CN106701631B - Streptomyces roseoflavus and application thereof - Google Patents
Streptomyces roseoflavus and application thereof Download PDFInfo
- Publication number
- CN106701631B CN106701631B CN201710044242.4A CN201710044242A CN106701631B CN 106701631 B CN106701631 B CN 106701631B CN 201710044242 A CN201710044242 A CN 201710044242A CN 106701631 B CN106701631 B CN 106701631B
- Authority
- CN
- China
- Prior art keywords
- fermentation
- streptomyces roseoflavus
- roseoflavus
- streptomyces
- strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/465—Streptomyces
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention provides a Streptomyces roseoflavus and application thereof, wherein the strain is Streptomyces roseoflavus (Streptomyces roseoflavus), the preservation number of the strain is CGMCC No.13416, and the strain is preserved in the common microorganism center of China Committee for culture Collection of microorganisms. The streptomyces roseoflavus and the microbial inoculum containing the streptomyces roseoflavus and/or the fermentation liquor thereof have obvious growth promotion effect on plants, can inhibit various pathogenic fungi, and have good growth promotion and prevention and control effects, so the streptomyces roseoflavus and the microbial inoculum have good application prospect.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to streptomyces roseoflavus and application thereof.
Background
In agricultural production, the use of chemical fertilizers increases the yield of crops, but also releases a large amount of harmful substances to the environment, pollutes soil, water sources and food, and poses great threats to human health and living environment. Meanwhile, in order to reduce the loss of plant diseases to agricultural production, a large amount of chemical pesticides are also applied for a long time in agricultural production, so that pesticide residues and environmental pollution are caused, the human health is finally harmed, the ecological balance is destroyed, and meanwhile, pathogenic bacteria generate serious drug resistance to various chemical bactericides. Biological control draws more and more importance to the world because of its advantages such as safety and non-toxicity to environment, ecology and human health. Therefore, in order to meet the needs of green agriculture and sustainable agricultural development, it is urgent to develop new fertilizer sources for replacing chemical fertilizers and biopesticides for replacing chemical pesticides.
Disclosure of Invention
In view of the problems existing in the technology, the invention provides the streptomyces roseoflavus and the application thereof, which have obvious growth promotion effect on plants, can inhibit various pathogenic fungi, and have good growth promotion and prevention and control effects, thereby having good application prospect.
The technical scheme for solving the technical problems is as follows:
the streptomyces roseoflavus is characterized by being streptomyces roseoflavus with the preservation number of CGMCC NO. 13416.
The strain has been preserved by China general Microbiological Culture Collection Center (CGMCC) at 2016, 12 and 02.
Furthermore, the streptomyces roseoflavus has a sequence shown in SEQ ID NO. 1.
The invention also provides a microbial inoculum which contains the streptomyces roseoflavus and/or the fermentation liquor of the streptomyces roseoflavus.
The invention also provides a preparation method of the microbial inoculum, which comprises the following steps:
1) inoculating the streptomyces roseoflavus of claim 1 into a Gao's 1 culture medium, culturing, and growing a bacterium block;
2) selecting the bacterium blocks in the step 1), inoculating the bacterium blocks into a fermentation culture medium, and fermenting to obtain a seed solution;
3) inoculating the seed liquid obtained in the step 2) into a fermentation medium, and fermenting to obtain a fermentation liquid.
Further, in step 1), the culture temperature was 28 ℃ and the culture time was 7 days.
Further, in the step 2), the fermentation mode is shaking fermentation, the rotating speed is 170rpm, the culture temperature is 28 ℃, and the culture time is 48 hours;
further, in the step 3), the inoculation amount is 10%, the culture temperature is 28 ℃, the fermentation time is 96 hours, and the rotating speed is 220 rpm;
further, the fermentation culture medium in the step 2) and the fermentation culture medium in the step 3) are both prepared according to the following formula: maltose 10%, yeast powder 4.0%, glucose 4.0%, and water in balance.
The culture medium and the culture conditions adopting the formula are favorable for normal growth of the strains, and the activity of the strains is maintained as much as possible, so that the yield and the stability of the growth-promoting and bacteriostatic metabolic products of the strains are improved.
