A herbicide *Storkia stolonifera* and its applications
Patent Information
- Application Number
- CN202611383413.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-09-08
- Publication Date
- 2026-10-20
Smart Images
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of agricultural biotechnology, specifically relating to a herbicide *Storkia stolonifera* and its applications. Background Technology
[0002] Weeds affect crop growth through resource competition, severely reducing crop yield and quality. Currently, the most common method for controlling weeds in the field is chemical herbicide control. However, long-term use of chemical herbicides can lead to herbicide resistance in weeds and cause environmental pollution. Biological control utilizes biocontrol microorganisms, including bacteria and fungi, as microbial herbicides to infect weeds, thereby achieving weed control in the field. Biological control is receiving increasing attention in the field of weed control due to its green, environmentally friendly, and agriculturally sustainable advantages.
[0003] Goosegrass (Eleusine indica (L.) Gaertn.), crabgrass (Digitaria sanguinalis (L.) Scop.), barnyard grass (Echinochloa crus-galli (L.) P. Beauv.), and foxtail grass (Setaria viridis (L.) P. Beauv.) are common field weeds, and there are reports of using biocontrol microorganisms to control them. Common fungi used for weed control include Fusarium, Alternaria, Botrytis, Stem mold, Anthracnose, and Helicobacter. Common bacteria used for weed control include Pseudomonas, Xanthomonas, and Flavobacterium.
[0004] Live herbicides are a relatively new subcategory of biopesticides, currently in the early stages of industrialization and promotion, with a market share far lower than that of chemical herbicides. Globally, more than ten fungal herbicides have been registered, with representative products including Devine (a palm fungus formulation) and Collego (a specific formulation of Colletotrichum gloeosporioides) from the United States. These products achieve over 90% control efficacy against specific target weeds, but their market penetration is slower than that of chemical herbicides, and their current overall market share is still less than 5%. Most marketed products are highly specific formulations, capable of controlling only one or a few target weeds, making it difficult to meet the comprehensive control needs of complex weed communities in farmland.
[0005] To address the industry pain points caused by the long-term and extensive use of chemical herbicides, such as weed resistance, pesticide residues, and damage to farmland ecosystems, and to ensure crop yield safety, reduce field control costs, and maintain the micro-ecological balance of farmland, there is an urgent need to develop microbial strains that can replace chemical herbicides for green weed control in agriculture. These strains have significant value in many aspects, including ecological security, industrial development, and scientific research innovation. Summary of the Invention
[0006] The purpose of this invention is to overcome the problems of herbicide resistance in weeds, pesticide residues, and ecological damage in existing technologies.
[0007] In this invention, strain C25 was obtained through isolation and screening. Colony morphology observation showed that C25 was flat and fluffy in potato dextrose medium, with white mycelia at the edges and dark green conidia. Genomic DNA extraction and ITS sequence PCR amplification of C25 were performed, and the ITS region sequence of strain C25 was searched using NCBI Blast (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi). The results showed that the ITS sequence of strain C25 had 100% similarity to *Talaromyces stollii*. Based on the combined morphological and molecular biological characteristics, strain C25 was identified as *Talaromyces stollii*.
[0008] Therefore, the present invention provides a *Talaromyces stollii* strain C25 with weed-controlling ability, classified and named *Talaromyces stollii*, which was deposited on July 29, 2026 at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province, with accession number GDMCC No:68748.
[0009] The present invention also provides a culture of the aforementioned Stollerium.
[0010] According to an embodiment of the present invention, the culture is prepared by inoculating the *Storkia stolonifera* into potato dextrose medium and culturing it. Specifically, the *Storkia stolonifera* is inoculated into potato dextrose medium and cultured at 23-33°C for 5-10 days; preferably, the *Storkia stolonifera* is inoculated into potato dextrose medium and cultured at 28°C for 7 days.
[0011] The present invention also provides a composition comprising the Stollerium stoloniferum and / or the culture thereof.
[0012] On the other hand, the present invention also provides the use of the *Storkia stolonifera*, the culture, or the composition, wherein the use includes at least one of the following:
[0013] (1) Application in weed control;
[0014] (2) Application in the preparation of weed control products.
[0015] Preferably, the weeds are selected from: goosegrass, crabgrass, foxtail grass, and barnyard grass.
[0016] Furthermore, the present invention provides a method for controlling weeds, wherein the method involves applying the *Storkia stolonifera*, the culture, or the composition to the weed plants or soil. Preferably, the weeds are selected from: goosegrass, crabgrass, foxtail grass, and barnyard grass.
[0017] Specifically, the method involves preparing a suspension of *Storkia spores*, which is then sprayed onto the leaves of weeds, the surface of plants, or by drip irrigation, broadcasting, or mixing into the soil. According to an embodiment of the invention, the method collects the spores from the culture.
