WO2024027266A1 - Bacillus thuringiensis biocontrol microbial agent for inhibiting reproduction of pyrausta nubilalis and preparation method therefor - Google Patents

Bacillus thuringiensis biocontrol microbial agent for inhibiting reproduction of pyrausta nubilalis and preparation method therefor Download PDF

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WO2024027266A1
WO2024027266A1 PCT/CN2023/093802 CN2023093802W WO2024027266A1 WO 2024027266 A1 WO2024027266 A1 WO 2024027266A1 CN 2023093802 W CN2023093802 W CN 2023093802W WO 2024027266 A1 WO2024027266 A1 WO 2024027266A1
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bacillus thuringiensis
bacillus
biocontrol agent
verticillium
reproduction
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PCT/CN2023/093802
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French (fr)
Chinese (zh)
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周义新
黄杰
蒋羽
刘琪
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湖北茂盛生物有限公司
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Priority to PCT/CN2023/093802 priority Critical patent/WO2024027266A1/en
Publication of WO2024027266A1 publication Critical patent/WO2024027266A1/en

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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
    • 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
    • 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
    • A01N63/38—Trichoderma
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P7/00—Arthropodicides
    • A01P7/04—Insecticides

Definitions

  • the present invention relates to the technical field of insect repellents, specifically a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer and its preparation method.
  • Corn borer also called corn borer, belongs to the order Lepidoptera and the family Boreridae. There are two types of corn borer in my country: Asian corn borer and European corn borer. They mainly damage corn, sorghum, millet, etc., and can also damage cotton, sugarcane, and hemp. , sunflower, rice, sugar beet, sugarcane beans and other crops are worldwide pests.
  • the object of the present invention is to provide a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer and its preparation method, so as to solve the problems raised in the above background technology.
  • the present invention provides the following technical solution: a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer and a preparation method thereof.
  • the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 15-25% , Metarhizium anisopliae 15-20%, Trichoderma viride 12-18%, Bacillus subtilis 10-15%, Beauveria bassiana 10-14%, and the balance is Verticillium cereus.
  • the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 15%, Metarhizium anisopliae 15%, Trichoderma viride 12%, Bacillus subtilis 10%, Beauveria bassiana 10% , the balance is Verticillium cereus.
  • the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 18%, Metarhizium anisopliae 17%, Trichoderma viride 14%, Bacillus subtilis 12%, Beauveria bassiana 12% , the balance is Verticillium cereus.
  • the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 22%, Metarhizium anisopliae 18%, Trichoderma viride 16%, Bacillus subtilis 14%, Beauveria bassiana 13% , the balance is Verticillium cereus.
  • the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 25%, Metarhizium anisopliae 20%, Trichoderma viride 18%, Bacillus subtilis 15%, Beauveria bassiana 14% , the balance is Verticillium cereus.
  • the Metarhizium anisopliae is ACCC30104
  • Trichoderma viride is ATCC 26802
  • Bacillus subtilis is ATCC605.
  • the Bacillus thuringiensis is selected from at least one of B.thuringiensis KCTC 1507, B.thuringiensis CVCM 1812 and B.thuringiensis DSM 5815.
  • the Beauveria bassiana is selected from at least one of B. bassiana ATCC 74040 and B. bassiana ATCC MYA-4886.
  • the Verticillium cereus is selected from at least one of L.lecanii VKM F-1199, L.lecanii NFCCI 0514 and L.lecanii IRAN 1822C.
  • a preparation method of a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer includes the following steps:
  • Beauveria bassiana and Verticillium cereus are respectively subjected to bacterial strain activation, seed liquid culture, liquid fermentation, temperature regulation and sporulation, and drying; Beauveria bassiana spore powder and Verticillium cereus spore powder are obtained;
  • Bacillus thuringiensis seed liquid into the expanded medium at an inoculum volume of 8%, and culture it at 30°C for 24 hours to obtain a Bacillus thuringiensis culture liquid;
  • Trichoderma viride seed liquid and the Bacillus subtilis seed liquid at a volume ratio of 1:2, then insert it into the expansion medium at an inoculum volume of 8% volume ratio, and culture it at 28°C for 24 hours to obtain Trichoderma viride-Bacillus subtilis spores.
  • Bacillus mixed culture solution Bacillus mixed culture solution;
  • a biological control agent is specially formulated.
