A kind of Streptomyces leucoderma and its application

Abstract

The present invention belongs to the field of microbial technology, and specifically relates to a Streptomyces albicans and its application. The present invention provides a Streptomyces albicans, and the deposit number of the Streptomyces albicans is: CGMCCNO.30852. The present invention also provides a method for preparing a biostimulant for preventing and treating continuous cropping disease using the Streptomyces albicans. The prepared biostimulant does not contain chemical components, can reduce the incidence of continuous cropping diseases, and has the function of promoting plant growth. The results of the embodiments show that on rice, the use of the biostimulant of the present invention can reduce the incidence of continuous cropping by 68%. On tomatoes, the use of the biostimulant of the present invention can reduce the incidence of continuous cropping by 30%. On cucumbers, the use of the biostimulant of the present invention can reduce the incidence of continuous cropping by 49% and increase the yield by 7%.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and in particular relates to Streptomyces albicans and applications thereof. Background Art

[0002] The European Biostimulant Industry Alliance was established in June 2011 and gave a definition of plant biostimulants in July 2012: Plant biostimulants are substances containing certain ingredients and microorganisms, which, when applied to plants or the rhizosphere, stimulate the natural processes of plants, including enhancing / benefiting nutrient absorption, nutritional efficacy, resistance to abiotic stress and crop quality, regardless of nutritional content.

[0003] As a rich marine biological resource, seaweed contains precursors of various bioactive molecules. After undergoing microbial fermentation, these precursors can be converted into functional substances that promote plant growth and enhance plant disease resistance.

[0004] Existing crop planting technology faces some challenges: continuous cropping diseases, that is, the disease problems caused by continuous planting of the same crop on the same piece of land, seriously affect the yield and quality of crops. This phenomenon is particularly serious in greenhouse crops, because the closed environment of the greenhouse exacerbates the spread and spread of diseases. In addition, in order to prevent and control these diseases, a large number of chemical drugs are used in agricultural production, which not only pollutes the environment, but also changes the natural properties of the soil, leading to soil compaction, acidification, and a decrease in water storage capacity and nutrient retention capacity, which in turn affects the food safety of grains and vegetables.

[0005] Long-term use of chemicals may also lead to an imbalance of beneficial microbial communities in the soil, further exacerbating the problem of soil degradation. Soil degradation not only affects the growth environment of crops, but also poses a threat to the sustainability of agricultural production. Therefore, how to reduce the occurrence of crop diseases in an eco-friendly way without sacrificing yield and quality has become a technical problem that needs to be solved urgently in the field of agricultural science and technology.

[0006] Researchers in this field have tried a variety of methods to solve this problem, including crop rotation, improving soil structure, and using biological control agents. Although these methods can alleviate the disease of continuous cropping to a certain extent, they often require high cost investment or have certain limitations in actual operation. Therefore, developing a new ecological agricultural technology that is both economical and efficient and can promote crop growth and improve disease resistance is of great significance for increasing crop yields, ensuring food safety, and achieving sustainable agricultural development. Summary of the invention

[0007] The purpose of the present invention is to provide a Streptomyces albicans and an application thereof. The Streptomyces albicans provided by the present invention can use seaweed as the main fermentation substrate to prepare a preparation that does not contain chemical components, can reduce the incidence of continuous cropping diseases, and has the function of promoting plant growth.

[0008] In order to achieve the above object, the present invention provides the following technical solutions:

[0009] The invention provides a Streptomyces albicans, and the preservation number of the Streptomyces albicans is: CGMCCNO.30852.

[0010] The above strains are classified and named as: Streptomyces albidoflavus

[0011] The name of the depository of the above-mentioned strains is: China General Microbiological Culture Collection Management Center, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, deposit date: June 3, 2024.

[0012] The invention also provides a biostimulant for preventing and treating the continuous cropping disease. The biostimulant is prepared by fermenting seaweed with Streptomyces alboflavinus.

[0013] The present invention also provides a method for preparing a biostimulant for preventing and treating continuous cropping disease, which comprises the following steps: using seaweed and inorganic salts as fermentation substrates, using Streptomyces albicans as a fermentation strain, extracting the fermentation liquid after liquid fermentation, and preparing the biostimulant.

