A zymovar strain for producing zymovar and application thereof

Abstract

This invention relates to a oryzanol-producing strain and its application, wherein the oryzanol-producing strain is Streptomyces. Streptomyces sp.YDZW-4702, the oryzanol-producing strain, has high oryzanol production activity and application value.

Description

Technical Field

[0001] This invention belongs to the field of agricultural technology, specifically relating to a strain of oryzanol that produces oryzanol and its application. Background Technology

[0002] Currently, several strains capable of producing oryzanol have been reported in existing technologies, but they generally suffer from two major shortcomings: firstly, the yield is generally low, making cost control difficult; secondly, the spectrum of action of the fermentation products is relatively narrow, resulting in limited efficacy against pests and diseases and failing to broadly cover the diverse needs of agricultural production. These key deficiencies at the strain level have restricted the application and promotion of oryzanol and related products. Summary of the Invention

[0003] To address the aforementioned technical problems, this application provides a oryzanol-producing strain, which exhibits high oryzanol yield and unexpected insecticidal activity.

[0004] According to one aspect of this application, a oryzanol-producing strain is provided, said oryzanol-producing strain being Streptomyces. Streptomyces sp. YDZW-4702;

[0005] The oryzanol-producing strain is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36558.

[0006] The Streptomyces Streptomyces sp. YDZW-4702 not only produces oryzanol, but its fermentation products also have a wide range of applications, which is conducive to promoting industrial development.

[0007] According to one aspect of this application, a oryzanol-producing strain is provided, said oryzanol-producing strain being Streptomyces. Streptomyces sp. YDZW-4702 satisfies any one or a combination of the following:

[0008] (1) Contains a molecular marker, the nucleotide sequence of which is shown in SEQ ID NO: 1;

[0009] (2) Contains a molecular marker, wherein the nucleotide sequence of the molecular marker has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher homology with the nucleotide sequence shown in SEQ ID NO: 1;

[0010] (3) The genomic DNA was amplified using the primers shown in SEQ ID NO: 2 and SEQ ID NO: 3. The amplification products were analyzed by agarose gel electrophoresis to obtain specific bands.

[0011] (4) The genomic DNA was amplified using the primers shown in SEQ ID NO: 2 and SEQ ID NO: 3. The amplified products were sequenced and analyzed to obtain the nucleotide sequence shown in SEQ ID NO: 1; or

[0012] The obtained nucleotide sequence has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher homology with the nucleotide sequence shown in SEQ ID NO: 1.

[0013] According to one aspect of this application, a molecular marker is provided, the nucleotide sequence of which is shown in SEQ ID NO: 1; or

[0014] The nucleotide sequence of the molecular marker has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher homology with the nucleotide sequence shown in SEQ ID NO: 1.

[0015] According to one aspect of this application, the use of the aforementioned molecular marker in identifying the aforementioned oryzanol-producing strains is provided.

[0016] According to one aspect of this application, a primer pair is provided, wherein the nucleotide sequence of the forward primer is shown in SEQ ID NO:2;

[0017] The nucleotide sequence of the reverse primer is shown in SEQ ID NO: 3.

[0018] According to one aspect of this application, the use of the above primer pair in identifying the above-mentioned oryzanol-producing strains is provided.

[0019] According to one aspect of this application, a method for identifying the above-mentioned oryzanol-producing strain is provided, characterized by comprising the following steps:

[0020] (1) Sequencing of the oryzanol-producing strain;

[0021] (2) Obtain a sequence containing a specific molecular marker, wherein the specific molecular marker satisfies any one or a combination of the following:

[0022] 1) The nucleotide sequence is as shown in SEQ ID NO: 1; and / or

[0023] 2) The nucleotide sequence has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher homology with the nucleotide sequence shown in SEQ ID NO: 1.

[0024] Optionally, the sequencing is selected from:

[0025] (1) Genome sequencing; and / or

[0026] (2) Sequencing of PCR products.

[0027] Optionally, the PCR product is obtained by amplification using the primers shown in SEQ ID NO: 2 and SEQ ID NO: 3.

[0028] According to one aspect of this application, the use of the above-described oryzanol-producing strain in the preparation of pesticide products is provided.

[0029] According to one aspect of this application, a pesticide product is provided which contains the fermentation product of the above-mentioned oryzanol-producing strain.

