CN114703080B - Streptomyces fuscoporia and application thereof - Google Patents

Abstract

The invention discloses a Streptomyces fuscoporia strainStreptomyces phaeogriseichromatogenes) The strain is preserved in China Center for Type Culture Collection (CCTCC) NO: m20211271. Experiments prove that the streptomyces fuscoporia JKTJ-3 has the antibacterial capability to various pathogenic fungi, has the characteristics of phosphate solubilizing, ferrite producing, lignin degrading and indoleacetic acid producing, has the capability of producing abundant secondary metabolites, has the growth promoting effect to various crop seedlings, and is an excellent strain for developing biological pesticides and microbial agents.

Description

Streptomyces fuscoporia and application thereof

Technical Field

The invention belongs to the technical field of microorganisms, and particularly relates to streptomyces fuscoporia and application thereof.

Background

Chemical pesticides are still an effective measure at present, but with the massive use of chemical pesticides, the problems of environmental pollution, pesticide residue, enhanced pathogen resistance, increased agricultural cost and the like are caused, and sustainable and healthy development of the agriculture is affected. Along with the increasing improvement of living standard of people, green prevention and control technology mainly comprising living microorganisms and produced metabolites is increasingly valued by various countries and scientific researchers, and becomes a hot spot for research in the field of pollution-free food and green food production.

Streptomyces belongs to an important microbial resource in actinomycetes, and is called a natural cell factory for drug synthesis, and can produce abundant natural secondary metabolites with biological activity. About two thirds of the antibiotics currently used are produced by this genus. The natural metabolite not only can be applied to antibiotics, but also comprises antitumor agents, pesticides, enzyme inhibitors, pigments, enzymes and the like, and has important significance in the fields of agriculture, medical treatment, food, chemical industry, environmental protection and the like. The reported secondary metabolites produced by streptomycete at home and abroad and functions thereof are as follows: streptomyces armoidis can produce a water-soluble compound with broad-spectrum bactericidal activity, and has inhibition effect on methicillin-resistant staphylococcus aureus; the secondary metabolite obtained by extracting streptomyces VITSTK7 with ethyl acetate can inhibit aspergillus fumigatus and can be used as a good choice of antifungal drugs; arasu et al obtain a polyketide with killing and inhibiting effects on cotton bollworm and Spodoptera litura larvae from Streptomyces AP-123 secondary metabolite; qin et al isolated endophyte from Jatropha curcas with the ability to produce siderophores, indoleacetic acid, phosphorus dissolving, etc., providing seedling height, root length and fresh weight; chen Qianqian A strain of Streptomyces aureofaciens with strong inhibition effect on phytophthora nicotianae, pythium aphanidermatum and Fusarium oxysporum is found in tobacco rhizosphere soil.

No report on the growth promotion effect of Streptomyces fuscoporia (Streptomyces phaeogriseichromatogenes) on pathogenic fungi and bacteria and on crops is found, and no functional prediction and analysis on the potential of the strain for producing secondary metabolites are found. The streptomycete JKTJ-3 for strongly inhibiting the strawberry anthracnose is obtained from the strawberry anthracnose, and then the bacterial strain is found to have an antibacterial effect on various pathogenic fungi and bacteria, has the growth promoting functions of generating indoleacetic acid, siderophores, dissolving phosphorus, degrading lignin and the like, and can effectively prevent and treat lettuce sclerotinia and watermelon damping-off; the potential of the strain for producing secondary metabolites is further predicted, and 5 new secondary metabolite synthesis gene clusters are found, so that the strain can become a good bacterial source for developing microbial pesticides and microbial fertilizers and guide the preparation of antibiotics.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention provides streptomyces fuscoporia (Streptomyces phaeogriseichromatogenes) and application thereof in inhibiting various disease fungi and bacteria and promoting plant growth.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

the applicant carries out biocontrol bacterium screening to obtain a strain of actinomycetes with strong antibacterial capability on strawberry anthracnose germs, the antibacterial spectrum is wide, the strain is identified as Streptomyces fuscoporia (Streptomyces phaeogriseichromatogenes) and named as Streptomyces fuscoporia JKTJ-3, and the strain is preserved in China center for type culture collection (address: university of Wuchang mountain of Wujia in Wuhan, hubei, city) on 10 months and 14 days in 2021 through morphological characteristics and 16S rRNA gene sequencing analysis, and the preservation number is CCTCC NO: m20211271.

