACGGCCTACCAAGGCGACGATCAGTAGCTGGTCTGAGAGGATGATCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGAGTGATGAAGgCCtTAGGGTTGTAAAGCTCTTTTGTCCGGGACGATAaTGACGGTACCGGAAGAATAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGGGCTAGCGTTGCTCGGAATCACTGGGCGTAAAGGGCGCGTAGGCGGCCGATTAAGTCGGGGGTGAAAGCCTGTGGCTCAACCACAGAATTGCCTTCGATACTGGTTGGCTTGAGACCGGAAGAGGACAGCGGAACTGCGAGTGTAGAGGTGAAATTCGTAGATATTCGCAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCCGGTTCTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATGCCAGCCGTTGGCCTGCTTGCAGGTCAGTGGCGCCGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGCARAAaCCTWACCATcCCCTTGACATGGCaTGTTACCTCGAGAGATCGGGGATCCTCTTCGRAGGCGTGCACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCACGTCCTTAGTTGCCATCATTCAGTTGGGCACTCTAGGGAGACTGCCGGTGATAAGCCGCGAGGAAGGTGTGGATGACGTCAAGTCCTCATGGCCCTTACGGGATGGGCTACACACGTGCTACAATGGCGGTGACAGTGGGACGCGAAACCGCGAGGTTGAGCAAATCCCCAAAAGCCGTCTCAGTTCGGATTGCACTCTGCAACTCGGGTGCATGAAGGCGGAATCGCTAGTAATCGTGGATCAGCACGCCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTTGGTCTTACCCGACGGCGCTGCGCCAACCGCAAGGGGGCAGGCGACCACGGTAGTCGCAC。

2. determination of seed germination growth promoting capability

In order to determine whether the strain YT2019B002 has the capability of promoting seed germination, a seed germination test is carried out by taking YT2019B002 fermentation liquor as a treatment group (YT2019B002) and an AMS culture medium as a control group (CK) (figure 4). The test result shows that the seeds of the treated group begin to germinate on the second day of culture, the germination time of the control group is later than that of the treated group, the germination begins on the third day, the germination rate of the control group on the third day is 39.39%, and the germination rate of the treatment group on the third day is 59.04%. The fifth day after inoculation, the germination rates and the seedling lengths after germination of all the different treatments are measured and counted, and the average germination rate of the CK control group is 47.74 percent, the average seedling length is 20.03 +/-0.02 mm, while the average germination rate of the YT2019B002 treatment group is 70.16 percent, the average seedling length is 25.29 +/-0.03 mm, and the germination rate and the average seedling length are both obviously higher than those of the control group. The bacterial strain YT2019B002 has the function of promoting the germination and growth of seeds.

3. Growth promotion test for potted plants

After the pepper seedlings are transplanted for 23d, the statistical results of all growth indexes show that fresh weight/dry weight, plant height, chlorophyll content and average true leaf number of the pepper seedlings treated by root irrigation (YT2019B002(G)) and spraying (YT2019B002(P)) by using YT2019B002 strain fermentation liquor are obviously higher than those of a control group treated by AMS culture solution; there was no significant difference between the growth indicators of the control of the two different treatment modes of the CK1 control group and the CK2 control group. In the treatment groups, the underground dry weight and the plant height of the pepper seedlings subjected to the 3-time spraying treatment in the YT2019B002(G) group are obviously higher than those of the one-time root irrigation treatment in the YT2019B002(P) group, and the average true leaf number is also large, so that the fermentation liquid of the strain YT2019B002 has the capability of obviously promoting the plant growth, and the multi-time spraying effect is better than that of the one-time root irrigation treatment (Table 2 and figure 5).

TABLE 2 growth indexes of pepper seedlings treated differently

a. b and c represent significant differences

Growth promoting mechanism of YT2019B002 strain

The results of the auxin IAA measurement show that standard curves y, 44.716x +0.812(R is 44.716x + 0.812) are obtained by plotting the standard products with the concentrations of 0, 0.2, 1.0, 2.0, 3.0, 6.0, 11.0, 20.0 and 45.0 mu g/mL and the absorbance values at 530nm²0.9985) (fig. 6a), the absorbance at 530nm of the treated group after 7d cultivation was 0.126 ± 0.009 with no added bacteria as a blank, indicating that the strain YT2019B002 has the function of secreting auxin IAA (fig. 6B), and the IAA concentration was 6.461 ± 0.401 μ g/mL according to the standard.

YT2019B002 was unable to grow on DF medium with ACC as the sole nitrogen source and also had no glucanase and siderophin activity as determined by plate.