Further, after the step 3), the method also comprises the step of centrifuging the fermentation liquor and taking the supernatant. For example: in specific operation, the rotating speed of the centrifugation can be 9500rpm, and the time of the centrifugation can be 10 min.
The invention also provides application of the streptomyces roseoflavus in promoting plant growth. The inventor unexpectedly discovers in research that the streptomyces roseoflavus can remarkably promote the growth of plants. The plant may be, but is not limited to, tomato, wheat, pepper, etc.
The invention also provides application of the streptomyces roseoflavus in inhibiting the growth of pathogenic fungi. The inventor unexpectedly discovers in research that the streptomyces roseoflavus can obviously inhibit the growth of pathogenic fungi, particularly the growth of hyphae of the pathogenic fungi. The pathogenic fungi can be any one or a mixture of any several of but not limited to tomato leaf mold, corn curvularia, ustilaginoidea virens, Lubao No.1 and rhizoctonia cerealis.
Drawings
FIG. 1 is a colony morphology of Streptomyces roseoflavus according to the present invention.
FIG. 2A and FIG. 2B are electron microscope scanning images of Streptomyces roseoflavus of the present invention under electron microscope conditions of different multiples, respectively.
FIG. 3 is a graph showing the results of phylogenetic tree analysis of Streptomyces roseoflavus according to the present invention.
FIG. 4 shows the results of tests for the inhibition of fungi, A for the test with sterile water and B for the test with the fermentation broth of Streptomyces roseoflavus (the broth of Streptomyces roseoflavus was added to the PDA medium in a proportion of 10% by volume).
FIGS. 5A, 5B and 5C are the results of the growth-promoting effect of Streptomyces roseoflavus of the present invention on tomatoes, wherein "CK" represents the irrigation of clear water for the control group, and "100X" represents the irrigation of diluted 100-fold fermentation broth of Streptomyces roseoflavus; fig. 5A is the result of the experiment for 15 days, and fig. 5B and 5C are the result of the experiment for 45 days.
FIG. 6A is the result of the test for verifying the growth promoting effect of Streptomyces roseoflavus of the present invention on wheat (growth diagram of wheat), wherein "CK" on the left is the test result of the control group treated with clear water, and "100X" on the right is the test result of the treatment after the fermentation broth of Streptomyces roseoflavus is diluted 100 times. FIG. 6B is a graph showing the results of experiments on the growth promoting effect of Streptomyces roseoflavus of the present invention on wheat (root growth of wheat), wherein "CK" on the left is the experimental result of the control group treated with clear water, and "100X" on the right is the experimental result of the treatment after the fermentation broth of Streptomyces roseoflavus is diluted 100 times.
FIGS. 7A to 7C are the results of experiments on the growth promoting effect of Streptomyces roseoflavus on Capsici fructus, wherein "CK" represents the irrigation of clear water in the control group, and "100X" represents the irrigation of diluted 100-fold fermentation broth of Streptomyces roseoflavus. Fig. 7A is the growth result for 15 days, fig. 7B is the growth result for 45 days, and fig. 7C is the growth result for 45 days.
Detailed Description
The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention.
The streptomyces roseoflavus provided by the invention has obvious growth promotion effect on tomatoes, hot peppers and wheat, can inhibit various pathogenic fungi, and has good growth promotion and prevention and control effects, so that the streptomyces roseoflavus has good application prospect.
The technical solution of the present invention is specifically described below.
The strain provided by the invention is streptomyces roseoflavus (Stremptomyyces roseoflavus), is separated from Qilian mountain area in Qinghai province by Zhang Cheng researchers, is preserved in the common microorganism center (CGMCC for short) of the China Committee for microbiological culture Collection center (CGMCC) in 2016, 12 months and 2 days, has the preservation number of CGMCC No.13416 and is named as NKZ-259. It has the following biological characteristics.
The following specifically describes the embodiments of the present invention with reference to specific examples.