  • Metarhizium anisopliae can penetrate into the body of pests through the body surface, reproduce in the pests' bodies, consume host nutrients and kill the pests; it can also produce Metarhizium anisopliae, which is Metarhizium anisopliae is a secondary metabolite produced during the metabolic process, which has contact killing and antifeeding effects on lepidopteran pests.
  • the two have the effect of synergistically eliminating corn borers.
  • the effect has significantly improved the control effect; because of its good effect and high stability, it can further reduce the number of pesticide applications, protect the environment, and is not easy to develop drug resistance, so it has great promotion and application value.
  • Figure 1 is a flow chart of the method of the present invention.
  • the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 15%, Metarhizium anisopliae 15%, Trichoderma viride 12%, Bacillus subtilis 10% , Beauveria bassiana 10%, the balance is Verticillium cereus;
  • Metarhizium anisopliae is ACCC30104
  • Trichoderma viride is ATCC 26802
  • Bacillus subtilis is ATCC605
  • Beauveria bassiana is selected from B.bassiana ATCC 74040
  • Verticillium cereus The bacteria were selected from L.lecanii VKM F-1199.
  • Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 18%, Metarhizium anisopliae 17%, Trichoderma viride 14%, Bacillus subtilis 12%, Beauveria bassiana 12%, the balance is wax Verticillium verticillium;
  • Metarhizium anisopliae is ACCC30104
  • Trichoderma viride is ATCC 26802
  • Bacillus subtilis is ATCC605
  • Bacillus thuringiensis is selected from B.thuringiensis CVCM 1812
  • Beauveria bassiana is selected from B.bassiana ATCC MYA-4886
  • the wax scale Verticillium was selected from L.lecanii NFCCI 0514.
  • Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 22%, Metarhizium anisopliae 18%, Trichoderma viride 16%, Bacillus subtilis 14%, Beauveria bassiana 13%, the balance is wax Verticillium verticillium;
  • Metarhizium anisopliae is ACCC30104
  • Trichoderma viride is ATCC 26802
  • Bacillus subtilis is ATCC605
  • Bacillus thuringiensis DSM 5815 is selected from B.bassiana ATCC 74040
  • Verticillium cereus The bacteria were selected from L.lecanii IRAN 1822C.
  • Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 25%, Metarhizium anisopliae 20%, Trichoderma viride 18%, Bacillus subtilis 15%, Beauveria bassiana 14%, the balance is wax Verticillium verticillium;
  • Metarhizium anisopliae is ACCC30104
  • Trichoderma viride is ATCC 26802
  • Bacillus subtilis is ATCC605
  • Bacillus thuringiensis is selected from the mixture of B.thuringiensis KCTC 1507
  • Coccidioides albicans B. bassiana was selected from the mixture of B. bassiana ATCC 74040 and B. bassiana ATCC MYA-4886
  • Verticillium cereus was selected from the mixture of L. lecanii VKM F-1199, L. lecanii NFCCI 0514 and L. lecanii IRAN 1822C.
  • a method for preparing a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer includes the following steps:
  • Beauveria bassiana and Verticillium cereus are respectively subjected to bacterial strain activation, seed liquid culture, liquid fermentation, temperature regulation and sporulation, and drying; Beauveria bassiana spore powder and Verticillium cereus spore powder are obtained;
  • Bacillus thuringiensis seed liquid into the expanded medium at an inoculum volume of 8%, and culture it at 30°C for 24 hours to obtain a Bacillus thuringiensis culture liquid;
  • Trichoderma viride seed liquid and the Bacillus subtilis seed liquid at a volume ratio of 1:2, then insert it into the expansion medium at an inoculum volume of 8% volume ratio, and culture it at 28°C for 24 hours to obtain Trichoderma viride-Bacillus subtilis spores.
  • Bacillus mixed culture solution Bacillus mixed culture solution;

Abstract

Provided are a Bacillus thuringiensis biocontrol microbial agent for inhibiting the reproduction of Pyrausta nubilalis and a preparation method therefor. The Bacillus thuringiensis biocontrol microbial agent comprises the following components by weight: 15-25% of Bacillus thuringiensis, 15-20% of Metarhizium anisopliae, 12-18% of Trichoderma viride, 10-15% of Bacillus subtilis, and 10-14% of Beauveria bassiana, the balance being Verticillium lecanii. The Bacillus thuringiensis biocontrol microbial agent kills Pyrausta nubilalis by means of biological control, which is different from traditional products, and is mainly formed by means of mixing a solution of Metarhizium anisopliae with a concentrated extraction solution, wherein Metarhizium anisopliae can penetrate into pests through the surface of the body, reproduce in the pests and consume the host's nutrition to kill the pests.