Use of streptomyces fuscoporia JKTJ-3 for inhibiting pathogenic fungi and bacteria: the pathogenic fungi comprise anthracnose, phoma nigrum, botrytis cinerea, globus hystericus, humic acid, verticillium, sclerotinia sclerotiorum, phoma cochinchinensis, phomopsis, rhizoctonia cerealis and fusarium; the pathogenic bacteria are Monilinia solani.

The application of Streptomyces fuscoporia JKTJ-3 or fermentation broth thereof in preventing and treating plant diseases including lettuce sclerotinia rot and watermelon damping-off.

Streptomyces fuscoporia JKTJ-3 has the capability of dissolving organic phosphorus and inorganic phosphorus, producing ferrite, degrading lignin and producing indoleacetic acid, has a growth promoting effect on plants (strawberries, cabbages, watermelons, cucumbers and corns), and is particularly favorable for forming root systems/root lumps of seedlings of plants, and improving plant height, stem thickness, biomass, leaf area, root system activity, chlorophyll and the like.

Compared with the prior art, the invention has the following advantages:

1. compared with the conventional chemical pesticides, the invention can reduce or replace the chemical pesticides to prevent and treat plant diseases, and has the characteristics of safety to people and livestock, high efficiency and environmental protection.

2. Compared with the existing fertilizer, the brown gray streptomyces (Streptomyces phaeogriseichromatogenes) JKTJ-3 provided by the invention has the functions of dissolving phosphorus, degrading lignin, producing indoleacetic acid and the like, improves the nutrient absorption of crops, promotes the growth of crops and realizes the weight-losing effect.

3. The application potential of the Streptomyces fuscoporia (Streptomyces phaeogriseichromatogenes) JKTJ-3 in disease prevention and growth promotion is reported for the first time, and a new bacterial source is provided for the development of microbial medicaments and microbial fertilizers.

4. The ability of Streptomyces fuscoporia (Streptomyces phaeogriseichromatogenes) JKTJ-3 to produce new secondary metabolites is predicted for the first time, and has guiding effect on the discovery of new natural products.

Drawings

FIG. 1 shows colony morphology of Streptomyces lividans JKTJ-3.

FIG. 2 shows the microscopic morphology of Streptomyces lividans JKTJ-3. The left panel shows the form of the aerial hyphae and spore chains of the JKTJ-3 and the right panel shows the form of spores.

FIG. 3 shows the inhibitory effect of Streptomyces lividans JKTJ-3 culture and fermentation product on the hyphae of strawberry anthracnose.

FIG. 4 shows the inhibition of pathogenic fungi by Streptomyces lividans JKTJ-3.

FIG. 5 shows the inhibition of Streptomyces lividans JKTJ-3 against Monascus nigra.

FIG. 6 shows that Streptomyces lividans JKTJ-3 has phosphate solubilizing, lignin degrading and ferrite producing properties. A is Meng Jinna organophosphorus medium, B is Meng Jinna inorganic phosphorus medium, C is CAS medium, and D is aniline blue-PDA medium.

FIG. 7 shows the indoleacetic acid production characteristics of Streptomyces lividans JKTJ-3.

FIG. 8 shows the growth promoting effect of Streptomyces lividans JKTJ-3 kernel culture on strawberry tissue culture seedlings by volatile gas.

FIG. 9 shows the growth promoting effect of Streptomyces lividans JKTJ-3 kernel culture on cabbage seedlings.

