Application of trichoderma xerophyllum to prevention and treatment of plant root-knot nematode
By using the fungal agent prepared by *Trichoderma xerophyte* YMF1.09953, the problems of soil residue and resistance of chemical nematicides have been solved, achieving efficient and green control of root-knot nematodes, especially significantly reducing the disease index in the case of complex diseases.
Patent Information
- Application Number
- CN202511390178.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-26
- Publication Date
- 2026-01-13
AI Technical Summary
Existing chemical nematicides pose problems such as soil residue, ecological risks, and drug resistance in the control of root-knot nematodes, making it difficult to effectively control nematode damage. Furthermore, traditional methods cannot effectively prevent the co-infection of nematodes with other pathogens.
The fungal agent prepared using *Trichoderma xerophyte* YMF1.09953 killed root-knot nematodes by fermenting the broth in a shaker. The fermentation broth had a mortality rate of over 90% against root-knot nematodes, and even after dilution, it still maintained a mortality rate of over 80%.
This provides a green and efficient control method that quickly kills nematodes, prevents larvae from invading plants, and significantly reduces plant disease indices, with better results than traditional chemical nematicides.
Smart Images
Figure CN121320105A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of microbial technology, specifically relating to the application of a strain of xerophytic Trichoderma in the control of plant root-knot nematodes. Background Technology
[0002] Plant root-knot nematodes are sessile, obligate endoparasitic nematodes that obtain nutrients from their hosts through feeding sites. These enlarged feeding sites cause root swelling, forming root knots that impair the plant's ability to absorb nutrients and water. Symptoms include weakened plant growth, wilting leaves, yellowing leaves, root deformities, low yield, and poor fruit quality. Root-knot nematodes have a very wide host range, including various vegetables such as cucumbers, tomatoes, peppers, and beans, as well as grains and oilseeds such as corn, potatoes, and soybeans. They are distributed in most parts of southern my country and some warmer regions in the north, and are found on almost every landmass in the world except Antarctica. Undoubtedly, parasitic nematodes cause significant losses to many temperate, subtropical, and tropical crops. The total annual economic loss to 37 important crops worldwide is estimated at US$358.24 billion, with estimated annual losses to 20 basic food crops at US$215.77 billion and to 17 important economic crops at US$142.47 billion. In addition, the proportion of plant nematode infestation in the growing areas of important crop species around the world is at a high level. For example, plant nematode diseases occur in 90% of banana growing areas worldwide.
[0003] Root-knot nematode disease and other diseases often occur together during crop cultivation. Root-knot nematodes play a crucial pioneering role in this process, creating numerous wounds in the roots. Pathogens then use these wounds to invade the host. The combined infection of multiple pathogens manifests as a variety of symptoms on the host, creating a synergistic effect where 1+1>2, significantly exacerbating the damage. An example is the interaction between *Ralstonia solanacearum*, the pathogen of tobacco bacterial wilt, and the southern root-knot nematode. With global warming, the increase in protected areas, continuous cropping, and the complexity of soil environments, root-related compound diseases have become commonplace, severely restricting agricultural production in my country.
[0004] Traditional control systems for root-knot nematodes have long relied on chemical nematicides. Taking abamectin as an example, these agents not only have significant problems such as long soil residue and potential ecological risks, but more seriously, long-term use has led to significant resistance in nematode populations, resulting in a continuous decline in control efficacy. Against this backdrop, environmentally friendly biological control technologies have gradually become a research hotspot.
[0005] The present invention aims to provide a strain of *Trichoderma xerophyte* with excellent nematode-attracting and killing ability, providing a new technical solution for the green and efficient control of root-knot nematodes. Summary of the Invention
[0006] The first objective of this invention is to provide the application of a strain of *Trichoderma xerophyte* in the control of plant root-knot nematodes. The second objective of this invention is to provide a fungal agent prepared based on the *Trichoderma xerophyte* YMF1.09953.