[0030] Optionally, the pesticide product is selected from any one of fermentation stock solution, fermentation dilution solution, fermentation concentrate, technical grade, mother drug, and formulation.

[0031] Optionally, the oryzanol content of the fermentation stock solution, fermentation dilution solution, or fermentation concentrate solution is 10-300000 μg / mL;

[0032] The oryzanol content of the technical grade and parent drug is 5-99%;

[0033] The preparation contains 0.01-30% oryzanol by mass.

[0034] Optionally, the dosage form of the formulation is selected from any one of powder, granules, oil, and seed treatment suspension.

[0035] Optionally, the dosage form of the formulation is selected from sustained-release formulations.

[0036] Optionally, the dosage form of the formulation is selected from suspensions.

[0037] Optionally, the formulation is selected from suspension seed coating agents.

[0038] Optionally, the dosage form of the formulation is selected from any one of seed treatment dispersible powder, seed treatment soluble powder, seed treatment liquid, seed treatment emulsion, and seed treatment microcapsule suspension.

[0039] Optionally, the dosage form of the formulation is selected from any one of the following: large granules, fine granules, microparticles, microcapsules, wettable powders, oil-dispersible powders, water-dispersible granules, emulsions, effervescent granules, dispersible tablets, effervescent tablets, sustained-release blocks, sustained-release tubes, sustained-release granules, soluble powders, soluble granules, soluble tablets, soluble solutions, aqueous solutions, soluble gels, spreading oils, ultra-low volume liquids, ultra-low volume microcapsule suspensions, emulsions, latexes, dispersible liquids, pastes, concentrated gels, water emulsions, oil emulsions, microemulsions, liposomes, microcapsule suspensions, oil suspensions, and suspension emulsions.

[0040] According to one aspect of this application, a method for preparing the above-mentioned pesticide product is provided, the method comprising the following steps:

[0041] Using Streptomyces Streptomyces The pesticide product was obtained by fermentation with sp. YDZW-4702.

[0042] Optionally, the preparation method includes the following steps:

[0043] (1) Utilizing Streptomyces Streptomyces Fermentation with sp. YDZW-4702 yields the fermentation stock solution, which is the pesticide product described above; or

[0044] The preparation method further includes the following steps:

[0045] (2) Further processing of the fermentation liquid to obtain any one of the following: fermentation dilution, fermentation concentrate, technical material, mother drug, or formulation, which is the pesticide product.

[0046] According to one aspect of this application, the use of the above-described strain, the above-described pesticide product, or the pesticide product obtained according to the above-described method in regulating plant growth or controlling plant diseases and pests is provided.

[0047] According to one aspect of this application, the use of the above-described strain, the above-described pesticide product, or the pesticide product obtained according to the above-described method in the preparation of plant growth regulators or plant disease and pest control agents is provided.

[0048] The positive and beneficial effects achieved by this application are as follows:

[0049] (1) Streptomyces provided in this application Streptomyces sp. YDZW-4702 has high activity in oryzanol production and can effectively help save energy and increase efficiency in oryzanol production.

[0050] (2) Streptomyces provided in this application Streptomyces sp. YDZW-4702 has a wider range of control effects in insecticides and fungicides, which is conducive to expanding the application scope of the product and promoting industrial development.

[0051] Preservation instructions:

[0052] Streptomyces Streptomyces sp. YDZW-47 was deposited at the China General Microbiological Culture Collection Center on November 10, 2025, and classified as Streptomyces. Streptomyces sp., with accession number CGMCCNo.36557.

[0053] Streptomyces Streptomyces sp. YDZW-4702 was deposited at the China General Microbiological Culture Collection Center on November 10, 2025, and classified as Streptomyces. Streptomyces sp., with accession number CGMCCNo. 36558. Attached Figure Description

[0054] Figure 1 Streptomyces Streptomyces Phylogenetic tree of sp. YDZW-47's 16S rDNA;

[0055] Figure 2 Streptomyces Streptomyces Phylogenetic tree of sp. YDZW-4702's 16S rDNA;

[0056] Figure 3 The HPLC chromatogram of oryzanol standard;

[0057] Figure 4 Streptomyces Streptomyces HPLC chromatogram of YDZW-47 fermentation broth;

[0058] Figure 5 Streptomyces Streptomyces HPLC chromatogram of the fermentation broth of sp. YDZW-4702;

[0059] Figure 6 The H-NMR spectrum of component 2;

[0060] Figure 7 The control effects of various strains on spider mites are shown in the graph.