FIG. 10 shows the growth promoting effect of Streptomyces lividans JKTJ-3 fermentation broth on maize seedlings.

FIG. 11 shows the control effect of Streptomyces lividans JKTJ-3 ferments on in vitro leaves of Brevibacterium lettuce.

FIG. 12 shows the control effect of Streptomyces fuscogenus JKTJ-3 fermentation product on Pythium type damping-off of watermelon.

FIG. 13 is a prediction of a novel terpene synthase type synthetic gene cluster of Streptomyces lividans JKTJ-3.

FIG. 14 is a prediction of a novel RiPP-like synthase type synthetic gene cluster of Streptomyces lividans JKTJ-3.

FIG. 15 is a synthetic gene cluster prediction of novel lanthipeptide of Streptomyces lividans JKTJ-3.

FIG. 16 is a synthetic gene cluster prediction of novel lanthopeptide-class-iii of Streptomyces lividans JKTJ-3.

Detailed Description

Example 1: isolation and identification of Streptomyces lividans JKTJ-3

1. In the process of separating strawberry anthracnose bacteria and screening biocontrol bacteria in 2021 month 1, a strain of contaminated streptomyces with strong inhibition effect on strawberry anthracnose bacteria is discovered. The spores were picked up and added to sterile water, diluted 10 times and plated to obtain a pure culture, numbered JKTJ-3. As shown in FIG. 1, purified Streptomyces producing yellow water-soluble pigments were obtained on ISP-2 plates, with aerial spore masses ranging from grey-white to grey-brown in color and matrix hyphae ranging from yellow to yellow-brown in color. Yellow water-soluble pigment is produced on Gaoshi culture No. 1, the aerial spore block is little or no, and the matrix hypha is yellow. The growth temperature is 12-45 ℃, the proper temperature is 24-40 ℃, and the proper pH is 4.5-7. The aerial hyphae, spore chains and spores of the JKTJ-3 were observed by scanning electron microscopy and are shown in fig. 2.

2. The physiological and biochemical index of the strain JKTJ-3 was measured, and the results are shown in Table 1.

TABLE 1 physiological and biochemical characteristics of strain JKYJ-3

+: a positive reaction; -: a negative reaction; weak positive response W

3. PCR amplification and sequencing are carried out on the 16S rRNA gene of the strain JKTJ-3, the obtained DNA fragment is connected with a T carrier by using a T carrier kit, positive clones are screened, and the positive clones are sent to a biological company for sequence sequencing, wherein the sequence is shown as SEQ ID NO. 1. According to the primer, the target fragment is selected, and sequence similarity comparison is carried out in EzbioCloud to obtain a result, and the strain JKTJ-3 has 99.4 percent of similarity with streptomyces brown gray producing fungus (Streptomyces phaeogriseichromatogenes). Based on morphological observation, physiological and biochemical assay results in combination with 16S rRNA gene sequencing and sequence alignment analysis results, the strain JKTJ-3 was identified as Streptomyces fuscogenus (Streptomyces phaeogriseichromatogenes). The applicant has submitted the Chinese typical culture collection to be preserved in 2021 for 10 months and 14 days, and the classification name is Streptomyces fuscoporia (Streptomyces phaeogriseichromatogenes) with the preservation number of CCTCC NO: M20211271.

Example 2: preparation of Streptomyces lividans JKTJ-3 culture and fermentation product

(1) Plate strain: PDA culture medium (fresh peeled potato 200g, glucose 20g, agar powder 13g, tap water 1000 ml). Culturing Streptomyces fuscoporia JKTJ-3 by streak culture on PDA plate, and culturing at 28deg.C for 7d to obtain first seed. Spores were scraped with 10ml of sterile water per dish, and 10 was prepared with sterile water ⁸ CFU/ml spore liquid.