Example 1 isolation and purification of Streptomyces roseoflavus (Stremptomyces roseoflavus NKZ-259)
The streptomyces roseoflavus (Stremptomyces roseoflavus NKZ-259) is obtained by separating from Qilian mountain soil in Qinghai province by adopting a dilution plate method and a plate marking method, and the separation and purification method comprises the following steps:
weighing 1g of soil sample, placing into a triangular flask with glass beads containing 99ml of sterile water, shaking for 30min, sucking 1ml of soil sample from the triangular flask with a pipette, adding into another test tube containing 9ml of sterile water, mixing, and making into 10-2、10-3、10-4Soil solutions of different dilutions. 100 μ L of the soil solution was spread on a Gao's 1 medium plate, 3 replicates were performed for each dilution, and incubated at 28 ℃ for 3 days. Selecting single bacterial colony, streaking on Gao's No.1 culture medium plate, observing the growth of bacterial colony regularly, purifying bacillus strain by plate streaking process, making slant mark and storing in 4 deg.c refrigerator.
Formula (g/L) of Gao's No.1 culture medium: 20.0 parts of soluble starch, 1.0 part of potassium nitrate, 0.5 part of dipotassium phosphate, 0.5 part of magnesium sulfate heptahydrate, 0.5 part of sodium chloride, 0.01 part of ferrous sulfate heptahydrate and 17 parts of agar. Weighing, heating, stirring, dissolving in distilled water, packaging in triangular flask, autoclaving at 121 deg.C, and pouring.
Example 2 measurement of bacteriostatic Activity of Streptomyces roseoflavus strain NK-259
1. Plate confrontation test
Culturing 17 pathogenic fungi (respectively tomato leaf mold, curvularia zeae, fusarium oxysporum, rhizopus persicae, black rot, green smut, poplar canker, corn microsporum, apple ring rot, grape black rot, apple rot, Lubao No.1, alternaria alternata, corn leaf blight, wheat gibberellic disease, rice blast and wheat sharp eyespot, all from agricultural antibiotic research group of Chinese academy of agricultural science) on PDA for 5 days, cutting small blocks with a scalpel, transferring to the center of PDA culture medium (Chinese called Potato glucose agar culture medium, English called Potato Dextrose agar medium), placing sterile oxford cups on two sides with equal distance to the pathogenic bacteria, adding fermentation liquor (28 ℃ for 96h) of Streptomyces roseoflavus NKZ-259 and sterile water, culturing at 25 ℃ for 3-9 days, the inhibitory effect of the NK-259 fermentation broth on the growth of pathogenic fungi was observed.
And (4) conclusion: streptomyces roseoflavus NKZ-259 has inhibitory effect on five of the above 17 pathogenic fungi, shown in FIG. 4A, including Phyllomyces lycopersici, Curvularia zeae, Ustilaginoidea virens, Ubah No.1, and Rhizoctonia cerealis.
2. Influence on hyphal growth
Inoculating Streptomyces roseoflavus NKZ-259 to Gaoshi No.1 culture medium, culturing at 28 deg.C for 7 days, and cutting 1cm with surgical knife2The mass was inoculated into the fermentation medium and fermented with shaking for 96h (28 ℃, 220 rpm). Then the fermentation liquor is centrifuged (10000r/min, 4 ℃), the supernatant is taken and filtered by a filter membrane of 0.22 μm for sterilization, and the sterile fermentation liquor is obtained.
The influence of the fermentation broth of Streptomyces roseoflavus NKZ-259 on the growth of hyphae of 5 pathogenic fungi (leaf mold of tomato, curvularia zeae, Ustilaginoidea virens, No.1 Lubao and Rhizoctonia cerealis) to be tested is detected by adopting a hypha growth rate method. Under the aseptic condition, 10mL of prepared fermentation liquor and 90mL of melted PDA culture medium are uniformly mixed, a plate culture medium containing the fermentation liquor is prepared in an aseptic culture dish, a culture medium without the fermentation liquor is used as a reference, 1 test pathogenic fungus cake with the diameter of 5mm is placed on the surface of each culture medium, each treatment is repeated for 3 times, after the culture is carried out for 7 days at the constant temperature of 25 ℃, the diameter of a bacterial colony is measured by a cross method, and the inhibition rate of hypha growth is calculated: the hypha growth inhibition ratio (control colony diameter-treated colony diameter)/control colony diameter × 100%. The results of the experiment are shown in FIG. 4B.
