KR19990026602A - New Acremonium sp. Use of MJN1 strain and its extract as pollution-free plant growth promoter - Google Patents

New Acremonium sp. Use of MJN1 strain and its extract as pollution-free plant growth promoter Download PDF

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KR19990026602A
KR19990026602A KR1019970048803A KR19970048803A KR19990026602A KR 19990026602 A KR19990026602 A KR 19990026602A KR 1019970048803 A KR1019970048803 A KR 1019970048803A KR 19970048803 A KR19970048803 A KR 19970048803A KR 19990026602 A KR19990026602 A KR 19990026602A
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서주원
정양조
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Abstract

본 발명은 고추, 양파, 마늘등 식물의 성장에 영향을 주는 신규의 Acremonium spp. 균주와 그 대사산물을 제공한다.The present invention is a novel Acremonium spp. Provide strains and their metabolites.

본 발명 균주는 밭 토양에서 분리 배양하여 균사체 및 배양배지 여과액에서 추출된 천연물질이 식물의 성장 촉진 효과를 나타내는 것을 실험적으로 확인하였다.The strain of the present invention was experimentally confirmed that the natural material extracted from the mycelia and culture medium filtrate separated and cultured in the field soil shows the effect of promoting the growth of the plant.

본 발명 신규의 균주는 Acremonium sp. MJN1으로 명명되었으며 균사체 추출물의 연속희석액의 처리를 통한 실험 및 실시예로 무공해 식물성장 촉진제로서의 신규의 용도를 제공한다.The novel strain of the present invention is Acremonium sp. Experiments and examples, named MJN1, through the treatment of serial dilutions of mycelium extracts, provide novel uses as pollution-free plant growth promoters.

Description

신규의 Ａｃｒｅｍｏｎｉｕｍ ｓｐ．ＭＪＮ１ 균주와 그 추출물의 무공해 식물성장 촉진제로서의 용도.Use of a novel AbcRem spore.MVN1 strain and its extract as a pollution-free plant growth promoter.

본 발명은 신규의 Acremonium sp. MJN1 균주와 그 추출물의 용도에 관한 것이다. 더 상세하게는 본 발명은 밭토양에서 분리 배양한 신규한 Acremonium sp. MJN1과 그 대사산물을 이용한 무공해 식물성장 촉진제로서의 용도에 관한 것이다.The present invention provides a novel Acremonium sp. It relates to the use of MJN1 strain and its extract. More specifically, the present invention is a novel Acremonium sp. It relates to the use as a pollution-free plant growth promoter using MJN1 and its metabolites.

본 발명은 식용작물의 증산에 직접 관련된다. 종래 식용작물의 수확 증대를 위하여 화학 합성 비료들이 제조되어 사용하여 왔다. 그러나, 과다한 화학 합성 비료의 사용은 토양의 산성화를 초래하고 따라서 작물의 내병성이 저하되어 살균제 또는 살충제등의 화학합성 농약을 반복적으로 증량하여 사용해야 하는 심각한 문제가 초래되어 왔다.The present invention relates directly to the production of edible crops. Chemical synthetic fertilizers have been prepared and used to increase the yield of conventional food crops. However, excessive use of chemical synthetic fertilizers has resulted in acidification of the soil and thus lowered the disease resistance of crops, which has caused a serious problem of repeatedly increasing the use of chemical synthetic pesticides such as fungicides or insecticides.

본 발명은 종래 전통적으로 실시하여온 작물 수확량 증진방법의 결함을 감안하여 안출된 것으로 신규 미생물과 그의 대사산물을 이용한 무공해 식물성장 촉진제로서의 용도를 제공함을 그 목적으로 한다.SUMMARY OF THE INVENTION The present invention has been made in view of the deficiency of the conventional crop yield enhancement method, and its object is to provide a use as a pollution-free plant growth promoter using new microorganisms and their metabolites.

본 발명의 다른 목적은 식물성 병원균이 아니며 동 균주으로부터 추출된 천연물질은 식물의 성장을 조절할 수 있는 무공해 식물성장 촉진제를 제공함에 있다.Another object of the present invention is not a plant pathogen, and the natural material extracted from the strain is to provide a pollution-free plant growth promoter that can control the growth of the plant.

본 발명의 또다른 목적은 본 발명 균주의 추출물을 작물체에 처리하는 경우 매우 적은 양으로도 식물의 성장을 촉진시키고 수확량을 증대시켜 현재 공해를 유발할 수 있는 화학비료 및 화학합성농약의 사용량을 현저히 줄이고 또 수확량을 증대할 수 있는 새로운 농법의 제공하는 데 있다.Another object of the present invention is to significantly reduce the amount of chemical fertilizers and chemical synthetic pesticides that can cause pollution by promoting the growth of plants and increasing the yield in very small amounts when the extract of the present invention is treated to crops It is also to provide a new farming method that can increase the yield.