(2) Wheat grain culture: soaking wheat grains with clear water overnight, removing soaking solution, adding tap water, boiling, frying, filtering, air drying, packaging into triangular flask, and sterilizing at 121deg.C for 60min. 1ml of 10 is taken ⁸ The spore liquid of CFU/ml is connected to wheat culture medium, and cultured for 10-15d at 28 ℃ to obtain the JKTJ-3 wheat culture.

(3) Liquid fermentation product: PDB liquid medium (fresh peeled potato 200)g, glucose 20g, tap water 1000 ml). 1ml of 10 is inoculated per 100ml of liquid medium ⁸ CFU/ml spore liquid was shake cultured at 28℃for 4d at 220rpm to prepare a fungus-containing fermentation broth. The sterilized fermentation broth was filtered with filter paper and then filtered with bacterial filter (diameter=45 μm) to prepare a sterilized fermentation broth. Thus obtaining two liquid fermentation products.

(4) Crude extract of fermentation product: the sterile fermentation broth was extracted by isovolumetric mixing with ethyl acetate, allowed to stand until the aqueous and organic phases were completely separated, then filtered, the organic phase was retained, rotary evaporated (50 ℃) until dry, and the dry material was dissolved with a small amount of methanol. Thus obtaining the crude extract of the fermented product.

Example 3: antibacterial effect of Streptomyces fuscoporia JKTJ-3 culture and fermentation product on strawberry anthracnose

The mycelium blocks of the strawberry anthracnose bacteria are taken by a puncher with the diameter of 8mm, placed in the center of a PDA flat plate, and simultaneously are streaked and cultured to produce brown gray streptomycete JKTJ-3 at a position 3cm away from the mycelium blocks, and the steps are repeated for 5 times. Culturing at 28deg.C, and measuring antibacterial size after 6 d.

3 holes are drilled at equal distance from the center of the PDA plate to 2.5cm, 100 μl of sterile fermentation product and crude extract of the fermentation product are respectively added into each hole, the center of the plate is connected with strawberry anthracnose hypha blocks with the diameter of 8mm, and the bacteria inhibition size is measured after culturing at 28 ℃ for 6 d.

As a result, as shown in FIG. 3, the culture and fermentation product of Streptomyces lividans JKTJ-3 have a very strong inhibitory effect on strawberry anthracnose.

Example 4: antibacterial effect of Streptomyces fuscoporia JKTJ-3 culture on various pathogenic fungi

Taking phoma tibioflavus, botrytis cinerea, melon coccidioides (melon), saprotifer, verticillium, sclerotium (capsicum), anthrax (capsicum), asparagus phoma, phomopsis, rhizoctonia, melon globus hystericus (watermelon) and fusarium colony edge mycelium blocks by using a puncher with the diameter of 8mm, placing the mycelium blocks in the circle center of a PDA flat plate, and culturing the streptomyces fuscoporia JKTJ-3 on the flat plate at a position 3cm away from the mycelium blocks by scribing. The antibacterial width was measured after 6d at 28℃and the results are shown in Table 2 and FIG. 4. As can be seen from Table 2 and FIG. 4, streptomyces fuscoporia JKTJ-3 has a broad antibacterial spectrum and a strong antibacterial effect.

TABLE 2 antibacterial breadth of Streptomyces fuscogenus JKTJ-3 against plant pathogenic fungi

Pathogenic bacteria	Antibacterial width (mm)	Pathogenic bacteria	Antibacterial width (mm)
				Phoma tibiof	21.5	Anthrax fungus (Pepper)	18.5
Botrytis cinerea	11	Asparagus cochinchinensis (Willd.) Makino	16.5
				Melon ball cavity fungus (melon)	18	Phomopsis (Fr.) Kummer	15.2
Pythium species	23.2	Rhizoctonia cerealis	21
				Verticillium sp	19	Melon ball cavity bacterium (watermelon)	15
Sclerotium (Pepper)	17.8	Fusarium (Fusarium)	12.1