Description

抑制玉米螟繁殖的苏云金杆菌生防菌剂及其制备方法Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer and its preparation method 技术领域Technical field
本发明涉及防虫剂技术领域，具体为抑制玉米螟繁殖的苏云金杆菌生防菌剂及其制备方法。The present invention relates to the technical field of insect repellents, specifically a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer and its preparation method.
背景技术Background technique
玉米螟，又叫玉米钻心虫，属于鳞翅目，螟蛾科，我国发生的玉米螟有亚洲玉米螟和欧洲玉米螟两种，主要为害玉米、高粱、谷子等，也能为害棉花甘蔗、大麻、向日葵、水稻、甜菜、甘蔗豆类等作物，属于世界性害虫。Corn borer, also called corn borer, belongs to the order Lepidoptera and the family Boreridae. There are two types of corn borer in my country: Asian corn borer and European corn borer. They mainly damage corn, sorghum, millet, etc., and can also damage cotton, sugarcane, and hemp. , sunflower, rice, sugar beet, sugarcane beans and other crops are worldwide pests.
现有技术中，防治玉米螟的药剂种类较多，但主要以化学农药为主。多年来应用菊酯类和有机磷类杀虫剂进行防治，喷药数量加大使其抗药性增强，防治难度大，同时带来环境污染问题。In the existing technology, there are many types of pesticides for preventing and controlling corn borer, but mainly chemical pesticides are used. For many years, pyrethroids and organophosphorus pesticides have been used for prevention and control. The increase in the number of sprays has increased the resistance to pesticides, making prevention and control difficult and causing environmental pollution problems.
发明内容Contents of the invention
本发明的目的在于提供抑制玉米螟繁殖的苏云金杆菌生防菌剂及其制备方法，以解决上述背景技术中提出的问题。The object of the present invention is to provide a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer and its preparation method, so as to solve the problems raised in the above background technology.
为实现上述目的，本发明提供如下技术方案：抑制玉米螟繁殖的苏云金杆菌生防菌剂及其制备方法，所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌15-25%、金龟子绿僵菌15-20%、绿色木霉12-18%、枯草芽孢杆菌10-15%、球孢白僵菌10-14%，余量为蜡蚧轮枝菌。In order to achieve the above object, the present invention provides the following technical solution: a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer and a preparation method thereof. The Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 15-25% , Metarhizium anisopliae 15-20%, Trichoderma viride 12-18%, Bacillus subtilis 10-15%, Beauveria bassiana 10-14%, and the balance is Verticillium cereus.
优选的，所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌15%、金龟子绿僵菌15%、绿色木霉12%、枯草芽孢杆菌10%、球孢白僵菌10%，余量为蜡蚧轮枝菌。Preferably, the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 15%, Metarhizium anisopliae 15%, Trichoderma viride 12%, Bacillus subtilis 10%, Beauveria bassiana 10% , the balance is Verticillium cereus.
优选的，所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌18%、金龟子绿僵菌17%、绿色木霉14%、枯草芽孢杆菌12%、球孢白僵菌12%，余量为蜡蚧轮枝菌。Preferably, the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 18%, Metarhizium anisopliae 17%, Trichoderma viride 14%, Bacillus subtilis 12%, Beauveria bassiana 12% , the balance is Verticillium cereus.
优选的，所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌22%、金龟子绿僵菌18%、绿色木霉16%、枯草芽孢杆菌14%、球孢白僵菌13%，余量为蜡蚧轮枝菌。Preferably, the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 22%, Metarhizium anisopliae 18%, Trichoderma viride 16%, Bacillus subtilis 14%, Beauveria bassiana 13% , the balance is Verticillium cereus.
优选的，所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌25%、金龟子绿僵菌20%、绿色木霉18%、枯草芽孢杆菌15%、球孢白僵菌14%，余量为蜡蚧轮枝菌。Preferably, the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 25%, Metarhizium anisopliae 20%, Trichoderma viride 18%, Bacillus subtilis 15%, Beauveria bassiana 14% , the balance is Verticillium cereus.