In conclusion, the strain YT2019B002 can promote plant growth by means of IAA production.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

SEQUENCE LISTING

<110> Guangdong province bioengineering research institute (Guangzhou sugar industry institute)

<120> high-efficiency growth-promoting methylobacterium strain and application thereof

<130>2020-08-05

<160>5

<170>PatentIn version 3.3

<210>1

<211>1380

<212>DNA

<213>Artificial Sequence

<400>1

cagcttacac atgcagtcga acgggcttct tcggaagtca gtggcagacg ggtgagtaac 60

acgtgggaac gtgcccttcg gttcggaata actcagggaa acttgagcta ataccggata 120

cgcccttatg gggaaaggtt tactgccgaa ggatcggccc gcgtctgatt agcttgttgg 180

tggggtaacg gcctaccaag gcgacgatca gtagctggtc tgagaggatg atcagccaca 240

ctgggactga gacacggccc agactcctac gggaggcagc agtggggaat attggacaat 300

gggcgcaagc ctgatccagc catgccgcgt gagtgatgaa ggccttaggg ttgtaaagct 360

cttttgtccg ggacgataat gacggtaccg gaagaataag ccccggctaa cttcgtgcca 420

gcagccgcgg taatacgaag ggggctagcg ttgctcggaa tcactgggcg taaagggcgc 480

gtaggcggcc gattaagtcg ggggtgaaag cctgtggctc aaccacagaa ttgccttcga 540

tactggttgg cttgagaccg gaagaggaca gcggaactgc gagtgtagag gtgaaattcg 600

tagatattcg caagaacacc agtggcgaag gcggctgtct ggtccggttc tgacgctgag 660

gcgcgaaagc gtggggagca aacaggatta gataccctgg tagtccacgc cgtaaacgat 720

gaatgccagc cgttggcctg cttgcaggtc agtggcgccg ctaacgcatt aagcattccg 780

cctggggagt acggtcgcaa gattaaaact caaaggaatt gacgggggcc cgcacaagcg 840

gtggagcatg tggtttaatt cgaagcaacg cgcaraaacc twaccatccc cttgacatgg 900

catgttacct cgagagatcg gggatcctct tcgraggcgt gcacacaggt gctgcatggc 960

tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc aacccacgtc 1020

cttagttgcc atcattcagt tgggcactct agggagactg ccggtgataa gccgcgagga 1080

aggtgtggat gacgtcaagt cctcatggcc cttacgggat gggctacaca cgtgctacaa 1140

tggcggtgac agtgggacgc gaaaccgcga ggttgagcaa atccccaaaa gccgtctcag 1200

ttcggattgc actctgcaac tcgggtgcat gaaggcggaa tcgctagtaa tcgtggatca 1260

gcacgccacg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca ccatgggagt 1320

tggtcttacc cgacggcgct gcgccaaccg caagggggca ggcgaccacg gtagtcgcac 1380

<210>2

<211>21

<212>DNA

<213>Artificial Sequence

<400>2

gcggcaccaa ctggggctgg t 21

<210>3

<211>20

<212>DNA

<213>Artificial Sequence

<400>3

gggcagcatg aagggctccc 20

<210>4

<211>20

<212>DNA

<213>Artificial Sequence

<400>4

agagtttgat cmtggctcag 20

<210>5

<211>22

<212>DNA

<213>Artificial Sequence

<400>5

tacggytacc ttgttacgac tt 22

Claims

1. The methylobacterium YT2019B002, wherein the deposited number of the methylobacterium YT2019B002 is GDMCC No: 61052.

2. the use of methylobacterium YT2019B002 in the preparation of auxin in claim 1.

3. Use of methylobacterium YT2019B002 in the preparation of plant growth regulating preparation according to claim 1.

4. The use of methylobacterium YT2019B002 in the preparation of fertilizer for promoting plant growth as claimed in claim 1.

5. The use of methylobacterium YT2019B002 in promoting plant growth as claimed in claim 1.

6. The use of claim 5, wherein the promoting plant growth comprises promoting plant seed germination.

7. The use of claim 5, wherein said promoting plant growth comprises promoting plant growth.

8. A biological agent characterized in that its active ingredient comprises Methylobacterium YT2019B002 as claimed in claim 1.

9. The method for culturing methylobacterium YT2019B002, which comprises the steps of: inoculating methylobacterium YT2019B002 into a culture medium, and culturing at 20-30 ℃; the culture medium is AMS culture medium.

10. A Methylobacillus YT2019B002 culture prepared by culturing the Methylobacillus YT2019B002 of claim 1.