Example 5: antibacterial effect of Streptomyces fuscoporia JKTJ-3 sterile fermentation broth on Gan Lanhei Monilinia putrescens

200 μl of 10 strength was uniformly spread on PDA plates with a sterile triangular spatula ⁸ CFU/ml of the cabbage black rot monospore bacillus liquid, then 3 holes are drilled at equal distance from the center of the PDA plate by 2.5cm, 100 μl of sterile fermentation liquid is added into each hole, and the mixture is cultured at 28 ℃ for 3 days, and then the antibacterial size is measured. As shown in FIG. 5, it can be seen that the aseptic fermentation broth of Streptomyces lividans JKTJ-3 has strong antibacterial effect on Gan Lanhei Monilinia solani.

Example 6: streptomyces lividans JKTJ-3 has the characteristics of phosphate dissolution, lignin degradation, and ferrite and indoleacetic acid production

And (3) respectively inoculating the Streptomyces fuscoporia JKTJ-3 to Meng Jinna organic phosphorus and inorganic phosphorus culture mediums, culturing for 7d at 28 ℃, and observing whether the edges of the colony have transparent rings or not to judge whether the colony has phosphate dissolving capability or not. Spot inoculated on a universal CAS detection plate, dark cultured at 28 ℃ for 2-7d, and whether the strain has the capacity of producing siderophores or not is judged according to whether the strain produces a distinct orange-yellow siderophore chelation ring on the plate. Spot inoculated on a universal aniline blue-PDA plate, dark culture at 28 ℃ for 2-7d, and judging whether the strain has lignin decomposing capability according to whether the blue color of the colony edge is faded. As shown in FIG. 6, streptomyces lividans JKTJ-3 has a phosphate-solubilizing function (FIG. 6-A, B), a ferrite-producing function (FIG. 6-C) and a lignin-degrading function (FIG. 6-D).

Taking 1ml of the mixture with the concentration of 10 ⁸ CFU/ml brown ash productionInoculating Streptomyces chromogenes JKTJ-3 spore liquid into PDB liquid culture medium, shaking at 220rpm at 28 ℃ for 24 hours, centrifuging the fermentation liquid at 4000rpm for 10 minutes, taking 100 mu L of supernatant, adding an equal volume of Salkowski colorimetric agent, rapidly darkening for 30 minutes, observing whether a chromogenic reaction exists, and primarily judging that the fermentation liquid has the capacity of producing indoleacetic acid (IAA) if the fermentation liquid shows red. Shake culture was performed at 220rpm and 28℃for 4d in PDB medium (200 mg/L of L-tryptophan was added). 1mL of the JKTJ-3 fermentation broth was centrifuged at 4000rpm for 10min, and the supernatant was taken, an equal volume of Salkowski colorimeter was added, and after rapid darkening for 30min, the absorbance of the sample was measured at OD 530. The IAA yield of the strain was determined according to a standard curve. As shown in FIG. 7 and Table 3, streptomyces lividans JKTJ-3 can produce indoleacetic acid.

TABLE 3 IAA content in the JKTJ-3 fermentation broths

Treatment of	IAA content
		JKTJ-3 (tryptophan free)	12.66mg/L
JKTJ-3 (containing 200mg/L tryptophan)	317.45mg/L

Example 7: promoting growth of strawberry tissue culture seedling by volatile gas produced by Streptomyces fuscoporia JKTJ-3 culture

Sterilized wheat grains were placed in a sterile uncapped 1.5ml centrifuge tube and inoculated at a concentration of 10. Mu.l ⁸ CFU/ml Streptomyces fuscoporia JKTJ-3 spore liquid is put into a strawberry tissue culture bottle, and the cover of the tissue culture bottle is tightly sealed by a sealing film to prevent the gas in the bottle from volatilizing. Measurement of strawberry tissue culture after 30d of cultureFresh weight of seedling, root number and root activity. The results are shown in Table 4 and FIG. 8, and the JKTJ-3 wheat grain culture produced volatile gas has growth promoting effect on strawberry tissue culture seedlings.

