WO2020197163A1 - Beauveria bassiana jef-462 ou beauveria bassiana jef-507 ayant un effet ant-bemisia tabaci, composition anti-bemisia tabaci le comprenant, et procédé anti-bemisia tabaci l'utilisant - Google Patents

Beauveria bassiana jef-462 ou beauveria bassiana jef-507 ayant un effet ant-bemisia tabaci, composition anti-bemisia tabaci le comprenant, et procédé anti-bemisia tabaci l'utilisant Download PDF

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WO2020197163A1
WO2020197163A1 PCT/KR2020/003749 KR2020003749W WO2020197163A1 WO 2020197163 A1 WO2020197163 A1 WO 2020197163A1 KR 2020003749 W KR2020003749 W KR 2020003749W WO 2020197163 A1 WO2020197163 A1 WO 2020197163A1
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jef
strain
bassiana
beauveria bassiana
tobacco powder
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PCT/KR2020/003749
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Korean (ko)
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김재수
백세현
김종철
김시현
이미롱
박소은
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전북대학교산학협력단
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • 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
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi

Definitions

  • the present invention relates to a Beauveria bassiana JEF-462 or Beauveria bassiana JEF-507 having a control effect on tobacco powder, a composition for controlling tobacco powder comprising the same, and a method for controlling tobacco powder using the same.
  • Tobacco powder causes damage by suction, and adults mediate a variety of viruses, and among them, tomato sulfide leaf curl virus (TYLCV) is a problem. Damage caused by the absorption includes inhibition of growth of crops, decrease in yield, fading and atrophy of leaves, and fallen leaves, and fruit coloring is irregular in branches and tomatoes. Larvae secrete nectar and cause soot disease, which inhibits photosynthesis in plants.
  • TYLCV tomato sulfide leaf curl virus
  • Korean Patent Publication No. 10-2016-0000537 discloses the control activity against the Boberia bassiana FG274 strain and its Pabam moth larvae
  • Korean Patent Publication No. 10-2016-0139521 discloses Control activity against Boberia bassiana SD15 strain and its spotted mites or peach aphids
  • Korean Patent Publication No. 10-2016-0084968 is a control activity against Boberia bassiana JEF007 strain and its saw-legged ant spermatozoa
  • domestic patent Publication No. 10-2011-0011239 is Boberia bassiana MaW1 strain and its control activity against blackheads or hairy toads
  • 10-2011-0094749 is Boberia bassiana MsW1 strain and its control activity against Northern Beards.
  • Korean Patent Publication No. 10-2015-0113255 discloses a control activity against water pests such as Boberia bassiana ERL 836 strain and apex or rice weevil.
  • water pests such as Boberia bassiana ERL 836 strain and apex or rice weevil.
  • new highly active microbial pesticides that can control tobacco dust with the same potency as chemical pesticides, and that are environmentally friendly and easy to handle and use.
  • the present inventors have made great efforts to discover new insect pathogenic fungi based on microorganisms that have a control effect on tobacco powder and to establish an effective tobacco powder management system while dramatically overcoming the resistance problem and environmental residual problem of existing synthetic pesticides. .
  • it confirmed the excellent insecticidal power of the tobacco powder of Beauveria bassiana JEF-462 or Beauveria bassiana JEF-507 strain, and completed the present invention.
  • the present invention was conceived to solve the above problems, and the present inventors confirmed the excellent insecticidal power of the Beauveria bassiana JEF-462 (KFCC11821P) or Beauveria bassiana JEF-507 (KFCC11822P) strain, and the present invention Completed.
  • An object of the present invention is to provide a Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) having a control effect on tobacco powder.
  • Another object of the present invention is to provide a composition for controlling tobacco powder comprising a Boberia bassiana JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain, a spore of the strain or a culture solution of the strain. will be.
  • Another object of the present invention is to provide a method for controlling tobacco powder by treating the composition for controlling tobacco powder on a pest, its habitat, a plant or soil to be protected from pests.
  • the present invention was conceived to solve the above problems, and the present inventors confirmed the excellent insecticidal power of the Beauveria bassiana JEF-462 (KFCC11821P) or Beauveria bassiana JEF-507 (KFCC11822P) strain, and the present invention Completed.
  • An object of the present invention is to provide a Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) having a control effect on tobacco powder.
  • Another object of the present invention is to provide a composition for controlling tobacco powder comprising a Boberia bassiana JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain, a spore of the strain or a culture solution of the strain. will be.
  • Another object of the present invention is to provide a method for controlling tobacco powder by treating the composition for controlling tobacco powder on a pest, its habitat, a plant or soil to be protected from pests.
  • Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain of the present invention is cultured in a solid medium containing millet to create spores, and the spore suspension is attached to tobacco powder larvae.
  • a mortality rate of 24% was confirmed in the untreated group, whereas the Beauveria bassiana JEF- 426 or JEF-507 (KFCC11822P) strains each showed a mortality rate of 100%, and also showed a higher mortality rate than other Boberia bassiana strains.
  • Boberia bassiana of the present invention (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain has the effect of controlling tobacco powder.
  • Figure 1 is a boberia bassiana ( Beauveria bassiana ) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain observed with the naked eye (Fig. ).
  • Figure 2a shows the base sequence of the ITS (Internal Transribed Spacer) of the Boberia bassiana ( Beauveria bassiana ) JEF-462 (KFCC11821P) strain.
  • Figure 2b shows the base sequence of the ITS (Internal Transribed Spacer) of the Boberia bassiana ( Beauveria bassiana ) JEF-507 (KFCC11822P) strain.
  • Figure 4 shows the results of the spore production of the Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain using an Italian millet.
  • Figure 5 is a mortality rate of tobacco powder (short repetition) ( Figure 5a) and Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain immersion treatment (Fig. Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) tobacco powder in which mycosis was formed by the strain (FIG. 5B) is shown.
  • Figure 6 is a mortality rate of tobacco powder (3 repetitions) (Fig. 6a) and Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain immersion treatment Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) tobacco powder in which mycosis was formed by the strain (Fig. 6b).
  • composition for controlling tobacco powder using a strain of Beauveria bassiana JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) according to the present invention provides excellent insecticidal power and is effective as a composition for controlling tobacco powder.
  • the present invention provides a strain of Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or Boberia bassiana (Beauveria bassiana) JEF-507 (KFCC11822P) having a control effect on tobacco powder having thrips control effect. can do.
  • 5 to 10 cm of the surface layer was collected in order to collect a soil sample for strain isolation.
  • About 1/4 of the collected soil samples were filled in a paper cup (JEF-462: Mindungsan, Jeongseon-gun, Gangwon, JEF-507: Palamsan, Jangseong-gun, Jeollanam-do).
  • Brown larvae (3-4 animals) were placed, and 1 ml of distilled water was supplied every day to maintain high humidity conditions. Infection was induced by supplying moisture to the brown mealworm larvae treated in the soil for 5-10 days. The strain was isolated from the mycosis of the larva.
  • the strain was isolated by culturing (25 ⁇ 2°C, dark condition) in 1/4 Sabouraud Dextrose Agar (1/4SDA; Difco, USA). A single colony was obtained through 2-3 separations. The acquired single colony was used to re-test the pathogenicity to the brown mealworm larva. The strain was re-tested to confirm pathogenicity and the gene was amplified using ITS primer (ITS1F: 5'-TCCGTAGGTGAACCTGCGG-3'/ ITS4R: 5'-TCCTCCGCTTATTGATATGC-3'). BLAST was performed using the amplified gene information. The isolated and identified insect pathogenic fungi were stored at -70°C with 15-20% glycerol stock for long-term storage.
  • the present inventors sequenced the strain obtained in Example 1 and finally identified it as a strain of Beauveria bassiana JEF-462 or JEF-507 (FIG. 2), and Korean microbial conservation Deposited with the Center on February 12, 2019 as deposit number KFCC11821P (Beauveria bassiana JEF-462) or KFCC11822P (Beauveria bassiana) JEF-507.
  • the present invention is for the control of tobacco powder, including the Boberia bassiana JEF-462 (KFCC11821P) or the Boberia bassiana JEF-507 (KFCC11822P) strain, spores of the strain or culture solution of the strain Composition can be provided.
  • the composition for controlling tobacco powder is preferably prepared through solid culture by inoculating the suspension of the strain on a grain medium, more preferably millet, rice, Italian millet, barley, It is to use a medium consisting of one or more grains selected from the group consisting of soybean, sorghum, crude, rice, and brown rice, of which it is most preferable to inoculate a millet medium and culture the solid (FIG. 4).
  • the solid culture is after inoculating the suspension of the strain in a medium, preferably incubating at 20 to 30° C. for 5 to 20 days, and then at room temperature conditions of 1 to It can be dried for 30 days, more preferably after incubation for 7 days at 25° C., and drying at room temperature for 1 day is preferable.
  • composition for controlling tobacco powder of the present invention contains spores of the strains of Beauveria bassiana JEF-462 (KFCC11821P) and/or JEF-507 (KFCC11822P), after the spores are attached to the host insect It germinates and causes mycosis, and an insecticidal effect appears. In addition, in addition to the tobacco powder directly in contact with this, even tobacco powder that does not directly contact the composition may exhibit a control effect due to the transmission of spores.
  • the composition for controlling tobacco powder of the present invention is preferably in a liquid, powder or granular form.
