KR101825966B1 - Protaetia brevitarsis fungal diseases controlling compositions - Google Patents

Protaetia brevitarsis fungal diseases controlling compositions Download PDF

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KR101825966B1
KR101825966B1 KR1020160060592A KR20160060592A KR101825966B1 KR 101825966 B1 KR101825966 B1 KR 101825966B1 KR 1020160060592 A KR1020160060592 A KR 1020160060592A KR 20160060592 A KR20160060592 A KR 20160060592A KR 101825966 B1 KR101825966 B1 KR 101825966B1
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박관호
최지영
남성희
김성현
곽규원
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Abstract

본 발명은 흰점박이꽃무지 진균병 방제용 조성물에 관한 것이다.
본 발명은, 흰점박이꽃무지 진균병 방제용 조성물로서, 락토바실러스 속(Lactobacillus sp .) 균주 또는 이의 배양액을 유효성분으로 포함하는 것이 특징이며, 바람직하게는 상기 락토바실러스 속 균주는 락토바실러스 플란타룸(Lactobacillus plantarum)인 것이 특징이다.
본 발명에 의해, 흰점박이꽃무지의 사육과정에서 유충에 다량으로 발생하는 진균병을 방제하는 효과를 갖는 흰점박이꽃무지 진균병 방제용 조성물이 제공된다.The present invention relates to a composition for controlling white spotted fungi.
The present invention relates to a composition for controlling white spotted fungi, which comprises a Lactobacillus sp . Strain or a culture thereof as an active ingredient. Preferably, the Lactobacillus sp . Strain is Lactobacillus sp . Room ( Lactobacillus plantarum ).
The present invention provides a composition for controlling a fungus of the spore mosaic virus having an effect of controlling a large amount of fungal diseases occurring in larvae in the course of rearing of white spotted flowers.

Description

흰점박이꽃무지 진균병 방제용 조성물{Protaetia brevitarsis fungal diseases controlling compositions}{Protaetia brevitarsis fungal diseases controlling compositions}

본 발명은 흰점박이꽃무지 진균병 방제용 조성물에 관한 것으로서, 보다 상세하게는 락토바실러스 속(Lactobacillus sp .) 균주를 유효성분으로 이용함으로써 흰점박이꽃무지의 사육과정에서 유충에 다량으로 발생하는 진균병을 방제하는 효과를 갖는 흰점박이꽃무지 진균병 방제용 조성물에 관한 것이다.The present invention relates to a composition for controlling white spotted fungi. More particularly, the present invention relates to a composition for controlling fungus caused by a large amount of fungi in larvae during the rearing of white spotted flowers by using a strain of Lactobacillus sp . The present invention relates to a composition for controlling fungi of white spotted flowers having an effect of controlling diseases.

국내 산업곤충 시장은 1990년 후반부부터 성장하기 시작하였으며 대상 곤충이 꿀벌, 화분매개충, 천적, 애완곤충 등으로 그 종이 다양화되고 곤충자원의 가치가 증대되었다. Domestic industrial insect market began to grow from the latter half of 1990, and the insect was diversified into bee, pollen, nemesis and pet insect, and the value of insect resource increased.

이와 관련하여 곤충을 식용화하기 위하여 다양한 기능성 및 영양원에 대한 연구가 추진되고 있으며 거저리, 장수풍뎅이 및 꽃무지는 현재 식품원료로 등재를 위해 연구를 가속화하고 있다.In this regard, studies on various functionalities and nutrients have been pursued in order to edify the insects, and the ducks, longevity beetles, and flower blooms have been accelerating their research to register as food raw materials.

최근 곤충(장수풍뎅이, 흰점박이꽃무지 및 갈색거저리)이 식품소재 및 동물사료로서 이용되고 산업적으로 곤충의 가치가 다양하고 커지면서 곤충을 사육하는 농가들이 증가하는 추세에 있다. Recently, insects (longevity beetles, white spotted flowers and brown goats) are used as food materials and animal feeds, and as the value of insects in industry varies and grows, there is an increasing trend of growing insects.

그러나 대량사육 등 사육규모가 증가할수록 곤충질병에 대한 문제가 커지고 있는 실정이므로 이를 예방하고 방제하기 위한 적절한 방법이 시급히 요구되고 있다. However, as the size of breeding such as mass rearing increases, the problem about insect diseases is increasing. Therefore, appropriate methods for preventing and controlling them are urgently required.

이에 관련 선행기술로는 등록특허 제10-1493426호(항진균 활성을 갖는 스트렙토마이세스 속 신균주 DUCC501 및 상기 신균주를 포함하는 유용곤충 질병 방제용 조성물) 및 공개특허 제10-2011-0092177호(미국부저병의 원인균에 대한 항균활성을 가지는 락토바실러스 플란타륨 YML 001, 상기 미생물을 함유하는 항균성 미생물 제제 및 상기 미생물을 이용한 방제방법)이 공지되어 있다. Prior art related to this is disclosed in Korean Patent No. 10-1493426 (DUCC501 strain of Streptomyces strain having antifungal activity and useful insect disease control composition including the new strain) and Published Patent Application No. 10-2011-0092177 Lactobacillus plantai YML 001 having antimicrobial activity against causative microorganisms of the United States of America, antimicrobial microorganism preparations containing the microorganisms and methods of controlling the microorganisms using the microorganisms are known.

그러나, 흰점박이꽃무지 진균병에 대한 EM 방제제 관련 연구는 미흡한 실정이다. However, studies on EM antiseptics against white spotted fungi have not been studied sufficiently.