TABLE 4 growth promoting effect of JKTJ-3 culture-produced volatile gas on strawberry tissue culture seedlings

Treatment of	Fresh weight/g	Number of roots/number of	Root system vitality/ug/(g.h)
				JKTJ-3	2.12±0.11	17.0±2.2	45.09±1.21
CK	1.45±0.21	12.2±1.9	20.00±0.97

Example 8: promoting growth of cucumber plug seedling by Streptomyces lividans JKTJ-3 sterile fermentation liquid

And respectively diluting the aseptic fermentation liquor of the streptomyces fuscoporia JKTJ-3 with aseptic water for 30 times, and mixing with a seedling substrate for standby according to the volume ratio of 1:20. Cucumber seeds were sterilized with 1% formaldehyde and sown in 50-hole trays. Each treatment was repeated 5 times, each as a plug. 7 plants are randomly selected every time after one month, the plant height, the stem thickness, the fresh weight and the dry weight are measured, and meanwhile, the TTC method and the spectrophotometry method are respectively adopted to measure the root system activity and the chlorophyll content.

As can be seen from tables 5 and 6, when the substrate is treated with the JKTJ-3 fermentation liquid, cucumber seedlings are grown, and the stem thickness, fresh weight, dry weight, root-crown ratio, strong seedling index, leaf area, root length, root volume, chlorophyll and root system activity of the cucumber seedlings are all higher than those of the control, the aseptic fermentation liquid of the Streptomyces fuscus JKTJ-3 has good growth promoting effect on cucumber plug seedlings.

TABLE 5 influence of the treatment of the substrate with the sterile fermentation broth of JKTJ-3 on the morphological index of cucumber seedlings

TABLE 6 influence of the treatment of the substrate with the sterile fermentation broth of JKTJ-3 on the leaf area, root system and physiological index of cucumber seedlings

Example 9: promoting growth of seedling of watermelon plug by using aseptic fermentation liquid of Streptomyces fuscoporia JKTJ-3

Example 9 the procedure was as in example 8. As can be seen from Table 7, the aseptic fermentation broth of the Streptomyces fuscoporia JKTJ-3 has good growth promoting effect on the plug seedlings of the watermelons, wherein the plant height, the stem thickness, the fresh weight, the dry weight, the root-to-crown ratio and the strong seedling index of the watermelon seedlings are higher than those of the control after the aseptic fermentation broth of the JFKTJ-3 is used for treating the substrate.

TABLE 7 Effect of JKTJ-3 sterile fermentation broth treatment Medium on watermelon seedling morphology index

Example 10: promoting growth of Streptomyces lividans JKTJ-3 wheat cultures on cabbage seedling

50kg of the culture of the JKTJ-3 wheat grains is added into each cubic matrix, and the mixture is uniformly mixed and then is put into a 90-type flowerpot for raising the seedlings of the cabbage. After 30d, the measurement index was similar to that of example 8. As can be seen from Table 8 and FIG. 9, the plant height, stem thickness, fresh weight, dry weight, and strong seedling index of the cabbage seedlings were all higher than those of the control, and it was found that Streptomyces fuscogenus JKTJ-3 wheat cultures had good growth promoting effect on the cabbage seedlings.