[0061] Figure 8 The agarose gel electrophoresis results are for the amplification products of SEQ ID NO: 2 and SEQ ID NO: 3. Detailed Implementation

[0062] For the purposes of this invention, unless otherwise stated, the terms used in this application have the following meanings:

[0063] The term "active ingredient" refers to the component in pesticide products that has toxic and killing activity against diseases, insects, and weeds, or that has a regulatory function on plant growth.

[0064] The term "raw material" refers to the product obtained during the production process, which consists of active ingredients and related impurities, and may include a small amount of additives if necessary.

[0065] The term "mother drug" refers to the product obtained during the production process, which consists of the active ingredient and related impurities, and may contain small amounts of necessary additives and appropriate diluents.

[0066] The term "formulation" refers to a stable product made from pesticide technical (mother drug) and suitable adjuvants, or processed by methods such as bio-fermentation and plant extraction.

[0067] The term "plant diseases and pests" refers to two types of disasters caused by biological factors during the growth and development of plants, specifically including diseases and pests. Diseases are caused by the infection of pathogenic organisms, resulting in infectious damage to plants, significantly hindering their growth and development, causing pathological changes in their internal and external structures, physiology, and tissue structure, leading to symptoms such as wilting and deformities. Pathogenic organisms include, but are not limited to, fungi and bacteria. Pests are caused by harmful organisms, resulting in damage to plants through feeding, boring, and other forms of damage, significantly hindering their growth and development, causing damaging changes in their external tissue structure, leading to symptoms such as notches and holes. Harmful organisms include, but are not limited to, insects and mites.

[0068] The term "plant growth regulator" refers to products that regulate plant development processes and alter the growth and development status of plants. Their effects include, but are not limited to, promoting or inhibiting germination, rooting, flower bud differentiation, flowering, fruiting, and leaf drop, enhancing crop resistance to adverse environments such as drought, low temperature, and salinity, and alleviating pesticide and fertilizer damage.

[0069] The reagents and solvents used in the following examples are commercially available or prepared using conventional methods known to those skilled in the art. Experimental methods in the following examples, unless otherwise specified, were performed according to conventional methods and conditions.

[0070] Preparation of PDA medium: Weigh 200 g of potato, wash, peel and chop it, add 800 ml of distilled water and boil for half an hour, filter with gauze, then add 20 g of glucose and 20 g of agar, and make up to 1000 ml with distilled water, and sterilize at 121℃ for 30 minutes.

[0071] Gao's No. 1 agar medium: 20g soluble starch, 0.5g NaCl, 1g KNO3, 0.01g FeSO4·7H2O, 0.5g K2HPO4, 0.5g MgSO4·7H2O, 20g agar, 1000mL distilled water, pH 7.2-7.4.

[0072] YMS medium: 10g soluble starch, 4g yeast extract, 10g malt extract, add purified water to make up to 1000ml, pH 7.0, 18g agar, sterilize at 121℃ for 20min.

[0073] Seed culture medium: 10g glucose, 10g sucrose, 15g fried soybean meal powder, 15g cottonseed meal powder, 2g CaCO3, 1000ml purified water, pH 7.0 before sterilization, filled into 25ml / 250ml Erlenmeyer flasks, sterilized at 121℃ for 20min.

[0074] Fermentation medium: 60g corn soluble starch, 20g glucose, 20g fried soybean meal powder, 20g cottonseed meal powder, 2g light calcium carbonate, 2g sodium chloride, 2g magnesium sulfate heptahydrate, 1000ml purified water, pH 6.0 before sterilization, filled into 25ml / 250ml Erlenmeyer flasks, sterilized at 121℃ for 20min.

[0075] The liquid chromatography detection method (HPLC) refers to "T / FJAASS 025—2025 Oryzanol Technical Grade".

[0076] Genomic DNA was extracted using a bacterial genomic DNA extraction kit from Tiangen Biotech Co., Ltd., following the instructions provided.