[0018] In the embodiments of the present invention, the disease index results of the control ability tests on four weeds—goosegrass, crabgrass, foxtail grass, and barnyard grass—were 90.6, 95.6, 87.8, and 73.1, respectively, indicating that the disease has a strong control ability against the four common weeds in the field.
[0019] The beneficial effects obtained by the present invention through the above technical solution include at least the following:
[0020] The *Storch bluegrass* strain provided by this invention offers a valuable microbial resource for developing biological control technologies for weeds, enhancing environmental management and agricultural ecological environment applications. It boasts advantages such as being green, environmentally friendly, and sustainable in agriculture. While the *Storch bluegrass* fungus *Purple Storch* exhibits herbicidal activity against various weeds including barnyard grass, crabgrass, and foxtail, the strain of this invention, *Storch bluegrass*, has not been previously reported to possess herbicidal activity. Furthermore, this strain exhibits activity against goosegrass, a previously unreported activity within the *Storch bluegrass* genus. This invention also enriches the variety of green pesticide products, and its potential applications include the discovery of novel herbicidal lead compounds in the secondary metabolites of weed-controlling fungi, providing new resource directions for new pesticide creation and contributing to the overall technological advancement of the agricultural microbial industry. Attached Figure Description
[0021] Figure 1 This is a photograph of the colony morphology of strain C25 after culturing at 28°C for 7 days on potato dextrose medium.
[0022] Figure 2 Phylogenetic tree of C25 strain was constructed to select different species of the genus Basilaria that have high ITS sequence similarity to strain C25.
[0023] Figure 3 Photos showing the weed control effect of C25 strain on goosegrass, crabgrass, foxtail grass and barnyard grass after 10 days. CK refers to the negative control, where no C25 strain was sprayed and only water was sprayed.
[0024] Strain preservation information:
[0025] The C25 strain of this invention was deposited on July 29, 2026, at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province, with accession number GDMCC No:68748 and classification name Talaromyces stollii. Detailed Implementation
[0026] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer as a result of the description. However, these embodiments are merely exemplary and do not constitute any limitation on the scope of protection defined by the claims of the present invention.
[0027] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that the upper and lower limits of the range and each intermediate value between them are specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.
[0028] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.
[0029] Unless otherwise specified, all reagents and materials used in the following examples were purchased from reputable chemical or biological reagent / material suppliers, and all reagents were of analytical grade.
[0030] The basic components of the culture medium used in the examples are as follows:
[0031] Potato glucose medium: 4g potato extract powder, 20g glucose, 15g agar, and distilled water to a final volume of 1L.
[0032] Example 1: Isolation of strain C25
[0033] Severely diseased *Eleusine indica* tissue was selected, surface-sterilized, and placed on potato dextrose agar. It was incubated at 28°C for 7 days. Subsequently, eight fungal strains with different colony morphologies, colors, and textures were isolated from the potato dextrose agar. A single colony of the most frequently isolated fungus was selected; the colony was grayish-green on the front and light grayish-brown on the back, named C25. C25 was purified by streak plating. The purified plates were incubated at 28°C. When single colonies appeared on the plates, streak plating was repeated. After three generations of purification, a pure culture of C25 was obtained for subsequent strain identification and weed control experiments.
[0034] Example 2: Identification of strain C25
[0035] The taxonomic status of strain C25 is mainly determined through morphological and molecular biological methods.
[0036] 1. Morphological identification
[0037] The C25 strain was inoculated into potato dextrose medium and incubated at 28°C for 7 days. Colony morphology was then observed. Figure 1 As shown in the figure. Observation of C25 colony morphology showed that C25 was flat and fluffy in potato dextrose medium, with white hyphae at the edges and dark green conidia.
[0038] 2. Molecular biological identification
[0039] (1) Extraction of C25 genomic DNA and PCR amplification of ITS sequence
[0040] The C25 strain was inoculated into potato dextrose medium and cultured at 28°C for 7 days before the bacterial cells were collected. Genomic DNA was then extracted from the C25 strain using a fungal genome extraction kit.
[0041] Using C25 genomic DNA as a template, ITS universal amplification primers were used.
[0042] PCR amplification was performed using a 25µL system, which mainly included 1µL each of upstream and downstream primers, 2µL of C25 genomic DNA, and a DNA polymerase mixture (containing DNA polymerase, Mg...). 2+ 12.5 µL of 4 types of dNTPs, PCR amplification buffer, etc. and 8.5 µL of ddH2O.
[0043] The PCR amplification program was as follows: 94℃ pre-denaturation for 3 min, followed by 94℃ denaturation for 30 s, 55℃ annealing for 30 s, and 72℃ extension for 1 min, for a total of 30 cycles. A final extension at 72℃ for 10 min was performed. The PCR amplification products were detected by agarose gel electrophoresis, and sequencing was performed after confirming the correct band positions.