  • the powder formulation refers to a powder form
  • the granular formulation refers to a form of grains or grains obtained by diluting a solid culture medium by 1 to 100 times the weight by using an extender, but is not limited thereto.
  • the strains of Beauveria bassiana JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) cause mycosis in adult cucumber thrips and exhibit excellent effects on tobacco powder.
  • the present invention can also provide a method for controlling tobacco powder by treating the above-described composition for controlling tobacco powder on pests, its habitat, plants or soil to be protected from pests.
  • composition for controlling tobacco powder can be prepared as a suspension.
  • composition for controlling tobacco powder may be subjected to immersion treatment, contact treatment, spray treatment, and most preferably immersion treatment.
  • the suspension of the present invention is preferably prepared by harvesting the spores of the cultured strain, which is a solid culture medium or composition for controlling tobacco powder, counting with a hemocytometer, and then diluting according to the treatment concentration using various surfactants. Do.
  • the number of spores may be 1x10 1 spores (conidia)/g to 1x10 10 spores (conidia)/g, preferably 1x10 3 spores (conidia) /g to 1x10 7 spores (conidia) / g may be, more preferably 1 ⁇ 10 7 spores (conidia) / ml is preferred.
  • immersion treatment in the tobacco powder means spraying 0.1 to 10 ml of suspension per 100 grains on the tobacco powder, and preferably immersion treatment with 1 ml of suspension. It is desirable. If the suspension is treated in excess of the above capacity, it is difficult to secure economic efficiency of control compared to the treatment amount, and if the suspension is treated less than the above capacity, it is difficult to obtain high control activity.
  • Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain spore suspension of the present invention was confirmed to show a high insecticidal effect when immersed in tobacco powder larvae (Fig. 5 And Figure 6).
  • Boberia bassiana (Beauveria bassiana) JEF-462 (KFCC11821P) or JEF-507 (KFCC11822P) strain showed a higher insecticidal effect of tobacco powder than other Boberia bassiana strains (FIGS. 5 and 6 ). .
  • the present inventors isolated insect pathogenic fungi from soil samples under the following conditions.
  • 5-10 cm of the surface layer was collected.
  • About 1/4 of the collected soil samples were filled in a paper cup (JEF-462: Mindungsan, Jeongseon-gun, Gangwon, JEF-507: Palamsan, Jangseong-gun, Jeollanam-do).
  • Brown larvae (3-4 animals) were placed, and 1 ml of distilled water was supplied every day to maintain high humidity conditions.
  • Infection was induced by supplying moisture to the brown mealworm larvae treated in the soil for 5-10 days.
  • the strain was isolated from the mycosis of the larva.
  • the strain was isolated by culturing (25 ⁇ 2°C, dark condition) in 1/4 Sabouraud Dextrose Agar (1/4SDA; Difco, USA). A single colony was obtained through 2-3 separations. The acquired single colony was used to re-test the pathogenicity to the brown mealworm larva. The strain was re-tested to confirm pathogenicity and the gene was amplified using ITS primer (ITS1F: 5'-TCCGTAGGTGAACCTGCGG-3'/ ITS4R: 5'-TCCTCCGCTTATTGATATGC-3'). BLAST was performed using the amplified gene information. The isolated and identified insect pathogenic fungi were stored in 15-20% glycerol and stored at -70°C for long-term storage.
  • the agar block in which Beauveria bassiana was cultured was transferred to Sabraud Dextrose Agar (SDA) and cultured (25° C., dark) for 7 days, and then observed with the naked eye and an optical microscope (400 times) (FIG. 1 ).
  • SDA Sabraud Dextrose Agar
  • FIG. 1 White hyphae, grayish white spores, and spore formation, which are typical of the strains of the genus Beauveria, were observed.
  • Example 2 DNA sequencing of the Beauveria bassiana strain of Example 1
  • the present invention identified the Beauveria bassiana of Example 1. Specifically, PCR was performed under the following conditions. 94°C for 5 minutes, 94°C for 60 seconds, 62°C for 30 seconds, and 72°C for 30 seconds were performed 30 times, and finally post maturation was performed at 72°C for 3 minutes. The sequence was carried out using the ITS primers in Table 1 below. PCR products were electrophoresed for 25 minutes at 140 V using 0.8% agarose gel and Eco dye. As a result of sequencing the finally obtained product, it was identified as B. bassiana (FIG. 2).
  • the present inventors named the Boberia bassiana of Example 1 as a strain, respectively, as Boberia bassiana JEF-462 (KFCC11821P) and Boberia bassiana JEF-507 (KFCC11822P). I did.
  • the ITS (internal transcribed spacer) sequences of the Beauveria bassiana JEF-462 strain and the Beauveria bassiana JEF-507 strain were respectively described in SEQ ID NO: 1 and SEQ ID NO: 2.
  • Spore suspension (1x10 7 conidia/ml) was prepared using B. bassiana JEF-462 or JEF-507 cultured in millet, and then immersed in tomato leaves with tobacco powdery larvae attached thereto. The experiment was performed while maintaining at 25°C, and the number of live insects and the number of dead insects were investigated from 3 to 5 days after treatment.
  • Example 5 The control effect of the strains of Beauveria bassiana JEF-462 and JEF-507 against tobacco powder (soaking treatment -3 repetitions)