[0014] Preferably, the seaweed is Sargassum, Zostera marina or Salvia thunbergii.

[0015] Preferably, the fermentation substrate also includes corn flour and soybean flour.

[0016] Preferably, the inorganic salt comprises magnesium sulfate and manganese sulfate.

[0017] Preferably, cellulase may be added to the fermentation broth.

[0018] Preferably, the fermentation broth is further filtered to obtain a filtrate, and an emulsifier is added to the filtrate.

[0019] Preferably, the emulsifier is xanthan gum.

[0020] Preferably, the method for preparing biostimulants also includes solid fermentation.

[0021] The present invention also provides the use of the micro-white yellow Streptomyces and / or biostimulant in preventing and controlling the continuous cropping disease.

[0022] Preferably, the use also includes promoting plant growth.

[0023] Preferably, the application object is rice, tomato or cucumber.

[0024] Beneficial effects of the present invention:

[0025] The biostimulant of the present invention has the following three important beneficial effects:

[0026] (1) Environmental friendliness and disease prevention: The biostimulant of the present invention does not contain chemical ingredients and is environmentally friendly. It can also significantly reduce the incidence of continuous cropping disease in various crops such as rice, tomatoes and cucumbers by 68%, 30% and 49% respectively, which can effectively reduce the use of chemical pesticides and promote sustainable agricultural development.

[0027] (2) Promoting plant growth and increasing yield: The biostimulant of the present invention can not only prevent and treat the disease of continuous cropping, but also promote plant growth. Its application in cucumber has increased the yield by 7%, which not only reduces the incidence of crops, but also directly increases economic benefits.

[0028] (3) Efficient preparation and wide application: The preparation method of the stimulant of the present invention is simple and is prepared using mature liquid fermentation technology, which ensures the stability of the product and the high yield of active factors. It also has wide applicability to crops, providing an efficient and economical solution for agricultural production. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0030] Figure 1 It is a process flow chart.

[0031] Figure 2 The rice growth conditions of the present invention (left) and the control group (right).

[0032] Figure 3 The tomato production of the present invention (left) and the control group (right).

[0033] Figure 4 The cucumber production of the present invention (left) and the control group (right). DETAILED DESCRIPTION

[0034] The invention provides a Streptomyces albicans.

[0035] The biological deposit information of Streptomyces albidoflavus of the present invention is: deposited in the China General Microbiological Culture Collection Center, and the deposit time is June 3, 2024. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, the unit abbreviation is CGMCC, the biological deposit number is CGMCC NO.30852, and the classification name is: Streptomyces albidoflavus.

[0036] The above strain, as shown in SEQ ID NO.1, is specifically:

[0037] 16S rRNA sequencing results:

[0038] 1acgaacgctg gcggcgtgcttaacacatgc aagtcgaacg atgaaccgctttcgggcggg

[0039] 61gattagtggc gaacgggtga gtaacacgtg ggcaatctgc cctgcactct gggacaagcc

[0040] 121ctggaaacgg ggtctaatac cggatatgac tgtccatcgc atggtggatg gtgtaaagct

[0041] 181ccggcggtgc aggatgagcc cgcggcctat cagcttgttg gtgaggtagt ggctcaccaa

[0042] 241ggcgacgacg ggtagccggc ctgagagggc gaccggccac actgggactg agacacggcc

[0043] 301cagactccta cgggaggcag cagtggggaatattgcacaatgggcgaaag cctgatgcag

[0044] 361cgacgccgcg tgagggatga cggccttcgg gttgtaaacc tctttcagca gggaagaagc

[0045] 421gaaagtgacg gtacctgcag aagaagcgcc ggctaactac gtgccagcag ccgcggtaat

[0046] 481acgtagggcg caagcgttgt ccggaattat tgggcgtaa gagctcgtag gcggcttgtc

[0047] 541acgtcggttg tgaaagcccg gggcttaacc ccgggtctgc agtcgatacg ggcaggctag

[0048] 601agttcggtag gggagatcgg aattcctggt gtagcggtga aatgcgcagatatcaggagg

[0049] 661aacaccggtg gcgaaggcgg atctctgggc cgatactgac gctgaggagc gaaagcgtgg

[0050] 721ggagcgaaca ggattagata ccctggtagt ccacgccgta aacggtgggc actaggtgtg

[0051] 781ggcaacattc cacgttgtcc gtgccgcagc taacgcatta agtgccccgc ctggggagta

[0052] 841cggccgcaag gctaaaactc aaaggaattg acggggcccc gcacaagcgg cggagcatgt

[0053] 901ggcttaattc gacgcaacgc gaagaacctt accaaggctt gacatacacc ggaaacgtct

[0054] 961ggagacaggc gcccccttgt ggtcggtgta caggtggtgc atggctgtcg tcagctcgtg

[0055] 1021tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc ttgtcccgtg ttgccagcag

[0056] 1081gcccttgtgg tgctggggac tcacgggaga ccgccggggt caactcggag gaaggtgggg

[0057] 1141acgacgtcaa gtcatcatgc cccttatgtc ttgggctgca cacgtgctac aatggccggt

[0058] 1201acaatgagct gcgataccgn gaggtggagc gaatctcaaa aagccggtct cagttcggat

[0059] 1261tggggtctgc aactcgaccc catgaagtcg gagtcgctag taatcgcagatcagcattgc

[0060] 1321tgcggtgaat acgttcccgg gccttgtaca caccgcccgt cacgtcacga aagtcggtaa

[0061] 1381cacccgaagc cggtggccca accccttgtg ggagggagct gtcgaaggtg ggactggcga

[0062] 1441ttgggacgaa gtcgtaacaa ggtagccgta ccggaagg

[0063] The Streptomyces albus used in the embodiment of the present invention is the aforementioned Streptomyces albus.

[0064] The invention provides a biostimulant for reducing the incidence of repeated cropping. The preparation method comprises: using seaweed and inorganic salt as fermentation substrates, using Streptomyces albicans as fermentation strains, and preparing the biostimulant through liquid fermentation, post-treatment and extraction processes.

[0065] The present invention does not specifically limit the type of seaweed, but preferably the seaweed is Sargassum, Zostera marina or Salvia thunbergii.

[0066] The fermentation substrate of the present invention may also include corn flour, soybean flour and the like.

[0067] The inorganic salt of the present invention can be magnesium sulfate, manganese sulfate,

[0068] The method for preparing the biostimulant of the present invention also includes solid fermentation.

[0069] The biostimulant of the present invention has the effects of reducing the incidence of repeated cropping and / or promoting plant growth.

[0070] The preparation method of the biostimulant of the present invention is as follows Figure 1 As shown, specifically including:

[0071] 1. Culture of bacteria

[0072] Solid culture medium for slant or plate: potato dextrose agar.

[0073] Culture conditions: temperature 28°C, relative humidity 80%, culture in the dark.

[0074] Cultivation time: 5 to 7 days

[0075] 2. Preparation of fermentation substrate: remove impurities from seaweed such as Sargassum, dry it, and crush it into 60-80 mesh; purchase corn flour, soybean flour, magnesium sulfate, and manganese sulfate from the market, and choose fresh, mold-free, food-grade raw materials.

[0076] Add 60-65kg of crushed seaweed, 13.5-25kg of cornmeal, 14-20.5kg of soybean flour, 0.9-1kg of magnesium sulfate, and 0.1-0.2kg of manganese sulfate into 1000L of water.

[0077] Mix the above raw materials and sterilize them; cool them down and set aside.

[0078] 3. Liquid fermentation conditions: temperature 28°C, stirring speed 80-600, pH 7.0-7.2, DO 40-95%.

[0079] Liquid fermentation post-treatment: plate and frame filter pressing, collect the filtrate, add emulsifiers (xanthan gum), homogenize and mix, and package. Cellulase can also be added before filter pressing.

[0080] 4. Prepare solid fermentation substrate: the proportions of each ingredient by weight are: 80% of Sargassum, Zostera marina or Salvia thunbergii, 15% of cornmeal, 4.95% of soybean meal or peptone, 0.03% of magnesium sulfate and 0.02% of manganese sulfate.

[0081] Solid fermentation conditions: temperature 29°C, relative humidity 80%, and protected from light.