TABLE 1
| Pathogenic fungi | Hypha growth inhibition ratio (%) |
| Leaf mold of tomato | 48.72±2.32 |
| Curvularia lunata (Fr.) Sing | 60.48±1.66 |
| Ustilaginoidea virens | 63.11±2.73 |
| Lubao No.1 | 40.00±1.13 |
| Rhizoctonia cerealis | 26.90±1.69 |
And (4) conclusion: as can be seen from Table 1, the Streptomyces roseoflavus NKZ-259 strain sterile fermentation broth has obvious inhibitory effects on the growth of hyphae of the above 5 test germs, and as shown in FIG. 4B, the inhibitory rates are 48.72%, 60.48%, 63.11%, 40.00% and 26.90%, respectively.
Example 3 morphological Observation of Streptomyces roseoflavus (Stremptomyces roseoflavus NKZ-259)
And (3) observing colony morphology: inoculating NKZ-259 strain on Gao's No.1 culture medium, culturing at 28 deg.C under inverted condition for 7 days, observing colony morphology, and finding that the colony is light pink and has small amount of white aerial hyphae (shown in FIG. 1);
and (3) observing by using an electron microscope: cutting a small amount of mycelium from the culture medium with a scalpel, and fixing in 2-4% glutaraldehyde fixing solution for 1Washing with 0.1M PBS buffer (pH7.2PBS) for 2 days, fixing with 1% OsO4 for 1-2 hr, washing with redistilled water for 30min, gradient-dehydrating with 30%, 50%, 70%, 80%, 90%, 95%, 100% ethanol for 15-20 min, treating with isoamyl acetate, and treating with CO2Drying the critical point, adhering the dried sample on a sample table, spraying gold on the surface of an ion sputtering instrument, scanning the sample under a scanning electron microscope after spraying gold, and observing the spore form to take a picture and record.
The spore filament is found to be straight or flexible through culture and observation, and the spore is oval or elliptical and has a smooth surface. (as shown in fig. 2A and 2B).
Example 4 identification of the 16s rRNA Gene sequence of Streptomyces roseoflavus strain NKZ-259
The strain is inoculated in YEME culture medium, after shaking culture (28 ℃, 220rpm) for 48h, 500 mu L of bacterial liquid is taken out to be put in an EP tube, centrifugation is carried out, supernatant is discarded, thalli are collected, DNA is extracted, and then PCR verification is carried out. The primers used for PCR amplification were as follows: a forward primer F: 5'-GCAGTCGAGCGGACAGAT-3', respectively; reverse primer R: 5'-AAGGAGGTGATCCAGCCGCA-3' are provided. The PCR reaction system comprises 2 XTaqPCRmastermix 12.5. mu.L, forward F primer 10.5. mu.L, reverse R primer 0.5. mu.L, bacterial liquid template DNA 1. mu.L, ddH2O10.5. mu.L, 25. mu.L in total. The PCR amplification condition is 94 ℃ for 5 min; 1min at 94 ℃; 30s at 51 ℃; 1min at 72 ℃ for 35 cycles; 10min at 72 ℃. The electrophoresis adopts agarose gel electrophoresis, nucleic acid dye is EB, and Maker is DL 2000. And finally, sending the PCR purified product to Shanghai biological engineering Co., Ltd for sequencing. And carrying out comparison analysis on related sequences in the obtained sequence NCBI GeneBank nucleic acid database. PCR amplification of 16SrRNA was performed using forward primer F and reverse primer R to obtain a fragment of approximately 1.4kb as shown in SEQ ID NO. 1. The similarity of the nucleotide sequence of the corresponding region of the sequence and the 16S rRNA gene of the streptomyces roseoflavus is 100 percent by comparison.
The obtained NKZ-25916s rRNA sequences of the strain were compared with 20 different neomycin 16s rRNA sequences in GenBank, and the extra-group sequence with larger difference was selected as Streptomyces coelicolor A3. The nucleotide and corresponding amino acid sequences were analyzed and aligned by DNAMAN version6.0(Lynnon Biosoft), and the full-length nucleotide sequences were compiled using Bioedit. MEGA (version 4.1) carries out ClustalW multi-sequence alignment and phylogenetic analysis; the evolutionary tree is constructed by a neighbor-join method, and bootstrap is set to 1000 times of repetition. The result shows that phylogenetic analysis of the obtained bacteria NKZ-259 is in the same branch with that of Streptomyces roseoflavus strain Men-myco-93-63, which indicates that the relativity is relatively close, and the inventor identifies the strain as Streptomyces roseoflavus (shown in figure 3) by combining morphological structure and physiological and biochemical characteristics.
