WO2017107872A1 - 一种微生物诱捕剂及应用 - Google Patents

一种微生物诱捕剂及应用 Download PDF

Info

Publication number
WO2017107872A1
WO2017107872A1 PCT/CN2016/110474 CN2016110474W WO2017107872A1 WO 2017107872 A1 WO2017107872 A1 WO 2017107872A1 CN 2016110474 W CN2016110474 W CN 2016110474W WO 2017107872 A1 WO2017107872 A1 WO 2017107872A1
Authority
WO
WIPO (PCT)
Prior art keywords
bottle
microbial
cgmcc
bacillus amyloliquefaciens
attractant
Prior art date
Application number
PCT/CN2016/110474
Other languages
English (en)
French (fr)
Inventor
何月秋
吴毅歆
何鹏搏
谢兰芬
杨发祥
Original Assignee
云南省微生物发酵工程研究中心有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 云南省微生物发酵工程研究中心有限公司 filed Critical 云南省微生物发酵工程研究中心有限公司
Publication of WO2017107872A1 publication Critical patent/WO2017107872A1/zh

Links

Classifications

    • 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
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/22—Bacillus
    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial 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
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus

Definitions

  • the invention belongs to the technical field of agricultural science pest control, and particularly relates to a microbial trapping agent and the application thereof in trapping male and female fruit flies and leaf flies.
  • Diptera is an Arthropod, Mandibulata, Insecta, and Pterygota. It is the second in the Insecta, next to Coleoptera and scales. The fourth largest target of the order Hymenoptera. The world is known for 85,000 species, globally distributed. More than 4,000 species are known in China. Diptera insects have a pair of wings and a pair of balance bars, including mosquitoes, flies, and crickets. Some species of the common Diptera, such as the larvae of the fly, leaf fly, fruit fly, and wheat mites, are important pests in agriculture. The genus Lepidoptera is harmful to many legumes; many species of the fruit fly family are harmful to citrus, pears, and peaches.
  • the fruit fly pest belongs to the family Diphritidae and is an important pest causing serious damage to fruit and vegetable crops.
  • the fruit fly family has known about 500 genera and more than 4,500 species. Among them, the group of harmful fruit flies of fruits and vegetables has 15 genera, 22 subgenus, and more than 150 species.
  • Bactrocera dorsalis Hendel
  • Bactrocera cucurbitae Coqil Lett
  • Bactrocera minax Enderlein
  • Hunan Hubei, Sichuan
  • Chongqing Guizhou, Fujian, Guangdong, Guangxi, Hainan, Yunnan, Taiwan and other provinces.
  • the host of the harmful fruit fly is very wide, involving a total of 24 families and 42 genera of economic fruit plants: 4 families and 16 genera of vegetable plants (wild host plants are not included).
  • Fruit fly pests have strong reproductive power and great harm.
  • the average fruit killing rate is 25-45%, and in severe cases it is as high as 80%, even in the end of production, which brings significant economic losses to fruit and vegetable crops, which seriously affects farmers' income and income.
  • the methods for controlling fruit fly pests mainly include agricultural control, chemical control, physical control and biological control.
  • agricultural control includes regular removal of insect-bearing fruits in plantations, fruit bagging and winter ploughing and smashing; chemical control measures are mainly spraying baits to kill fruit flies; physical control measures are combined with fruit fly traps and baits.
  • the traps used to control fruit flies are mainly chemical synthetic traps, such as cue lure, methyleugenol and the like. Because agricultural control measures are time-consuming and labor-intensive, the effect is not good; chemical control not only makes the fruit fly resistant, but also harmful to humans and animals, and is prone to environmental pollution such as pesticide residues; in practice, the neutral trap has volatilization. Fast, short-lasting period, frequent medications, etc., can not achieve the control purpose of controlling the rate of detection, and currently the most widely used trapping agent A The eugenol only attracts males and the like, and its application is limited.
  • Sweet and sour liquid is time-consuming, laborious, and costly; estrogen can only induce males, because males can perform multiple matings, the control effect is often not ideal; protein traps can attract females and males, and the effect is good.
  • the current protein attractant in foreign markets is pure animal protein, and the production cost is high.
  • Liriomyza sativae Blanchard belongs to the genus Liriomyza of the genus Agromyzidae, commonly known as the genus Liriomyza, map worm, ghost plot, etc. It is a dangerous pest of the latent leaves. China was first discovered in Sanya City, Hainan City in December 1993. Later, Liriomyza sativae gradually spread northward. In 1994, it occurred in Kunming, Yunnan province. In 1995, it appeared in Shanghai and Nanjing. In 1996, it broke out in Shandong and Beijing. The disaster occurred in Henan and Shanxi in 1997. In 1998, it became a disaster in the northeast and Inner Mongolia. Except Mongolia, it was distributed throughout the country. It has more than 200 host plants, among which vegetables are most seriously affected by solanaceous fruits (except sweet peppers and peppers), beans, melons, and umbelliferae.
  • an object of the present invention is to provide a novel microbial trap suitable for trapping fruit flies and fruit fly pests.
  • a microbial trap whose active ingredient is a fermentation broth of Bacillus amyloliquefaciens YN2010-KC CGMCC No. 5722.
  • the activated Bacillus amyloliquefaciens YN2010-KC CGMCC No. 5722 is inoculated at a dose of 5-10%.
  • the seed culture medium is inoculated into a seed culture medium at a culture temperature of 35 ° C, a rotation speed of 120 rpm / min, shaken for 36 to 48 hours, to obtain a first-stage seed liquid, the seed culture medium is a liquid LB medium, pH 7.0 ⁇ 7.2;
  • the first-stage seed liquid is transferred to the secondary seed culture medium in the seed tank, at 28-37 ° C, the rotation speed is 100 rpm/min, the ventilation volume is 1000 L/h, and the tank pressure is maintained.
  • the culture is carried out for 36 to 48 hours under conditions of 0.02 to 0.03 MPa to obtain a secondary seed liquid;
  • the secondary seed culture medium is composed of the following components of the following mass fraction: corn flour 0.3 to 0.5%, soybean powder 1 to 1.5%, and sucrose 0.2.
  • the secondary seed solution is inoculated into the fermentation medium in the fermenter at 28-37 ° C, the rotation speed is 100 rpm / min, the aeration volume is 10000 L / h, and the tank pressure is maintained at 0.02 -
  • the culture broth is cultured at 0.03 MPa for 48-72 h.
  • the fermentation broth is The fermentation broth of Bacillus amyloliquefaciens YN2010-KC CGMCC No. 5722, which is the same as the secondary seed culture medium of step (2).
  • the present invention has the following beneficial effects:
  • the special odor emitted by the product of the invention can induce fruit flies and fruit flies to feed, can be male and female, has a long action period, can be used for one-time feeding within two weeks, and has low cost and strong competitiveness.
  • the trapping of Citrus sinensis 23.71% of the fruit flyer attractant and 18.81% of the methyl eugenol.
  • the females induced by the B-Rite flies attractant increased by 37.78%, which was 30 times higher than that induced by methyl eugenol.
  • the trapping of the citrus fruit fly 64.43% more than the methyl eugenol, the number of trapped citrus fruit fly females increased by 33.25 times than the methyl eugenol.
  • the product of the invention does not contain any chemical toxic agents, does not kill or kill any other non-target organisms, and is an environmentally friendly fruit fly trap.
  • the product of the invention can completely replace the chemical insecticide, greatly reducing the amount of chemical pesticides, thereby reducing the negative impact of chemical pesticides on natural enemy insects, and has no drug resistance.
  • the product of the invention can improve the quality and yield of agricultural products, has obvious ecological safety, ecological benefits and is simple to use.
  • the Bacillus amyloliquefaciens YN2010-KC according to the present invention was deposited on January 12, 2012 in the General Microbiology Center of China Microbial Culture Collection Management Committee (CGMCC), and the deposit number is CGMCC No. 5722, and in 2012. It was found to be alive on January 12th.
  • the address of the general microbiology center No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, China, postcode: 100101.
  • CGMCC General Microbiology Center of China Microbial Culture Collection Management Committee
  • accession number is CGMCC No.5722 Bacillus amyloliquefaciens YN2010-KC
  • a microbial trap provided by the present invention will be described in detail with reference to the embodiments, but the present invention is not limited thereto. Those skilled in the art can improve the process parameters and change the specific excipient types, etc., and still belong to the present. The scope of protection of the invention. The examples are not conventionally described in the conventional methods.
  • the LB solid medium and the PSA medium (potato sucrose agar medium) were thawed, poured into a sterilized culture dish, and cooled to prepare LB and PSA plates.
  • a strain obtained by separation and purification was named as YN2010-KC by dilution plate separation and plate scribing.
  • strain YN2010-KC was cultured on LB medium and cultured at 37 °C for 36 hours for morphological observation.
  • the cells were rod-shaped, end-circle, peri-flagellate, motility, and produced oval spores; in LB and PSA plates.
  • the colony is round, pale brown, the colony is slightly darker in central color, raised, and the surface is dry and wrinkled. It is basically consistent with the morphological characteristics of Bacillus described in "Handbook of Common Bacterial System Identification" (edited by Dong Xiuzhu et al., Science Press, 2001). It is preliminarily judged that strain YN2010-KC belongs to Bacillus sp.
  • strains were positive for Gram stain, spore staining, starch hydrolysis, glucose utilization, citrate utilization, V-P reaction, contact enzyme reaction, nitrate reduction, and gelatin liquefaction.
  • Embodiment 2 A method for preparing a microbial trap of the present invention comprises the following steps:
  • Bacillus amyloliquefaciens YN2010-KC CGMCC No. 5722 preserved at -80 ° C was inoculated on liquid LB medium and cultured at 35 ° C for 24 h to obtain activated Bacillus amyloliquefaciens YN2010- KC CGMCC No. 5722, the activated Bacillus amyloliquefaciens YN2010-KC CGMCC No. 5722 was inoculated into the seed culture medium at a culture temperature of 35 ° C and a rotation speed of 120 rpm/min. , shaking the bed for 36h, to obtain a first-level seed liquid, the seed medium is a liquid LB medium, pH 7.0 ⁇ 7.2;
  • a secondary seed solution is obtained;
  • a secondary seed solution is obtained;
  • the secondary seed medium is composed of the following mass fraction components: corn flour 0.5%, soybean powder 1%, sucrose 0.5%, peptone 0.3%, fish meal 0.1%, MgSO 4 ⁇ 7H 2 O 0.02%, CaCO 3 0.4%, (NH 4 ) 2 SO 4 0.03%, the balance is water, pH 7.0-7.2; the secondary seed culture medium is 121 Sterilize for 30 min at °C, and cool down to 30 °C, and connect to the first-stage seed solution;
  • the secondary seed solution was inoculated into the fermentation medium in the fermenter at 35 ° C, the rotation speed was 100 rpm / min, the aeration volume was 10000 L / h, and the tank pressure was maintained at 0.02 - 0.03 MPa.
  • the fermented liquid was packed in medical plastic bottles when the number of viable bacteria of Bacillus amyloliquefaciens YN2010-KC CGMCC No. 5722 was ⁇ 3 ⁇ 10 9 cfu/ml. That is, the microbial trap is the same as the secondary seed medium of the step (2).
  • the microbial trap of the present invention is prepared by the method for preparing the microbial trap described in Example 2.
  • Example 3 Comparative experiment of microbial traps and other attractants of the present invention on trapping of Citrus
  • the trapping effect test was carried out, together with the "Gruit" Drosophila attractant, (a protein bait, commercially available), methyl eugenol (attribute attractant, commercially available) and water ( Blank control) for comparison.
  • the dosage of the microbial trap of the invention is 50 ml/bottle
  • the dosage of the fruit rot extract is 50 ml/bottle
  • the amount of methyl eugenol is 3 ml/bottle.
  • each of the plastic bottles was filled with one of the above-mentioned attractants, and the bottle cap was screwed, and four holes were uniformly distributed on the wall of the bottle 2 cm from the liquid level in the bottle, and the holes had a hole diameter of 5 mm.
  • the plastic bottle was suspended 0.5-1.5 meters from the citrus land.
  • the number of citrus fruit flies (head) induced two days later is shown in Table 1. It can be seen that the number of microbial traps of the present invention is higher than that of the fruit fly attractant and methyl eugenol, and the female rate is as high as 51.67%.
  • the number of microbial traps of the present invention is 23.71% higher than that of the fruit fly attractant, and 18.81% higher than the methyl eugenol.
  • the female insect attractant of the present invention increased by 37.78% compared with the fruit extractor, and the female induced by methyl eugenol increased 30 times.
  • Table 1 The attracting test of the microbial trap of the present invention on Bactrocera dorsalis
  • Example 4 The trapping test of the citrus fruit fly by the microbial trap of the present invention
  • the fruit fly trapping test was carried out at Nanxi Farm, Hekou County, Yunnan province, and the test reagent: the microbial trap of the present invention.
  • the plastic bottle containing the reagent and the operation are the same as in the third embodiment.
  • the dosage of the microbial trap of the present invention is 50 ml/bottle, and the plastic bottle containing the reagent is suspended at 0.5-1.5 m from the citrus land at noon on July 5, 2014.
  • the amount of insects in each bottle was investigated in the following day, 2 days and 3 days. See Table 2, it can be seen that the fermenting agent of the present invention produces volatile protein and other compounds to attract fruit flies to eat, and the effect is remarkable, and the male and female are tempted.
  • the citrus fruit fly test was carried out in the citrus garden of Mengzi County, Yunnan province, and compared with methyl eugenol.
  • the amount of the microbial trap of the present invention is 50 ml/bottle, and the amount of methyl eugenol is 3 ml/vial.
  • the plastic bottle containing each reagent and the operation were the same as in Example 3.
  • the plastic bottle containing the reagent was suspended at 0.5-1.5 meters from the citrus land at noon on December 4, 2014, 1 day, 2 days and 3 days after the feeding.
  • the amount of insects in each bottle is shown in Table 3. It can be seen that the fermentation of the microbial traps of the present invention produces volatile protein and other compounds to induce the consumption of the citrus fruit fly, except that the amount of insects on the first day is lower than the sex attractant methyl eugenol. In addition, the effect of trapping the big fruit fly at other times is better than the sex attractant methyl eugenol, and it is male and female. Within three days, the microbial trap of the present invention trapped 64.43% more than the eugenol, and the trapped larvae of the citrus fruit increased by 33.25 times than the methyl eugenol.
  • the trapping effect test was carried out on the tomato and vegetable base in Mengzi County, Yunnan province, and the same field was compared with the 30 ⁇ 30cm yellow plate.
  • Each yellow plastic bottle was filled with 50 ml of the microbial trap of the present invention, and the cap was screwed, and four holes were uniformly distributed on the wall of the bottle at a distance of 2 cm from the liquid surface of the bottle, and the holes had a pore diameter of 5 mm.
  • the field was released.
  • the plastic bottles were hung from 0.5 to 0.8 meters above the ground, 8 plastic bottles were suspended per acre, and the adjacent plastic bottles were separated by 5 to 10 m.
  • the yellow plates were suspended from the ground.
  • Example 7 The microbial trap of the present invention for the trapping of male and female populations of Liriomyza sativae
  • Example 8 The microbial trapping agent of the present invention is capable of trapping the number of male and female males

Abstract

本发明公开一种微生物诱捕剂及应用。该诱捕剂的活性成分为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722发酵液，有效活菌数含量≧3×109cfu/ml。该诱捕剂散发出的特殊气味能引诱实蝇、果蝇和美洲斑潜蝇取食，雌雄同捕，作用周期长，成本低，不含有任何化学有毒药剂，不杀死和不杀伤任何其他非靶标生物，属环境友好型生物诱捕剂。对柑橘小实蝇的诱捕：比对照增加18.81～23.71％，比对照诱到的雌虫增加37.78％～30倍。对柑橘大实蝇的诱捕：比对照增加64.43％，诱捕雌虫数增加33.25倍。对美洲斑潜蝇的诱捕：雌虫率占55～60％。

Description

一种微生物诱捕剂及应用 技术领域
本发明属于农业科学害虫控制技术领域，具体涉及一种微生物诱捕剂，以及在诱捕雌雄果实蝇、斑潜蝇中的应用。
背景技术
双翅目(Diptera)是节肢动物门(Arthropoda)、有颚亚门(Mandibulata)、昆虫纲(Insecta)、有翅亚纲(Pterygota)的1目，是昆虫纲中仅次于鞘翅目、鳞翅目、膜翅目的第四大目。世界已知85000种，全球分布。中国已知4000余种。双翅目昆虫有一对翅膀和一对平衡棒，包括蚊、蝇、虻。常见双翅目一些类群如种蝇、叶潜蝇、果实蝇、麦瘿蚊等的幼虫，都是农业的重要害虫。潜蝇科为害多种豆科植物；实蝇科的许多种类为害柑橘、梨、桃等。
实蝇类害虫属于双翅目实蝇科(Tephritidae)，是对果蔬类作物造成严重损失的一类重要害虫。实蝇科迄今为止已知约500属，4500余种，其中，果蔬类有害实蝇的类群达15属、22亚属、150余种，我国以柑橘小实蝇[Bactrocera dorsalis(Hendel)]，瓜实蝇[Bactrocera cucurbitae(Coqil Lett)]，柑橘大实蝇[Bactrocera minax(Enderlein)]等较为常见，主要分布于湖南、湖北、四川、重庆、贵州、福建、广东、广西、海南、云南、台湾等省。有害实蝇的寄主范围很广，涉及经济水果类植物共计24科、42个属：蔬菜类植物4科、16属(野生寄主植物未包括在内)。实蝇类害虫繁殖力强、危害性大。在实蝇分布或流行区，寄主果实被害率平均为25-45％，严重时高达80％，甚至绝产，给果蔬类作物带来了重大经济损失，严重影响了农民增产创收。
目前，防治实蝇类害虫的方法主要有农业防治、化学防治、物理防治和生物防治等。其中农业防治包括定期清除种植园内的带虫果实、果实套袋和冬季翻耕灭蛹等；化学防治措施主要是喷施毒饵杀灭实蝇；物理防治措施采用实蝇诱捕剂和毒饵相结合的技术，目前，用于防治实蝇的诱捕剂主要是化学合成诱捕剂，如引诱酮(cue lure)、甲基丁香酚(methyleugenol)等。由于农业防治措施费时费工，效果不好；化学防治不仅容易使实蝇产生抗药性，而且对人、畜有害，易产生农药残留等环境污染的问题；在实际应用中性诱捕剂又存在挥发快、持效期短、用药频繁等问题，达不到控制蛀果率的防治目的，而且目前应用最广泛诱捕剂甲 基丁香酚只引诱雄虫等等，其应用受到限制。
国内外针对实蝇有糖醋液、雌性激素、蛋白质三大类诱剂。糖醋液费时、费力，成本较高；雌性激素只能诱雄虫，由于雄虫能行多次交配，防效常不太理想；蛋白质类诱捕剂可以引诱雌雄虫，效果好。但目前国外市场上的蛋白质诱剂为纯净动物蛋白，生产成本高。
美洲斑潜蝇(Liriomyza sativae Blanchard)属于双翅目潜蝇科(Agromyzidae)斑潜蝇属(Liriomyza)，俗称蔬菜斑潜蝇、地图虫、鬼画符等，是潜叶为害的危险性害虫。我国最早于1993年12月在海南省三亚市首次发现，随后美洲斑潜蝇逐渐向北扩散，1994年在云南昆明发生为害，1995年在上海、南京等地出现，1996年在山东、北京暴发成灾，1997年在河南、山西出现，1998年在东北、内蒙古成灾，除西藏外，全国均有分布。它的寄主植物多达200余种，其中蔬菜以茄果类(甜椒、辣椒除外)、豆类、瓜类、伞形花科等受害最重。
美洲斑潜蝇成虫为小型蝇类，体黑色，体长1.3～2.3毫米，寿命8.7～12.6天，每雌一生产卵70～125枚，孵化率高达90％以上。幼虫乳白色至淡黄色，长3毫米左右，年繁殖15～20代。冬季进入日光温室内，可四季为害，主要有2次为害高峰，分别出现在5～6月和9～10月，年世代可达20代以上。成虫、幼虫均可为害，雌成虫在寄主叶片上刺孔并在其中取食和产卵，幼虫在叶片表皮下孵化后潜食叶肉或叶柄，产生不规则的蛇形白色虫道，破坏叶片叶绿素，影响光合作用，严重时叶片枯黄脱落甚至整株枯死；苗期造成毁苗。危害严重田块受害株100％，叶片受害70％。盛产期西葫芦，在发现危害后，7天左右吃尽叶肉，15天左右毁灭。幼虫和成虫通过取食还可传播某些病毒病。
目前，随着耕作制度的变化，持续暖冬现象和露地及保护地蔬菜面积的不断扩大，潜叶蝇已成为多种蔬菜上危害特大、防治特困难的灾害性害虫之一，全年发生世代多而重叠，传播蔓延快，大田春末夏初秋季保护地全年发生尤其严重。蔬菜的保护地栽培为蔬菜斑潜蝇提供了良好的越冬场所。该虫不但可在大棚内越冬，而且可在棚内发生危害，成为露地蔬菜田的重要虫源。因此,搞好大棚内的防治可有效地防止该虫在揭棚后向露地蔬菜扩散危害。
因该虫世代重叠现象严重，抗药性强，生产中较难防治。目前，生产上对美洲斑潜蝇的防治仍主要是以化学防治为主，为了解决多年来化学农药带来的食品安全问题，应利用生物防治措施来控制虫害，但生物防治技术还没有真正在生产 上应用和发挥效益，寻找针对性强，防效高，以及对人、畜安全，没有环境污染的一种新诱剂迫在眉睫。
发明内容
针对现有技术的需要，本发明的目的是提供一种适用于诱捕实蝇和果蝇类害虫、可雌雄同捕的新型微生物诱捕剂。
为实现上述发明目的，本发明的技术方案如下：
1.一种微生物诱捕剂，其活性成分为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的发酵液。
2.根据技术方案1所述的一种微生物诱捕剂，所述解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的发酵液通过以下方法制备得到：
(1)将解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722活化后，按5～10％接种量，将活化的所述解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722接种到种子培养基，在培养温度35℃、转速120rpm/min条件下，摇床培养36～48h，得到一级种子液，所述种子培养基为液体LB培养基，pH 7.0～7.2；
(2)按10～12％接种量，将一级种子液转接到种子罐内的二级种子培养基，在28～37℃，转速100rpm/min，通气量为1000L/h，罐压保持在0.02～0.03MPa条件下培养36～48h，得到二级种子液；所述二级种子培养基由以下质量分数的以下成分组成：玉米粉0.3～0.5％，黄豆粉1～1.5％，蔗糖0.2～0.5％，蛋白胨0.1～0.3％，鱼粉0.1～0.3％，MgSO4·7H2O 0.02～0.04％，CaCO3 0.4～0.8％，(NH4)2SO40.03～0.06％，余量为水，pH 7.0～7.2；
(3)按10～12％接种量，将二级种子液接种到发酵罐内的发酵培养基，在28～37℃，转速100rpm/min，通气量为10000L/h、罐压保持在0.02～0.03MPa条件下培养48～72h，培养后的发酵液中解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的有效活菌数≧3×109cfu/ml时，该发酵液即为所述解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的发酵液，所述发酵培养基与步骤(2)所述二级种子培养基相同。
3.技术方案1或2所述的微生物诱捕剂在防治实蝇类害虫的应用。
4.技术方案1或2所述的微生物诱捕剂在防治美洲斑潜蝇的应用。
5.根据技术方案3所述的微生物诱捕剂在防治实蝇类害虫的应用，所述的应用是将所述微生物诱捕剂装入塑料瓶内，每瓶的装入量为50～100ml，旋紧瓶盖，在距离瓶内液面1～3cm的瓶壁上均布4个孔，所述各孔的孔径为4～6mm；将塑料瓶悬挂于离地面0.5～1.5米处，每亩悬挂塑料瓶8～10个，相邻的塑料瓶间隔5～7m；每隔20～30天向塑料瓶内添加一次所述微生物诱捕剂25～50ml。
6.根据技术方案3所述的微生物诱捕剂在防治实蝇类害虫的应用，所述的应用是将所述微生物诱捕剂装入塑料瓶内，每瓶的装入量为50～100ml，旋紧瓶盖，在距离瓶内液面1～3cm的瓶壁上均布4个孔，所述各孔的孔径为7～8mm；将塑料瓶悬挂于离地面0.5～1.5米处，每亩悬挂塑料瓶8～10个，相邻的塑料瓶间隔5～7m；每隔20～30天向塑料瓶内添加一次所述微生物诱捕剂25～50ml。
与现有技术相比，本发明具有以下有益效果：
1、本发明产品散发出的特殊气味能引诱实蝇和果蝇取食，可雌雄同捕，作用周期长，一次性投料可以两周内持效，且成本低，具有较强的竞争力。对柑橘小实蝇诱捕：比果瑞特果蝇引诱剂增加23.71％，比甲基丁香酚增加18.81％。比果瑞特果蝇引诱剂诱到的雌虫增加37.78％，比甲基丁香酚诱到的雌虫增加30倍。对柑橘大实蝇诱捕：比甲基丁香酚增加64.43％，诱捕的柑橘大实蝇雌虫数量比甲基丁香酚增加33.25倍。对美洲斑潜蝇的诱捕：雌虫率占55～60％，产生了预料不到的技术效果。甲基丁香酚诱不到斑潜蝇，只能用糖醋液为对照，而后者也能诱到雌雄虫，只是总虫量比本产品低很多。
2、本发明产品中不含有任何化学有毒药剂，不杀死和不杀伤任何其他非靶标生物，属于环境友好型果实蝇诱捕剂。
3、本发明产品完全可以取代化学杀虫药剂，大大减少化学农药用量，从而减少化学农药对天敌昆虫的负面影响，无抗药性。
4、本发明产品可提高农产品质和产量，对生态安全，生态效益显著，使用简单。
本发明所述的解淀粉芽孢杆菌Bacillus amyloliquefaciens YN2010-KC于2012年1月12日保藏在中国微生物菌种保藏管理委员会普通微生物中心(简称：CGMCC)，保藏号为CGMCC No.5722，并于2012年1月12日检测为存活。该普通微生物中心地址：中国北京市朝阳区北辰西路1号院3号，邮编:100101。
中国微生物菌种保藏管理委员会普通微生物中心(简称：CGMCC)
地址：中国北京市朝阳区北辰西路1号院3号，邮编:100101
保藏日期：2012年1月12日
保藏号为CGMCC No.5722解淀粉芽孢杆菌Bacillus amyloliquefaciens YN2010-KC
具体实施方式
下面结合实施例对本发明提供的一种微生物诱捕剂进行详细说明，但并不对本发明构成限制，本领域技术人员在此基础上改进工艺参数，改变具体的辅料类型等替换和改动，依旧属于本发明的保护范围。各实施例无特殊说明的为常规方法。
实施例1  解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC的获得、鉴定和保藏
1、本发明解淀粉芽孢杆菌YN2010-KC的获得
1.1材料
采自云南农业大学后山农场玉米根际周围的土壤。
1.2培养基制备
将LB固体培养基和PSA培养基(马铃薯蔗糖琼脂培养基)融化，分别倒入灭菌培养皿，冷却，制成LB和PSA平板。
1.3分离培养
采用稀释平皿分离法和平板划线分离，将分离纯化所得的一菌株命名为YN2010-KC。
2、解淀粉芽孢杆菌YN2010-KC的鉴定
2.1、形态学鉴定
取菌株YN2010-KC在LB培养基上培养，37℃培养36小时，进行形态观察鉴定，菌体为杆状、端圆，周生鞭毛、具运动性，产生椭圆形芽孢；在LB和PSA平板上菌落圆形、淡茶褐色、菌落中央颜色略深、隆起、表面干燥有褶皱。与《常见细菌系统鉴定手册》(东秀珠等编著，科学出版社，2001年)中描述的芽孢杆菌属形态特征基本一致，初步判断菌株YN2010-KC属于芽孢杆菌(Bacillus sp.)。
2.2、生理生化鉴定
菌株革兰氏染色、芽孢染色、淀粉水解、葡萄糖利用、柠檬酸盐利用、V-P反应、接触酶反应、硝酸盐还原、明胶液化等均呈阳性。
3、解淀粉芽孢杆菌YN2010-KC的保藏
经上述鉴定表明所分离获得的菌株为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)，其代号为YN2010-KC，所述的解淀粉芽孢杆菌Bacillus amyloliquefaciens YN2010-KC于2012年1月12日保藏在中国微生物菌种保藏管理委员会普通微生物中心(简称：CGMCC)，保藏号为CGMCC No.5722，并于2012年1月12日检测为存活。该普通微生物中心地址：中国北京市朝阳区北辰西路1号院3号，邮编:100101。
实施例2  本发明微生物诱捕剂的制备方法，包括以下步骤：
(1)将-80℃保存的解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722接种在液体LB培养基上，35℃培养24h，得活化的解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722，按10％接种量，将活化的所述解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722接种到种子培养基，在培养温度35℃、转速120rpm/min条件下，摇床培养36h，得到一级种子液，所述种子培养基为液体LB培养基，pH 7.0～7.2；
(2)按10％接种量，将一级种子液转接到种子罐内的二级种子培养基，在35℃，转速100rpm/min，通气量为1000L/h，罐压保持在0.02～0.03MPa条件下培养36～48h，得到二级种子液；得到二级种子液；所述二级种子培养基由以下质量分数的成分组成：玉米粉0.5％，黄豆粉1％，蔗糖0.5％，蛋白胨0.3％，鱼粉0.1％，MgSO4·7H2O 0.02％，CaCO3 0.4％，(NH4)2SO4 0.03％，余量为水，pH7.0～7.2；所述二级种子培养基于121℃灭菌30min，待降温至30℃，接入一级种子液；
(3)按10％接种量，将二级种子液接种到发酵罐内的发酵培养基，在35℃，转速100rpm/min，通气量为10000L/h、罐压保持在0.02～0.03MPa条件下培养48～72h，培养后的发酵液中解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的有效活菌数≧3×109cfu/ml时，将发酵液用医用 塑料瓶包装，即为所述微生物诱捕剂，所述发酵培养基与步骤(2)所述二级种子培养基相同。
本发明微生物诱捕剂通过实施例2所述的微生物诱捕剂的制备方法制备得到。
实施例3  本发明微生物诱捕剂与其它诱剂对柑橘小实蝇诱捕的对比试验
在云南省元江县柑橘园内进行诱捕效果试验，同时与“果瑞特”果蝇引诱剂，(属蛋白饵剂，市售)、甲基丁香酚(属性诱剂，市售)和水(空白对照)进行对比。本发明微生物诱捕剂用量50ml/瓶，果瑞特果蝇引诱剂用量50ml/瓶，甲基丁香酚用量3ml/瓶。
每个塑料瓶装上述一种引诱剂，旋紧瓶盖，在距离瓶内液面2cm的瓶壁上均布4个孔，所述各孔的孔径为5mm。于2015年5月16日中午，将塑料瓶悬挂于离柑橘地0.5-1.5米处。两天后诱到柑橘小实蝇数量(头)见表1。可见，本发明微生物诱捕剂诱虫数量均高于果瑞特果蝇引诱剂和甲基丁香酚，且雌虫率高达51.67％。本发明微生物诱捕剂诱虫数量比果瑞特果蝇引诱剂增加23.71％，比甲基丁香酚增加18.81％。本发明微生物诱捕剂比果瑞特果蝇引诱剂诱到的雌虫增加37.78％，比甲基丁香酚诱到的雌虫增加30倍。
表1  本发明微生物诱捕剂对柑橘小实蝇引诱试验
Figure PCTCN2016110474-appb-000001
实施例4  本发明微生物诱捕剂对柑橘果实蝇诱捕试验
在云南省河口县南溪农场进行果实蝇诱捕试验，试验试剂：本发明微生物诱捕剂。装试剂的塑料瓶及操作与实施例3相同，本发明微生物诱捕剂用量50ml/瓶，于2014年7月5日中午装有试剂的塑料瓶悬挂于离柑橘地0.5-1.5米处，自投料后1天、2天和3天调查各瓶中虫量见表2，可见，本发明诱捕剂发酵产生挥发性蛋白质类等化合物引诱果实蝇食用，效果显著，且雌雄同诱。
表2  本发明微生物诱捕剂诱捕果实蝇(柑橘大实蝇、小实蝇、番石榴实蝇、瓜实蝇等)的效果
Figure PCTCN2016110474-appb-000002
实施例5  本发明微生物诱捕剂对柑橘大实蝇防治效果的对比试验
在云南省蒙自县柑橘园内进行诱捕柑橘大实蝇试验，同时与甲基丁香酚进行对比。本发明微生物诱捕剂用量50ml/瓶，甲基丁香酚用量3ml/瓶。
装各试剂的塑料瓶及操作与实施例3相同，于2014年12月4日中午将装有试剂的塑料瓶悬挂于离柑橘地0.5-1.5米处，自投料后1天、2天和3天调查各瓶中虫量见表3，可见，用本发明微生物诱捕剂发酵产生挥发性蛋白质类等化合物引诱柑橘大实蝇食用，除第一天诱虫量低于性诱剂甲基丁香酚外，其它时间内诱捕大实蝇的效果均优于性诱剂甲基丁香酚，且雌雄同诱。三天内，本发明微生物诱捕剂诱捕柑橘大实蝇数量比甲基丁香酚增加64.43％，诱捕的柑橘大实蝇雌虫数量比甲基丁香酚增加33.25倍。
表3  本发明微生物诱捕剂诱捕柑橘大实蝇的效果
Figure PCTCN2016110474-appb-000003
实施例6  本发明微生物诱捕剂与黄板田间诱虫对比试验
在云南省蒙自县番茄蔬菜基地上进行诱捕效果试验，同田与30×30cm黄板进行了对比。每个黄色塑料瓶装50ml本发明微生物诱捕剂，旋紧瓶盖，在距离瓶内液面2cm处的瓶壁上均布4个孔，所述各孔的孔径为5mm。于2015年6 月16日中午进行田间释放，将塑料瓶悬挂于距地面0.5～0.8米高，每亩悬挂塑料瓶8个，相邻的塑料瓶间隔5～10m；在同田，将黄板悬挂于距地面0.5-0.8米高，每亩悬挂黄板8个，相邻的黄板间隔5～10m。每两天后诱到美洲斑潜蝇数量(头)见表4。可见，本发明微生物诱捕剂平均每天诱虫数量均高于黄板，8天内，本发明微生物诱捕剂诱虫1988头，黄板诱虫379头，是黄板诱虫量的5.2倍。
表4  本发明微生物诱捕剂和黄板对美洲斑潜蝇引诱的对比试验
Figure PCTCN2016110474-appb-000004
实施例7  本发明微生物诱捕剂对美洲斑潜蝇雌雄数量诱捕试验
在云南省江川县牛魔村蔬菜基地进行豇豆美洲斑潜蝇诱捕试验，将50ml本发明微生物诱捕剂直接加入外壁贴有黄色胶带塑料瓶中，旋紧瓶盖，在距离瓶内液面2cm处的瓶壁上均布4个孔，所述各孔的孔径为5mm。于2015年2月3日中午悬挂于距离地面0.5～0.8米高，自投料后1天、2天和3天调查各瓶中虫量见表5，可见，用本发明微生物诱捕剂发酵产生的挥发性蛋白质类等化合物引诱美洲斑潜蝇食用，效果显著，且雌雄同诱，雌虫率达55～60％。
表5  本发明微生物诱捕剂捕雌雄美洲斑潜蝇统计
Figure PCTCN2016110474-appb-000005
实施例8  本发明微生物诱捕剂对橘小实雌雄数量诱捕试验
在云南省元江县芒果地进行诱捕橘小实蝇试验，将50ml本发明微生物诱捕剂直接加入透明塑料瓶中，旋紧瓶盖，在距离瓶内液面2cm处的瓶壁上均布4个孔，所述各孔的孔径为7-8mm。于2015年5月3日中午悬挂于距离地面0.5～1.5米高，自投料后1天、2天和3天调查各瓶中虫量见表6，可见，用本发明微生 物诱捕剂发酵产生的挥发性蛋白质类等化合物引诱橘小实食用，效果显著，且雌雄同诱，雌虫率达55.86～63.40％。
表6  本发明微生物诱捕剂捕雌雄橘小实统计
Figure PCTCN2016110474-appb-000006

Claims (6)

  1. 一种微生物诱捕剂，其特征在于：其活性成分为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的发酵液。
  2. 根据权利要求1所述的一种微生物诱捕剂，其特征在于，所述解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的发酵液通过以下方法制备得到：
    (1)按5～10％接种量，将活化的所述解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722接种到种子培养基，在培养温度35℃、转速120rpm/min条件下，摇床培养36～48h，得到一级种子液，所述种子培养基为液体LB培养基，pH 7.0～7.2；
    (2)按10～12％接种量，将一级种子液转接到种子罐内的二级种子培养基，在28～37℃，转速100rpm/min，通气量为1000L/h，罐压保持在0.02～0.03MPa条件下培养36～48h，得到二级种子液；所述二级种子培养基由以下质量分数的成分组成：玉米粉0.3～0.5％，黄豆粉1～1.5％，蔗糖0.2～0.5％，蛋白胨0.1～0.3％，鱼粉0.1～0.3％，MgSO4·7H2O 0.02～0.04％，CaCO3 0.4～0.8％，(NH4)2SO40.03～0.06％，余量为水，pH 7.0～7.2；
    (3)按10～12％接种量，将二级种子液接种到发酵罐内的发酵培养基，在28～37℃，转速100rpm/min，通气量为10000L/h、罐压保持在0.02～0.03MPa条件下培养48～72h，培养后的发酵液中解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的有效活菌数≧3×109cfu/ml时，该发酵液即为所述解淀粉芽孢杆菌(Bacillus amyloliquefaciens)YN2010-KC CGMCC No.5722的发酵液，所述发酵培养基与步骤(2)所述二级种子培养基相同。
  3. 权利要求1或2所述的微生物诱捕剂在防治实蝇类害虫的应用。
  4. 权利要求1或2所述的微生物诱捕剂在防治美洲斑潜蝇的应用。
  5. 根据权利要求3所述的应用，其特征在于：所述的应用是将所述微生物诱捕剂装入塑料瓶内，每瓶的装入量为50～100ml，旋紧瓶盖，在距离瓶内液面1～3cm的瓶壁上均布4个孔，所述各孔的孔径为4～6mm；将塑料瓶悬挂于离地面0.5～1.5米处，每亩悬挂塑料瓶8～10个，相邻的塑料瓶间隔5～7m；每隔20～30天向塑料瓶内添加一次所述微生物诱捕剂25～50ml。
  6. 根据权利要求3所述的应用，其特征在于：所述的应用是将所述微生物 诱捕剂装入塑料瓶内，每瓶的装入量为50～100ml，旋紧瓶盖，在距离瓶内液面1～3cm的瓶壁上均布4个孔，所述各孔的孔径为7～8mm；将塑料瓶悬挂于离地面0.5～1.5米处，每亩悬挂塑料瓶8～10个，相邻的塑料瓶间隔5～7m；每隔20～30天向塑料瓶内添加一次所述微生物诱捕剂25～50ml。
PCT/CN2016/110474 2015-12-21 2016-12-16 一种微生物诱捕剂及应用 WO2017107872A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201510958305.8 2015-12-21
CN201510958305.8A CN105410032B (zh) 2015-12-21 2015-12-21 一种微生物诱捕剂及应用

Publications (1)

Publication Number Publication Date
WO2017107872A1 true WO2017107872A1 (zh) 2017-06-29

Family

ID=55488953

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2016/110474 WO2017107872A1 (zh) 2015-12-21 2016-12-16 一种微生物诱捕剂及应用

Country Status (2)

Country Link
CN (1) CN105410032B (zh)
WO (1) WO2017107872A1 (zh)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105410032B (zh) * 2015-12-21 2018-04-24 云南省微生物发酵工程研究中心有限公司 一种微生物诱捕剂及应用
GB2551121B (en) * 2016-06-02 2021-01-06 Fungialert Ltd In situ detection of pathogens
CN107593620B (zh) * 2017-09-30 2024-03-22 河南农业大学 用于黑水虻生长繁殖的生物砖及其制作和使用方法
CN108377819A (zh) * 2018-02-05 2018-08-10 云南省烟草公司普洱市公司 一种防治烟草黑胫病的方法
CN109320368A (zh) * 2018-11-08 2019-02-12 北京泰克美高新技术有限公司 防治斑潜蝇的生物制剂及其制备方法
CN111513083A (zh) * 2019-04-22 2020-08-11 泰山医学院 一种新型家蝇特异性长效细菌引诱剂及其制备方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1997276A (zh) * 2004-03-11 2007-07-11 巴伦西亚理工大学 用于防治实蝇科昆虫的包含特异性引诱剂的食物诱饵
CN101589728A (zh) * 2009-06-30 2009-12-02 福建农林大学 橘小实蝇蛋白引诱剂及其制备方法
CN102754665A (zh) * 2012-07-17 2012-10-31 华中农业大学 异常汉逊酵母菌在制备柑橘实蝇类引诱饵剂中的应用
CN104371959A (zh) * 2014-11-18 2015-02-25 吴山功 生物除臭灭蝇复合菌剂及其应用
CN104498040A (zh) * 2014-11-10 2015-04-08 杨凌绿都生物科技有限公司 解淀粉芽胞杆菌作为土壤改良剂的应用
CN105410032A (zh) * 2015-12-21 2016-03-23 云南省微生物发酵工程研究中心有限公司 一种微生物诱捕剂及应用

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101416641B (zh) * 2008-09-12 2011-08-17 云南农业大学 防治十字花科根肿病的生物制剂及其应用
CN101584346A (zh) * 2008-11-19 2009-11-25 南京林业大学 一种高效毒杀植物寄生线虫生物制剂的制备方法
CN102154156B (zh) * 2010-12-28 2013-02-13 南京农业大学 能防治番茄青枯病或根结线虫的拮抗菌及其微生物有机肥料
CN102952768B (zh) * 2012-11-05 2015-03-11 云南省烟草农业科学研究院 一种芽孢杆菌及其菌剂和制备方法与应用
CN105519582B (zh) * 2015-12-21 2018-11-13 云南省微生物发酵工程研究中心有限公司 一种防治烟草番茄根结线虫制剂和应用
CN105494445A (zh) * 2015-12-21 2016-04-20 云南农业大学 一种防治辣椒枯萎病制剂和应用
CN105385638B (zh) * 2015-12-21 2019-01-04 云南省微生物发酵工程研究中心有限公司 一种微生物解磷制剂及其制备方法和应用

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1997276A (zh) * 2004-03-11 2007-07-11 巴伦西亚理工大学 用于防治实蝇科昆虫的包含特异性引诱剂的食物诱饵
CN101589728A (zh) * 2009-06-30 2009-12-02 福建农林大学 橘小实蝇蛋白引诱剂及其制备方法
CN102754665A (zh) * 2012-07-17 2012-10-31 华中农业大学 异常汉逊酵母菌在制备柑橘实蝇类引诱饵剂中的应用
CN104498040A (zh) * 2014-11-10 2015-04-08 杨凌绿都生物科技有限公司 解淀粉芽胞杆菌作为土壤改良剂的应用
CN104371959A (zh) * 2014-11-18 2015-02-25 吴山功 生物除臭灭蝇复合菌剂及其应用
CN105410032A (zh) * 2015-12-21 2016-03-23 云南省微生物发酵工程研究中心有限公司 一种微生物诱捕剂及应用

Also Published As

Publication number Publication date
CN105410032A (zh) 2016-03-23
CN105410032B (zh) 2018-04-24

Similar Documents

Publication Publication Date Title
Epsky et al. History and development of food-based attractants
WO2017107872A1 (zh) 一种微生物诱捕剂及应用
CN106544278B (zh) 一种用于蟑螂生物防治的病原真菌
CN108018242B (zh) 一种粘质沙雷氏菌及其分离方法和应用
CN102021122B (zh) 一种高效杀虫真菌及其用途
CN106359454A (zh) 一种短稳杆菌农药的制备方法
CN103783030B (zh) 一种捕食性天敌昆虫绿步甲与绿僵菌组合防治害虫的方法
CN107779408A (zh) 一种用于防治金龟子的球孢白僵菌及其微生物菌剂
CN102657235B (zh) 含厚孢轮枝菌和淡紫拟青霉的杀线虫组合物及其制备方法
CN108575456B (zh) 玉米螟的生物防治方法
CN107926986A (zh) 一种松毛虫质型多角体病毒和白僵菌杀虫悬浮剂及其制备方法和应用
CN102907458B (zh) 生物灭蚊剂及其制备方法
US9526240B2 (en) Sprayable dispersed starch-based bioplastic formulation to control pests
CN111004724B (zh) 一种对朱红毛斑蛾幼虫有高致病力的白僵菌菌株及其应用
CN104988092B (zh) 一种苏云金芽孢杆菌、制备方法及应用
CN103981108A (zh) 一种用于防治韭菜迟眼蕈蚊的球孢白僵菌及其应用
CN115161214A (zh) 一种防控地下害虫的生物有机肥、制备方法和应用
Abada et al. Effect of combination among bioagents, compost and soil solarization on management of strawberry Fusarium wilt
CN113424837A (zh) 一种白僵菌与植物源挥发物联用的玉米螟防控方法
CN112708565A (zh) 一种蝗虫微孢子虫株系以及制备和使用方法
CN104982461A (zh) 捕食螨与蜡蚧轮枝菌联合应用的生物农药及其制备、应用方法
CN110583701A (zh) 白僵菌和细菌复配杀草地贪夜蛾的药剂
CN105039208B (zh) 一种蜡状芽孢杆菌、制备方法及应用
CN114586584B (zh) 一种基于全周期生物系统的甜椒绿色轻简控害方法
CN116064291B (zh) 一种霍氏肠杆菌en-314t及其在茶谷蛾幼虫期的生物防治方法与应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16877683

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 10.10.2018)

122 Ep: pct application non-entry in european phase

Ref document number: 16877683

Country of ref document: EP

Kind code of ref document: A1