본 발명의 목적은 흰점박이꽃무지의 사육과정에서 유충에 다량으로 발생하는 진균병을 방제하는 효과를 갖는 흰점박이꽃무지 진균병 방제용 조성물을 제공하는데 있다.It is an object of the present invention to provide a composition for controlling the fungus of the spider mite, which has an effect of controlling a large amount of fungal diseases occurring in larvae during the rearing of white spotted flowers.

본 발명이 이루고자 하는 기술적 과제들은 이상에서 언급한 기술적 과제들로 제한되지 않으며, 언급되지 않은 다른 기술적 과제들은 아래의 기재로부터 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에게 명확하게 이해될 수 있을 것이다.It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.

상기 목적을 달성하기 위한 본 발명은, 흰점박이꽃무지 진균병 방제용 조성물로서, 락토바실러스 속(Lactobacillus sp .) 균주 또는 이의 배양액을 유효성분으로 포함하는 것이 특징이며, 바람직하게는 상기 락토바실러스 속 균주는 락토바실러스 플란타룸(Lactobacillus plantarum)인 것이 특징이다. In order to achieve the above object, the present invention is a composition for controlling white spotted fungi, characterized by comprising a Lactobacillus sp . Strain or a culture thereof as an active ingredient, and preferably the Lactobacillus sp. The strain is characterized by being Lactobacillus plantarum .

상기 진균은 메타리지움 아니소플리애(Metarhizium anisopliae)인 것이 특징이다.The fungus is Metarhizium < RTI ID = 0.0 > anisopliae ).

또 다른 본 발명의 흰점박이꽃무지 진균병 방제용 조성물로는, 락토바실러스 플란타룸(Lactobacillus plantarum) 균주 또는 이의 배양액, 바실러스 서브틸리스 서브스페이스(Bacillus subtilis subsp) 균주 또는 이의 배양액 및, 사카로마이세스 세레비지애(Sacharomyces cerevisiae) 균주 또는 이의 배양액을 유효성분으로 포함하는 것이 특징이다.Another composition of the present invention for controlling white spotted fungi is a Lactobacillus plantarum strain or a culture thereof, a Bacillus subtilis subspecies ( Bacillus subtilis subsp. subsp ) or a culture thereof, and Sacharomyces < (R) > cerevisiae strain or a culture thereof as an active ingredient.

이러한 상기 흰점박이꽃무지 진균병 방제용 조성물은 흰점박이꽃무지 진균병의 기주곤충에 주입하여 흰점박이꽃무지 진균병을 방제하는 것이 특징이다.The composition for controlling fungicides of the present invention is characterized in that it is injected into host insects of a white spotted flower fungus to control a white spotted fungus.

본 발명에 의해, 흰점박이꽃무지의 사육과정에서 유충에 다량으로 발생하는 진균병을 방제하는 효과를 갖는 흰점박이꽃무지 진균병 방제용 조성물이 제공됨으로써, 곤충병원균의 사전예방이 가능해져 농가 소득원 감소를 줄일 수 있음과 동시에 미생물을 이용함으로써 안전하고 건강한 흰점박이꽃무지 생산이 가능하게 되어 농가의 소득향상이 기대된다.The present invention provides a composition for controlling fungal diseases of white spotted flowers, which has an effect of controlling a large amount of fungal diseases occurring in larvae in the process of rearing white spotted flowers, thereby enabling prevention of insect pathogens, It is possible to produce a safe and healthy white spotted flower by using microorganisms and it is expected that the income of farmers can be improved.

도 1은 메타리지움 아니소플리애(Metarhizium anisopliae)에 각 EM 균주별로 도포한 후, 15일 경과 후 메타리지움 아니소플리애(Metarhizium anisopliae) 진균 억제정도를 나타낸 도면이다.
도 2는 3령 흰점박이꽃무지의 혈림프 내 EM 균주별 Injection 실험결과를 나타낸 도면이다.Figure 1 is a graphical representation of the < RTI ID = 0.0 > Metarhizium & Anisopliae ) was applied to each EM strain, and after 15 days, Metarhizium anisopliae ) fungi.
FIG. 2 is a graph showing the results of injection experiments of EM strains in the hemolymph of the third generation white spotted flowers.

이하, 본 발명을 상세하게 설명하며, 본 발명의 요지를 불필요하게 흐릴 수 있는 공지 기능 및 구성에 대한 상세한 설명은 생략한다.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail, and a detailed description of known functions and configurations that may unnecessarily obscure the gist of the present invention will be omitted.

본 발명에 따른 흰점박이꽃무지 진균병 방제용 조성물은 락토바실러스 속(Lactobacillus sp .) 균주 또는 이의 배양액을 유효성분으로 포함하는 것이 특징이며, 보다 바람직하게는 상기 락토바실러스 속 균주는 락토바실러스 플란타룸(Lactobacillus plantarum)인 것이 특징이다. The composition for controlling white spotted fungi according to the present invention is characterized by comprising a Lactobacillus sp . Strain or a culture thereof as an active ingredient. More preferably, the Lactobacillus sp . Strain is Lactobacillus sp . Room ( Lactobacillus plantarum ).