TABLE 8 growth promoting effect of JKTJ-3 wheat culture treatment substrate on cabbage seedlings

Treatment of	Plant height/cm	Stem thickness/mm	Fresh weight/g on ground	Underground fresh weight/g	Dry weight/g on ground	Dry weight/g under ground	Total dry weight/g	Index of strong seedlings
									JKTJ-3	4.19	9.04	17.49	2.66	2.09	0.27	2.36	0.39
CK	3.03	8.30	11.90	2.08	1.45	0.18	1.63	0.32

Example 11: promoting growth of corn seedling by treating substrate with Streptomyces lividans JKTJ-3 bacteria-containing fermentation liquid

Example 11 the procedure was as in example 8. As can be seen from Table 9 and FIG. 10, the seedling cultivation of corn is carried out after the substrate is treated by the JKTJ-3 bacteria-containing fermentation liquor, and the plant height, the stem thickness, the fresh weight, the dry weight, the root cap ratio, the strong seedling index, the chlorophyll and the root system activity of the corn seedlings are all higher than those of the control, so that the seedling cultivation of the Streptomyces fuscoporia JKTJ-3 bacteria-containing fermentation liquor corn plug has good growth promoting effect.

TABLE 9 growth promoting effect of JKTJ-3 fermentation broth treatment substrates on maize seedlings

Example 12: control effect of Streptomyces fuscoporia JKTJ-3 sterile fermentation liquor on Sclerotinia lettuce

The sterilized toilet paper was laid in a tray and wetted with sterilized water. And then taking 5-6 pieces of lettuce seedling leaves, cleaning and sucking the leaves, spraying a JKTJ-3 sterile fermentation liquid, and placing the leaves in a tray. Punching sclerotium fungus mycelium blocks along the edge of fungus filaments by using a puncher with the diameter of 5mm, picking the mycelium blocks, inoculating the mycelium blocks to the middle part of lettuce leaves and deviating from the main vein, tightly attaching the mycelium-containing surfaces to the leaves, covering the bottom of the leaves by using sterile toilet paper after inoculation, covering and moisturizing by using a preservative film, placing the leaves in a climatic chamber with the temperature of 22 ℃ and the relative humidity of 95% for dark culture, measuring the size of a disease spot after 3d, and calculating the relative prevention and treatment effect according to the following formula. The results are shown in Table 10 and FIG. 11, and the relative control effect of the JKTJ-3 ferment on the in vitro leaves of the Lactuca sativa is up to 83.9%.

Table 10 relative control of the JKTJ-3 ferments against Lactuca sativa in vitro leaves

Example 13: fusarium brown producing streptomyces JKTJ-3 bacteria-containing fermentation liquor for preventing and treating pythium watermelon damping-off

1kg of Streptomyces fuscoporia JKTJ-3 bacteria-containing fermentation broth is added per cubic meter of matrix, the mixture is uniformly mixed, then the mixture is put into a 90-percent flowerpot, 1 percent formaldehyde sterilized watermelon seeds are sown, and the watermelon seeds are covered with a film at 30 ℃ for moisturizing and accelerating germination. After the seeds arch the soil, the flower pot is inoculated with a pythium mycelium block with the diameter of 6mm, and the relative humidity is controlled to be more than 95 percent by continuously coating a film at 28 ℃ for moisture preservation. Counting the number of seedlings after sowing for 3d, counting the number of disease seedlings after 5-7d, and calculating the control effect. As can be seen from Table 11 and FIG. 12, the JKTJ-3 bacteria-containing fermentation broth improves the emergence rate and the seedling rate of the watermelons, and the damping-off prevention effect of the watermelons reaches 94.7%.

TABLE 11 control of Pythium type damping-off of watermelon by JKTJ-3 fermentation product

Embodiment case 14: analysis of secondary metabolite produced by Streptomyces lividans JKTJ-3