따라서, 본발명은 상기 목적을 달성하기 위하여 밭토양으로부터 신규한 미생물 균주를 순수 분리하여 배양하는 공정;Therefore, the present invention is a step of purely separating and cultivating a novel microbial strain from the field soil to achieve the above object;

본 발명 균주의 형태학적, 생리학적, 생화학적 및 배양학적 특성을 조사분류 동정하는 공정; 대사물질을 추출하는 공정 및; 식물성장촉진 시험공정으로 구성된다. 이하, 본 발명의 구체적인 구성 및 작용을 실험 및 실시예를 들어 설명한다.Investigating and identifying the morphological, physiological, biochemical and culture characteristics of the strains of the present invention; Extracting metabolites; It consists of a plant growth promotion test process. Hereinafter, the specific configuration and operation of the present invention will be described with reference to experiments and examples.

실험예 1Experimental Example 1

본 발명 균주의 분리 배양Isolation of the Strain of the Invention

본 발명 균주는 충남 금산지역의 밭토양에서 순수 분리하고 분리한 균주를 본 발명 합성배지에서 배양하였다. 합성 배지조성은 글리세린 15g/L, 펩톤 6g/L, 흑설탕 15g/L, NaCl 5g/L, FeSO40.0015g/L, CuSO40.02g/L, KH2PO40.3g/L, MnSO4(0.2% 용액) 0.1mL/L, MgSO40.05g/L 로 하였다. 한천배지에서 25℃로 배양하면 균사체가 mat를 형성하면서 성장하며 배지상에 암색의 확산성 물질을 생성하였다.The strain of the present invention was purely isolated from the field soil of the Geumsan region of Chungnam and cultured in the synthetic medium of the present invention. Synthetic medium composition was glycerin 15g / L, peptone 6g / L, brown sugar 15g / L, NaCl 5g / L, FeSO 4 0.0015g / L, CuSO 4 0.02g / L, KH 2 PO 4 0.3g / L, MnSO 4 ( 0.2% solution) 0.1 mL / L, MgSO 4 0.05g / L. When cultured at 25 ℃ in agar medium, mycelium grows to form a mat to produce a dark diffuse material on the medium.

실험예 2Experimental Example 2

본 발명 균주의 분류 동정Classification Identification of Strains of the Invention

본 발명 균주를 버기스 매뉴얼에 의해 조사분류한 결과 진균류에 속하는 Acremonium spp 속 균주로 분류되었다. 생육 최적 온도는 20℃이며 최저 온도는 4℃, 최고 온도는 35℃였다. 또 생육 최적 pH는 7.0이었다. 본발명 균주는 대기중의 유리 질소를 고정하지 못하였으며 탄소원으로 1개의 탄소원자를 가진 화합물은 사용하지 못하였다. 본 발명 균주가 이용하는 탄소원으로는 슈크로스, 글루코스 기타 1개 이상의 탄소원자를 가진 탄수화물이었다. 본 발명 균주는 유기 합성균으로 호기성이며 산화효소와 카탈라제 친화균이었다. 따라서 본 발명 균주는 호기적 발효 균임을 확인하였다.The strains of the present invention were classified into strains of the genus Acremonium spp belonging to the fungus as a result of the investigation and classification using the bugis manual. The optimum temperature for growth was 20 ° C, the lowest temperature was 4 ° C, and the highest temperature was 35 ° C. The optimum pH for growth was 7.0. The strain of the present invention did not fix free nitrogen in the atmosphere and did not use a compound having one carbon atom as a carbon source. The carbon source used by the strain of the present invention was sucrose, glucose, and a carbohydrate having one or more carbon atoms. The strain of the present invention was an organic synthetic bacterium, aerobic and oxidase and catalase affinity. Therefore, the strain of the present invention was confirmed to be aerobic fermentation bacteria.

본 발명의 균주는 Acremonium sp. MJN1으로 명명하고 1997년 9월 22일 생명공학연구소에 KCTC 0385BP로 기탁하였다.The strain of the present invention is Acremonium sp. It was named MJN1 and deposited on September 22, 1997 as KCTC 0385BP at the Biotechnology Research Institute.

실험예 3Experimental Example 3

식물 성장 촉진 물질의 추출 및 성장 촉진 활성 측정Measurement of Extraction and Growth Promoting Activity of Plant Growth Promoter

본 발명 균주를 상기 실험예1의 배지에서 15일 이상, 20℃, 200rpm의 교반 속도로 진탕배양한 후 배양액을 제거하고 균사체만을 70% 에탄올로 처리하여 48시간 200rpm으로 교반후 균사체를 제거한 에탄올층만을 회수하였다.After culturing the strain of the present invention for 15 days or more in a culture medium of Experimental Example 1 at a stirring speed of 20 ° C. and 200 rpm, the culture medium was removed and treated with only 70% ethanol for 48 hours and stirred at 200 rpm for 48 hours to remove the mycelium. Was recovered.

회수된 추출액을 박막 크로마토 그래프법을 이용하여 전개후 Rf 값 0.78이상의 상위 물질만을 분리하였다.The recovered extract was developed using thin layer chromatography to separate only the upper substance with an Rf value of 0.78 or more.

본 발명 균주에서 추출한 활성물질은 상추종자와 볍씨를 이용하여 발아시험을 실시하여 검증하여 본결과, 상추의 하배축은 대조구에 비해 40%이상 생장을 증진 시켰고, 볍씨에서는 발아율을 20% 뿌리의 건조중량을 35%이상 증진 시켰다.The active substance extracted from the strain of the present invention was verified by germination test using lettuce seed and rice seed. As a result, the lower hypocotyl of lettuce improved growth by more than 40% compared to the control, and the germination rate of rice seed was 20% dry weight of root. Increased by more than 35%.

본 발명의 진균균주와 그 추출물의 실제 사용예는 다음과 같다.Examples of practical use of the fungal strains and extracts of the present invention are as follows.

사용실시예1Example 1

참외와 고추에 대한 본 발명 Acremonium sp. MJN1 균주 추출물의 농도에 따른 활성도 측정실험Inventive Acremonium sp. Activity measurement experiment according to the concentration of MJN1 strain extract

본 발명 균주 추출물의 식용 작물에 미치는 생육의 영향을 알아보기 위해 식물의 생리학적 변화를 측정하였다. 상기 식물에 대하여 본 발명 균사로부터 추출된 물질을 처리하고 1개월 후에 광합성, 증산작용, Stomatal Conductance, CO2농도에 따른 광합성율의 변화를 LI-6200 portable photosynthesis system (Li-Cor, Inc, Lincoln, NE, USA)을 이용하여 측정하였다.Physiological changes of plants were measured to determine the effect of growth on the edible crop of the strain extract of the present invention. Changes in photosynthesis rate according to photosynthesis, transpiration, Stomatal Conductance, and CO 2 concentration were measured one month after treatment of the material extracted from the mycelium of the present invention with respect to the plant LI-6200 portable photosynthesis system (Li-Cor, Inc, Lincoln, NE , USA).

엽록소의 함량은 0.5g의 잎을 채취해 2mm 정도의 길이로 잘라 5mL의 N,N-Dimethylformamide를 넣어 24시간 4℃의 암소에 보관하고 엽록소를 추출한후 이 용액의 흡광도를 UV/Vis Spectrophotometer (Model 4050, LKB)로 측정하여 계산하였다.The content of chlorophyll is 0.5g of leaves, cut into 2mm length and 5mL of N, N-dimethylformamide is added and stored in the cow at 4 ℃ for 24 hours. After chlorophyll extraction, the absorbance of this solution is measured by UV / Vis Spectrophotometer (Model 4050, LKB).

생산량은 상기 식물의 수확기에 열매를 수확하여 총 열매수와 생중량을 측정하였다. 상기 사용실시예에 대한 실험결과는 표1과 표2에 나타내었다.Yields were harvested at the harvest time of the plant to measure the total fruit number and fresh weight. Experimental results for the use examples are shown in Table 1 and Table 2.

참외에 식물 성장촉진 물질을 처리한 결과 10-3과 10-5의 희석액을 처리한 개체구의 광합성률은 대조구에 비해 각각 7.4%, 7.1%가 증가하였다. 엽록소 함량은 희석정도에 따라 6∼10% 정도 증가하였다. 기공컨덕턴스에 있어서는 10-3, 10-4, 10-5희석액을 처리한 식물이 대조구에 비해 각각 20%, 38%, 50%가 증가하였다. 결실량은 대조구 식물이 11개인데 비해 10-3, 10-5희석액을 처리한 식물은 15개, 14개로 각각 36%와 27%가 증가하였다.In addition, the photosynthesis rate of 10 -3 and 10 -5 dilutions increased by 7.4% and 7.1%, respectively. Chlorophyll content increased by 6-10% depending on the degree of dilution. In pore conductance, plants treated with 10 -3 , 10 -4 , and 10 -5 dilutions increased 20%, 38%, and 50%, respectively, compared to the control. Deletion amount was increased to 36% and 27%, respectively, compared to 11 control plants, 15 and 14 plants treated with 10 -3 and 10 -5 dilutions.

고추에서 엽록소 함량은 10-5희석액에서 대조구에 비해 20%높게 나타났다. 기공컨덕턴스는 10-4희석액을 처리한 개체들에서 대조구에 비해 49%가 증가하였으며 10-5희석액을 처리한 개체구들은 약 8%정도가 증가하였다. 각 개체구들의 길이는 대조구 개체들의 평균 길이가 78cm 인데 비해 10-3, 10-5희석액을 처리한 개체들의 평균 길이는 각각 90cm와 87cm로 15.3%와 11.5%가 증가하였다. 고추의 결실량은 대조구가 평균 35g의 고추를 맺은데 비해 10-3희석액을 처리한 개체들은 48g의 결실을 맺어 각각 35%의 증산효과를 보였다.Chlorophyll content in red pepper was 20% higher than that in the control at 10 -5 dilution. Pore conductance increased 49% in the 10 -4 dilution group compared to the control, and increased by 8% in the 10 -5 dilution group. The average length of the control populations was 78cm, whereas the average length of the individuals treated with 10 -3 and 10 -5 dilutions increased by 15.3% and 11.5% to 90cm and 87cm, respectively. The average amount of red pepper was 35g in the control group, whereas those treated with 10 -3 dilutions resulted in 48g of deletion.

주 : Pn : 광합성률(단위: μmolCO2m-2s-1)Pn: Photosynthesis rate (μmolCO2m-2s-1)

Chl : 엽록소 함량(단위: mg/g)Chl: Chlorophyll content in mg / g

Cs : 기공 컨덕턴스(단위: ㎝/s)Cs: pore conductance (unit: cm / s)

Products : 총 결실량 (단위: 개수)Products: Total fruit loss (unit: number)

주 : Pn : 광합성률 (단위: μmolCO2m-2s-1)Pn: Photosynthesis rate (Unit: μmolCO2m-2s-1)

Chl : 엽록소 함량(단위: mg/g)Chl: Chlorophyll content in mg / g

Cs : 기공 컨덕턴스(단위: ㎝/s)Cs: pore conductance (unit: cm / s)

Growth : 길이생장(단위: cm)Growth: Length growth (unit: cm)

Products : 총 결실량(단위: 개수)Products: Total fruit loss (unit: number)

사용실시예 2Example 2

본 발명 Acremonium sp. MJN1 균주 추출물의 마늘과 양파에 대한 포장시험Acremonium sp. Field Test of Garlic and Onion of MJN1 Strain Extracts

본 발명 균주의 추출물을 10-3에서10-5까지 희석하여 엽면시비하였다. 대상식물인 마늘과 양파는 96년 10월에 파종하여 월동한 것을 사용하였으며 97년 4월에 상기 후보균주와 후보물질을 처리한후 마늘의 경우 5월에 광합성율, 증산율 등을 측정하여 효과를 확인 하였고 6월 중순에 처리구와 무처리구 전량을 수확하여 생중량을 측정하고 각각에 대하여 분산분석등을 통한 통계처리로 효과를 분석한 결과는 표3과 같다.The extract of the strain of the present invention was diluted from 10 -3 to 10 -5 to foliar fertilization. Garlic and onion, which are the target plants, were sown in October 1996 and winterized. After treating the candidate strain and candidates in April 97, garlic was measured by measuring photosynthesis rate and transpiration rate in May. In mid-June, all the treated and untreated areas were harvested to measure the fresh weight, and the results of the statistical analysis through variance analysis were shown in Table 3.

주 : Pn : 광합성률(단위: μmolCO2m-2s-1),Pn: photosynthesis rate (unit: μmolCO2m-2s-1),

Cs : 기공 컨덕턴스 (단위: cm/s),Cs: pore conductance (unit: cm / s),

Tr : 증산작용 (단위: mg H2O/m2/sec)Tr: transpiration (unit: mg H 2 O / m 2 / sec)

표 3에서 보는 바와 같이 본 발명 균주(MJN1) 추출물중 TLC Rf 값 0.95의 10-3희석액에서 대조구에 비해 50%정도 광합성률이 증가했다.As shown in Table 3, the photosynthesis rate of the strain of the present invention (MJN1) was increased by 50% compared to the control in the 10-3 dilution of TLC Rf value 0.95.