[0077] Specific amplification was performed using PrimeSTAR® Max DNA Polymerase, a rapid PCR polymerase from Takara Bio Inc., and the amplification system is as follows:

[0078]

[0079] The amplification procedure is as follows:

[0080]

[0081] The Streptomyces strains used in this application are:

[0082]

[0083] Example 1: Isolation and Identification of Strains

[0084] 1. Strains Isolation

[0085] 10 g of soil sample from the State-owned Gu Nong Farm in Changtai District, Zhangzhou City, Fujian Province was dissolved in 90 mL of sterile water and serially diluted with sterile water to 100 times and 1000 times. An appropriate amount of the 1000-fold dilution was spread on Gao's No. 1 agar plates and incubated at 28°C for 3 days. Single colony A was screened from numerous single colonies for further purification and culture. Under sterile conditions, the spores of the single colony purified and cultured on Gao's No. 1 agar were scraped and transferred to Erlenmeyer flasks containing fermentation medium and incubated at 30°C and 200 rpm for 5 days to obtain the fermentation broth for later use.

[0086] Using the same method, single colony B was screened from numerous single colonies isolated from soil samples at the southern foot of Yaoshan Mountain in Pucheng County, Shaanxi Province, and fermentation broth was prepared for later use.

[0087] 2. Strain identification

[0088] The 16S rDNA identification method was used to identify the above single colonies A and B. The specific identification steps are as follows:

[0089] (1) Genomic DNA extraction: Single colony A and single colony B were activated and cultured on PDA medium plates, and then genomic DNA was extracted.

[0090] (2) 16S rDNA amplification: Using genomic DNA as a template, 16S rDNA fragments were amplified from genomic DNA using universal 16S rDNA primers (27F sequence as shown in SEQ ID NO: 4 and 1492R sequence as shown in SEQ ID NO: 5). The amplification products were sent to Beijing Liuhe BGI Genomics Co., Ltd. for analysis.

[0091] (3) 16S rDNA sequence analysis: The 16S rDNA sequence of the single colony A obtained from sequencing was compared with the 16S rDNA sequences in GenBank using BLAST. The 16S rDNA sequences of the top ten strains were selected, and a phylogenetic tree was constructed using Mega 5.0 software. The results showed that the 16S rDNA sequences of single colony A and GenBank were compared with those of GenBank. Streptomyces sp. strain D101 clustered on one branch (see...) Figure 1 The 16S rDNA sequence of a single colony B obtained from sequencing was compared with the 16S rDNA sequences in GenBank using BLAST. The 16S rDNA sequences of the top ten strains were selected, and a phylogenetic tree was constructed using Mega 5.0 software. The results showed that single colony B and... Streptomyces sp strain FJL194 , Streptomyces decoyicus strain NRRL 2666 Streptomyces ossamyceticus strain NBRC 13983 clustered on one branch (see...) Figure 2).

[0092] Based on the above identification results, single colony A and single colony B were identified as belonging to the genus Streptomyces. Streptomyces sp., of which single colony A was named Streptomyces. Streptomyces sp. YDZW-47, a single colony B was named Streptomyces. Streptomyces sp. YDZW-4702. Deposited at the China General Microbiological Culture Collection Center, Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Streptomyces. Streptomyces sp. YDZW-47 has the accession number CGMCC No. 36557; Streptomyces Streptomyces sp. YDZW-4702 has the accession number CGMCC No.36558.

[0093] Example 2 Identification of Active Substances

[0094] 1. Liquid chromatography detection

[0095] Streptomyces prepared in Example 1 Streptomyces sp. YDZW-47, Streptomyces Streptomyces sp.YDZW-4702, Streptomyces globosum ( Streptomyces caniferus The fermentation broth of NEAU6 was analyzed by liquid chromatography, and the active substances were identified using oryzanol standards. The HPLC chromatogram of the standards is shown below. Figure 3 As shown, the peak time of the standard oryzanol was 4.491; Streptomyces... Streptomyces The HPLC chromatogram of the YDZW-47 fermentation broth is as follows: Figure 4 As shown, component 1 has a peak elution time of 4.544; Streptomyces. Streptomyces The HPLC chromatogram of the fermentation broth of sp. YDZW-4702 is as follows: Figure 5 As shown, component 2 has a peak elution time of 4.469. Based on the localization analysis of the above oryzanol standard, the results indicate that both component 1 and component 2 are oryzanol.

[0096] 2. Identification of the structure of the active ingredient

[0097] Separation, concentration, and drying were performed on Streptomyces using gel column chromatography. Streptomyces sp. YDZW-47, Streptomyces Streptomyces Components 1-2 were obtained from the fermentation broth of sp. YDZW-4702. After mass spectrometry (MS) and nuclear magnetic resonance (NMR) analysis, it was finally determined that components 1-2 were both oryzanol.