[0082] Post-processing of solid fermentation: drying at low temperature (<50°C), crushing into 60-80 mesh fineness, and packaging.

[0083] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0084] The production processes, experimental methods or detection methods involved in the embodiments of the present invention, unless otherwise specified, are all conventional methods in the prior art, and their names and / or abbreviations are all conventional names in the field, and are very clear and unambiguous in the relevant application fields. Technical personnel in the field can understand the conventional process steps based on the names and apply the corresponding equipment, and implement them according to conventional conditions or the conditions recommended by the manufacturer.

[0085] The various instruments, equipment, raw materials or reagents used in the embodiments of the present invention are not particularly limited in terms of their sources, and are all conventional products that can be purchased through regular commercial channels, or can be prepared according to conventional methods well known to those skilled in the art.

[0086] Example 1

[0087] Liquid fermentation substrate: 65kg Sargassum (crushed), 13.5kg cornmeal, 20.5kg soybean flour, 0.9kg magnesium sulfate, 0.1kg manganese sulfate, 1000L primary reverse osmosis purified water.

[0088] For laboratory seed production, potato glucose medium is used as the slant solid culture medium. After sterilization, the Streptomyces alboflavin strain taken out of the refrigerator is inoculated with aseptic operation and cultured at 28°C for 3 to 5 days. The surface of the culture medium is observed to grow light yellow to light brown colonies.

[0089] The fermentation substrate was put into the fermentation tank and mixed evenly, and sterilized at 121° C. for 40 min to obtain a liquid fermentation medium.

[0090] The liquid fermentation medium was cooled to 28°C, the colonies on the surface of the solid medium were washed off with sterile water under aseptic operation, and then inoculated into the liquid medium by flame method.

[0091] The liquid fermentation conditions were: temperature 28°C, natural pH, rotation speed 280 rpm, and culture time 5 days.

[0092] Fermentation is complete when the liquid becomes viscous and no foreign bacteria are found under microscopic examination.

[0093] Add 1.5‰ (w / v) cellulase to the fermentation broth and stir evenly at a speed of 120 rpm until the viscosity of the fermentation broth decreases significantly, which takes about 4 to 6 hours.

[0094] The fermentation broth was filtered through a plate and frame filter press and the filtrate was retained.

[0095] Add 0.5‰ (w / v) xanthan gum to the filtrate, homogenize under a pressure of 60 MPa, mix thoroughly to obtain a biostimulant mixture, and store it in a low temperature and dark place after packaging.

[0096] Select two mu of rice fields with a long-term continuous cropping: plant spacing 5 inches, row spacing 8 inches, 25 days old seedlings. Apply the above mixture to the soil at the root of the rice at a dosage of 2 kg per mu, once a month during the planting season. The rice not treated with this mixture was used as a control. The growth condition is as follows Figure 2 shown.

[0097] The incidence of rice in the example and the control during the planting process was recorded, and the incidence of each was calculated at harvest. Incidence (%) = (number of diseased plants / number of investigated plants) × 100

[0098] The number of tillers in the control example and the example plots was counted 40 days after transplantation, as shown in Table 1.

[0099] Table 1 Comparison between implementation site and control site

[0100] Grouping Plant density (approximately / mu) Survey strain Average tillering number Disease strain Embodiment 10000 1000 7.6 89 Comparative 10000 1000 7 278

[0101] After calculation, the morbidity rate of the control plot was 27.8%, and the morbidity rate of the embodiment plot was 8.9%, which was reduced by 68%. The number of tillers in the control plot was 7.0, and the number of tillers in the embodiment plot was 7.6, which was increased by 8.6%. The control group was significantly smaller in plant height than the embodiment group, and the root system was not well developed ( Figure 2 ).

[0102] Example 2

[0103] Liquid fermentation substrate: 60kg Sargassum (crushed), 25kg cornmeal, 14kg soybean flour, 0.9kg magnesium sulfate, 0.1kg manganese sulfate, 1000L primary reverse osmosis purified water.

[0104] The preparation method of the biostimulant mixed solution is the same as that of Example 1 except that the composition of the fermentation substrate is different.