설명하면, 본 발명의 발명자들은 흰점박이꽃무지의 사육과정에서 유충에 다량으로 발생하는 진균병을 방제하기 위하여 EM(Effective Micro-organism)을 이용하여 식용곤충으로 사육이 많이 되고 있는 흰점박이꽃무지에 자주 나타나는 메타리지움 아니소플리애(Metarhizium anisopliae)도 방제하면서 인체에 해롭지 않은 사육을 도모하고, 사육농가의 피해를 줄이기 위하여 본 발명을 개발하게 된 것이다.The inventors of the present invention have found that, in order to control a large amount of fungal diseases occurring in larvae during the rearing process of white spotted flowers, it is possible to use the effective micro-organism (EM) Metarhizium , which frequently appears in anisopliae ) to prevent harm to the human body, and to reduce the damage of the farming house.

즉, 여러 균주들을 대상으로 흰점박이꽃무지 유충 사육시 흰점박이꽃무지 진균병(fungal pathogen)을 일으키는 원인균인 메타리지움 아니소플리애(Metarhizium anisopliae)에 대해 항균활성을 확인하며, 상기 항균활성을 나타내는 균주들을 각각 고체배지에서 배양하여 얻은 포자를 이용하거나, 또는 액체배지에서 배양하여 얻은 배양액을 이용하여 흰점박이꽃무지 진균병(fungal pathogen)에 의해 흰점박이 유충이 폐사되는 것을 방제하는 것이다.In other words, the antimicrobial activity of Metarhizium anisopliae , which is a causative agent of fungal pathogen of white spotted spider mite , was observed in a variety of strains , and the antimicrobial activity Is inhibited by the fungal pathogen of the white spotted spider mite using a culture obtained by culturing each strain in a solid medium or in a liquid medium.

이때, 항균활성을 나타내는 상기 균주들은 락토바실러스 플란타룸(Lactobacillus plantarum) 균주, 바실러스 서브틸리스 서브스페이스(Bacillus subtilis subsp) 균주 및, 사카로마이세스 세레비지애(Sacharomyces cerevisiae) 균주인 것으로, 상기 균주들이 처리된 사육환경에서는 진균이 전혀 발생되지 않으며, 상기 EM이 처리되지 않은 사육상자에는 진균이 발생되어 모든 유충이 폐사됨을 확인하였다. The strains exhibiting antimicrobial activity may be selected from the group consisting of Lactobacillus plantarum strain, Bacillus subtilis subsp strain, and Sacharomyces cerevisiae strain. S. cerevisiae strain, and it was confirmed that no fungi were generated in the environment where the strains were treated, and that all the larvae were killed due to the occurrence of fungi in the breeding box not treated with the EM.

그 중, 상기 락토바실러스 플란타룸(Lactobacillus plantarum) 균주가 EM으로 시중에서 쉽게 구할 수 있고, 곤충병원균으로 작용하지 않으며 인체에 무해하므로 식용곤충의 진균방제에 유익한 것으로 판단되어, 이를 유효성분으로 포함하는 흰점박이꽃무지 진균병 방제용 조성물을 제공하게 된다. Among them, the Lactobacillus plantarum strain can be easily obtained in the market by EM, does not act as an insect pathogen, and is harmless to human body. Therefore, it is considered to be useful for controlling fungi of edible insects, The present invention provides a composition for controlling white spotted fungi.

특히, 상기 락토바실러스 플란타룸(Lactobacillus plantarum) 균주의 포자 농도는 1×107 내지 1×109 CFU/㎖가 되도록하여 흰점박이꽃무지 진균병 방제효과를 나타내게 한다.Particularly, the spore concentration of the Lactobacillus plantarum strain is adjusted to 1 × 10 7 to 1 × 10 9 CFU / ml to exhibit the effect of controlling the white spotted fungus.

이때, 상기 포자농도가 상기 농도범위보다 적을 경우 흰점박이꽃무지 진균병에 대한 방제효능이 떨어지게 되며, 상기 농도범위보다 많을 경우에는 과도한 포자함량으로 인해 곤충에 주입이 용이하지 않아 혈강주사처리로 인한 방제방법이 어렵게 된다.If the spore concentration is lower than the concentration range, the control effect against the white spider mushroom fungus disease becomes poor. If the spore concentration is higher than the concentration range, the insect is not easily injected due to the excessive spore content. The control method becomes difficult.

또한, 본 발명에 따른 흰점박이꽃무지 진균병 방제용 조성물은 상기 균주의 배양액을 유효성분으로 사용하는데, 이때 상기 배양액 제조시 배양은 25~28℃전후하여 7~15일간 배양하는 것이 바람직하다. 이는 7일 미만으로 배양하는 경우에는 항진균성 물질을 충분히 생성하지 못해 방제효과가 미미하고, 15일을 초과하여 배양하는 경우에는 생산원가의 상승에 비하여 생산효과가 낮기 때문이다. 그러나, 필요에 따라 배양기간을 조절할 수도 있다.In addition, the composition for controlling fungicidal disease of white spotted flowers according to the present invention uses a culture medium of the above strain as an active ingredient, wherein the culture is preferably cultured at about 25 to 28 ° C for 7 to 15 days. This is because, when cultivated for less than 7 days, the effect of controlling is insufficient due to insufficient generation of antifungal substances, and when cultivating for more than 15 days, production effect is lower than the increase of production cost. However, the incubation period may be adjusted as needed.

