CN115197853B - Endophyte Epicoccum thailandicum LF-28 strain and application thereof - Google Patents
Endophyte Epicoccum thailandicum LF-28 strain and application thereof Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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Abstract
The application discloses an endophyte Epicoccum thailandicum LF-28 strain and application thereof, and the preservation number is CGMCC No.40134. The strain can be used in various prevention and treatment scenes of fusarium oxysporum, can effectively protect susceptible plants of fusarium oxysporum and promote plant growth, thereby further improving the prevention and treatment effect of the plants on fusarium oxysporum.
Description
Technical Field
The application relates to the technical field of endophytes, in particular to an endophyte Epicoccum thailandicumLF-28 strain and application thereof.
Background
Root rot is one of the most serious soil-borne diseases of ginseng plants, and the perennial loss of pseudo-ginseng caused by root rot reaches 5-20%; the serious loss is 70 percent, and the sustainable development of the ginseng industry and the important barrier of ecological construction are seriously affected. The traditional chemical control is difficult to control, and is easy to cause soil dysbiosis, pesticide residues and environmental pollution, and the biological microbial inoculum which is safe, pollution-free, low in cost and high in efficiency can be used for controlling root rot, so that the edible safety of traditional Chinese medicinal materials can be ensured. Therefore, screening antagonistic strains of pathogenic bacteria and developing an antibacterial agent are hot spots in the current research on preventing and treating root rot of ginseng plants.
Endophytes are bacteria, fungi or actinomycetes growing in healthy plant roots, stems, leaves and other tissues, and the addition of the endophytes does not cause obvious diseases of host plants, and are important components of a plant micro-ecological system. The plant endophyte has widely distributed antibacterial active strain, can produce various antibacterial products and becomes antagonistic bacteria of plant pathogenic bacteria under certain conditions. How to derive the effect of the herba sophorae flavescentis and the grassleaf sweelflag rhizome extracts on the growth of fusarium oxysporum is researched, and the results show that the herba sophorae flavescentis and the grassleaf sweelflag rhizome extracts can inhibit the growth of pathogenic bacteria hyphae and the occurrence of banana root rot.
Endophytes promote the growth and development and yield of host plants by biosoliding nitrogen and regulating plant hormones. Therefore, screening based on endophyte antagonistic bacteria and research and development of biocontrol bacteria are feasible methods and approaches for preventing and treating root rot.
However, the existing strain for preventing and treating the root rot of the ginseng plant has single species, and the effect is reduced after long-term use, so that the endophytic strain with the specific preventing and treating effect on the root rot of the ginseng plant is needed.
Disclosure of Invention
The application provides an endophyte Epicoccum thailandicumLF-28 strain and application thereof, which are used for solving the technical problem that the endophyte capable of preventing and treating ginseng root rot is lacking in the prior art.
The application provides an endophyte Epicoccum thailandicumLF-28 strain with a preservation number of CGMCC No.40134.
The strain can be used in various prevention and treatment scenes of fusarium oxysporum, can effectively protect susceptible plants of fusarium oxysporum and promote plant growth, thereby further improving the prevention and treatment effect of the plants on fusarium oxysporum.
The application also provides application of the endophytic epipoccus strain LF-28 in preventing and controlling plant fungal diseases, wherein the plant fungi are Fusarium oxysporum.
Preferably, the fungus is Fusarium oxysporum of the genus Panax;
preferably, the fungus is Fusarium oxysporum of root rot of pseudo-ginseng;
preferably, the fungus is present in the soil of notoginseng continuous cropping.
The application also provides application of the endophytic epicoccum strain LF-28 in promoting plant growth, wherein fermentation liquor of the endophytic epicoccum strain LF-28 contains indoleacetic acid and gibberellin.
Preferably, the indoleacetic acid content in the fermentation broth of the endophytic epicoccum strain LF-28 is more than 20 mg.L -1 The method comprises the steps of carrying out a first treatment on the surface of the Gibberellin content greater than 15 mg.L -1 。
After the strain is fermented, the obtained fermentation liquor contains the substances with obvious effect of promoting plant growth, can effectively improve the growth condition of ginseng plants, is particularly suitable for the growth condition of planting pseudo-ginseng in continuous cropping soil, is beneficial to improving the yield of planting pseudo-ginseng on the continuous cropping soil, and has no toxic effect on plants.
In another aspect of the present application there is provided a biological agent comprising an endophytic epicoccum strain LF-28 as described above; the biological agent is used for preventing and controlling plant fungal diseases and/or promoting plant growth.
Preferably, a bacterial solution and/or a bacterial cell comprising said Epicoccus endophyte LF-28.
Preferably, the plant fungus is fusarium oxysporum;
preferably, the fungus is Fusarium oxysporum of the genus Panax;
preferably, the fungus is Fusarium oxysporum of root rot of pseudo-ginseng;
preferably, the fungus is present in the soil of notoginseng continuous cropping.
The obtained biological preparation contains the strain and can be used for preventing and treating fusarium oxysporum capable of infecting various plants. Is especially suitable for preventing and controlling fusarium oxysporum infected with ginseng plants. Because the notoginseng plant is easier to infect the root rot fusarium oxysporum relative to the ginseng plant, the strain has better control effect on the root rot fusarium oxysporum of the notoginseng plant. The biological agent can effectively solve the problem that the continuous cropping soil can not continuously plant the pseudo-ginseng, and is beneficial to enlarging the planting area of the pseudo-ginseng.
In another aspect of the present application, there is provided a fermentation process of an Epicoccus endophyte LF-28 as described above, comprising the steps of:
culturing endophytic epicoccum strain LF-28 under fermentation conditions, and separating to obtain fermentation liquor;
the fermentation condition is that the fermentation is carried out for 24 to 60 hours at the temperature of 24 to 26 ℃, and the rotating speed of a culture device is 0 to 120r min -1 The method comprises the steps of carrying out a first treatment on the surface of the The initial pH of the fermentation environment was 7.0; the strain is adopted for fermentation under the conditions, and the viable bacteria content is 1 multiplied by 10 8 ～1×10 9 cfu/mL broth. Can be used as raw materials to prepare the biocontrol microbial agent.
Preferably, the fermentation condition is that the fermentation is carried out for 60 hours at 26 ℃, and the rotating speed of the culture device is 120r min -1 The method comprises the steps of carrying out a first treatment on the surface of the The initial pH of the fermentation environment was 7.0. The fermentation is the most preferable condition, and fermentation can be carried out according to the condition to obtain fermentation liquor with the highest viable bacteria content.
In another aspect of the present application, there is provided a method for controlling a fungal plant disease, comprising the steps of: applying a biological agent comprising the above before planting the plant and during the growth of the plant;
preferably, the application method is at least one of spraying, root irrigation and drip irrigation with a liquid containing the biological agent.
The biological agent obtained in the application at least contains the fermentation liquor of the strain, and the obtained fermentation liquor can be used according to a common method in the prior art, for example, after the fermentation liquor is prepared into liquid with the concentration of 1-5% of the mass of the biological agent, the biological agent is sprayed to carry out foliar application, and soil application is carried out through root irrigation and drip irrigation, so that the use effect is achieved.
The beneficial effects that this application can produce include:
1) Under the aseptic condition, the epicoccum strain provided by the application adopts separated and purified endophytes, antagonistic strains are screened through a flat plate counter test, morphology and molecular biological identification are carried out, a single factor test is preferred to be the strain fermentation condition, and the control effect of the microbial inoculum is verified through a potting test. The biocontrol bacteria for antagonizing the root rot of the pseudo-ginseng are screened from endophytes separated from pseudo-ginseng plants, and the effect of resisting the root rot of the ginseng is verified under a potting test through fermentation culture, so that the foundation is laid for developing a specific microbial agent for the ginseng plants.
2) The epiphyte strain provided by the application has the bacteriostasis rate of 61.94% on fusarium oxysporum, has a colony diameter of 2.00cm and shows stronger bacteriostasis activity, can secrete growth regulating hormone gibberellin and indoleacetic acid required by plant growth, can effectively improve the problems of soil impoverishment and germ content excess after continuous cropping of pseudo-ginseng, and is particularly suitable for improving the continuous cropping soil of pseudo-ginseng.
The preservation information of the strain used in the application is as follows: endophytes Epicoccum thailandicum LF-28 are deposited in: china general microbiological culture Collection center (CGMCC) of national institute of culture Collection of microorganisms (CGMCC) No. 1, no. 3, north Chen West Lu, chaoyang, beijing. The preservation date is: 2022, 4, 21; the number is: CGMCC No.40134. The class of biological material is named Epicoccum thailandicum.
Drawings
FIG. 1 is a photograph of colony morphology and a photograph of microscopic observation of endophytes Epicoccum thailandicumLF-28 provided herein; wherein a) is a schematic diagram of a colony photo of an endophytic epicoccum strain LF-28; b) A partial magnification micrograph of a colony grown on the endophytic epicoccum strain LF-28;
FIG. 2 is a schematic diagram of the ITS sequence homology phylogenetic tree of strain LF-28 provided herein;
FIG. 3 is a schematic diagram showing the results of the experiment of antagonizing F.tumefaciens by strain LF-28 provided in the present application; wherein a) is a blank group plate culture result photo; b) Photographs of the plate culture results obtained for the experimental group inoculated with strain LF-28 provided herein;
FIG. 4 is a schematic diagram of fermentation condition screening of strain LF-28 provided herein; wherein A is the relation curve of the fermentation time and the viable count of the strain LF-28; b is a relation curve of the fermentation temperature and the viable count of the strain LF-28; c is a relation curve of the rotating speed of the strain LF-28 fermentation device and the viable count; d is a relation curve of initial pH value and viable count of the strain LF-28 fermentation; and (3) injection: different lowercase letters represent significance differing by up to 0.05 level.
Detailed Description
For the purposes of making the objects, technical solutions and advantages of the embodiments of the present application more clear, the technical solutions of the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is apparent that the described embodiments are some of the embodiments of the present application, but not all of the embodiments. The components of the embodiments of the present application, which are generally described and illustrated in the figures herein, may be arranged and designed in a wide variety of different configurations.
Accordingly, the following detailed description of the embodiments of the present application, provided in the accompanying drawings, is not intended to limit the scope of the application, as claimed, but is merely representative of selected embodiments of the application. All other embodiments, based on the embodiments herein, which would be apparent to one of ordinary skill in the art without undue burden are within the scope of the present application.
Technical means which are not described in detail in the application and are not used for solving the technical problems of the application are all arranged according to common general knowledge in the field, and various common general knowledge arrangement modes can be realized.
Examples
Reagents not described in detail in the examples below are commercially available products.
The pathogenic bacteria of root rot are fusarium oxysporum Fusarium oxysporum, which are isolated from the laboratory and stored in the strain. The test ginseng seeds were supplied by Shengshi Baicao pharmaceutical Co.
Reagent consumable: 200g of potato, 20g of glucose (national medicine group chemical Co., ltd., batch No. 20180327) and 13g of agar (national medicine group chemical Co., ltd., batch No. 20180722).Spin Kit for Soil kit (MPBIO Co., lot 20200731), fungus Universal primer (Beijing qing biosciences Co., lot 20200824), phenol-cotton blue lactate staining solution (Fuzhou Feijing biosciences Co., lot 20200526).
Test instrument: DL-CJ-1 NP-II sterile console (Beijing Dongbian Harr instruments Co., ltd.), FR-2L type three-mesh fluorescence microscope (Beijing century science instruments Co., ltd.), 2720 type polymerase chain reaction instrument (Applied in the United states)Company), DYY-8C electrophoresis apparatus (Beijing Liuyi Instrument Co., ltd.), DH420 type electrothermal incubator (Beijing Kogyo Yongxing Instrument Co., ltd.).
EXAMPLE 1 endophyte Epicoccum thailandicumLF-28 Source and isolation
The endophyte is separated from roots, stems and leaves of a Yunnan pseudo-ginseng sample.
The endophyte separation comprises the following steps:
the endophytic fungi are cultivated by a pure cultivation method. The separation and purification method comprises the following steps: fresh and healthy roots, stems and leaves of pseudo-ginseng are taken, washed by running water, then subjected to surface disinfection treatment, the tissue is cut into pieces by a sterilized scalpel, inoculated in a PDA culture medium after being cut into 0.5cm, cultured at 25 ℃, bacterial spots are grown, different bacterial colonies are respectively picked into the PDA culture medium, and the bacterial colonies are gradually separated and purified until a purified single bacterial strain, namely endophyte Epicoccum thailandicumLF-28, is obtained.
The isolated strains were found to be: the academy of China academy of microbiology research in North Chen West Lu No. 1, 3, the Korean region of Beijing, is provided. The preservation date is: 2022, 4, 21; the number is: CGMCC No.40134. The class of biological material is named Epicoccum thailandicum.
Strain morphology and molecular characterization
The endophyte LF-28 grows faster during the culture period, white hypha grows in one day of culture at 28 ℃, the mycelium is finer and flocculent, the colony is round, and after 3 days of culture, the single hypha forms a gray black round colony with a clean edge on the culture medium (figure 1). Under a microscope, the large conidium of the endophyte LF-28 is sickle-shaped or long column-shaped, and the conidium peduncles are slender. The strain was compared with reference to the morphological characteristics of fungi listed in the fungus identification handbook, and initially identified as Epicoccum.
The sequence of the strain LF-28 is shown in the attached sequence table, the sequence similarity with Epicoccum thailandicum ICMP 17673 (accession number NR_ 166015.1) is 99.22% after ITS sequence alignment, and the strain LF-28 is identified as Epicoccum thailandicum in the same branch of a phylogenetic tree (figure 2, combined with the morphological characteristics of the strain).
Example 2 screening of root rot antagonistic bacteria of endophytes Epicoccum thailandicumLF-28 antagonistic strains of pathogenic bacteria Fusarium oxysporum were screened by plate counter method, specific procedures are described (Gu Pengli, shen Shuo, hu Yingjie. Screening, identification and biocontrol factor first detection [ J ]. Fujian agricultural journal, 2020,35 (12): 1336-1345.) [8] of a strain of Bacillus subtilis antagonistic bacteria of potato. Fusarium oxysporum cakes are taken out by a puncher with the diameter of 1.00cm and transferred to a new PDA culture medium, meanwhile, endophytic fungi are cultured at 4 points at a position 2.50cm away from the center of the PDA solid culture medium, and strains with antagonism are screened. PDA plates inoculated with Fusarium oxysporum mycelium pellet alone served as controls. Culturing in an incubator at 28 ℃ for 5 days, repeating each strain for 3 times, measuring the colony diameter of fusarium oxysporum in a culture dish, and calculating the bacteriostasis rate. Antibacterial ratio = (control colony diameter-treated colony diameter)/(control colony diameter-petiole diameter) ×100%.
The obtained results:
through a plate counter test, the strain LF-28 (Epicoccum thailandicum) has strong antibacterial activity on Fusarium oxysporum, the diameter of a colony obtained in the inhibition test is 2.00cm, and the antibacterial rate is 61.94% (shown in FIG. 4 and Table 1).
TABLE 1 bacterial strain LF-28 (Epicoccum thailandicum) antibacterial Rate (%)
As can be seen from Table 1 and FIG. 4, the strain provided by the application has an obvious inhibition effect on Fusarium oxysporum, the growth of the fungus is effectively inhibited, and the colony diameter is obviously reduced compared with the CK group.
EXAMPLE 3 identification of endophytes Epicoccum thailandicumLF-28
Mycelium and spores with good growth condition are picked up to be made into a temporary loading tablet, and the shapes of mycelium forms, spore shapes, spore diaphragms, diaphragm numbers and the like are observed under a microscope, and the morphological identification is carried out according to fungus identification handbook [9] (Wei Jingchao. Fungus identification handbook [ M ]. Shanghai: shanghai science and technology Press, 1979:143.).
Morphological results:
the endophyte LF-28 grows faster during culture, white mycelium grows out after one day of culture at 28 ℃, the mycelium is finer and flocculent, the colony is round, after 3d of culture, the single mycelium forms a gray black round colony with a neat edge on a culture medium (figure 1), and the diameter of the colony is 7.0cm.
Under a microscope, the large conidium of the endophyte LF-28 is sickle-shaped or long column-shaped, and the conidium peduncles are slender.
The strain was compared with reference to the morphological characteristics of fungi listed in the fungus identification handbook, and initially identified as Epicoccum.
Spin KitThe for oil (MPBIO) kit extracts total DNA of endophytic fungi. Amplification of ITS2 rRNA Gene fragment Using fungal Universal primer ITS1/ITS2 (GARDES M, BRUNS TD. ITS primers with enhanced specificity for basidiomycetes-application to the identification of mycorrhizae and rusts [ J ]].Mol Ecol,1993,(22):113-118.)[10]. Sequencing results were aligned with sequences in NCBI database, homology alignment analysis was performed using Blast on-line tool, and phylogenetic tree was constructed using MEGA7.0 software, the results of which are shown in FIG. 2.
Results:
the sequence of the strain LF-28 is shown in a sequence table, the sequence similarity with Epicoccum thailandicum ICMP 17673 (accession number NR_ 166015.1) is 99.22% after ITS sequence alignment, and the strain LF-28 is identified as Epicoccum thailandicum by combining with the strain shape characteristics in the same branch of a phylogenetic tree (figure 2).
Example 4 screening of endophytes Epicoccum thailandicumLF-28 for suitable fermentation conditions
The single factor test investigated the effect of incubation time, shake flask rotation speed, temperature and initial pH on strain fermentation. The number of viable bacteria was counted by plate dilution coating method, reference to "microbiology experiment" Shen Ping, chen Xiangdong. Microbiology experiment [ M ]. 5 th edition. Beijing: higher education Press, 2018:40-41.) [11], 3 replicates per group.
The conditions for a single experiment are shown in fig. 4 on the abscissa.
Screening fermentation time: the fermentation temperature of each experiment is 25 ℃ and the rotation speed of the culture device is 100r min -1 The pH value is 7.0;
the fermentation temperature screening is as follows: the fermentation time of each experiment is 48 hours, and the rotating speed of the culture device is 100r min -1 The pH value is 7.0;
when the rotational speed of the culture device is screened: the fermentation time of each experiment is 48 hours, the fermentation temperature is 25 ℃, and the pH value is 7.0;
at initial pH screening: the fermentation time of each experiment is 48 hours, the fermentation temperature is 25 ℃, and the rotating speed of the culture device is 100 r.min -1 。
Fermentation condition screening results:
the number of viable bacteria in the microbial inoculum Epicoccum thailandicum increased with the increase of the culture time (24 to 60 hours), and when the culture time was longer than 60 hours, the number of viable bacteria increased slowly, so that 60 hours was the optimal fermentation time (FIG. 4A).
The viable count of the microbial inoculum increases with the increase of the temperature (24-26 ℃), and when the temperature is higher than 26 ℃, the viable count of the microbial inoculum is stabilized and then the viable count of the microbial inoculum is reduced, and the optimal culture temperature is selected as 26 ℃ (FIG. 4B).
Epicoccum thailandicum the number of viable cells increases with the increase of the rotational speed (0 to 120 r.min) -1 ) When the rotating speed is greater than 120 r.min -1 After that, the viable count tends to be stable, so 120 r.min is selected -1 The flask fermentation speed was optimized for this (FIG. 4C).
The viable count of the inoculum increased with increasing pH and then decreased, and Epicoccum thailandicum viable count reached a maximum when pH was 7.0, so 7.0 was chosen as its optimal initial fermentation pH (FIG. 4D).
The results indicate that Epicoccum thailandicum suitable initial fermentation conditions are: epicoccum thailandicum suitable initial fermentation conditions are: 60h,26 ℃,120 r.min -1 ，pH 7.0。
Example 5 detection of growth-promoting Effect of biocontrol agent of endophyte Epicoccum thailandicumLF-28
The optimal fermentation conditions obtained by the screening in example 4 were repeated for three times, and the obtained fermentation broth was used as the biocontrol agent samples 1 to 3. Method for measuring gibberellin and indoleacetic acid contents in bacterial manure by high performance liquid chromatography according to Ma Zhenyu, lv Zhihua, jiang Tingfu, wang Yuangong. Analytical test techniques and apparatus, 2005 (03): 174-177. The methods disclosed in the foregoing patent publication detect the contents of indoleacetic acid (IAA) and gibberellin (GA 3) in the resulting fermentation broth.
IAA yield in fermentation broth of strain LF-28 (Epicoccum thailandicum) was 36.84 mg.L -1 ，GA 3 Yield of 18.56 mg.L -1 。
TABLE 2 IAA and GA in LF-28 fermentation broths of Strain (Epicoccum thailandicum) 3 Content of (3)
From the above, the fermentation broth obtained after the strain LF-28 is fermented contains a certain amount of regulating components for promoting plant growth, which is beneficial to improving the growth of plants in soil using the fermentation broth and can effectively overcome the problem of nutrient deficiency in the soil of continuous cropping of pseudo-ginseng.
Example 6 Effect verification of endophyte Epicoccum thailandicumLF-28
The experimental method comprises the following steps:
and (3) filling the sterilized humus soil into a plastic seedling raising tray, screening ginseng seeds with consistent buds and surface sterilization, and sowing the ginseng seeds on the seedling raising tray. And (3) pouring sterile water after sowing, and carrying out illumination treatment for 8 hours per day to ensure the normal emergence and growth of ginseng. Sowing for 60d, and after the ginseng seedlings emerge, keeping healthy and uniform-sized seedling groups for testing. The test consisted of 3 treatments.
The biocontrol microbial inoculum used in the following treatment is a fermentation broth obtained after fermentation under the optimal fermentation conditions selected in example 4.
The treatment is as follows: (1) a negative control group; (2) a positive control group; (3) inoculation of biocontrol bacteria + pathogenic bacteria (inoculation of pathogenic bacteria 2 weeks after inoculation of biocontrol bacteria). Root irrigation (Liu Min, hu Chenyun, ding Molong, etc.) dynamic study of the growth of two endophytes in ginseng and root zone soil [ J ]]Modern world science and technology-traditional Chinese medicine, 2014,16 (4): 790-795.) [12]Inoculating Ginseng radix, and regulating concentration of pathogenic bacteria and biocontrol bacteria to 1×10 8 cfu/mL, inoculation amount was 50. Mu.L, roots of ginseng plants were beaten along the rhizome with a pipette gun and 10mL of sterile water was irrigated per group. After 28d inoculation, the growth and incidence rate of ginseng seedlings under different treatments are counted, and the disease index is graded and referred to Ma Fengru (Ma Fengru, xing Yunzhang, wang Yunqiu, etc. the investigation method of ginseng rust and rot is primarily described in J]Chinese medicine science and technology, 1983, (5): 9-10, 15-16) [13 ]]. Each group was biologically replicated with 36 ginseng seedlings.
Experimental results:
the results of processing the experimental data obtained are shown in Table 1:
incidence = number of disease/total number of plants x 100%
Disease index = Σ (disease-level plant tree×representative series)/(plant number sum×highest representative series value) ×100
Control effect = (control disease index-treatment disease index)/control disease index x 100%
After 28d inoculation of the Epicoccum thailandicum biocontrol microbial agent provided by the application, the incidence rate (63.11%) and the disease index (54.89%) of the root rot of the ginseng seedlings are obviously reduced, and the plant height (39.14%), the leaf length (25.73%) and the fresh weight (41.17%) of the ginseng seedlings are obviously promoted.
TABLE 3 Effect of microbial inoculum on control and growth of root rot of Ginseng
| Group of | Morbidity/% | Index of disease condition | Plant height/cm | Leaf length/cm | Fresh weight/g |
| Negative control group | / | / | 5.34 | 2.06 | 0.17 |
| Positive control group | 98.09 | 41.50 | / | / | / |
| Inoculant group | 36.19 | 18.72 | 7.43 | 2.59 | 0.24 |
According to the prior art, since the pseudo-ginseng plant is easier to infect fusarium oxysporum, compared with the ginseng plant, the obtained Epicoccum thailandicum biocontrol microbial inoculum is applied to pseudo-ginseng seedlings under the same conditions, and the result of reducing the incidence rate of the pseudo-ginseng seedlings is similar to that of the ginseng seedlings, and is not described herein.
The endophyte isolated in the application is identified as Epicoccum thailandicum, and the strain has the functions of inhibiting fusarium oxysporum and promoting plant growth. The optimal fermentation conditions of the strain are as follows: 60h,26 ℃,120 r.min -1 pH 7.0, the diameter of the antibacterial colony of the strain is 2.00cm, the antibacterial rate is 61.54%, and IAA and GA are contained in the fermentation liquor 3 The yields of (C) were 36.84mg/L and 8.56mg/L, respectively.
The potting test result shows that 28 days after inoculation of the biocontrol microbial inoculum containing Epicoccum thailandicum fermentation liquid, the incidence rate (63.11%) and the disease index (54.89%) of the root rot of the ginseng seedlings are obviously reduced, and the plant height (39.14%), the leaf length (25.73%) and the fresh weight (41.17%) of the ginseng seedlings are obviously promoted.
The endophyte Epicoccum thailandicum can inhibit fusarium oxysporum and promote plant growth, provides a new alternative strain for preventing and treating the root rot of the medical plant, and provides a material for the development of microbial agents of the medical plant.
Although the present application has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described above, or equivalents may be substituted for elements thereof, and any modifications, equivalents, improvements and changes may be made without departing from the spirit and principles of the present application.
<110> Chinese traditional medicine institute, wenshan Miao Xiang Notoginseng radix science and technology Co., ltd
<120> endophyte Epicoccum thailandicum LF-28 strain and application thereof
<160> 1
<210> 1
<211> 217
<212> DNA
<213> artificial sequence
<400> 1
ggtgaactgc ggaaggatca ttacctagat ttgtaagctt tgcctgctat ctcttaccca 60
tgtcttttga gtaccttacg tttcctcggt gggttcgccc accgattgga caaatttaaa 120
ccctttgccc ttgaaatcaa cgtctgaaaa aacttaatag ttacaacttt caaccacgga 180
tctcttggtt ctggcatcga tgaagaacgc agccttg 217
Claims (5)
1. A biological agent comprising an endophytic strain of the genus EpicoccusEpicoccum thailandicum LF-28 with a preservation number of CGMCC No.40134; the endophytic epicoccum strainEpicoccum thailandicumThe content of indoleacetic acid in the fermentation liquor of LF-28 is more than 20 mg/L, and the content of gibberellin is more than 15 mg/L.
2. The biological agent of claim 1, wherein the method of fermentative preparation of the biological agent comprises the steps of: culturing endophytic epicoccum strains under fermentation conditionsEpicoccum thailandicumAfter LF-28, obtaining fermentation liquor; culturing for 24-60 hours at the temperature of 24-26 ℃ under the fermentation condition, wherein the rotating speed of a culture device is 0-120 r/min; the initial pH of the fermentation environment was 7.0.
3. The biological agent of claim 1, wherein the endophytic epicoccum strain is preparedEpicoccum thailandicumThe fermentation condition of the fermentation liquid of LF-28 is 60h cultivated at 26 ℃, and the rotating speed of the cultivation device is 120 r/min; the initial pH of the fermentation environment was 7.0.
4. A method for controlling fungal plant diseases, comprising the steps of: applying the biological agent of any one of claims 1-3 before planting plants and during plant growth; the fungi are Fusarium oxysporum of root rot of pseudo-ginseng existing in the pseudo-ginseng continuous cropping soil; the plant is ginseng.
5. The method of claim 4, wherein the method of applying is at least one of spraying, root irrigation, and drip irrigation with a liquid comprising the biological agent.
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