[0098] The H-NMR spectrum of component 2 is as follows Figure 6 As shown.

[0099] The above results indicate that Streptomyces Streptomyces sp. YDZW-47, Streptomyces Streptomyces sp.YDZW-4702 can produce oryzanol.

[0100] Example 3: Determination of oryzanol fermentation level

[0101] (1) Streptomyces Streptomyces sp. YDZW-47, Streptomyces Streptomyces sp. YDZW-4702, Streptomyces globosum ( Streptomyces caniferus NEAU6 was inoculated onto YMS plates and cultured at 28°C for 10 days;

[0102] (2) Scrape the colony spores on the plate in step (1) and inoculate them into the seed culture medium. Under the conditions of 25℃, 200rpm and 48 hours of culture, culture on a shaker until the pH of the seed liquid is 7.0 and the mycelial concentration is 20% to complete the seed liquid culture.

[0103] (3) Inoculate the seed liquid into the fermentation medium at a volume ratio of 10%, and then place it in a shaker and culture it under the conditions of 25℃ and 200rpm for 200h to obtain the fermentation liquid.

[0104] Streptomyces were detected using high-performance liquid chromatography (HPLC). Streptomyces sp. YDZW-47, Streptomyces Streptomyces sp. YDZW-4702, Streptomyces globosum ( Streptomyces caniferus The fermentation broth of NEAU6 was tested for the content of oryzanol. The results showed that the content of oryzanol in the three fermentation broths was 3998 μg / ml, 4316 μg / ml and 3913 μg / ml, respectively.

[0105] The results above show that Streptomyces Streptomyces The fermentation level of sp. YDZW-4702 was significantly higher than that of Streptomyces. Streptomyces sp. YDZW-47 and Streptomyces simulans ( Streptomyces caniferus NEAU6.

[0106] Example 4 Other Activity Verification

[0107] 1. Different strains of Ralstonia solanacearum Ralstonia solanacearum Inhibitory effect

[0108] Take 1 mL of the Streptomyces prepared in Example 3. Streptomyces sp. YDZW-47, Streptomyces Streptomycessp. YDZW-4702, Streptomyces globosum ( Streptomyces caniferus Filter the NEAU6 fermentation broth to remove the bacterial cells, dilute it 100 times, and set it aside.

[0109] Take 200 mL of the thawed and cooled culture medium to 48 ℃~50 ℃, and add 1 mL of 1×10 -8 A CFU / mL suspension of Ralstonia solanacearum was shaken well, poured into a petri dish, and allowed to solidify to obtain a bacterial layer.

[0110] Take a petri dish containing a bacterial layer, add three sterile filter paper discs, and drop the above diluted solution onto each disc. Repeat three times. Incubate the petri dish at 28°C for 48 hours. After incubation, measure the diameter of the inhibition zone using the cross-sectional method and calculate the average value.

[0111] The statistical results are shown in the table below:

[0112]

[0113] 2. Control effects of different strains on spider mites

[0114] The Streptomyces prepared in Example 3 were used respectively Streptomyces sp. YDZW-47, Streptomyces Streptomyces sp. YDZW-4702, Streptomyces globosum ( Streptomyces caniferus Filter the NEAU6 fermentation broth to remove the bacterial cells, dilute it 100 times, and set it aside.

[0115] Sixty adult spider mites of uniform size were collected using a brush and placed in a potted white kidney bean plant with two true leaves. The mites were allowed to move freely for 5 hours, and the number of live mites was counted. The diluted solution was then evenly sprayed onto both sides of the kidney bean leaves using a handheld sprayer until the leaves were moist but no liquid droplets were dripping. The treated potted plants were then placed in a greenhouse environment with a temperature of 26±1℃, humidity of 60%-70%, and a light intensity (L:D) of 16h:8h. Water was used as a blank control. Each treatment was repeated three times. The number of live mites was surveyed after 7 days, and the control effect was calculated. The calculation formula is:

[0116] Mite reduction rate (%) = (Number of live mites before application - Number of live mites after application) / Number of live mites before application * 100;

[0117] Control efficacy (%) = (reduction rate of live mites in the treatment - reduction rate of live mites in the blank control) / (100 - reduction rate of live mites in the blank control) * 100.