优选的，所述金龟子绿僵菌为ACCC30104，绿色木霉为ATCC 26802，枯草芽孢杆菌为ATCC605。Preferably, the Metarhizium anisopliae is ACCC30104, Trichoderma viride is ATCC 26802, and Bacillus subtilis is ATCC605.
优选的，所述苏云金芽孢杆菌选自B.thuringiensis KCTC 1507、B.thuringiensis CVCM 1812和B.thuringiensis DSM 5815中的至少一种。Preferably, the Bacillus thuringiensis is selected from at least one of B.thuringiensis KCTC 1507, B.thuringiensis CVCM 1812 and B.thuringiensis DSM 5815.
优选的，所述球孢白僵菌选自B.bassiana ATCC 74040和B.bassiana ATCC MYA-4886中的至少一种。Preferably, the Beauveria bassiana is selected from at least one of B. bassiana ATCC 74040 and B. bassiana ATCC MYA-4886.
优选的，所述蜡蚧轮枝菌选自L.lecanii VKM F-1199、L.lecanii NFCCI 0514和L.lecanii IRAN 1822C中的至少一种。Preferably, the Verticillium cereus is selected from at least one of L.lecanii VKM F-1199, L.lecanii NFCCI 0514 and L.lecanii IRAN 1822C.
一种抑制玉米螟繁殖的苏云金杆菌生防菌剂制备方法，所述制备方法包括以下步骤：A preparation method of a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer, the preparation method includes the following steps:
分别将球孢白僵菌和蜡蚧轮枝菌经菌种活化、种子液培养、液体发酵、调温发孢、干燥；即得到球孢白僵菌孢子粉和蜡蚧轮枝菌孢子粉；Beauveria bassiana and Verticillium cereus are respectively subjected to bacterial strain activation, seed liquid culture, liquid fermentation, temperature regulation and sporulation, and drying; Beauveria bassiana spore powder and Verticillium cereus spore powder are obtained;
将金龟子绿僵菌种子液按照10%体积比的接种量接入到扩大培养基中，28℃培养36h，得到金龟子绿僵菌培养液；Add the Metarhizium anisopliae seed liquid into the expanded medium at an inoculum volume of 10%, and culture it at 28°C for 36 hours to obtain the Metarhizium anisopliae culture solution;
将苏云金芽孢杆菌种子液按照8%体积比的接种量接入到扩大培养基中，30℃培养24h，得到苏云金芽孢杆菌培养液；Add the Bacillus thuringiensis seed liquid into the expanded medium at an inoculum volume of 8%, and culture it at 30°C for 24 hours to obtain a Bacillus thuringiensis culture liquid;
将绿色木霉种子液和枯草芽孢杆菌种子液按照1:2的体积比混合，然后按照8%体积比的接种量接入到扩大培养基中，28℃培养24h，得到绿色木霉-枯草芽孢杆菌混合培养液；Mix the Trichoderma viride seed liquid and the Bacillus subtilis seed liquid at a volume ratio of 1:2, then insert it into the expansion medium at an inoculum volume of 8% volume ratio, and culture it at 28°C for 24 hours to obtain Trichoderma viride-Bacillus subtilis spores. Bacillus mixed culture solution;
将球孢白僵菌孢子粉和蜡蚧轮枝菌孢子粉、苏云金芽孢杆菌培养液、绿色木霉-枯草芽孢杆菌混合培养液按照2:1:3的体积比混合均匀，得到复合液体菌剂。Mix Beauveria bassiana spore powder, Verticillium cereus spore powder, Bacillus thuringiensis culture medium, and Trichoderma viride-Bacillus subtilis mixed culture medium in a volume ratio of 2:1:3 to obtain a composite liquid inoculant .