[0044] (3) Phylogenetic analysis
[0045] A Blast search of the ITS region sequence of strain C25 was performed using NCBI (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi), revealing a 100% similarity between the ITS sequence of strain C25 and that of *Talaromyces stollii*. Phylogenetic trees were then constructed using different *Talaromyces* species that showed high similarity to the ITS sequence of strain C25. For example... Figure 2 As shown, phylogenetic analysis indicates that strain C25 is most closely related to *T. stollii*.
[0046] Based on the morphological and molecular biological characteristics of strain C25, strain C25 was identified as *Talaromyces stollii*. Strain C25 was deposited at the Guangdong Provincial Microbial Culture Collection Center, accession number GDMCC No: 68748.
[0047] Example 3: Control ability of strain C25 against four weeds
[0048] Herbicidal ability of four weeds—goosegrass, crabgrass, foxtail, and barnyard grass—was tested. Seeds of all four weeds were soaked in a 1000 mg / L gibberellin solution for 24 hours to break dormancy. After rinsing with water, the seeds were evenly scattered in petri dishes lined with three layers of filter paper. Once the coleoptiles emerged, they were transplanted into square pots (8.5 cm × 8.5 cm) containing a mixture of seedling substrate and organic fertilizer (3:1). The pots were then placed in a greenhouse with a light intensity of L / D = 12 h / 12 h, a temperature of (28 ± 5) ℃, a relative humidity of (60 ± 20)%, and a 200 μmol / L chlorine dioxide concentration. -2 s -1 Temperature in darkness: (20±4)℃, relative humidity: (50±20)%.
[0049] Strain C25 was inoculated into potato dextrose medium and cultured at 28°C for 7 days. Spores were then collected and a spore suspension was prepared. The C25 spore suspension was sprayed onto four weeds, and after 10 days of moist cultivation, the disease incidence of the four weeds was investigated and the disease index was calculated. Spraying with water served as a control.
[0050] The severity of disease on each weed was graded from 0 to 4: Grade 0, healthy plant, no visible lesions; Grade 1, lesion area less than 25%; Grade 2, lesion area 26-50%; Grade 3, lesion area 51-75%; Grade 4, lesion area exceeding 75% of leaf area. Disease was investigated and the disease index was calculated. The disease index calculation formula is: [Σ(number of diseased leaves × corresponding disease grade value) / (total number of investigated leaves × highest disease grade value)] × 100.
[0051] The results showed that, Figure 3 As shown, compared with the negative control CK (no C25 strain spore suspension), the disease severity of goosegrass, crabgrass, and foxtail grass was very high after spraying with C25 strain spore suspension. When the disease severity of goosegrass, crabgrass, and foxtail grass was investigated on the 10th day, it was found that the disease severity of most goosegrass, crabgrass, and foxtail grass was grade 4. According to statistics, the disease index of goosegrass, crabgrass, and foxtail grass after spraying with C25 was 90.6, 95.6, and 87.8, respectively.
[0052] C25 strain exhibited slightly lower herbicidal activity against barnyard grass than against goosegrass, crabgrass, and foxtail. Compared to the negative control (CK), the disease index of barnyard grass was 73.1 10 days after inoculation with C25 strain spore suspension. This indicates that strain C25 possesses strong control capabilities against all four weeds—goosegrass, crabgrass, foxtail, and barnyard grass—with the best efficacy against goosegrass, crabgrass, and foxtail.
Claims
1. A species of *Talaromyces stollii*, characterized in that, The accession number is: GDMCC No:68748.
2. The culture of Stollerium as described in claim 1.
3. The culture as described in claim 2, characterized in that, The culture is prepared by inoculating the Stollerium stoloniferum into potato dextrose medium and culturing it.
4. The culture as described in claim 3, characterized in that, The culture was prepared by inoculating the Stollerium stoloniferum into potato dextrose medium and culturing it at 28°C for 7 days.
5. A composition, characterized in that, The composition contains *Storzella stolonifera* as described in claim 1 and / or the culture as described in any one of claims 2 to 4.
6. The use of *Storkia stolonifera* as claimed in claim 1, the culture as claimed in any one of claims 2 to 4, or the composition as claimed in claim 5, characterized in that, The application includes at least one of the following: (1) Application in weed control; (2) Application in the preparation of weed control products; The weeds were selected from: goosegrass, crabgrass, foxtail grass, and barnyard grass.
7. A method for controlling weeds, characterized in that, The Stolpas bacterium of claim 1, the culture of any one of claims 2 to 4, or the composition of claim 5 are applied to weed plants or soil; The weeds were selected from: goosegrass, crabgrass, foxtail grass, and barnyard grass.
8. The method as described in claim 7, characterized in that, The spores of *Storbazium* as described in claim 1 are prepared into a suspension, which is then sprayed onto the leaves of weeds, the surface of plants, or drip-irrigated, broadcast, or mixed into the soil.
9. The method as described in claim 8, characterized in that, The spores are collected from the culture according to any one of claims 2 to 4.