[0007] The first objective of this invention is achieved as follows: *Trichoderma xerophytes* (… Trichoderma xerophilum The application of YMF1.09953 is in the control of plant root-knot nematodes. The xerophytic Trichoderma YMF1.09953 is deposited at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC) under the number GDMCC No. 66977, with a deposit date of September 17, 2025. The deposit address is No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province.
[0008] The second objective of this invention is achieved by providing a fungal agent prepared based on the aforementioned *Trichoderma xerophyte* YMF1.09953, the preparation method of which is as follows: 1) Transfer the strain of *Trichoderma xerophyte* YMF1.09953 into seed culture medium, and then culture it on a shaker at 25-30℃ and 150-200 r / min for 24-36 h to obtain the fermentation seed liquid; 2) Inoculate the fermentation seed liquid into the fermentation medium and culture it on a shaker at 25-30℃ and 150-200 r / min for 72-96 hours. The resulting fermentation liquid is the target inoculum.
[0009] The beneficial effects of this invention are as follows: This invention, through experimental verification, demonstrates that *Trichoderma xerophyte* YMF1.09953 has a significant lethal effect on root-knot nematodes. Its fermentation broth exhibits a mortality rate exceeding 90% against root-knot nematodes, and even after a 50-fold dilution, it maintains a mortality rate of over 80%. It acts rapidly, quickly killing nematodes within 12 hours and effectively preventing larvae from invading plants. This invention provides a new technical solution for the green and efficient control of root-knot nematodes, showing promising application prospects in the field of plant root-knot nematode control. Attached Figure Description
[0010] Figure 1 The colony morphology and strain characteristics of *Trichoderma xerophyte* YMF1.09953 of this invention are shown in the figures (AB represents the colony morphology of *Trichoderma xerophyte* YMF1.09953; DI represents the spore attachment morphology of *Trichoderma xerophyte* YMF1.09953; J and K represent the molecular spore morphology of *Trichoderma xerophyte* YMF1.09953). Figure 2 This is the phylogenetic tree of the xerophytic Trichoderma YMF1.09953 of this invention; Figure 3The following figures illustrate the mortality of nematodes after treatment with the fermentation broth of *Trichoderma xerophyte* YMF1.09953 of this invention: Figure A shows the overall morphology of naturally dead nematodes under a 40x microscope; Figure B shows a magnified detail of naturally dead nematodes under a 100x microscope; Figure C shows the overall morphology of nematodes dead under the influence of the fermentation broth under a 40x microscope; and Figure D shows a detail of nematodes dead under the influence of the fermentation broth under a 100x microscope. Detailed Implementation
[0011] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments, but this does not limit the present invention in any way. Any modifications or improvements made based on the teachings of the present invention shall fall within the protection scope of the present invention.
[0012] This invention provides the application of *Trichoderma xerophyte* YMF1.09953, specifically its application in the control of plant root-knot nematodes. The *Trichoderma xerophyte* is deposited at the Guangdong Provincial Center for Microbial Culture Collection, with the number GDMCC No. 66977, located at No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province.
[0013] The application uses metabolites produced by the culture of *Trichoderma xerophyte* YMF1.09953 to kill plant root-knot nematodes.
[0014] The application involves preparing a formulation of *Trichoderma xerophyte* YMF1.09953 to control plant root-knot nematodes.
[0015] Furthermore, the present invention provides a fungal agent prepared based on the aforementioned *Trichoderma xerophyte* YMF1.09953.
[0016] Furthermore, the preparation method of the bacterial agent is as follows: 1) Transfer the strain of *Trichoderma xerophyte* YMF1.09953 into seed culture medium, and then culture it on a shaker at 25-30℃ and 150-200 r / min for 24-36 h to obtain the fermentation seed liquid; 2) Inoculate the fermentation seed liquid into the fermentation medium and culture it on a shaker at 25-30℃ and 150-200 r / min for 72-96 hours. The resulting fermentation liquid is the target inoculum.
[0017] Further, in step 1), the seed culture medium is PDB culture medium.
[0018] Further, in step 2), the fermentation medium is PDB medium.
[0019] Furthermore, the present invention provides the application of the aforementioned microbial agent in the control of plant root-knot nematodes.
[0020] Example 1: Isolation, purification and identification of strains Strain strain isolation methods: Take 10g of soil sample from the karst rocky desertification area of Shilin County, Yunnan Province, China, add it to 90ml of sterile water containing glass beads, shake at 220 rpm for 1 hour, let stand for 2 minutes, and then take the supernatant for serial dilution (10). - ¹ to 10 -4 Afterwards, 100 μL of each diluted solution was spread onto RBA medium (1 g potassium dihydrogen phosphate, 10 g glucose, 0.5 g magnesium sulfate heptahydrate, 20 g agar, 5 g peptone, 3.3 ml 1% Bengal red aqueous solution, diluted to 1 L with water. Autoclaved at 121°C for 20 min) containing antibiotics (streptomycin 40 mg / L, ampicillin 30 mg / L). The medium was incubated at 25°C for 5–7 days, with daily observation. Morphologically typical colonies were selected and transferred to PDA medium (200 g potato, 20 g glucose, 18 g agar, diluted to 1 L with water. Autoclaved at 121°C for 20 min) for purification to obtain *Trichoderma xerophyte* YMF1.09953. The colony morphology is shown in the figure. Figure 1 .
[0021] Molecular biological identification ( Figure 2 The identification results showed that strain YMF1.09953 was a new species of *Trichoderma xerophyte*, and its ITS sequence, Rpb2 sequence and tef sequence are shown in SEQ ID No. 1-3, respectively.
[0022] Example 2: Preparation of Fermentation Broth 1. Seed Culture: The pure culture YMF1.09953 from the plate in Example 1 was transferred to multiple 250 mL Erlenmeyer flasks containing 100 mL of PDB culture (200 g potato juice boiled, 20 g crushed corn juice boiled, 10 g sucrose, 10 g glucose, 0.5 g dipotassium hydrogen phosphate, 0.2 g magnesium sulfate heptahydrate, and water to a final volume of 1 L. Autoclaving at 121℃ for 20 min). The cultures were then incubated on a shaker at 27℃±1℃ and 160 r / min for 36 h to obtain the fermentation seed culture.
[0023] 2. Fermentation culture: The fermentation seed liquid was inoculated into PDB medium at a rate of 5%, and 150 mL of the liquid was placed in a 250 mL Erlenmeyer flask. The mixture was cultured on a shaker at 27 °C and 165 r / min for 136 hours to obtain the fermentation broth.
[0024] Experimental Example 1: Test of the lethal activity of fermentation broth against nematodes in Example 2 Experimental Methods: An experimental group and a blank control group were set up. 700 µL of fermentation broth and PDB liquid medium were added to cell culture dishes in each group, followed by 20 µL of a suspension of 80-100 second-instar Southern root-knot nematodes. The mixture was thoroughly mixed and incubated at 22°C. The number of live and dead nematodes was recorded under a stereomicroscope every 30 minutes. Nematodes were considered dead if they exhibited stiffness and showed no response to needle stimulation. The nematode mortality rate and corrected mortality rate were calculated. Each experiment was repeated 5 times. Results are shown in Table 1.
[0025]
[0026]
[0027] Table 1. Results of lethal activity of the fermentation broth prepared in Example 2 against Southern root-knot nematodes.
[0028] As shown in Table 2, the fermentation broth prepared in Example 2 had a 12-hour lethality of over 90% against Southern root-knot nematodes.
[0029] The fermentation broth prepared in Example 2 was diluted 50 times and then subjected to nematode lethality testing according to the above method. The test results are shown in Table 2.
[0030] Table 2. Results of lethal activity of the fermentation broth dilution (×50) prepared in Example 2 against Southern root-knot nematodes.