与现有技术相比，本发明的有益效果是：Compared with the prior art, the beneficial effects of the present invention are:
本发明提出的抑制玉米螟繁殖的苏云金杆菌生防菌剂及其制备方法，通过生物防治的手段进行玉米螟的消杀，其中特殊配制了生物防治剂，此有别于传统的产品，主要是利用金龟子绿僵菌液与浓缩浸提液混合而成，金龟子绿僵菌能通过体表穿透进入害虫体内，在害虫体内繁殖消耗寄主营养使害虫致死；还能产生绿僵菌素，此为绿僵菌在代谢过程中产生的一种次生代谢产物，对鳞翅目害虫具有触杀作用和拒食作用，然后配合以中草药成分提取而得的浓缩浸提液，两者具有协同消灭玉米螟的效果，显著提高了防治效果；因其效果好、稳定性高，又能进一步降低施药次数，保护了环境，还不易产生耐药性，极具推广应用价值。 The Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer and its preparation method proposed by the present invention kills corn borer through biological control means, in which a biological control agent is specially formulated. This is different from traditional products, mainly It is made by mixing Metarhizium anisopliae liquid and concentrated extract. Metarhizium anisopliae can penetrate into the body of pests through the body surface, reproduce in the pests' bodies, consume host nutrients and kill the pests; it can also produce Metarhizium anisopliae, which is Metarhizium anisopliae is a secondary metabolite produced during the metabolic process, which has contact killing and antifeeding effects on lepidopteran pests. It is then combined with a concentrated extract extracted from Chinese herbal ingredients. The two have the effect of synergistically eliminating corn borers. The effect has significantly improved the control effect; because of its good effect and high stability, it can further reduce the number of pesticide applications, protect the environment, and is not easy to develop drug resistance, so it has great promotion and application value.​
附图说明Description of the drawings
图1为本发明方法流程图。Figure 1 is a flow chart of the method of the present invention.
具体实施方式Detailed ways
为了使本发明的目的、技术方案进行清楚、完整地描述，及优点更加清楚明白，以下结合附图对本发明实施例进行进一步详细说明。应当理解，此处所描述的具体实施例是本发明一部分实施例，而不是全部的实施例，仅仅用以解释本发明实施例，并不用于限定本发明实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。In order to clearly and completely describe the purpose and technical solution of the present invention, and make the advantages more clear, the embodiments of the present invention will be further described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described here are part of the embodiments of the present invention, rather than all embodiments. They are only used to explain the embodiments of the present invention and are not used to limit the embodiments of the present invention. Those of ordinary skill in the art will not All other embodiments obtained without creative efforts belong to the scope of protection of the present invention.
实施例一Embodiment 1
请参阅图1，本发明提供一种技术方案：苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌15%、金龟子绿僵菌15%、绿色木霉12%、枯草芽孢杆菌10%、球孢白僵菌10%，余量为蜡蚧轮枝菌；Please refer to Figure 1. The present invention provides a technical solution: the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 15%, Metarhizium anisopliae 15%, Trichoderma viride 12%, Bacillus subtilis 10% , Beauveria bassiana 10%, the balance is Verticillium cereus;
其中，金龟子绿僵菌为ACCC30104，绿色木霉为ATCC 26802，枯草芽孢杆菌为ATCC605，苏云金芽孢杆菌选自B.thuringiensis KCTC 1507，球孢白僵菌选自B.bassiana ATCC 74040，蜡蚧轮枝菌选自L.lecanii VKM F-1199。Among them, Metarhizium anisopliae is ACCC30104, Trichoderma viride is ATCC 26802, Bacillus subtilis is ATCC605, Bacillus thuringiensis KCTC 1507, Beauveria bassiana is selected from B.bassiana ATCC 74040, and Verticillium cereus The bacteria were selected from L.lecanii VKM F-1199.
实施例二Embodiment 2
苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌18%、金龟子绿僵菌17%、绿色木霉14%、枯草芽孢杆菌12%、球孢白僵菌12%，余量为蜡蚧轮枝菌；Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 18%, Metarhizium anisopliae 17%, Trichoderma viride 14%, Bacillus subtilis 12%, Beauveria bassiana 12%, the balance is wax Verticillium verticillium;
其中，金龟子绿僵菌为ACCC30104，绿色木霉为ATCC 26802，枯草芽孢杆菌为ATCC605，苏云金芽孢杆菌选自B.thuringiensis CVCM 1812，球孢白僵菌选自B.bassiana ATCC MYA-4886，蜡蚧轮枝菌选自L.lecanii NFCCI 0514。Among them, Metarhizium anisopliae is ACCC30104, Trichoderma viride is ATCC 26802, Bacillus subtilis is ATCC605, Bacillus thuringiensis is selected from B.thuringiensis CVCM 1812, Beauveria bassiana is selected from B.bassiana ATCC MYA-4886, and the wax scale Verticillium was selected from L.lecanii NFCCI 0514.