[0105] A greenhouse with a high incidence of tomato diseases in previous years was selected as the test site. Drip irrigation was used and the mixture was applied at a dosage of 1.5 kg per mu. Tomatoes without the mixture were used as controls. Figure 3 shown.

[0106] The incidence rate was calculated in the same manner as in Example 1. The yield was also calculated, as shown in Table 2.

[0107] Table 2 Comparison of Example and Control

[0108] Grouping Plant density (approximately / mu) Survey strain Disease strain Output(kg) Embodiment 2500 250 30 9130 Comparative 2500 250 55 8600

[0109] According to calculations, the morbidity rate in greenhouses using this mixed solution was reduced by 45% and the yield was increased by 6%.

[0110] Example 3

[0111] The preparation method of the fermentation broth is the same as that in Example 1.

[0112] The fermentation liquid of Example 1 is mixed with a solid fermentation substrate (80% by weight of Sargassum, Zostera marina or Salvia thunbergii, 15% by weight of cornmeal, and 0.5% by weight of soybean meal or peptone) at an inoculation rate of 10% by weight of the solid fermentation substrate, and the mixture is dispensed into a plastic or metal frame with holes.

[0113] The solid fermentation material is cultured and fermented at room temperature of 29° C., relative humidity of 80%, and in the dark, during which the humidity is maintained between 75% and 85% by a humidifier.

[0114] After 7 days of solid culture, the material was dried at low temperature (<50°C).

[0115] The dried material is crushed into a powder of 60-80 mesh, which is a biostimulant powder product and is stored in a light-proof, cool warehouse.

[0116] The above powder product was used in greenhouse cucumber experiments at a dosage of 1 kg per mu. The same example and comparative example were set up. Figure 4 The morbidity statistics are shown in Table 3.

[0117] Table 3 Comparison of Example and Control

[0118] Grouping Plant density (approximately / mu) Survey strain Disease strain Output(kg) Embodiment 3000 300 38 15500 Comparative 3000 300 75 14500

[0119] The incidence rate and yield were calculated in the same manner as in Examples 1 and 2.

[0120] According to calculation, the incidence rate of the embodiment is reduced by 49% and the output is increased by 7% compared with the comparison ratio.

[0121] From the above examples, it can be seen that the biostimulant provided by the present invention can significantly reduce the incidence of continuous cropping disease in various crops such as rice, tomatoes and cucumbers, reducing them by 68%, 30% and 49% respectively. In addition, the biostimulant of the present invention can not only prevent and treat continuous cropping disease, but also promote plant growth. The application of the biostimulant in rice increases the number of tillers by 8.6%, and the application of the biostimulant in tomatoes increases the yield by 6%. The application of the biostimulant in cucumbers increases the yield by 7%.

[0122] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing a biostimulant for preventing and treating continuous cropping disease, characterized in that: The biostimulant is produced by Streptomyces albicans ( Streptomyces albidoflavus ) is prepared by fermenting seaweed, and the preservation number of the Streptomyces albicans is: CGMCC NO.30852; The preparation method of the biostimulant specifically comprises the following steps: S1. Culture of bacteria: Slant or plate solid culture medium: potato dextrose agar medium; Culture conditions: temperature 28°C, relative humidity 80%, culture in the dark; Cultivation time: 5~7 days; S2. Preparation of fermentation substrate: remove impurities from seaweed, dry it, and crush it to 60-80 mesh; add 60-65 kg of crushed seaweed, 13.5-25 kg of cornmeal, 14-20.5 kg of soybean flour, 0.9-1.0 kg of magnesium sulfate, and 0.1-0.2 kg of manganese sulfate into 1000 L of water; mix well and sterilize; cool down and set aside; S3, liquid fermentation conditions: temperature 28 °C, stirring speed 80-600 rpm, pH 7.0-7.2, DO 40-95%; Post-treatment of liquid fermentation: plate and frame filter press, collect the filtrate, add emulsifier xanthan gum, mix homogenously, and package.

2. The preparation method according to claim 1, characterized in that: The preparation method further comprises adding cellulase before the plate and frame filter pressing.

3. The use of the biostimulant prepared by the preparation method according to claim 1 in preventing and controlling plant continuous cropping diseases and / or promoting plant growth, characterized in that: The plant is rice, tomato or cucumber.

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