CN114317328B - Streptomyces aureomycin and application thereof - Google Patents
Streptomyces aureomycin and application thereof Download PDFInfo
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- CN114317328B CN114317328B CN202111487735.8A CN202111487735A CN114317328B CN 114317328 B CN114317328 B CN 114317328B CN 202111487735 A CN202111487735 A CN 202111487735A CN 114317328 B CN114317328 B CN 114317328B
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- streptomyces
- ginseng
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- aureomycin
- aureofaciens
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The streptomyces aureomycin provided by the invention has strong inhibition activity on the main root rot fungi of pseudo-ginseng, namely rust rot fungi, can effectively reduce the occurrence of the rust rot of pseudo-ginseng when being applied to the root of pseudo-ginseng as a microbial inoculum, and improves the emergence rate of pseudo-ginseng planted in continuous cropping soil and the fresh weight of a single plant, thereby having extremely high economic value.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to streptomyces aureomycin capable of inhibiting activity of pseudo-ginseng root rot fungi and application thereof.
Background
Notoginseng radix [ Panax notoginseng (Burk.) is a plant of Panax of Araliaceae, and has effects of promoting blood circulation, removing blood stasis, relieving swelling and pain. Modern pharmacological and clinical researches show that notoginseng has very good curative effect on cardiovascular and cerebrovascular diseases. At present, the application of pseudo-ginseng is very wide, statistics show that more than 1300 enterprises only take pseudo-ginseng as raw materials in China, 3207 production lots are obtained, more than 400 products are produced, wherein the number of Chinese patent medicine varieties exceeds 300, and almost all Chinese medicine pharmaceutical enterprises are covered.
The continuous cropping obstacle (consecutive monoculture problem) is a phenomenon that when the same or similar crops are continuously cultivated in the same land, even under normal cultivation and management conditions, the vigour becomes weak, the yield is reduced, the quality is reduced, and the pest and disease damage becomes worse (Zhang Chongyi, 2009, etc.). At present, the research on continuous cropping obstacle in China is mainly focused on wheat, corn, sorghum, soybean, eggplant, cucumber and the like serving as grain crops, and the research on continuous cropping obstacle of medicinal plants is relatively few. In the conventional cultivated medicinal plants, about 70% of root (rhizome) medicinal materials are contained (Zhang Aihua, etc., 2011). However, most root (rhizome) medicinal plants have continuous cropping obstacle phenomenon during cultivation, such as rhizoma Atractylodis, rehmanniae radix, ginseng radix, notoginseng radix, radix Pseudostellariae, coptidis rhizoma, radix Angelicae sinensis, etc. (Xia Pinhua, etc., 2010). With the advancement of standardized cultivation of medicinal plants and the continuous cropping obstacles becoming a technical bottleneck in the process of cultivation of medicinal plants (Guo Lanping, etc., 2006).
As with other ginseng medicinal plants, the planting of pseudo-ginseng has extremely strong earth-repellent property and has serious continuous cropping obstacle. It is generally considered that plots planted with pseudo-ginseng require more than 10 years of continuous planting of crops such as corn, upland rice, alfalfa, etc., to be planted again (Jian Zaiyou, etc., 2009). According to the early investigation of the subject group, the pseudo-ginseng can be planted again in some plots even with rotation intervals of up to 30 years. The driving of market demands and the temporary lack of continuous cropping obstacles effectively overcome the defect that the proper land of the traditional production area of pseudo-ginseng is gradually tense, the planting area is continuously extended, and the trafficability of medicinal material production is difficult to guarantee.
