KR20130056585A - Plant growth promotion by using bacterial strains isolated from roots of miscanthus sacchariflorus - Google Patents
Plant growth promotion by using bacterial strains isolated from roots of miscanthus sacchariflorus Download PDFInfo
- Publication number
- KR20130056585A KR20130056585A KR1020110122257A KR20110122257A KR20130056585A KR 20130056585 A KR20130056585 A KR 20130056585A KR 1020110122257 A KR1020110122257 A KR 1020110122257A KR 20110122257 A KR20110122257 A KR 20110122257A KR 20130056585 A KR20130056585 A KR 20130056585A
- Authority
- KR
- South Korea
- Prior art keywords
- strain
- plant growth
- present
- promoting
- strains
- Prior art date
Links
Images
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
-
- 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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
- A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Dentistry (AREA)
- Virology (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Microbiology (AREA)
- Environmental Sciences (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dispersion Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Toxicology (AREA)
- General Engineering & Computer Science (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
본 발명은 물억새(Miscanthus sacchariflorus) 뿌리로부터 분리한 미생물을 이용한 식물 생장 촉진방법에 관한 것으로, 구체적으로 본 발명은 억새뿌리로부터 분리한 투메바실러스 속(Tumebacillus sp.) 균주, 바실러스 속(Bacillus sp.) 균주, 아그로박테리움 속(Agrobacterium sp.) 균주 및 라시니바실러스 속(Lysinibacillus sp.) 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 식물 생장 촉진용 미생물 제제에 관한 것이다.
The present invention is Miscanthus sacchariflorus ) relates to a method for promoting plant growth using microorganisms isolated from the roots, and specifically, the present invention relates to a genus of Tumebacillus isolated from a silver grass root. sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp.) It relates to a microbial agent for promoting plant growth containing one or more strains selected from the group consisting of strains, or cultures thereof as an active ingredient.
농업에서 비료는 작물의 수량을 결정짓는 중요한 농자재이다. 그러나 지나치게 많이 사용할 경우, 하천수의 부영양화, 지하수의 수질오염, 토양양분의 불균형, 및 작물의 생육불량 등의 원인으로 작용하여 작물의 수량감소는 물론 작물의 품질하락을 야기할 수 있다. 따라서 농업생산성을 높이기 위해 과다하게 사용되고 있는 비료나 농약 때문에 자연환경과 사람의 건강이 크게 위협받고 있는 현시점에서 생산성을 유지하여 농가의 소득을 증대시키고 소비자들이 건강하게 살 권리도 충족시킬 수 있는 유일한 대안은 환경친화적인 농법의 개발에 있다고 할 수 있다.In agriculture, fertilizers are an important source of agricultural crops. However, when used too much, it may cause eutrophication of river water, water pollution of groundwater, imbalance of soil nutrients, and poor growth of crops, which may lead to a decrease in crop yield and a decrease in crop quality. Therefore, the only alternative that can increase the income of farmers and satisfy the right of consumers to live healthy in the present time when the natural environment and human health are seriously threatened by the fertilizer or pesticide that is excessively used to increase agricultural productivity. Can be said to develop environmentally friendly farming methods.
그러나 산업의 발달 및 다양화로 인해 농경지 면적이 감소하고 있기 때문에 작물의 생산성을 높이기 위한 비료의 사용은 불가피한 것이며, 따라서 기존의 화학비료보다 미생물을 이용한 친환경적인 기능성 농자재의 개발을 위한 관심이 높아지고 있다.However, the use of fertilizer to increase the productivity of crops is inevitable due to the decrease in agricultural land area due to the development and diversification of the industry. Therefore, there is a growing interest in developing eco-friendly functional agricultural materials using microorganisms rather than conventional chemical fertilizers.
이러한 관심으로 인해 화학비료의 사용량은 1990년대 초를 정점으로 하여 지속적으로 감소하는 추세에 있으며, 1997년 이후에는 저농도의 화학비료 개발 및 공급 등과 같은 시비량 감축유도 및 퇴비, 유기질 비료의 사용 확대로 화학비료의 사용량이 더욱 감소하고 있다. 화학비료의 사용량은 1990년대 초반 1,104천 톤, 1995년에 954천 톤, 2000년에 801천 톤으로 감소하였고, 2005년에는 이보다 더욱 감소하였으며, 이에 반해 퇴비 및 유기질비료의 사용량은 점차 증가되고 있는 추세이다. 그러나 아직도 국내의 화학비료 사용량은 OECD 국가 중 높은 수준이다.
Due to this interest, the consumption of chemical fertilizers has been continuously decreasing since the early 1990s, and since 1997, chemical fertilizers have been reduced by inducing fertilization, composting, and using organic fertilizers. The use of fertilizers is falling further. The consumption of chemical fertilizers decreased to 1,104 thousand tons in the early 1990s, 954 thousand tons in 1995 and 801 thousand tons in 2000, and even more in 2005, while the consumption of compost and organic fertilizers is increasing. It is a trend. However, domestic chemical fertilizer use is still the highest among OECD countries.