实施例三Embodiment 3
苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌22%、金龟子绿僵菌18%、绿色木霉16%、枯草芽孢杆菌14%、球孢白僵菌13%，余量为蜡蚧轮枝菌；Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 22%, Metarhizium anisopliae 18%, Trichoderma viride 16%, Bacillus subtilis 14%, Beauveria bassiana 13%, the balance is wax Verticillium verticillium;
其中，金龟子绿僵菌为ACCC30104，绿色木霉为ATCC 26802，枯草芽孢杆菌为ATCC605，苏云金芽孢杆菌选自B.thuringiensis DSM 5815，球孢白僵菌选自B.bassiana ATCC 74040，蜡蚧轮枝菌选自L.lecanii IRAN 1822C。Among them, Metarhizium anisopliae is ACCC30104, Trichoderma viride is ATCC 26802, Bacillus subtilis is ATCC605, Bacillus thuringiensis DSM 5815, Beauveria bassiana is selected from B.bassiana ATCC 74040, and Verticillium cereus The bacteria were selected from L.lecanii IRAN 1822C.
实施例四Embodiment 4
苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌25%、金龟子绿僵菌20%、绿色木霉18%、枯草芽孢杆菌15%、球孢白僵菌14%，余量为蜡蚧轮枝菌；Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 25%, Metarhizium anisopliae 20%, Trichoderma viride 18%, Bacillus subtilis 15%, Beauveria bassiana 14%, the balance is wax Verticillium verticillium;
其中，金龟子绿僵菌为ACCC30104，绿色木霉为ATCC 26802，枯草芽孢杆菌为ATCC605，苏云金芽孢杆菌选自B.thuringiensis KCTC 1507、B.thuringiensis CVCM 1812和B.thuringiensis DSM 5815的混合物，球孢白僵菌选自B.bassiana ATCC 74040和B.bassiana ATCC MYA-4886的混合物，蜡蚧轮枝菌选自L.lecanii VKM F-1199、L.lecanii NFCCI 0514和L.lecanii IRAN 1822C中的混合物。Among them, Metarhizium anisopliae is ACCC30104, Trichoderma viride is ATCC 26802, Bacillus subtilis is ATCC605, Bacillus thuringiensis is selected from the mixture of B.thuringiensis KCTC 1507, B.thuringiensis CVCM 1812 and B.thuringiensis DSM 5815, and Coccidioides albicans B. bassiana was selected from the mixture of B. bassiana ATCC 74040 and B. bassiana ATCC MYA-4886, and Verticillium cereus was selected from the mixture of L. lecanii VKM F-1199, L. lecanii NFCCI 0514 and L. lecanii IRAN 1822C.
实施例五Embodiment 5
一种抑制玉米螟繁殖的苏云金杆菌生防菌剂制备方法，所述制备方法包括以下步骤：A method for preparing a Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer. The preparation method includes the following steps:
分别将球孢白僵菌和蜡蚧轮枝菌经菌种活化、种子液培养、液体发酵、调温发孢、干燥；即得到球孢白僵菌孢子粉和蜡蚧轮枝菌孢子粉；Beauveria bassiana and Verticillium cereus are respectively subjected to bacterial strain activation, seed liquid culture, liquid fermentation, temperature regulation and sporulation, and drying; Beauveria bassiana spore powder and Verticillium cereus spore powder are obtained;
将金龟子绿僵菌种子液按照10%体积比的接种量接入到扩大培养基中，28℃培养36h，得到金龟子绿僵菌培养液；Add the Metarhizium anisopliae seed liquid into the expanded medium at an inoculum volume of 10%, and culture it at 28°C for 36 hours to obtain the Metarhizium anisopliae culture solution;
将苏云金芽孢杆菌种子液按照8%体积比的接种量接入到扩大培养基中，30℃培养24h，得到苏云金芽孢杆菌培养液；Add the Bacillus thuringiensis seed liquid into the expanded medium at an inoculum volume of 8%, and culture it at 30°C for 24 hours to obtain a Bacillus thuringiensis culture liquid;
将绿色木霉种子液和枯草芽孢杆菌种子液按照1:2的体积比混合，然后按照8%体积比的接种量接入到扩大培养基中，28℃培养24h，得到绿色木霉-枯草芽孢杆菌混合培养液；Mix the Trichoderma viride seed liquid and the Bacillus subtilis seed liquid at a volume ratio of 1:2, then insert it into the expansion medium at an inoculum volume of 8% volume ratio, and culture it at 28°C for 24 hours to obtain Trichoderma viride-Bacillus subtilis spores. Bacillus mixed culture solution;
将球孢白僵菌孢子粉和蜡蚧轮枝菌孢子粉、苏云金芽孢杆菌培养液、绿色木霉-枯草芽孢杆菌混合培养液按照2:1:3的体积比混合均匀，得到复合液体菌剂。Mix Beauveria bassiana spore powder, Verticillium cereus spore powder, Bacillus thuringiensis culture medium, and Trichoderma viride-Bacillus subtilis mixed culture medium in a volume ratio of 2:1:3 to obtain a composite liquid inoculant .
尽管已经示出和描述了本发明的实施例，对于本领域的普通技术人员而言，可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型，本发明的范围由所附权利要求及其等同物限定。Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art will understand that various changes, modifications, and substitutions can be made to these embodiments without departing from the principles and spirit of the invention. and modifications, the scope of the invention is defined by the appended claims and their equivalents.

Claims (10)

  1. 抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌15-25%、金龟子绿僵菌15-20%、绿色木霉12-18%、枯草芽孢杆菌10-15%、球孢白僵菌10-14%，余量为蜡蚧轮枝菌。The Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer is characterized in that: the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 15-25%, Metarhizium anisopliae 15-20%, Green Wood mold 12-18%, Bacillus subtilis 10-15%, Beauveria bassiana 10-14%, and the balance is Verticillium cereus.
  2. 根据权利要求1所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌15%、金龟子绿僵菌15%、绿色木霉12%、枯草芽孢杆菌10%、球孢白僵菌10%，余量为蜡蚧轮枝菌。 The Bacillus thuringiensis biocontrol agent for inhibiting the reproduction of corn borer according to claim 1, characterized in that: the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 15%, Metarhizium anisopliae 15% , Trichoderma viride 12%, Bacillus subtilis 10%, Beauveria bassiana 10%, and the balance is Verticillium cereus.​
  3. 根据权利要求1所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌18%、金龟子绿僵菌17%、绿色木霉14%、枯草芽孢杆菌12%、球孢白僵菌12%，余量为蜡蚧轮枝菌。The Bacillus thuringiensis biocontrol agent for inhibiting the reproduction of corn borer according to claim 1, characterized in that: the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 18%, Metarhizium anisopliae 17% , Trichoderma viride 14%, Bacillus subtilis 12%, Beauveria bassiana 12%, and the balance is Verticillium cereus.
  4. 根据权利要求1所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌22%、金龟子绿僵菌18%、绿色木霉16%、枯草芽孢杆菌14%、球孢白僵菌13%，余量为蜡蚧轮枝菌。The Bacillus thuringiensis biocontrol agent for inhibiting the reproduction of corn borer according to claim 1, characterized in that: the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 22%, Metarhizium anisopliae 18% , Trichoderma viride 16%, Bacillus subtilis 14%, Beauveria bassiana 13%, and the balance is Verticillium cereus.
  5. 根据权利要求1所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述苏云金杆菌生防菌剂包括以下重量比组分：苏云金芽孢杆菌25%、金龟子绿僵菌20%、绿色木霉18%、枯草芽孢杆菌15%、球孢白僵菌14%，余量为蜡蚧轮枝菌。The Bacillus thuringiensis biocontrol agent for inhibiting the reproduction of corn borer according to claim 1, characterized in that: the Bacillus thuringiensis biocontrol agent includes the following weight ratio components: Bacillus thuringiensis 25%, Metarhizium anisopliae 20% , Trichoderma viride 18%, Bacillus subtilis 15%, Beauveria bassiana 14%, and the balance is Verticillium cereus.
  6. 根据权利要求1所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述金龟子绿僵菌为ACCC30104，绿色木霉为ATCC 26802，枯草芽孢杆菌为ATCC605。The Bacillus thuringiensis biocontrol agent for inhibiting the reproduction of corn borer according to claim 1, characterized in that: Metarhizium anisopliae is ACCC30104, Trichoderma viride is ATCC 26802, and Bacillus subtilis is ATCC605.
  7. 根据权利要求1所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述苏云金芽孢杆菌选自B.thuringiensis KCTC 1507、B.thuringiensis CVCM 1812和B.thuringiensis DSM 5815中的至少一种。The Bacillus thuringiensis biocontrol agent for inhibiting the reproduction of corn borer according to claim 1, characterized in that: the Bacillus thuringiensis is selected from at least one of B.thuringiensis KCTC 1507, B.thuringiensis CVCM 1812 and B.thuringiensis DSM 5815. A sort of.
  8. 根据权利要求1所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述球孢白僵菌选自B.bassiana ATCC 74040和B.bassiana ATCC MYA-4886中的至少一种。The Bacillus thuringiensis biocontrol agent for inhibiting the reproduction of corn borer according to claim 1, characterized in that: the Beauveria bassiana is selected from at least one of B. bassiana ATCC 74040 and B. bassiana ATCC MYA-4886 .
  9. 根据权利要求1所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂，其特征在于：所述蜡蚧轮枝菌选自L.lecanii VKM F-1199、L.lecanii NFCCI 0514和L.lecanii IRAN 1822C中的至少一种。The Bacillus thuringiensis biocontrol agent for inhibiting the reproduction of corn borer according to claim 1, characterized in that: the Verticillium cereus is selected from the group consisting of L.lecanii VKM F-1199, L.lecanii NFCCI 0514 and L.lecanii IRAN At least one of 1822C.
  10. 一种如权利要求1-9任意一项所述的抑制玉米螟繁殖的苏云金杆菌生防菌剂的抑制玉米螟繁殖的苏云金杆菌生防菌剂制备方法，其特征在于：所述制备方法包括以下步骤：A method for preparing the Bacillus thuringiensis biocontrol agent that inhibits the reproduction of corn borer according to any one of claims 1 to 9, characterized in that: the preparation method includes the following step:
    分别将球孢白僵菌和蜡蚧轮枝菌经菌种活化、种子液培养、液体发酵、调温发孢、干燥；即得到球孢白僵菌孢子粉和蜡蚧轮枝菌孢子粉；Beauveria bassiana and Verticillium cereus are respectively subjected to bacterial strain activation, seed liquid culture, liquid fermentation, temperature regulation and sporulation, and drying; Beauveria bassiana spore powder and Verticillium cereus spore powder are obtained;
    将金龟子绿僵菌种子液按照10%体积比的接种量接入到扩大培养基中，28℃培养36h，得到金龟子绿僵菌培养液；Add the Metarhizium anisopliae seed liquid into the expanded medium at an inoculum volume of 10%, and culture it at 28°C for 36 hours to obtain the Metarhizium anisopliae culture solution;
    将苏云金芽孢杆菌种子液按照8%体积比的接种量接入到扩大培养基中，30℃培养24h，得到苏云金芽孢杆菌培养液；Add the Bacillus thuringiensis seed liquid into the expanded medium at an inoculum volume of 8%, and culture it at 30°C for 24 hours to obtain a Bacillus thuringiensis culture liquid;
    将绿色木霉种子液和枯草芽孢杆菌种子液按照1:2的体积比混合，然后按照8%体积比的接种量接入到扩大培养基中，28℃培养24h，得到绿色木霉-枯草芽孢杆菌混合培养液；Mix the Trichoderma viride seed liquid and the Bacillus subtilis seed liquid at a volume ratio of 1:2, then insert it into the expansion medium at an inoculum volume of 8% volume ratio, and culture it at 28°C for 24 hours to obtain Trichoderma viride-Bacillus subtilis spores. Bacillus mixed culture solution;
    将球孢白僵菌孢子粉和蜡蚧轮枝菌孢子粉、苏云金芽孢杆菌培养液、绿色木霉-枯草芽孢杆菌混合培养液按照2:1:3的体积比混合均匀，得到复合液体菌剂。Mix Beauveria bassiana spore powder, Verticillium cereus spore powder, Bacillus thuringiensis culture medium, and Trichoderma viride-Bacillus subtilis mixed culture medium in a volume ratio of 2:1:3 to obtain a composite liquid inoculant .
PCT/CN2023/093802 2023-05-12 2023-05-12 Bacillus thuringiensis biocontrol microbial agent for inhibiting reproduction of pyrausta nubilalis and preparation method therefor WO2024027266A1 (en)

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