Example 5 fermentation Process of Streptomyces roseoflavus strain NKZ-259
The formula of the culture medium is as follows: maltose 10%, yeast powder 4.0%, glucose 4.0%, and water in balance. The percentages in the formula are mass percentages.
And (3) fermentation process:
(1) coating Streptomyces roseoflavus NKZ-259 with sterile toothpick, inoculating on Gao's No.1 culture medium, and culturing at 28 deg.C for 7 days;
(2) cutting 1cm with scalpel2Inoculating the strain blocks into a fermentation medium, and performing shake fermentation for 48h (28 ℃, 170 rpm);
(3) inoculating the seed liquid of the second step into a fermentation culture medium according to the inoculation amount of 10% by volume, and performing shake fermentation for 96h (the fermentation temperature is 28 ℃, and the rotation speed is 220 rpm).
Example 6 measurement of growth promoting Effect of Streptomyces roseoflavus strain NKZ-259
(1) Tomato growth promotion test method:
the test is carried out in nutrient soil by adopting plug planting, the matrix is prepared from the nutrient soil and vermiculite according to the ratio of 9:1, and the nutrient soil and the vermiculite are placed in the plug. Spreading 2-3 tomato seeds in each hole, covering the seeds with a small amount of nutrient soil, irrigating a proper amount of water, covering with a preservative film, and removing the preservative film when the seedlings grow out of the soil and grow 2 cotyledons. The test was divided into two groups, the test group was irrigated with 100-fold diluted NKZ-259 fermentation broth, the control group was irrigated with clear water, each treated 20 seedlings, and the procedure was repeated 3 times. Culturing in a greenhouse at 25 deg.C under relative humidity of 60%, growing for about 7 days, irrigating with bacteria liquid when tomato seedlings grow 2 true leaves, irrigating roots every 10 days, and irrigating with clear water at other times. After 3 times of root irrigation, namely when the tomato seedlings grow to 45 days, carefully digging out the tomato seedlings, washing away soil at the roots, and measuring the growth indexes of the tomato seedlings: root length, plant height, fresh weight, dry weight.
TABLE 2 promotion of tomato growth by fermentation broth seed soaking of strain NKZ-259
From table 2 and fig. 5A, 5B and 5C, it can be seen that the root irrigation with the fermentation broth of strain NKZ-259 has an obvious growth promoting effect on tomato seedlings, tomato seedlings after root irrigation with the fermentation broth grow rapidly, have dense leaves, thick stems, especially developed root systems, and significantly increased root length, compared with the control group, the root length is increased by 39.23%, the plant height is increased by 5.62%, the fresh weight is increased by 11.70%, and the dry weight is increased by 31.82%.
(2) Wheat seed soaking test
Firstly, soaking wheat seeds for 24 hours by respectively using fermentation liquor of NKZ-259 and clear water, then respectively sowing the wheat seeds in glass culture dishes, spreading two layers of filter paper at the bottom of each culture dish, placing 15 seeds in each culture dish, and repeating the steps for 3 times. Diluting the fermentation liquor by 100 times, irrigating the fermentation liquor and clear water every day to ensure that the filter paper is wet, culturing in a greenhouse at 25 ℃ under the relative humidity of 60%, and measuring the root length, plant height, fresh weight and dry weight of the wheat after 15 days.
TABLE 3 promotion of wheat growth by fermentation broth seed soaking of strain NKZ-259
As can be seen from Table 3 and FIGS. 6A and 6B, the seed soaking test shows that the fermentation liquor of the strain NKZ-259 can obviously promote the growth of wheat, the root length of the wheat seedlings soaked in the fermentation liquor is increased by 32.50%, the plant height is increased by 23.79%, the fresh weight is increased by 46.93%, and the dry weight is increased by 37.03% compared with the control group. The wheat true leaves soaked by the fermentation liquor appear earlier than those of the control group, the leaves are large, the petioles are thick, the root system is developed, and the growth is more uniform and vigorous.