[0118] The growth of spider mites on the leaves of each treatment is as follows: Figure 7 As shown, the specific statistical data is as follows:

[0119]

[0120] The above results indicate that Streptomyces Streptomyces sp. YDZW-4702 has a significant control effect on spider mites, while Streptomyces simonii (…) Streptomyces caniferus NEAU6, Streptomyces Streptomyces sp. YDZW-47 has a weak effect on spider mites, indicating that Streptomyces... Streptomyces sp. YDZW-4702 has unique insecticidal activity.

[0121] Example 5 Streptomyces Streptomyces SCAR molecular marker analysis of sp. YDZW-4702

[0122] Streptomyces Streptomyces sp. YDZW-47, Streptomyces Streptomyces sp. YDZW-4702, Streptomyces lilacinus Hainan variant ( Streptomyces lavendulae var. hainanensis) UV-11, Streptomyces hainanensis ( Streptomyces lavendulae var.hainanensis new var.) 46#, Streptomyces Streptomyces sp. ZS-Ⅱ-D190515, Streptomyces MKL-2 ( Streptomyces microaureaus MKL-2), Streptomyces simonii ( Streptomyces caniferus NEAU6, Streptomyces StreptomycesGenomic DNA from eight Streptomyces strains sp. 2356 was used as templates. Twenty random primers were used (TGCCGAGCTG (SEQ ID NO: 6), AGTCAGCCAC (SEQ ID NO: 7), AATCGGGCTG (SEQ ID NO: 8), AGGGGTCTTG (SEQ ID NO: 9), GTTTCGCTCC (SEQ ID NO: 10), CATCCCCCCTG (SEQ ID NO: 11), CTGCTGGGAC (SEQ ID NO: 12), GTGAGGCGTC (SEQ ID NO: 13), CCGCATCTAC (SEQ ID NO: 14), GTGTGCCCCA (SEQ ID NO: 15), GGACCCAACC (SEQ ID NO: 16), TTGGCACGGG (SEQ ID NO: 17), AGCGCCATTG (SEQ ID NO: 18), GGTGCGGGAA (SEQ ID NO: 19), GTGACATGCC (SEQ ID NO: 20), TGCGCCCTTC (SEQ ID NO: 21), GGACTGCAGA (SEQ ID NO: 22), GGACTGCAGA (SEQ ID NO: 2356 ... Streptomyces (SEQ ID NO: 22), CTCACCGTCC (SEQ ID NO: 23), GGTGATCAGG (SEQ ID NO: 24), CTGAGACGGA (SEQ ID NO: 25) were randomly amplified. The amplified products were analyzed by agarose gel electrophoresis. Streptomyces The specific band of sp. YDZW-4702 was cut, recovered, and sequenced to obtain the sequence of the specific band.

[0123] Primers were designed based on the obtained unique band sequences (forward primer as shown in SEQ ID NO: 2; reverse primer as shown in SEQ ID NO: 3). Genomic DNA from the above eight Streptomyces strains was used as templates for amplification. The amplification products were analyzed by agarose gel electrophoresis, and the results are as follows: Figure 8 As shown, only Streptomyces Streptomyces sp. YDZW-4702 amplified a specific band (lane 1 in the figure), while the other 7 strains failed to amplify specific bands, verifying that primers (SEQ ID NO: 2 and SEQ ID NO: 3) can specifically identify Streptomyces. Streptomyces sp. YDZW-4702.

[0124] SEQ ID NO: 2 and SEQ ID NO: 3 specifically amplified Streptomyces StreptomycesThe specific band obtained from sp. YDZW-4702 was recovered and sequenced, and the sequence is shown in SEQ ID NO: 1.

[0125] Blast alignment of the specific band sequence shown in SEQ ID NO: 1 in GenBank revealed no identical or similar sequences among all publicly available Streptomyces strains listed in NCBI (No significant similarity found). This further demonstrates that the specific sequence shown in SEQ ID NO: 1 can be used to identify Streptomyces. Streptomyces sp. YDZW-4702.

[0126] The above results indicate that primers SEQ ID NO: 2 and SEQ ID NO: 3 are effective for identifying Streptomyces. Streptomyces The specific primers for strain YDZW-4702, and the PCR amplification product sequence, as shown in SEQ ID NO: 1, are for Streptomyces. Streptomyces The SCAR molecular marker for sp. YDZW-4702. Primers SEQ ID NO: 2 and SEQ ID NO: 3, as well as the specific band sequence (SEQ ID NO: 1), can be rapidly and effectively identified. Streptomyces sp.YDZW-4702.