Sequencing the genome of Streptomyces lividans JKTJ-3 using PacBio platform and Illumina PE150 platform at Tianjin Nor He information technologies Inc., querying and predicting the cluster of secondary metabolite synthetic genes of the strain using the anti SMASH database, two new clusters of synthetic genes encoding terpene synthases types were found, the core region of the first cluster of synthetic genes of terpene synthases type being about 19.1kb and the second about 21.7kb, predicting that the new terpene secondary metabolites might exert inhibitory effects on pathogenic bacteria, pests, weeds, etc., as shown in FIG. 13. A novel ribosome-like dependent synthetic gene cluster (RiPP-like) with a core region of about 11.3kb, as shown in fig. 14; a new lasspoptide type synthetic gene cluster, the core region of which is approximately 21.3kb, as shown in fig. 15; a novel synthetic gene cluster of lanthipatide-class-iii, the core region of which is approximately 22.7kb, is shown in FIG. 16. Predicting a series of biological activities such as receptor antagonism, enzyme inhibition and antibacterial activity of the new synthetic gene cluster tool of the last three. None of these five synthetic gene clusters were included in the biosynthetic gene cluster databases MIBIG and NCBI. Therefore, the strain has the potential of being used for preparing biocontrol bactericides and plant growth-promoting bacteria, and simultaneously suggests that the strain can be used for synthesizing antibiotics.

Sequence listing

<110> academy of agricultural sciences in Wuhan City

<120> Streptomyces fuscoporia and application thereof

<160> 1

<170> SIPOSequenceListing 1.0

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<213> Artificial sequence (Artificial Sequence)

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tgcagtcgaa cgatgaagcc cttcggggtg gattagtggc gaacgggtga gtaacacgtg 60

ggcaatctgc cctgcactct gggacaagcc ctggaaacgg ggtctaatac cggatatgac 120

catcttgggc atccttgatg gtgtaaagct ccggcggtgc aggatgagcc cgcggcctat 180

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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 acgtcgattg tgaaagctcg gggcttaacc 540

ccgagtctgc agtcgatacg ggctagctag agtgtggtag gggagatcgg aattcctggt 600

gtagcggtga aatgcgcaga tatcaggagg aacaccggtg gcgaaggcgg atctctgggc 660

cattactgac gctgaggagc gaaagcgtgg ggagcgaaca ggattagata ccctggtagt 720

ccacgccgta aacggtggga actaggtgtt ggcgacattc cacgtcgtcg gtgccgcagc 780

taacgcatta agttccccgc ctggggagta cggccgcaag gctaaaactc aaaggaattg 840

acgggggccc gcacaagcgg cggagcatgt ggcttaattc gacgcaacgc gaagaacctt 900

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caggtggtgc atggctgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg 1020

agcgcaaccc ttgtcccgtg ttgccagcag gcccttgtgg tgctggggac tcacgggaga 1080

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ttgggctgca cacgtgctac aatggccggt acaatgagct gcgataccgt gaggtggagc 1200

gaatctcaaa aagccggtct cagttcggat tggggtctgc aactcgaccc catgaagtcg 1260

gagtcgctag taatcgcaga tcagcattgc tgcggtgaat acgttcccgg gccttgtaca 1320

caccgcccgt cacgtcacga aagtcggtaa cacccgaagc cggtggccca accccttgtg 1380

ggaggga 1387

Claims (5)

1. Streptomyces griseus producingStreptomyces phaeogriseichromatogenes) The JKTJ-3 is characterized by being preserved in China Center for Type Culture Collection (CCTCC) NO: M20211271.

2. The use of streptomyces fuscoporia JKTJ-3 according to claim 1 for inhibiting pathogenic fungi, wherein the pathogenic fungi are anthrax, phoma nigrum, botrytis cinerea, globus citrulli, pythium, verticillium, sclerotinia sclerotiorum, phoma asparagi, phomopsis, rhizoctonia solani or fusarium.

3. Use of streptomyces fuscoporia JKTJ-3 according to claim 1 for inhibiting pathogenic bacteria, characterized in that said pathogenic bacteria is monospora nigroca.

4. The use of Streptomyces lividans JKTJ-3 or a fermentation broth thereof for controlling plant diseases according to claim 1, wherein the plant diseases are lactuca sativa sclerotinia or watermelon damping-off.

5. Use of streptomyces fuscoporia JKTJ-3 or a fermentation broth thereof according to claim 1 for promoting plant growth.