The cultivation period of the pseudo-ginseng is generally 3 years, and the seedling stage is 1 year; after transplanting, the seedlings need to grow for 2 years again, namely, continuous cropping is carried out for two years, wherein the seedlings grow for 1 year after transplanting and are called as two years and seven years, and the seedlings grow for 2 years and are called as three years and seven years. In production, seven plants of two years and seven years planted after continuous cropping are serious in pest and disease damage and low in plant survival rate, and the method is one of the expression characteristics of the continuous cropping obstacle of pseudo-ginseng. On the other hand, if the seedlings of the pseudo-ginseng are transplanted again (continuous cropping) on the land after the harvest of seven years, the incidence rate of the pseudo-ginseng reaches more than 90 percent, and almost no healthy plants exist after one year, namely the seedling is another expression characteristic of continuous cropping obstacle of the pseudo-ginseng. Thus, notoginseng radix shows typical continuous cropping obstacle characteristics-serious disease. The main diseases in the continuous cropping process of pseudo-ginseng include root rot, black spot, powdery mildew, damping off, epidemic disease and the like (Wang Chaoliang and the like, 2000). The incidence rate of root rot can reach 50%, and the root rot is a nearly destructive disease which damages the growth of the root system of pseudo-ginseng (wandering, 2009). Therefore, development of a technical scheme capable of inhibiting diseases and continuous cropping obstacles of pseudo-ginseng is urgently needed, and economic value of planting pseudo-ginseng in unit soil is improved.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides streptomycete and application thereof, which are used for solving the problems of serious diseases and reduced yield and quality caused by continuous cropping obstacles in pseudo-ginseng planting in the prior art. In order to solve the technical problems, the invention specifically provides the following technical scheme:
in a first aspect of the present invention, there is provided streptomyces aureomycin (Streptomyces aureofaciens) 90372, the preservation information of which is as follows: the preservation number is CGMCC No.22755; the classification is named: streptomyces aureomycin (Streptomyces aureofaciens); the preservation unit is China general microbiological culture Collection center; the preservation address is Beijing city, the North Chen Xiyu No. 1 and 3, the post code: 100101.
in a second aspect of the present invention, there is provided a microbial agent for inhibiting a primary root rot fungus of pseudo-ginseng, the microbial agent comprising the above streptomyces aureofaciens (Streptomyces aureofaciens) 90372. In a preferred embodiment, the viable count of streptomyces aureomycin (Streptomyces aureofaciens) 90372 in the microbial inoculum is 10 6 ～10 8 cfu/mL, most preferably, the viable count of streptomyces aureomycin (Streptomyces aureofaciens) 90372 in the microbial inoculum is 10 8 cfu/mL。
The third aspect of the invention provides an application of streptomyces aureofaciens (Streptomyces aureofaciens) 90372 or a microbial inoculum prepared from the streptomyces aureofaciens in resisting rust rot of pseudo-ginseng.
In one embodiment, the above-described use is directed to reducing the occurrence of root rot in pseudo-ginseng.
In a preferred embodiment, the above application comprises applying streptomyces aureomycin (Streptomyces aureofaciens) 90372 to root soil of planted panax notoginseng, wherein the number of viable streptomyces aureomycin (Streptomyces aureofaciens) 90372 is 10 6 ～10 8 cfu/mL, most preferably, the viable count of Streptomyces aureofaciens (Streptomyces aureofaciens) 90372 at the time of administration is 10 8 cfu/mL。
In a fourth aspect, the invention provides an application of streptomyces aureofaciens (Streptomyces aureofaciens) 90372 or a microbial inoculum prepared from the streptomyces aureofaciens in promoting the growth of pseudo-ginseng.
In one embodiment, the above application refers to inhibiting the occurrence of a pseudo-ginseng continuous cropping obstacle.
In another embodiment, the above application refers to improving emergence rate and fresh weight of the individual plants of the pseudo-ginseng.
In a preferred embodiment, the above application comprises applying streptomyces aureomycin (Streptomyces aureofaciens) 90372 to the root soil of the planted panax notoginsengIn the soil, the number of viable bacteria of streptomyces aureofaciens (Streptomyces aureofaciens) 90372 is 10 when the composition is applied 6 ～10 8 cfu/mL, most preferably, the viable count of Streptomyces aureofaciens (Streptomyces aureofaciens) 90372 at the time of administration is 10 8 cfu/mL。
Compared with the prior art, the invention has the following beneficial effects:
the streptomyces aureomycin (Streptomyces aureofaciens) provided by the invention can grow by using dextrin, D-galactose, N-acetyl-D-glucosamine, alpha-D-glucose, pectin and D-gluconic acid as carbon sources; can grow in an environment with pH=5, pH=6, or containing 1% NaCl; the compound pesticide composition has strong inhibition activity on sodium lactate, rifamycin SV, minocycline, lincolmycin, guanidine hydrochloride, tetrazole violet, tetrazole blue, nalidixic acid, potassium tellurite, aztreonam and sodium butyrate, and has strong inhibition activity on main root rot fungi-rust rot fungi of pseudo-ginseng, can effectively reduce the occurrence of rust rot of pseudo-ginseng when being applied to root of pseudo-ginseng as a microbial inoculum, and can improve the emergence rate and the fresh weight of single plant of pseudo-ginseng planted in continuous cropping soil, thereby having extremely high economic value.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate the invention and together with the embodiments of the invention, serve to explain the invention. In the drawings:
fig. 1 is a colony chart of streptomyces aureofaciens 90372 provided in example 1 of the present invention.