(3) The pepper growth promotion test method comprises the following steps:
the test is carried out in nutrient soil by adopting plug planting, the matrix is prepared from the nutrient soil and vermiculite according to the ratio of 9:1, and the nutrient soil and the vermiculite are placed in the plug. Spreading 2-3 pepper seeds in each hole, covering the seeds with a small amount of nutrient soil, irrigating a proper amount of water, covering with a preservative film, and removing the preservative film when the seedlings grow out of the soil and grow 2 cotyledons. The test was divided into two groups, the test group was irrigated with 10-fold diluted NKZ-259 fermentation broth, the control group was irrigated with clear water, each treated 20 seedlings, and the procedure was repeated 3 times. Culturing in a greenhouse at 25 deg.C under relative humidity of 60%, irrigating fermentation liquid when pepper seedlings grow 2 true leaves, irrigating roots every 10 days, and irrigating clear water at other times. After 3 times of root irrigation, namely when the pepper seedlings grow to about 45 days, carefully digging out the pepper seedlings, washing away soil at the roots, and measuring the growth indexes of the pepper seedlings: root length, plant height, fresh weight, dry weight.
TABLE 4 growth promoting effect of fermentation broth seed soaking of strain NKZ-259 on Capsici fructus
As can be seen from table 4 above and fig. 7A, 7B and 7C, root irrigation with the fermentation broth of strain NKZ-259 has an obvious growth promoting effect on pepper seedlings, pepper seedlings after root irrigation with the fermentation broth have developed root systems, grow rapidly, have thick leaves and thick stems, and the plant height of the pepper seedlings is increased by 16.92% compared with that of a control group, the root length is increased by 44.15%, the fresh weight is increased by 63.08%, and the dry weight is increased by 38.89%.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
<110> institute of plant protection of Chinese academy of agricultural sciences
<120> Streptomyces roseoflavus and application thereof
<160>1
<210>1
<211>1413
<212>DNA
<213> Artificial sequence
<220>
<221>16s rRNA Gene
<400>1
tgcagtcgaa cgatgaagcc cttcggggtg gattagtggc gaacgggtga gtaacacgtg 60
ggcaatctgc ccttcactct gggacaagcc ctggaaacgg ggtctaatac cggataacac 120
tctctctcgc atgggagagg gttgaaagct ccggcggtga aggatgagcc cgcggcctat 180
cagctagttg gtgaggtaga agctcaccaa ggcgacgacg ggtagccggc ctgagagggc 240
gaccggccac actgggactg agacacggcc cagactccta cgggaggcag cagtggggaa 300
tattgcacaa tgggcgaaag cctgatgcag cgacgccgcg tgagggatga cggccttcgg 360
gttgtaaacc tctttcagca gggaagaagc gaaagtgacg gtacctgcag aagaagcgcc 420
ggctaactac gtgccagcag ccgcggtaat acgtagggcg caagcgttgt ccggaattat 480
tgggcgtaaa gagctcgtag gcggcttgtc acgtcggttg tgaaagcccg gggcttaacc 540
ccgggtctgc agtcgatacg ggcaggctag agttcggtag gggagatcgg aattcctggt 600
gtagcggtga aatgcgcaga tatcaggagg aacaccggtg gcgaaggcgg atctctgggc 660
cgatactgac gctgaggagc gaaagcgtgg ggagcgaaca ggattagata ccctggtagt 720
ccacgccgta aacggtggga actaggtgtg ggcgacattc cacgtcgtcc gtgccgcagc 780
taacgcatta agttccccgc ctggggagta cggccgcaag gctaaaactc aaaggaattg 840
acgggggccc gcacaagcgg cggagcatgt ggcttaattc gacgcaacgc gaagaacctt 900
accaaggctt gacatacacc ggaaagcatc agagatggtg ccccccttgt ggtcggtgta 960
caggtggtgc atggctgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg 1020
agcgcaaccc ttgtcccgtg ttgccagcaa gccccttcgg gggtgttggg gactcacggg 1080
agaccgccgg ggtcaactcg gaggaaggtg gggacgacgt caagtcatca tgccccttat 1140
gtcttgggct gcacacgtgc tacaatggcc ggtacaatga gctgcgatac cgcgaggtgg 1200
agcgaatctc aaaaagccgg tctcagttcg gattggggtc tgcaactcga ccccatgaag 1260
tcggagtcgc tagtaatcgc agatcagcat tgctgcggtg aatacgttcc cgggccttgt 1320
acacaccgcc cgtcacgtca cgaaagtcgg taacacccga agccggtggc ccaacccctt 1380
gtgggaggga gctgtcgaag gtgggactgg cga 1413
Claims (7)
1. The streptomyces roseoflavus is characterized by being streptomyces roseoflavus with the preservation number of CGMCC NO. 13416.