또한, 본 발명에 따른 흰점박이꽃무지 진균병 방제용 조성물은 흰점박이꽃무지 진균병(fungal pathogen)을 일으키는 원인균인 메타리지움 아니소플리애(Metarhizium anisopliae)에 대해 항균활성을 나타내는 락토바실러스 플란타룸(Lactobacillus plandrl tarum) 균주 또는 이의 배양액, 바실러스 서브틸리스 서브스페이스(Bacillus subtilis subsp) 균주 또는 이의 배양액 및, 사카로마이세스 세레비지애(Sacharomyces cerevisiae) 균주 또는 이의 배양액 모두를 포함하기도 한다. In addition, the composition for controlling fungicidal disease of white spotted flowers according to the present invention is useful as a fungicidal agent against Metarhizium anisopliae causing fungal pathogens of white spotted flowers, Lanta Room ( Lactobacillus plandrl tarum ) or a culture thereof, Bacillus subtilis subspaces ( Bacillus subtilis subsp ) or a culture thereof, and a Sacharomyces cerevisiae strain or a culture thereof.

이때, 상기 락토바실러스 플란타룸(Lactobacillus plantarum) 균주의 포자 농도는 1×107 내지 1×109 CFU/㎖이며, 상기 바실러스 서브틸리스 서브스페이스(Bacillus subtilis subsp) 균주의 포자 농도는 1×107 내지 1×109 CFU/㎖이며, 상기 사카로마이세스 세레비지애(Sacharomyces cerevisiae) 균주의 포자 농도는 1×107 내지 1×109 CFU/㎖가 되도록하여 흰점박이꽃무지 진균병 방제효과를 나타내게 한다. 그러나, 상기 포자농도는 한정되는 것은 아니며, 병원균의 감염정도 및 주변 환경 조건 등에 따라 유효농도를 조절하여 사용할 수 있다. At this time, the spore concentration of the Lactobacillus plantarum strain is 1 × 10 7 to 1 × 10 9 CFU / ml, the Bacillus subtilis subspaces ( Bacillus subtilis subsp ) strain is 1 × 10 7 to 1 × 10 9 CFU / ml, and the Sacharomyces S. cerevisiae strain is adjusted to a concentration of 1 × 10 7 to 1 × 10 9 CFU / ml to exhibit the effect of controlling the white spotted fungus. However, the spore concentration is not limited, and the effective concentration may be adjusted depending on the degree of infection of the pathogens and environmental conditions.

또한, 상기 락토바실러스 플란타룸 균주의 배양액과 바실러스 서브틸리스 서브스페이스 균주의 배양액과 사카로마이세스 세레비지애 균주의 배양액은 1 ~ 3 : 1 : 1 ~ 2로 이루어질 경우 우수한 흰점박이꽃무지 진균병 방제효과를 나타내게 된다. In addition, when the culture medium of the Lactobacillus plantarum strain, the culture medium of the Bacillus subtilis subsp. Strain and the culture medium of the Saccharomyces cerevisiae strain is 1: 3: 1: 1 to 2, excellent white spotted flowers Thereby exhibiting the fungal disease control effect.

이는 상기 혼합비율을 벗어날 경우 생산원가의 상승에 비하여 흰점박이꽃무지 진균병 방제효과가 높게 나타나지 않은 바, 상기 비율로 이루어지는 것이 적합하다.This is because, when the mixing ratio is out of the above range, the effect of controlling the white foxtail fungus disease is not high as compared with the increase of the production cost, and it is preferable that the ratio is the above ratio.

이렇게 제조된 상기 흰점박이꽃무지 진균병 방제용 조성물은 흰점박이꽃무지 진균병의 기주곤충 즉, 흰점박이꽃무지 유충에 직접 주입하여 흰점박이꽃무지 진균병을 방제하게 한다. 이때 상기 주입방식은 7일에 한번씩 흰점박이꽃무지 유충 1마리당 1×105 내지 1×107/50㎕로 혈강주사하는 것이 적합하다.The thus-prepared composition for controlling fungicidal disease of white spotted flowers is directly injected into a host insect of a white spotted flower fungus, that is, a white spotted fly larva, thereby controlling a white spotted fungal fungus. In this case, the injection system is adapted to scan a first hyeolgang maridang white spotted flower plain larvae Every 7 days 1 × 10 5 to 1 × 10 7 / 50㎕.

이하에서는 실시예를 들어 본 발명에 관하여 더욱 상세하게 설명할 것이나, 이들 실시예는 단지 설명의 목적을 위한 것으로 본 발명의 보호 범위를 제한하고자 하는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to examples, but these examples are for illustrative purposes only and are not intended to limit the scope of protection of the present invention.

<실시예 1> 시중에서 구할 수 있는 EM을 사용하여 In vivo상태에서 흰점박이꽃무지 2령 및 3령 유충 진균 방제 확인<Example 1> The in vivo state of the white spotted flowers using the commercially available EM was confirmed in the second and third instar fungi control

진균이 잘 발생되는 환경을 조성(사육상자 밀폐, 진균 성장 적합 온도조성) 한 후 EM을 처리한 군과 처리하지 않은 군을 비교하여 진균피해를 확인하였다.The fungus damage was confirmed by comparing the EM treated group with the untreated group after establishing an environment in which fungi were well generated (breeding box sealing, fungus growth suitable temperature composition).

1. 실험방법1. Experimental Method

1) In vivo로 2령 흰점박이꽃무지 균을 처리하는 방법1) In vivo treatment of 2nd instar white spider mite

곤충사육농가와 유사한 환경을 조성한 군과 연구실에서 개별사육을 통한 환경군으로 나누어 실험하였다. The experiment was divided into two groups, one group which had similar environment to insect farmers and the other group which had individual breeding in laboratory.

우선, 녹강균이 오염된 톱밥과 물 또는 EM(1×107CFU/㎖)을 3:1의 부피(톱밥 300g+물/ EM 100ml)로 섞어서 30일간 먹여서 실험하였다. 곤충사육농가와 유사한 환경군을 조성하기 위해 농가에서 주로 사육에 사용하고 있는 400×270×230mm 투명 플라스틱 사육 상자에 2령, 3령 흰점박이꽃무지 그룹을 각각 10두씩 넣어 사육하였고, 각 3반복을 하였다. First, the sawdust contaminated with green rust bacterium was mixed with water or EM (1 × 10 7 CFU / ml) in a volume of 3: 1 (sawdust 300 g + water / EM 100 ml) for 30 days. In order to create an environment group similar to that of insect farmers, 10 pairs of 2 nd and 3 rd old white spotted flowers were placed in a 400 × 270 × 230 mm transparent plastic breeding box, Respectively.

플라스틱 사육 상자는 control의 경우 환기가 되는 조건과 밀폐조건으로 나누어 실험하였고, EM 대신 3차 증류수를 톱밥에 넣어 섞어주었다. The plastic breeding box was divided into ventilation condition and airtight condition for control, and the third distilled water instead of EM was mixed in sawdust.

환기조건은 사육 상자에 아무런 조치를 하지 않았고, 밀폐조건은 사육상자 입구에 비닐랩을 씌우고 여러 개의 구멍을 내어 환기가 되는 것을 최소한으로 막아 진균이 잘 자라는 조건을 만들었다. The ventilation conditions did not take any action on the breeding box, and the sealing conditions were such that a plastic wrap was put on the entrance of the breeding box and multiple holes were made to prevent ventilation by minimizing the fungal growth.

또한, 사육농가에서는 주로 비닐하우스나 가건물에서 온도 습도 제어 없이 자연환경으로 사육하는 경우가 많으므로 곤충사육농가와 유사환경조건에서 온도를 제어하지 않고 사육을 하였다. Also, in breeding farms, breeding is usually done in a greenhouse or cabinets without control of temperature under the same environmental conditions as those of insect farmers, because they are often kept in a natural environment without temperature and humidity control.

연구실에서 개별사육을 하는 방법으로는 지름 100mm 플라스틱 페트리디쉬에 흰점박이꽃무지 각 1두씩 60g의 톱밥에 20ml의 각각의 EM을 넣고 잘 섞은 후 28~29℃를 유지하여 사육하였다. As a method of individual breeding in the laboratory, 20 ml of each EM was added to 60 g of sawdust each of 1 spotted white spotted flowers in a plastic petridish dish having a diameter of 100 mm, and the mixture was kept at 28 to 29 ° C for breeding.

EM의 종류는 A균: 고초균(바실러스 서브틸리스 서브스페이스)+효모균(사카로마이세스 세레비지애)+유산균(락토바실러스 플란타룸), B균: 광합성균, C균: 질화세균이며 각각의 농도는 1×107colony/ml로 처리하였다. EM은 10일 간격으로 처리를 해 주었다. 그리고 30일 뒤에 흰점박이꽃무지의 Mortality를 확인하였다.The types of EM are A bacteria: Bacillus subtilis subspaces + yeast bacteria (Saccharomyces cerevisiae) + lactic acid bacteria (lactobacillus planta), B bacteria: photosynthesis bacteria, C bacteria: nitrifying bacteria Was treated with 1 x 10 &lt; 7 &gt; colony / ml. EM was treated at intervals of 10 days. After 30 days, the mortality of white spotted flowers was confirmed.

2) Control과 처리 균 구: A, B, C 균2) Control and treated bacteria: A, B, C

Control: 3차 증류수 처리Control: 3rd Distilled Water Treatment

A균: 고초균(바실러스 서브틸리스 서브스페이스)+효모균(사카로마이세스 세레비지애)+유산균(락토바실러스 플란타룸)= 1 ml(1×107colony함유) + 1 ~ 2ml(1×107colony함유) + 1 ~ 3 ml(1×107colony함유)1 ml (containing 1 × 10 7 colony) + 1 to 2 ml (1 × 10 7 colony) + Bacteria (Bacillus subtilis subspace) + Yeast (Saccharomyces cerevisiae) + Lactobacillus (Lactobacillus plantarum) 10 7 colony containing) + 1 to 3 ml (containing 1 × 10 7 colony)

B균: 광합성균B bacteria: photosynthetic bacteria

C균: 질화세균C bacteria: nitrifying bacteria

3) 처리 환경: 자연온도(곤충 사육농가 환경과 같음)3) Processing environment: Natural temperature (same as insect farm environment)

3.1) 자연온도 (사육 상자에 넣어 사육, 농가에서 키우는 방식)3.1) Natural temperature (breeding in a breeding box, farming method)

사육상자 규격: 400×270×230mm에 15-30두 사육(10두×3반복)Breeding box Specification: 400 × 270 × 230mm to 15-30 breeding (10 × 2 × 3 repeats)

3.1.1.) 자연 온도(낮 25℃/ 밤 20℃) 2령 1개월 후(밀폐: 랩을 씌우고 구멍을 내어 산소 공급)3.1.1.) Natural temperature (day 25 ° C / night 20 ° C) 2 months after 1 month (sealed: wrapped and punctured to supply oxygen)

3.1.2.) 곤충사육농가 환경 온도(28-29℃ 유지)에서 녹강균에 오염된 톱밥으로 3령 흰점박이꽃무지 사육 (밀폐: 랩을 씌우고 구멍을 내어 산소 공급 또는 환기)3.1.2.) Insect Breeding In the farmhouse environment temperature (28-29 ℃), the third piece of sawdust contaminated with rust-bacillus is the white spotted bloom plain raising (sealing: lapping and oxygenation or ventilation)

3.2.) 실험실 사육 환경(1두씩 페트리디쉬에 개별 사육) 온도(29-30℃ 유지)에서 녹강균에 오염된 톱밥으로 2령과 3령 흰점박이꽃무지 사육3.2.) Laboratory Breeding Environment (Individual Breeding in Petrified Dishes) At the temperature (maintained at 29-30 ℃), sawdust contaminated with rust bacillus was used as the second and third white spotted flowers.

2. 실험결과2. Experimental results

2령Second generation Mortality(%)Mortality (%) 3령Third generation Mortality(%)Mortality (%) control(환기)control (ventilation) all dieall die 100100 all dieall die 100100 control(밀폐)control all dieall die 100100 all dieall die 100100 A균A fungus 10두 생존10 Both Survival 6767 23두 생존23 Two survival 23,323,3 B균B bacteria 2두 생존Two living together 93.393.3 17두 생존17 Two survivors 43.343.3 C균C bacteria 5두 생존5 Both Survival 83.383.3 23두 생존23 Two survival 23.323.3

상기 표 1에 나타나 있듯이, 밀폐된 사육환경에서 진균이 잘자라는 온도인 20~25℃로 맞추어 유충을 사육한 결과, EM처리를 한 경우 진균이 밀폐된 환경에서도 진균이 성장하지 못하는 것을 확인하였으며, 특히, 흰점박이꽃무지에서 A균 처리 시 다른 균보다 Mortality 떨어짐을 확인하였다.As shown in Table 1, when the larvae were raised at 20-25 ° C, which is a temperature at which the fungus grows well in an enclosed rearing environment, it was confirmed that the fungi did not grow even in the environment where the fungi were enclosed by the EM treatment, Especially, it was confirmed that the mortality of the A spp.

또한, 2령 또는 3령의 흰점박이꽃무지를 29~30℃ 온도를 일정유지한 실험실에서 사육한 결과 하기 표 2와 같이 나타났다.In addition, the second or third white spotted flowers were raised in a laboratory maintained at a temperature of 29 to 30 ° C as shown in Table 2 below.

2령Second generation Mortality(%)Mortality (%) 3령Third generation Mortality(%)Mortality (%) controlcontrol 20두->14두(1두 진균병 발생, 3두 번데기)20 Duplicates -> 14 Duplicates (1 Duplicate Fungal Disease, 3 Duplicates) 1010 20두->18두 생존Twenty to two -> Two to survive 3030 A균A fungus 20두->17두(0두 진균병 발생, 3두 번데기)Twenty - two> 17 (two fungal infections, three two pups) 1010 20두->18두 생존Twenty to two -> Two to survive 1515 B균B bacteria 20두->15두(2두 진균병 발생, 2두 번데기)Twenty-two to fifteen (two two fungal infections, two pupa) 1010 20두->18두 생존Twenty to two -> Two to survive 2525 C균C bacteria 20두->16두(3두 진균병 발생, 16두 번데기)Twenty - two to sixteen (three two fungal infections, sixteen) 55 20두->19두 생존20 to 20 -> Two to survive 2525

상기 표 2에 나타나 있듯이, A균 EM(고초균, 유산균, 효모균)이 포함된 톱밥을 먹인 유충이 가장 진균병이 적게 발생되었으므로, 이를 토대로 상기 균들에 대한 in vitro 실험을 진행하였다.As shown in the above Table 2, since the fungi fed with sawdust containing A microorganism EM (Bacillus subtilis, Lactobacillus, Yeast) were the least fungal diseases, the in vitro experiment was conducted on the microbes based on them.

<실시예 2> 시중에서 구할 수 있는 EM을 사용하여 In vitro상태에서 흰점박이꽃무지 2령 및 3령 유충 진균 방제 확인<Example 2> Examination of second and third instar fungi of white spotted flowers in vitro using commercially available EM

1. 실험방법1. Experimental Method

Metarhizium anisopliae (KACC40969)에 EM의 A균에 포함된 고초균(Bacillus subtilis subsp), 유산균(Lactobacillus plantarum), 효모균(Sacharomyces cerevisiae)를 각각 1 plate에 처리하여 진균 억제 효과를 측정하였다. Ma가 잘 자랄 수 있는 SDA(Saudbourd Agar)배지에 고초균(바실러스 서브틸리스 서브스페이스), 효모균(사카로마이세스 세레비지애), 유산균(락토바실러스 플란타룸)을 각각 농도 1×107 colony/ml (OD 600nm로 0.4~0.6, 1ml)를 200㎕를 배지에 도말한 후, Ma를 페트리디쉬 중앙에 seeding 하였다. 각 실험은 3반복 하였으며, 4주 뒤에 진균이 자란 표면적을 측정하여 진균 억제 정도를 확인 하였다. Metarhizium Bacillus subtilis subsp., Lactobacillus plantarum , and Sacharomyces cerevisiae , which were included in EM bacteria, were treated with 1 plate of anisopliae (KACC40969) to measure the fungicidal effect. (Bacillus subtilis subspace), yeast (Saccharomyces cerevisiae), and lactic acid bacteria (Lactobacillus plantarum) were added to SDA (Saudbourg Agar) medium to which Ma could grow well at a concentration of 1 × 10 7 colony / ml (0.4 to 0.6 at 1 at 600 nm, 1 ml) was seeded onto the medium, and Ma was seeded at the center of the petri dish. Each experiment was repeated three times. After 4 weeks, the surface area of the fungi was measured to confirm the degree of fungal inhibition.

2. 실험결과2. Experimental results

상기 실험결과, 도 1 및 하기 표 3과 같이 나타났다.The results of the experiment are shown in Fig. 1 and Table 3 below.

15일15th 30일30 days ControlControl 5 cm5 cm 6.67 cm6.67 cm Lactobacillus Lactobacillus plantarumplantarum 1.5 cm1.5 cm 4.7 cm4.7 cm Bacillus Bacillus subtilissubtilis subspsubsp 1 cm1 cm 6.16 cm6.16 cm SacharomycesSacharomyces cerevisiaecerevisiae 1 cm1 cm 2.5 cm2.5 cm 락스Rocks N/AN / A 6.83 cm6.83 cm

상기 표 3 및 도 1에 나타나 있듯이, Metarhizium anisopliae (KACC40969)에 EM의 A균에 포함된 고초균(바실러스 서브틸리스 서브스페이스), 효모균(사카로마이세스 세레비지애), 유산균(락토바실러스 플란타룸) 모두 진균 억제효과를 나타냄을 확인하였다. As shown in Table 3 and Figure 1, Metarhizium anisopliae (KACC40969) of Bacillus subtilis include in the A strains of EM (Bacillus subtilis subspace), yeast (Saccharomyces as MY process three Levy jiae), lactic acid bacterium (Lactobacillus Planta room) were confirmed to indicate the fungal inhibitory effect .

<실시예 3> 3령 흰점박이꽃무지의 혈림프 내 EM 균 Injection 실험<Example 3> Injection experiment of EM bacteria in the hemolymph of the third generation white spotted flower

1. 실험방법1. Experimental Method

Injection실험은 3령 흰점박이꽃무지를 72시간 굶긴 후에 Negative Control로 3차증류수, Positive Control로 Nutrient broth, MRS broth, R2A broth를 1두에 50㎕씩 injection하였다. Injection experiments were performed by injecting 50 μl of Nutrient broth, MRS broth, and R2A broth with 3 rd distilled water using Negative Control, Positive Control,

Bacillus subtilis subsp, Lactobacillus plantarum, Sacharomyces cerevisiae를 1×107/50㎕를 BD ultra-fine 인슐린 주사기(31게이지, 6mm)를 사용하여 injection하였다. Bacillus subtilis subsp, was injection to the Lactobacillus plantarum, Sacharomyces cerevisiae to 1 × 10 7 / 50㎕ using BD ultra-fine insulin syringe (31 gauge, 6mm).

Injection한 뒤 1두씩 지름 100mm 원형 플라스틱 페트리디쉬에 발효톱밥을 넣어준 후 7일 뒤 Mortality를 측정하였다.After injection, fermented sawdust was placed in a 100 mm diameter round plastic petri dish and the mortality was measured 7 days later.

2. 실험결과 2. Experimental results

상기 실험결과 도 2 및 하기 표 4와 같이 나타났다. The results of the experiment are shown in FIG. 2 and Table 4 below.

Injection subjectInjection subject 3령 Mortality(%)Third Mortality (%) ControlControl 1010 Nutrient Broth InjectionNutrient Broth Injection 3.33.3 MRS Broth InjectionMRS Broth Injection 2020 R2A Broth InjectionR2A Broth Injection 3.33.3 Lactobacillus plantarum Injection Lactobacillus plantarum Injection 6.76.7 Bacillus subtilis subsp . Injection Bacillus subtilis subsp . Injection 23.323.3 Sacharomyces cerevisiae Injection Sacharomyces cerevisiae Injection 1010

도 2 및 상기 표 4에 나타나 있듯이, in vitro 실험에서 Lactobacillus plantarum, Bacillus subtilis subsp ., Sacharomyces cerevisiae 모두 Metarhizium anisopliae를 억제하는 효과가 나타났다.As shown in FIG. 2 and Table 4, in vitro experiments showed that Lactobacillus plantarum, Bacillus subtilis subsp ., Sacharomyces cerevisiae All showed an inhibitory effect on Metarhizium anisopliae .

그러나, 흰점박이꽃무지 혈강 주사(1×107/50㎕ injection)를 하였을 때 Mortality는 Lactobacillus plantarum의 Mortality가 가장 적게 나타났다.However, when the white-spotted flower solid hyeolgang injection (1 × 10 7 / 50㎕ injection ) Mortality Mortality is that of Lactobacillus plantarum it was the least.

또한 B . subtilis subsp .와 S. cerevisiae는 Reference에서 흰점박이꽃무지에서 pathogenic하다고 알려진 바는 없으나 다른 곤충 종에서 pathogen으로 분류되었고 혈림프에 injection을 하였을 경우도 B. subtilis subsp.의 경우 mortality가 비교적 높아 곤충의 표피의 상처를 통해 감염이 될 경우 치사 가능성이 있어 배제하였다. 따라서 L. plantarum(유산균)이 EM으로 시중에서 쉽게 구할 수 있고, 인체에 무해하므로 식용곤충의 진균방제에 유익할 것으로 사료된다.Also B. subtilis subsp . And S. cerevisiae are not known to be pathogenic in the white spotted flowers of Reference, but they are classified as pathogen in other insect species and B. subtilis subsp. The mortality was relatively high, so that it was possible to die if the infection was caused by the wound of the epidermis of the insect. Therefore, it is considered that L. plantarum (lactic acid bacteria) can be easily obtained in the market by EM and harmless to humans, so it is beneficial to control fungus of edible insects.

상기의 본 발명은 바람직한 실시예를 중심으로 살펴보았으며, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자는 본 발명의 본질적 기술 범위 내에서 상기 본 발명의 상세한 설명과 다른 형태의 실시예들을 구현할 수 있을 것이다. 여기서 본 발명의 본질적 기술범위는 특허청구범위에 나타나 있으며, 그와 동등한 범위 내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined by the appended claims. It will be possible. The scope of the present invention is defined by the appended claims, and all differences within the scope of the claims are to be construed as being included in the present invention.

Claims (10)

락토바실러스 속(Lactobacillus sp.) 균주 또는 이의 배양액을 유효성분으로 포함하는 것을 특징으로,
메타리지움 아니소플리애(Metarhizium anisopliae)로부터 발병되는 흰점박이꽃무지 진균병 방제용 조성물.
A strain of Lactobacillus sp. Or a culture thereof as an active ingredient,
( EN ) Composition for controlling fungi of white spotted flowers caused by Metarhizium anisopliae .
제 1항에 있어서,
상기 락토바실러스 속 균주는 락토바실러스 플란타룸(Lactobacillus plantarum)인 것을 특징으로 하는,
메타리지움 아니소플리애(Metarhizium anisopliae)로부터 발병되는 흰점박이꽃무지 진균병 방제용 조성물.
The method according to claim 1,
Characterized in that the strain of the genus Lactobacillus is Lactobacillus plantarum .
( EN ) Composition for controlling fungi of white spotted flowers caused by Metarhizium anisopliae .
제 1항에 있어서,
상기 락토바실러스 속 균주의 포자 농도가 1×107 내지 1×109 CFU/㎖인 것을 특징으로 하는,
메타리지움 아니소플리애(Metarhizium anisopliae)로부터 발병되는 흰점박이꽃무지 진균병 방제용 조성물.
The method according to claim 1,
Wherein the spore concentration of the strain of the genus Lactobacillus is 1 x 10 &lt; 7 &gt; to 1 x 10 &lt; 9 &gt;
( EN ) Composition for controlling fungi of white spotted flowers caused by Metarhizium anisopliae .
삭제delete 락토바실러스 플란타룸(Lactobacillus plantarum) 균주 또는 이의 배양액, 바실러스 서브틸리스 서브스페이스(Bacillus subtilis subsp) 균주 또는 이의 배양액 및, 사카로마이세스 세레비지애(Sacharomyces cerevisiae) 균주 또는 이의 배양액을 유효성분으로 포함하는 것을 특징으로,
메타리지움 아니소플리애(Metarhizium anisopliae)로부터 발병되는 흰점박이꽃무지 진균병 방제용 조성물.
A Lactobacillus plantarum strain or a culture thereof, a Bacillus subtilis subsp strain or a culture thereof, and a Sacharomyces cerevisiae strain or a culture thereof, as active ingredients Characterized in that,
( EN ) Composition for controlling fungi of white spotted flowers caused by Metarhizium anisopliae .
제 5항에 있어서,
상기 락토바실러스 플란타룸(Lactobacillus plantarum) 균주의 포자 농도가 1×107 내지 1×109 CFU/㎖인 것이 특징인,
메타리지움 아니소플리애(Metarhizium anisopliae)로부터 발병되는 흰점박이꽃무지 진균병 방제용 조성물.
6. The method of claim 5,
Wherein the spore concentration of the Lactobacillus plantarum strain is 1 x 10 7 to 1 x 10 9 CFU / ml.
( EN ) Composition for controlling fungi of white spotted flowers caused by Metarhizium anisopliae .
제 5항에 있어서,
상기 바실러스 서브틸리스 서브스페이스(Bacillus subtilis subsp) 균주의 포자 농도가 1×107 내지 1×109 CFU/㎖인 것이 특징인,
메타리지움 아니소플리애(Metarhizium anisopliae)로부터 발병되는 흰점박이꽃무지 진균병 방제용 조성물.
6. The method of claim 5,
Wherein the spore concentration of the Bacillus subtilis subsp . Strain is 1 x 10 7 to 1 x 10 9 CFU / ml.
( EN ) Composition for controlling fungi of white spotted flowers caused by Metarhizium anisopliae .
제 5항에 있어서,
상기 사카로마이세스 세레비지애(Sacharomyces cerevisiae)의 포자 농도가 1×107 내지 1×109 CFU/㎖인 것이 특징인,
메타리지움 아니소플리애(Metarhizium anisopliae)로부터 발병되는 흰점박이꽃무지 진균병 방제용 조성물.
6. The method of claim 5,
Wherein the spore concentration of said Sacharomyces cerevisiae is from 1 x 10 7 to 1 x 10 9 CFU / ml.
( EN ) Composition for controlling fungi of white spotted flowers caused by Metarhizium anisopliae .
삭제delete 제 1항 내지 제 3항 및, 제 5항 내지 제 8항 중 어느 한항의 흰점박이꽃무지 진균병 방제용 조성물 유효량을 흰점박이꽃무지 진균병의 기주곤충에 주입하는 것을 특징으로 하는,
메타리지움 아니소플리애(Metarhizium anisopliae)로부터 발병되는 흰점박이꽃무지 진균병 방제방법.An effective amount of the composition for controlling fungicidal disease of white spotted flowers of any one of claims 1 to 3 and 5 to 8 is injected into a host insect of a white spotted fungus.
A method for the prevention of fungi of white spotted flowers caused by Metarhizium anisopliae .
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