Fig. 2 is a gram staining chart of streptomyces aureofaciens 90372 provided in example 1 of the present invention.
FIG. 3 shows the effect of Streptomyces aureofaciens 90372 on fresh weight of Notoginseng radix.
Detailed Description
The invention will be further illustrated with reference to specific examples. It should be understood that the detailed description and specific examples are intended for purposes of illustration only and are not intended to limit the scope of the invention.
The specific techniques or conditions are not identified in the examples and are described in the literature in this field or are carried out in accordance with the product specifications. The reagents or equipment used were conventional products available for purchase through regular channels, with no manufacturer noted.
The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the examples described below, unless otherwise specified, are all commercially available products.
Example 1: isolation of strains and identification of physiological and biochemical functions
The Streptomyces sp is separated from healthy Notoginseng radix rhizosphere soil by adopting a plate separation method, is numbered 90372, and has the following specific results: the bacteria are gram-negative bacteria, chain-shaped, yellow bacterial colony, round and raised edge. The method is characterized in that alpha-D-glucose, D-galactose, D-mannitol, vinegar bamboo peach mycin, rifamycin SV, L-aspartic acid, L-glutamic acid, L-arginine, L-histamine, lincolamycin, guanidine hydrochloride, tetradecsodium sulfate, D-gluconic acid, mucic acid, quinic acid, vancomycin, tetrazolium blue, tetrazolium violet, citric acid, L-malic acid, gamma-aminobutyric acid and sodium bromate are utilized, and the method accords with the morphological, physiological and biochemical characteristics of streptomycete. The strain is preserved in China general microbiological culture collection center (CGMCC) at the year of 2021, 6 and 23, with the preservation number of CGMCC No.22755; the classification is named: streptomyces aureomycin (Streptomyces aureofaciens); the preservation address is Beijing city, the North Chen Xiyu No. 1 and 3, the post code: 100101.
the 90372 strain was examined for its ability to utilize 71 carbon source substrates on a Biolog Gen III microplate and its sensitivity to 23 chemicals, and the measurement results are shown in Table 1:
TABLE 1
+: indicating a positive reaction; -: indicating a negative reaction.
As is clear from Table 1, strain 90372 was able to grow using dextrin, D-galactose, N-acetyl-D-glucosamine, alpha-D-glucose, pectin, and D-gluconic acid as carbon sources; can grow in an environment with pH=5, pH=6, or containing 1% NaCl; is insensitive to sodium lactate, rifamycin SV, minocycline, lincolmycin, guanidine hydrochloride, tetrazolium violet, tetrazolium blue, nalidixic acid, potassium tellurite, aztreonam, sodium butyrate.
Example 2 test of bacteriostatic ability of Streptomyces aureofaciens 90372
In this example, 4 species of rust rot fungi (Cylindrocarpon destructans) were supplied from plant pathology laboratory, university of Yunnan agricultural university, plant protection institute.
The detection method comprises the following steps:
the inhibition activity of streptomyces aureofaciens 90372 on the growth of the mycelium of the test rust rot fungi is measured by adopting a plate counter culture method. Preparing bacterial cakes on a super clean workbench by using a puncher with the diameter of 5mm, transferring the bacterial cakes to the center of a PDA culture medium flat plate, inoculating the separated and purified streptomyces aureofaciens to a position 2.5cm away from pathogenic bacteria by using a sterile toothpick, setting 10 times of repetition, taking the flat plate with the pathogenic bacteria cakes as a control, culturing at the constant temperature of 26 ℃, measuring the bacterial colony diameter when the control bacterial colony grows to 2/3 of the culture medium, and calculating the bacteriostasis rate according to the following test. Antibacterial ratio (%) = [ (control colony radius-counter culture colony radius)/control colony radius ] ×100%.
Wherein, the formula of the PDA culture medium is as follows: 200g of potato, 20g of glucose, 18g of agar powder and 1000mL of water.
As shown in Table 2, the inhibition capacities of streptomyces aureofaciens 90372 on 4 kinds of test rust rot fungi are different, and the inhibition rate is up to more than 70%. The strain has strong inhibition activity on the pseudo-ginseng main root rot fungus rust rot fungus.