[0127] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A strain for producing oryzanol, characterized in that, The oryzanol-producing strain is Streptomyces. Streptomyces sp. YDZW-4702; The oryzanol-producing strain is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36558.

2. The oryzanol-producing strain according to claim 1, characterized in that, The oryzanol-producing strain is Streptomyces. Streptomyces sp. YDZW-4702 satisfies any one or a combination of the following: (1) Contains a molecular marker, the nucleotide sequence of which is shown in SEQ ID NO: 1; (2) Genomic DNA was amplified using the primers shown in SEQ ID NO: 2 and SEQ ID NO:

3. The amplification products were analyzed by agarose gel electrophoresis to obtain specific bands. (3) The genomic DNA was amplified using the primers shown in SEQ ID NO: 2 and SEQ ID NO:

3. The amplified products were sequenced and analyzed, and the nucleotide sequence obtained was shown in SEQ ID NO:

1.

3. The use of the oryzanol-producing strain according to any one of claims 1-2 in the preparation of pesticide products.

4. A pesticide product, characterized in that, It contains the fermentation product of the oryzanol-producing strain according to any one of claims 1-2.

5. The pesticide product according to claim 4, characterized in that, The pesticide product is selected from any one of the following: fermentation stock solution, fermentation dilution solution, fermentation concentrate, technical grade pesticide, mother drug, and formulation.

6. The pesticide product according to claim 5, characterized in that, The oryzanol content of the fermentation stock solution, fermentation dilution solution, or fermentation concentrate solution is 10-300000 μg / mL. The oryzanol content of the technical grade and parent drug is 5-99%; The preparation contains 0.01-30% oryzanol by mass.

7. The pesticide product according to claim 5, characterized in that, The dosage form of the preparation is selected from any one of powder, granules, oil, and seed treatment suspension.

8. The pesticide product according to claim 5, characterized in that, The dosage form of the preparation is selected from sustained-release formulations.

9. The pesticide product according to claim 5, characterized in that, The dosage form of the preparation is selected from suspensions.

10. The pesticide product according to claim 5, characterized in that, The formulation is selected from suspension seed coating agents.

11. The pesticide product according to claim 5, characterized in that, The dosage form of the formulation is selected from any one of seed treatment dispersible powder, seed treatment soluble powder, seed treatment liquid, seed treatment emulsion, and seed treatment microcapsule suspension.

12. The pesticide product according to claim 5, characterized in that, The dosage form of the formulation is selected from any one of the following: large granules, fine granules, microparticles, microcapsules, wettable powders, oil-dispersible powders, water-dispersible granules, emulsions, effervescent granules, dispersible tablets, effervescent tablets, sustained-release blocks, sustained-release tubes, sustained-release granules, soluble powders, soluble granules, soluble tablets, soluble solutions, aqueous solutions, soluble gels, spreading oils, ultra-low volume liquids, ultra-low volume microcapsule suspensions, emulsions, latexes, dispersible liquids, pastes, concentrated gels, water emulsions, oil emulsions, microemulsions, liposomes, microcapsule suspensions, oil suspensions, and suspension emulsions.

13. A method for preparing the pesticide product according to any one of claims 4-12, characterized in that, The preparation method includes the following steps: Using the Streptomyces as described in claim 1 Streptomyces The pesticide product was obtained by fermentation with sp. YDZW-4702.

14. The preparation method according to claim 13, characterized in that, The preparation method includes the following steps: (1) Using the Streptomyces as described in claim 1 Streptomyces Fermentation with sp. YDZW-4702 yields the fermentation stock solution, which is the pesticide product described above; or The preparation method further includes the following steps: (2) Further processing of the fermentation liquid to obtain any one of the following: fermentation dilution, fermentation concentrate, technical material, mother drug, or formulation, which is the pesticide product.

15. Use of the strain according to any one of claims 1-2, the pesticide product according to any one of claims 4-12, or the pesticide product obtained by the method according to any one of claims 13-14 in regulating plant growth or controlling plant diseases and pests.

16. Use of the strain according to any one of claims 1-2, the pesticide product according to any one of claims 4-12, or the pesticide product obtained by the method according to any one of claims 13-14 in the preparation of plant growth regulators or plant disease and pest control agents.

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