2. A microbial inoculum comprising the strain of Streptomyces roseoflavus and/or the fermentation broth of Streptomyces roseoflavus according to claim 1.
3. A method for preparing the microbial agent of claim 2, comprising the steps of:
1) inoculating the streptomyces roseoflavus of claim 1 into a Gao's 1 culture medium, culturing, and growing a bacterium block;
2) selecting the bacterium blocks in the step 1), inoculating the bacterium blocks into a fermentation culture medium, and fermenting to obtain a seed solution;
3) inoculating the seed liquid obtained in the step 2) into a fermentation medium, and fermenting to obtain a fermentation liquid.
4. The method for preparing the microbial inoculum according to claim 3, wherein in the step 1), the culture temperature is 28 ℃ and the culture time is 7 days; in the step 2), the fermentation mode is shaking fermentation, the rotating speed is 170rpm, the culture temperature is 28 ℃, and the culture time is 48 hours; in the step 3), the inoculation amount is 10%, the culture temperature is 28 ℃, the fermentation time is 96h, and the rotating speed is 220 rpm; the fermentation culture medium in the step 2) and the step 3) has the following formula: maltose 10%, yeast powder 4.0%, glucose 4.0%, and water in balance.
5. The method for preparing the microbial inoculum according to claim 3 or 4, further comprising a step of centrifuging the fermentation broth to collect the supernatant after the step 3).
6. The use of a strain of Streptomyces roseoflavus according to claim 1 for promoting the growth of plants, wherein the plants are selected from any one of tomato, wheat and pepper.
7. The use of a strain of Streptomyces roseoflavus according to claim 1 for inhibiting the growth of pathogenic fungi, wherein the pathogenic fungi are selected from any one or a mixture of any two of Phyllomycete lycopersici, Curvularia zeae, Ustilaginoidea, Rubao No.1 and Rhizoctonia cerealis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710044242.4A CN106701631B (en) | 2017-01-19 | 2017-01-19 | Streptomyces roseoflavus and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710044242.4A CN106701631B (en) | 2017-01-19 | 2017-01-19 | Streptomyces roseoflavus and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106701631A CN106701631A (en) | 2017-05-24 |
| CN106701631B true CN106701631B (en) | 2020-06-05 |
Family
ID=58909312
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710044242.4A Active CN106701631B (en) | 2017-01-19 | 2017-01-19 | Streptomyces roseoflavus and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106701631B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108192829B (en) * | 2018-01-13 | 2021-03-02 | 鲁东大学 | Streptomyces luteorusand biological control preparation and application thereof |
| CN108486184A (en) * | 2018-03-22 | 2018-09-04 | 中国农业科学院植物保护研究所 | The identification and application of rose yellow streptomycete NKZ-259 secondary metabolites IAA |
| CN110122484A (en) * | 2019-05-22 | 2019-08-16 | 中国农业科学院植物保护研究所 | Powder-type rose yellow streptomycete NKZ-259 and its to tomato growth-promoting method |
| CN114521479B (en) * | 2022-03-14 | 2023-05-26 | 河南科技大学 | Two-stage tobacco seedling raising method based on microbial growth promotion and air whole root |
| CN115956502A (en) * | 2022-09-26 | 2023-04-14 | 河南科技大学 | Application of streptomyces roseoflavus LN33 in aspect of promoting growth and development of tobacco |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050072341A (en) * | 2004-01-06 | 2005-07-11 | 학교법인고려중앙학원 | Streptomyces roseoflavus ls-a24 strain effective against plant pathogenic fungi and fungicides produced from it |
| CN1957711A (en) * | 2005-11-02 | 2007-05-09 | 河北农业大学 | Biologic bactericide and preparation method |
| CN105145634A (en) * | 2015-08-17 | 2015-12-16 | 河南科技学院 | Bactericide containing Streptomyces eurocidicus fermentation product as well as preparation method and application of bactericide |
-
2017