본 발명 균주 추출물의 TLC분리 형태인 두 물질을 양파와 마늘에 처리하여 각각의 생중량을 측정한 결과는 표4와 같다. 추출물중 Rf 값 0.82의 처리는 마늘에서 60%의 생중량을 대조구에 비해 증가시키는 것으로 나타났다.The results of measuring the fresh weight of each of the two substances in the TLC separation form of the strain of the present invention by treating onion and garlic are shown in Table 4. Treatment with an Rf value of 0.82 in the extract increased the fresh weight of 60% in garlic compared to the control.

본 발명 균주 추출물중 Rf 값 0.95의 경우 양파와 마늘에서 모두 40%이상의 성장촉진효과를 나타냈다. 따라서 본 발명 균주 추출물은 양파와 마늘에 모두 현저한 성장촉진 효과가 나타났다.In the extract of the present invention, the Rf value of 0.95 showed a growth promoting effect of more than 40% in both onion and garlic. Therefore, the strain extract of the present invention showed a significant growth promoting effect on both onion and garlic.

표 3.에서 측정된 마늘의 광합성량 측정과 비교해 보면, 본 발명 균주 추출물중 Rf 값 0.95의 10-3희석액에서 광합성률이 50% 증가하였고 생중량 측정결과도 10-3희석액에서 이와 일치하는 52%의 성장촉진효과를 나타냈다. 따라서, 마늘의 경우 본 발명균주 추출물에 포함된 물질에 의해 증가된 광합성 작용이 식물 성장을 유지하는 에너지로 사용되기보다는 거의 모두 생체의 중량을 늘리는 방향으로 작용하였던 것으로 추정된다.Compared to the photosynthesis amount of garlic measured in Table 3, the photosynthesis rate was increased by 50% in the 10-3 dilution of Rf value 0.95 in the strain extract of the present invention, and the result of the gravimetric weight was 52% of the equivalent in the 10-3 dilution. It showed a growth promoting effect. Therefore, in the case of garlic, it is assumed that the photosynthetic action increased by the material contained in the strain of the present invention acted in almost all directions of increasing the weight of the living body rather than being used as energy for maintaining plant growth.

이상 실험 및 사용실시예에서 입증된 바와같이, 본 발명 Acremonium sp. MJN1 균주의 추출물은 소량의 사용으로도 식물(참외, 고추, 마늘, 양파등)에 대하여 그 성장을 촉진하고 생산량을 증대시킬 수 있으므로 무공해 농업 및 화훼산업상 매우 유용한 발명인 것이다.As demonstrated in the above experiments and use examples, the present invention Acremonium sp. Extract of the MJN1 strain is a very useful invention for pollution-free agriculture and flower industry because it can promote the growth and increase the production for plants (melon, pepper, garlic, onion, etc.) even with a small amount.

Claims (3)

신규의 진균 균주 Acremonium sp. MJN1 균주(KCTC 0385BP)와 그 균주 추출물New fungal strain Acremonium sp. MJN1 strain (KCTC 0385BP) and its strain extract 신규의 진균 균주 Acremonium sp. MJN1 또는 그 균주 추출물의 무공해 식물성장촉진제로서의 용도.New fungal strain Acremonium sp. Use of MJN1 or its strain extracts as pollution-free plant growth promoter. 신규의 진균 균주 Acremonion sp. MJN1 또는 그 균주 추출물을 사용하는 식물성장촉진 방법.New fungal strain Acremonion sp. Plant growth promoting method using MJN1 or its strain extract.
KR1019970048803A 1997-09-25 1997-09-25 Novel acremonium sp mjn1 and extract thereof for its use of plant growth regulation KR100225254B1 (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20060058232A (en) * 2004-11-24 2006-05-30 서주원 The effects of adenosine or its analogues on various antibiotic production and the developmental method of new pesticide using them
CN115369042A (en) * 2021-05-18 2022-11-22 中国科学院微生物研究所 Acremonium 320 and application thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20060058232A (en) * 2004-11-24 2006-05-30 서주원 The effects of adenosine or its analogues on various antibiotic production and the developmental method of new pesticide using them
CN115369042A (en) * 2021-05-18 2022-11-22 中国科学院微生物研究所 Acremonium 320 and application thereof
CN115369042B (en) * 2021-05-18 2024-04-05 中国科学院微生物研究所 Acremonium 320 and application thereof

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