TABLE 2
EXAMPLE 3 promotion of emergence of Notoginseng radix seed by Streptomyces aureofaciens 90372
1. Preparation of bacterial liquid
Activating streptomyces aureomycin 90372 on KB solid culture medium, picking single colony, shaking culturing in KB liquid fermentation culture medium at 160rpm and 28 ℃ for 48h, and diluting the fermentation liquid to OD 600 =0.3 for subsequent field rooting.
Wherein the KB solid medium has the following formula per 1L of medium: 20g peptone, 10mL glycerol, 1.5gK 2 HPO 4 ·3H 2 0，1.5g MgSO 4 ·7H 2 O,17g agar, 1000mL distilled water, pH 7.0.
2. Influence of Streptomyces 90372 on emergence of Notoginseng radix seed
The test arrangement is carried out in a pseudo-ginseng planting shed of a farm in a river of a seek. Part of the pseudo-ginseng continuous cropping soil is subjected to heat sterilization at 121 ℃ for 20min, and the other part of the pseudo-ginseng continuous cropping soil is not treated. All the continuous cropping soil is respectively placed into flowerpots, 12 pseudo-ginseng seeds are planted in each flowerpot, and each treatment is repeated 5 times. Sampling and investigation are carried out three months later, and the emergence rate of the plants is measured and recorded.
As a result, the results shown in Table 3 were obtained, and the emergence rate of Notoginseng radix was increased after root irrigation with Streptomyces aureofaciens 90372 under soil sterilization/non-sterilization conditions.
TABLE 3 Effect of Streptomyces 90372 on emergence of Notoginseng seeds
3. Influence of Streptomyces 90372 on fresh weight of Notoginseng radix
Planting Notoginseng radix seeds in a flowerpot filled with sterilized soil, 12 plants per bottle, and inoculating suspension of streptomyces aureofaciens 90372 into the flowerpot, wherein the content of streptomyces aureofaciens 90372 viable bacteria in the suspension is 10 6 ～10 8 cfu/mL, 25mL per bottle, with the same volume of liquid KB medium added as a control, after 3 months of culture under natural conditions in each treatment 5 pots, the individual fresh weight of the recorded plants was measured.
As a result, as shown in FIG. 3, the average fresh weight of the Notoginseng radix was increased after root irrigation of Streptomyces aureofaciens 90372 in sterilized soil.
It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.
Claims (10)
1. The streptomyces aureomycin (Streptomyces aureofaciens) 90372 is characterized in that the preservation number of the strain is CGMCC No.22755, the preservation unit is China general microbiological culture Collection center, the preservation address is North Chen Xiyu No. 1, 3 in the Korean region of Beijing, and the postal code is 100101.
2. A microbial agent for inhibiting main root rot fungi of pseudo-ginseng, characterized in that the microbial agent comprises streptomyces aureofaciens (Streptomyces aureofaciens) 90372 according to claim 1.
3. The microbial agent according to claim 2, wherein the number of viable bacteria of streptomyces aureomycin (Streptomyces aureofaciens) 90372 in the microbial agent is 10 6 ～10 8 cfu/mL。
4. The microbial preparation according to claim 3, wherein the number of viable bacteria of streptomyces aureomycin (Streptomyces aureofaciens) 90372 in the microbial preparation is 10 8 cfu/mL。
5. Use of streptomyces aureomycin (Streptomyces aureofaciens) 90372 according to claim 1 or a microbial agent according to any one of claims 2-4 for combating rust rot of notoginseng.
6. The use according to claim 5, wherein the use is to reduce the occurrence of root rot of pseudo-ginseng.
7. Use of streptomyces aureomycin (Streptomyces aureofaciens) 90372 according to claim 1 or a microbial agent according to any one of claims 2-4 for promoting the growth of pseudo-ginseng.
8. The use according to claim 7, wherein the use is for inhibiting the occurrence of a pseudo-ginseng continuous cropping obstacle.
9. The use according to claim 7, wherein the use is to increase the emergence rate and the fresh weight of the individual plants of pseudo-ginseng.
10. The use according to any one of claims 5 to 9, wherein the use comprises applying streptomyces aureomycin (Streptomyces aureofaciens) 90372 to the root soil of planted pseudo-ginseng, the number of viable streptomyces aureomycin (Streptomyces aureofaciens) 90372 being 10 6 ～10 8 cfu/mL。
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| US5527526A (en) * | 1993-06-30 | 1996-06-18 | Idaho Research Foundation, Inc. | Use of streptomyces bacteria to control plant pathogens |
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