- 2017-01-19 CN CN201710044242.4A patent/CN106701631B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050072341A (en) * | 2004-01-06 | 2005-07-11 | 학교법인고려중앙학원 | Streptomyces roseoflavus ls-a24 strain effective against plant pathogenic fungi and fungicides produced from it |
| CN1957711A (en) * | 2005-11-02 | 2007-05-09 | 河北农业大学 | Biologic bactericide and preparation method |
| CN105145634A (en) * | 2015-08-17 | 2015-12-16 | 河南科技学院 | Bactericide containing Streptomyces eurocidicus fermentation product as well as preparation method and application of bactericide |
Non-Patent Citations (2)
| Title |
|---|
| Isolation and Antifungal and Antioomycete Activities of Staurosporine from Streptomyces roseoflavus Strain LS-A24;HEE JIN PARK et al.，;《Journal of Agricultural and Food Chemistry》;20060322;第54卷(第8期);第3041-3046页 * |
| 链霉菌F-1013的种类鉴定及其生理活性分析;林梅;《中国优秀博硕士学位论文全文数据库（硕士） 农业科技辑》;20061215(第12期);摘要，第7页第2.2节，第8页表4-3，第14页第2.1节，表5-1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106701631A (en) | 2017-05-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111304132B (en) | Microbial agent YF beneficial to growth of saline-alkali soil corns and application thereof | |
| CN111172080B (en) | Bacillus belgii and application thereof | |
| CN106701631B (en) | Streptomyces roseoflavus and application thereof | |
| CN112899201B (en) | Bacillus belgii, application thereof and method for preventing and treating banana wilt | |
| CN112175888B (en) | Bacillus belgii Hsg1949 and application thereof | |
| CN110628687B (en) | Streptomyces 5017 and application thereof in antagonism of phytopathogens | |
| CN113201474B (en) | Bacillus subtilis TBWR1, application thereof and obtained control agent | |
| CN105199996B (en) | A kind of bacillus amyloliquefaciens and its application for preventing graw mold of tomato | |
| CN112094768B (en) | Bacillus licheniformis ZF480 and application thereof | |
| CN105176894B (en) | A kind of bacillus amyloliquefaciens and its microbial bacterial agent of prevention graw mold of tomato | |
| CN108148794A (en) | A kind of the bacillus subtilis DYr3.3 and preparation method and application of broad-spectrum antibacterial activity | |
| CN107699526B (en) | Actinomycete strain for preventing and treating gray mold and application thereof | |
| CN111778173B (en) | Bacillus subtilis Pro1A2, microbial inoculum and preparation method thereof, and application of bacillus subtilis Pro1A2 in cultivation of melons | |
| CN109112069B (en) | Biocontrol endophytic fungus and application thereof | |
| CN105567600A (en) | Pathogen verticillium antagonistic bacterium and application thereof | |
| CN110317747B (en) | Bacillus amyloliquefaciens JT68 and application thereof in prevention and treatment of tea anthracnose | |
| CN113755389B (en) | Bacillus bailii and application thereof | |
| CN108641989A (en) | One plant of Methylotrophic bacillus and its application | |
| CN105754891B (en) | A kind of crop disease substance Antagonistic Actinomycetes and its screening and application | |
| CN108220211B (en) | Acinetobacter oleophilic NMB17 and application thereof in plant disease control | |
| CN116676234A (en) | Salt-tolerant bacillus JK-25, microbial inoculum, preparation method and application thereof | |
| CN115197853B (en) | Endophyte Epicoccum thailandicum LF-28 strain and application thereof | |
| CN115873770A (en) | Bacillus belgii and application thereof in prevention and treatment of tomato diseases | |
| CN105524874A (en) | Bacillus methylotrophicus strain NKG-1 and plant growth promoting method | |
| CN116240126A (en) | Multifunctional bacillus belgium SB10 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Zhang Kecheng Inventor after: Shi Liming Inventor after: Ge Beibei Inventor after: Liu Binghua Inventor after: Zhao Wenjun Inventor after: Xu Jun Inventor before: Zhang Kecheng Inventor before: Shi Liming Inventor before: Ge Beibei Inventor before: Liu Binghua Inventor before: Zhao Wenjun |
|
| CB03 | Change of inventor or designer information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |