CN120137800A - A kind of Mortierella longispora and application thereof - Google Patents
A kind of Mortierella longispora and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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Abstract
本发明公开了一种长孢被孢霉及其应用，所述长孢被孢霉(Mortierella elongata)的菌株号为JSAFC 247，于2024年12月09日保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏编号为CGMCC No.41703。本申请提供的长孢被孢霉菌株对铜绿丽金龟幼虫致病力强，环保无污染、不易产生抗药性，可作为一种活体生物农药，在铜绿丽金龟的生物防治中具有非常强的应用潜力。
The present invention discloses a long-spore ...
Description
Technical Field
The invention belongs to the technical field of microorganisms, and relates to mortierella longifolia and application thereof.
Background
The tortoise (Anomala corpulenta) belongs to the family Coleoptera, the family Porphyraceae, is an important underground pest, is widely distributed in China, has trails in various places, is common in environments such as orchards, farmland greenbelts, forest belts and the like, the adult and the larva are used as the harm, the adult bites the tree and the leaf part of the crop, the scarcity is caused, the larva bites the seed just germinated at the root part of the crop, the seedling is lacked, the ridge is broken, or the root part and the root system of the plant stem are bitten, so that the plant is dead.
The traditional control method of the cudweed is mainly based on chemical pesticide control, and the chemical pesticide has the advantages of quick response, obvious effect, inhibition of pest population density in a short time and protection of agriculture and forestry, but in long-term benefit, the large-scale use of the pesticide generates serious 3R problem (pesticide residue, drug resistance and rampant of pests), kills natural enemies and other pollinating insects, and damages ecological balance, self-repairing capability of the earth and other problems are not ignored. Along with the concept of ecological health development, research and application of pest biological control technology are increasingly paid attention to. Biological control is a trend of pest control method in the future, can solve a series of problems (high toxicity, environmental pollution, natural enemies killing, drug resistance, residues and the like) caused by chemical pesticides, uses pathogenic microorganisms, sex attractant, food attractant, natural enemies of insects, plant source pesticides and the like for pest control, develops a precise, efficient, safe and environment-friendly green control new technology, and research and application of biocontrol bacteria become a hotspot in the field of biological control. The main application research is mainly focused on beauveria bassiana and metarhizium anisopliae, and the development and the utilization of other pest biocontrol microbial inoculum resources are less.
Disclosure of Invention
The invention aims to provide mortierella (Mortierella elongata) and application thereof in controlling copper green scarves.
The invention provides a mortierella (Mortierella elongata), the strain number of the mortierella (Mortierella elongata) is JSAFC 247, and the mortierella is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of 41703 at the year 2024 and the month 12 and 09.
Further, the nucleotide sequence of the ITS gene of mortierella longifolia is shown as SEQ ID NO. 1.
The invention also provides a biocontrol microbial inoculum, and the active ingredient of the biocontrol microbial inoculum is the mortierella longifolia (Mortierella elongata).
The invention also provides application of the mortierella (Mortierella elongata) and the biocontrol microbial inoculum in controlling the cuvettes.
Further, the application is specifically by spraying with a spore suspension of Mortierella elongata (Mortierella elongata) JSAFC 247 on the larvae of the Pacific mossambica or on plants fed by the larvae of the Pacific mossambica.
Further, the preparation method of the spore suspension comprises (1) taking the strain of biocontrol mortierella JSAFC, picking hypha with inoculating needle under aseptic condition, inoculating sterilized PDA culture medium, culturing for 5 days, (2) scraping hypha under aseptic condition, washing hypha with aseptic water and filtering with gauze, and (3) adjusting spore concentration with aseptic water under aseptic condition.
Further, the culture conditions in the step (1) are 25 ℃, 75% humidity and dark culture.
Further, the spore suspension had a concentration of 10 8/ml.
Compared with the prior art, the application has the remarkable advantages that 1 mortierella (Mortierella elongata) is obtained by separating and purifying soil of Jiangsu Jurong. The strain can be used as a biocontrol strain, has higher mortality rate to the copper green scarab, is environment-friendly and nontoxic, can obviously inhibit the survival activity of the copper green scarab, and has great significance for developing biocontrol bacteria for preventing and controlling the copper green scarab in the future.
Drawings
FIG. 1:Mortierella elongata JSAFC 247 shows colony morphology.
The phylogenetic tree of fig. 2:Mortierella elongata JSAFC 247.
FIG. 3:Mortierellaelongata JSAFC 247 effect on the growth of P.aeruginosa larvae.
Detailed Description
The technical scheme of the invention is further described below with reference to the accompanying drawings.
As used herein, the term "biocontrol bacteria" refers to beneficial microorganisms, which can control plant diseases, mainly bacteria, fungi, actinomycetes.
Example 1:Mortierellaelongata JSAFC 247 isolation and identification
1. Mortierellaelongata JSAFC 247 separation of 247
The soil of Jiangsu Jurong is taken, JSAFC 247 is obtained by separation, and the colony is characterized in that the colony is cultured on a PDA flat-plate culture medium, quickly grows in a rose shape, and the surface of the colony is light yellow.
2. Mortierella elongata JSAFC 247 molecular biological identification
(1) Genomic DNA of strain JSAFC 247 was extracted using TIANGEN company's kit.
(2) The ITS genes of the genomic DNA were amplified separately. The DNA was amplified using a reaction volume of 30. Mu.L, containing 15. Mu.L of 2X EASYTAQ PCR Supermix (+dyne), 1. Mu.L (10. Mu.M) each of the primer pair ITS1 (SEQ ID NO.2: 5'-CTTGGTCATTTAGAGGAAGTAA-3') & ITS4 (SEQ ID NO.3: 5'-TCCTCCGCTTATTGATATGC-3'), 2. Mu.L of template DNA and 11. Mu.L of ddH 2 O, under the following PCR amplification procedure conditions of 94℃pre-denaturation for 5min, 94℃denaturation for 30s,54℃annealing for 45s,72℃extension for 60s for 35 cycles, and finally 72℃extension for 10min.
(3) The amplified PCR product is subjected to 1% agarose gel electrophoresis, and then observed under ultraviolet lamp, and the PCR product with target band is sent to Nanjing qing department Biotechnology Co., ltd for sequencing, wherein the sequencing result is that ITS gene sequence of strain JSAFC 247 is SEQ ID NO.1(GAAGTAAAAAATCGTAACAAGGTTTCCGTAGGTGAACCTGCGGAAGGATCATTC ATAATAAGTGTTTTATGGCACTTTTTAAATCCATATCCACCTTGTGTGCAATGTCAGTCGATCTTCTTCATGGAGATCGACCAAACATCAACCTTTATTTTTTAACTCTTTGTCTGAAAAATATTATGAATAAACAATTCAAAATACAACTTTCAACAACGGATCTCTTGGCTCTCGCATCGATGAAGAACGCAGCGAAATGCGATACGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCATATTGCGCTCTTTGGTATTCCGAAGAGCATGCTTGTTTGAGTATCAGTAAACACCTCAAAGCTTTTGGATTTTTTAATCGAAAAGCTTTGGACTTGAGCAATCCCAACACCAATCTTTTTGAGATCGGTGGCGGGTTGCTTGAAATGCAGGTGCAGCTGGACATTCTCCTGAGCTAAAAGCATATTCATTTAGTCCCGTCAAACGGATTATTACTTTTGCTGCAGCTAACATAAAGGGAGTTTGACCGTATTGGCTGACTGATGCAGGATTTCACAAGGGTCGGCAACGATTCTTGTTAAACTCGATCTCAAATCAAGTAAGACTACCCGCTGAACTTAAGCATATCAAAAGGCGGAG).
(4) The SEQ ID NO.1 sequences were aligned on the NCBI database website and phylogenetic trees were established, see FIG. 2. The comparison results are shown in Table 1. From the results shown in Table 1, it was estimated that the biocontrol bacterium JSAFC 247,247 of the present invention was Mortierella elongata (Mortierellaelongata), and was named Mortierella elongata JSAFC 247,247, and the strain was deposited at China general microbiological culture Collection center (CGMCC) No.41703, classification No. Mortierellaelongata JSAFC 247,247, and the deposit address was China academy of sciences of China, no. 3, north Star, chaoyang, beijing, at 2024, and month 09.
TABLE 1SEQ ID NO.1 sequence alignment
Example 2:Mortierella elongata JSAFC 247 determination of pathogenicity of larvae of P.aeruginosa
1. Preparation of spore suspension
(1) 1 Mycelium block of strain Mortierella elongata JSAFC mm in diameter was inoculated into 9cm diameter petri dish (containing 15ml PDA medium) with a puncher, and cultured in the dark at 25deg.C and 75% humidity for 7 days.
(2) Mortierella elongata JSAFC 247 cultured for 7 days was sterilized, the mycelia were scraped off, washed with sterilized water and filtered with gauze, and then spore concentration was adjusted to 10 8/mL.
2. Inoculation of test insects
(1) The larvae of the copper green scarab beetles (collected from the part of the white town Si Zhuang Cuntian in Jurong City) are bred in a artificial climatic box (the temperature is 26+/-1 ℃ and the humidity is 80 percent, and the light-dark ratio is 14:10) until the larvae are 2 years old for standby.
(2) The insecticidal activity of JSAFC 247,247 against the copper green scarab beetles was measured using sterilized ultrapure water as a negative control.
(3) The spore suspension of JSAFC strain 247 is prepared, the larvae of the copper green scarlet beetles are taken, each group is provided with 3 repeated areas, each area is provided with 20 heads, each head is dipped for 30s, the heads are placed on sterile filter paper, the heads are crawled and dried to moisture on the body, then the heads are placed into a feeding box containing sterile soil, sterilized feed is fed daily, the infection and death conditions of the larvae are observed and recorded every day, and the continuous observation is carried out for 28 days.
The experimental results are shown in table 3 and fig. 3.
TABLE 3 mortality of copper green scarves after JSAFC A247 inoculation (%)
As can be seen from the data in Table 3, the larva mortality rate of the treated group is obviously higher than that of the control group after 28 days of inoculation, the JSAFC strain in the invention has the mortality rate of more than 98% on the copper green scara, the death peak period is 14-20 days, and dead insects appear after 3 days of inoculation. The strain JSAFC 247,247 of the invention has good killing effect on the larvae of the copper green scarab beetles.
In conclusion, JSAFC 247,247 provided by the invention provides a biological control resource for controlling the larvae of the copper green scarab beetles, and has the characteristics of strong toxicity, high insecticidal speed and the like. Meanwhile, the problems of pesticide residue, environmental pollution and the like generated by controlling pests by using chemical pesticides are solved, and the pesticide has good development potential and production value.
Claims (8)
1. The mortierella (Mortierella elongata) is characterized in that the mortierella (Mortierella elongata) has a strain number of JSAFC 247 and is preserved in China general microbiological culture collection center (CGMCC) with a preservation number of 41703 at a date of 09 of 12 months of 2024.
2. Mortierella (Mortierella elongata) according to claim 1, characterized in that the nucleotide sequence of ITS gene of mortierella is shown in SEQ ID No. 1.
3. The biocontrol microbial agent is characterized in that the active ingredient of the biocontrol microbial agent is mortierella longifolia (Mortierella elongata) according to any one of claims 1-2.
4. The application of mortierella (Mortierella elongata) of any one of claims 1-2 and the biocontrol microbial inoculum of claim 3 in controlling copper green scarab beetles.
5. The use according to claim 4, characterized in that it is carried out by spraying a spore suspension of mortierella elongata (Mortierella elongata) JSAFC 247 according to any one of claims 1-2 onto the larvae of the tortoise plastron, or onto the plants fed by the larvae of the tortoise plastron.
6. The method of claim 5, wherein the spore suspension is prepared by (1) taking the strain of Mortierella elongata JSAFC, picking up mycelia with an inoculating needle under aseptic condition, inoculating to sterilized PDA culture medium, culturing for 5 days, (2) scraping mycelia under aseptic condition, washing mycelia with aseptic water and filtering with gauze, and (3) adjusting spore concentration with aseptic water under aseptic condition.
7. The use according to claim 6, wherein the culture conditions in step (1) are 25 ℃, 75% humidity, dark culture.
8. The use according to claim 5, characterized in that the concentration of the spore suspension is 10 8/mL.
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| CN202510371081.4A CN120137800A (en) | 2025-03-27 | 2025-03-27 | A kind of Mortierella longispora and application thereof |
| CN202511513443.5A CN121427691A (en) | 2025-03-27 | 2025-10-22 | Mortierella elongata and application thereof |
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| CN202511513443.5A Pending CN121427691A (en) | 2025-03-27 | 2025-10-22 | Mortierella elongata and application thereof |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107760610A (en) * | 2017-09-29 | 2018-03-06 | 中国科学院南京土壤研究所 | One plant long spore Mortierella and its application |
| WO2020023699A1 (en) * | 2018-07-25 | 2020-01-30 | Flagship Pioneering Innovations V, Inc. | Compositions and related methods for agriculture |
| CN117946876A (en) * | 2024-02-04 | 2024-04-30 | 河南农业大学 | A Chaetomium truncatulum F8 strain and its application |
| CN118879506A (en) * | 2024-07-15 | 2024-11-01 | 北京林业大学 | Polysporus cucurbitae and its application |
| CN119614390A (en) * | 2025-01-10 | 2025-03-14 | 江苏农林职业技术学院 | Pochorus chlamydosporium Mortierella and application thereof |
| CN119614395A (en) * | 2025-01-10 | 2025-03-14 | 江苏农林职业技术学院 | Engella elongata and application thereof |
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- 2025-03-27 CN CN202510371081.4A patent/CN120137800A/en active Pending
- 2025-10-22 CN CN202511513443.5A patent/CN121427691A/en active Pending
Patent Citations (7)
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| CN107760610A (en) * | 2017-09-29 | 2018-03-06 | 中国科学院南京土壤研究所 | One plant long spore Mortierella and its application |
| WO2020023699A1 (en) * | 2018-07-25 | 2020-01-30 | Flagship Pioneering Innovations V, Inc. | Compositions and related methods for agriculture |
| CN112739214A (en) * | 2018-07-25 | 2021-04-30 | 旗舰创业创新五公司 | Compositions and related methods for use in agriculture |
| CN117946876A (en) * | 2024-02-04 | 2024-04-30 | 河南农业大学 | A Chaetomium truncatulum F8 strain and its application |
| CN118879506A (en) * | 2024-07-15 | 2024-11-01 | 北京林业大学 | Polysporus cucurbitae and its application |
| CN119614390A (en) * | 2025-01-10 | 2025-03-14 | 江苏农林职业技术学院 | Pochorus chlamydosporium Mortierella and application thereof |
| CN119614395A (en) * | 2025-01-10 | 2025-03-14 | 江苏农林职业技术学院 | Engella elongata and application thereof |
Non-Patent Citations (2)
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| MICHAEL J DILEGGE ET AL;: "A novel approach to determine generalist nematophagous microbes reveals Mortierella globalpina as a new biocontrol agent against Meloidogyne spp. nematodes", 《SCI REP》, vol. 9, no. 1, 17 May 2019 (2019-05-17), pages 1 - 9 * |
| 裘智杰 等;: "被孢霉在可持续农业生产中的应用研究进展", 《江苏农业学报》, vol. 40, no. 4, 31 December 2024 (2024-12-31), pages 762 - 768 * |
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