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Abstract

La présente invention concerne : Beauveria bassiana JEF-462 (KFCC11821P) ou Beauveria Bassiana JEF-507 (KFCC11822P) ayant un effet anti-Bemisia tabaci; une composition anti-Bemisia tabaci le comprenant; et un procédé anti-Bemisia tabaci l'utilisant.
PCT/KR2020/003749 2019-03-22 2020-03-19 Beauveria bassiana jef-462 ou beauveria bassiana jef-507 ayant un effet ant-bemisia tabaci, composition anti-bemisia tabaci le comprenant, et procédé anti-bemisia tabaci l'utilisant WO2020197163A1 (fr)

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KR10-2019-0032711 2019-03-22
KR1020190032711A KR102162105B1 (ko) 2019-03-22 2019-03-22 담배가루이에 방제효과를 갖는 Beauveria bassiana JEF-462 또는 Beauveria bassiana JEF-507, 이를 포함하는 담배가루이 방제용 조성물 및 이를 이용한 담배가루이 방제방법

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114403163A (zh) * 2022-01-26 2022-04-29 合肥高尔生命健康科学研究院有限公司 一种防治大棚茄子烟粉虱的方法、及球孢白僵菌的培养方法
WO2023202608A1 (fr) * 2022-04-22 2023-10-26 浙江泰达作物科技有限公司 Souche de beauveria bassiana à haute résistance aux rayonnements ultraviolets ainsi que son procédé de mutagenèse directionnelle et son utilisation

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KR102395628B1 (ko) 2021-09-06 2022-05-06 안동대학교 산학협력단 신규 미생물 보베리아 바시아나 aad16 균주와 이를 이용한 미생물 제제 및 살충방법

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114403163A (zh) * 2022-01-26 2022-04-29 合肥高尔生命健康科学研究院有限公司 一种防治大棚茄子烟粉虱的方法、及球孢白僵菌的培养方法
WO2023202608A1 (fr) * 2022-04-22 2023-10-26 浙江泰达作物科技有限公司 Souche de beauveria bassiana à haute résistance aux rayonnements ultraviolets ainsi que son procédé de mutagenèse directionnelle et son utilisation

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