[0031] As shown in Table 2, the fermentation broth prepared in Example 2, after being diluted 50 times, had a mortality rate of over 80% on second-instar larvae of Southern Root-knot Nematode after 12 hours of exposure.
[0032] Experimental Example 2: Field Trial 1. Test Methods This experiment used tomatoes infected with the southern root-knot nematode as the test crop and was conducted in yellow soil conditions. The experimental plots covered 100 square meters and included seven treatments: experimental groups 1-5 (treated with different concentrations of the fermentation broth prepared in Example 2), a positive control group (2% abamectin suspension), and a blank control group (water). Each group had 50 tomato seedlings planted at a spacing of 50 cm, and the plots were planned using a randomized block design. At transplanting, each seedling was irrigated with 50 ml of the corresponding pesticide at the base. During the experiment, routine field management (including watering and fertilization) was maintained consistently across all treatments.
[0033] Experimental Group 1: 10 ml / plant of fermentation broth prepared in Example 2 + 40 ml / plant of water Experimental Group 2: 20 ml / plant of fermentation broth prepared in Example 2 + 30 ml / plant of water Experimental Group 3: 30 ml / plant of fermentation broth prepared in Example 2 + 20 ml / plant of water Experimental Group 4: 40 ml / plant of fermentation broth prepared in Example 2 + 10 ml / plant of water Experimental group 5: 50 ml / strain of fermentation broth prepared in Example 2 Positive control group: 20 ml / strain of 2% abamectin suspension + 30 ml / strain of water Control group: 50ml water / plant 2. Survey Methods After all tomatoes were harvested, the incidence rate, disease index, and relative control effect were investigated.
[0034]
[0035]
[0036]
[0037]
[0038] Table 3 Disease Grading Criteria
[0039] 3. Experimental Results Table 4. Field application efficacy of the fermentation broth prepared in Example 2 against root-knot nematodes.
[0040] As shown in Table 4, the incidence rate, root necrosis rate and disease index of tomatoes in experimental groups 1-5 were significantly lower than those in the positive control group. This indicates that the fermentation liquid prepared in Example 2 of this invention has a better control effect on field nematodes than abamectin and has broad application prospects in the field of plant root-knot nematode control. It is suitable for further industrial development and promotion.
Claims
1. Xerophytic Trichoderma ( Trichoderma xerophilum Application of YMF1.09953 in the control of plant root-knot nematodes. The xerophytic Trichoderma is deposited at the Guangdong Provincial Center for Microbial Culture Collection, with the number GDMCC No: 66977, and the deposit address is No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province.
2. The application according to claim 1, characterized in that, The application uses metabolites produced by the culture of *Trichoderma xerophyte* YMF1.09953 to kill plant root-knot nematodes.
3. The application according to claim 1, characterized in that, The application involves preparing a formulation of *Trichoderma xerophyte* YMF1.09953 to control plant root-knot nematodes.
4. A fungal agent prepared based on the *Trichoderma xerophyte* YMF1.09953 as described in claim 1.
5. The fungal agent prepared from *Trichoderma xerophyte* YMF1.09953 according to claim 4, characterized in that, The preparation method of the bacterial agent is as follows: 1) Transfer the strain of *Trichoderma xerophyte* YMF1.09953 into seed culture medium, and then culture it on a shaker at 25-30℃ and 150-200 r / min for 24-36 h to obtain the fermentation seed liquid; 2) Inoculate the fermentation seed liquid into the fermentation medium and culture it on a shaker at 25-30℃ and 150-200 r / min for 72-96 h. The resulting fermentation liquid is the target inoculum.
6. The fungal agent prepared from *Trichoderma xerophyte* YMF1.09953 according to claim 5, characterized in that, In step 1), The seed culture medium is PDB medium.
7. The fungal agent prepared from *Trichoderma xerophyte* YMF1.09953 according to claim 6, characterized in that, In step 2), The fermentation medium is PDB medium.
8. The application of the microbial agent according to claim 4 in the control of plant root-knot nematodes.