에틸렌가스는 식물 내에서 호르몬의 일종으로서 식물의 성숙을 촉진하는데, 대표적인 예로 과실의 숙도, 씨앗의 발아, 유묘(어린 모종) 생장, 잎과 꽃잎의 이탈, 기관 노화, 스트레스, 병원성에 대한 반응 등 식물의 전 생활사에 있어서도 중요한 역할을 담당하고 있는 것으로 알려져 있다. 에틸렌은 식물 종과 세포 타입에 의존적으로 생장을 촉진하기도 하고 저해시키기도 하는 것으로 알려져 있는데, 저수위의 에틸렌은 뿌리 신장을 촉진하고 뿌리의 빠른 생장에 영향을 미치지만, 고수위의 에틸렌은 오히려 뿌리 신장을 억제하는 것으로 밝혀진 바 있다.Ethylene gas is a hormone in plants that promotes the maturation of plants. Typical examples include fruit ripening, seed germination, seedling growth, leaves and petals leaving, organ aging, stress, and response to pathogenicity. It is known to play an important role in the whole life history of plants. Ethylene is known to promote and inhibit growth depending on plant species and cell type. Low ethylene promotes root elongation and affects rapid growth of roots, whereas high ethylene inhibits root elongation. It has been found to do.
식물이 스트레스를 받으면 에틸렌 생합성에 의해 에틸렌의 전구체인 ACC의 생성량이 증가하고 이로 인해 식물체 내의 에틸렌 농도가 높아져 식물 생장을 저해한다. 스트레스를 받은 식물에 ACC는 ACC 탈아미노화 효소(ACC-deaminase)를 처리한 경우에, ACC가 가수분해되어 스트레스 에틸렌의 생성농도가 낮아져 식물 생장에 긍정적인 영향을 미치는 것으로 보고된바 있다(Penrose DM & Glick BR, Can J Microbiol 47:368-372, 2001). 또한, ACC 탈아미노화 효소를 생성하는 PGPB(plant growth promoting bacteria)를 처리한 식물체에 ACC를 처리하자 스트레스 에틸렌의 생성량이 무처리군에 비해 적게 나타났었다(Glick BR et al., J Theor Biol 190:63-68, 1998).When plants are stressed, ethylene biosynthesis increases the amount of ACC that is a precursor of ethylene, thereby increasing the concentration of ethylene in the plant and inhibiting plant growth. In the stressed plants, ACC has been reported to have a positive effect on the plant growth due to the hydrolysis of ACC, resulting in a decrease in the production of stress ethylene when ACC deaminase is treated (Penrose). DM & Glick BR, Can J Microbiol 47: 368-372, 2001). In addition, when ACC was treated to plants treated with plant growth promoting bacteria (PGPB) that produced ACC deamination enzymes, the production of stress ethylene was lower than that of the untreated group (Glick BR et al., J Theor Biol). 190: 63-68, 1998).
아울러, 근권 미생물로부터 생성되는 고농도의 IAA(indole acetic acid)에 의해 무화과 나무에서 ACC가 생성되고, 이렇게 생성된 식물 내부의 ACC는 외부로 배출되어 외부와의 균형을 맞춰 유지된다. 이때, 외부에서 연속적으로 PGPB에 의해 ACC가 가수분해되면 식물 내 ACC 생성량이 줄어들어서, 스트레스 에틸렌의 생합성량은 줄어들게 되는 것으로 보고된바 있다(Arshd M et al., Trends in Biotechnol. 25:356-362, 2007).
In addition, ACC is generated from the fig tree by a high concentration of indole acetic acid (IAA) generated from the root zone microorganisms, and the ACC inside the plant is discharged to the outside to maintain balance with the outside. At this time, when ACC is hydrolyzed continuously by PGPB from outside, it has been reported that the amount of ACC produced in plants decreases, thereby reducing the biosynthesis of stress ethylene (Arshd M et al., Trends in Biotechnol. 25: 356-). 362, 2007).
식물 호르몬으로서 인돌아세트산 또는 옥신이라고 불리는 IAA, 인돌부틸산(IBA) 및 앱시스산(ABA) 등이 있는데, 옥신인 IAA는 식물이 씨에서 발아하여 생장하는 데에는 필요하며, 특히 줄기의 신장에 관여하는 식물 생장 호르몬의 일종이며, 상기 앱시스산은 종자의 발생, 휴면(dormancy) 또는 가뭄, 고농도의 염분 및 저온 등의 환경 스트레스에 대한 반응 등 다양한 생리적 반응과 관련된 식물 호르몬으로서, 광합성 산물을 발달 중에 있는 종자 쪽으로 수송하고 저장 단백질의 합성을 촉진하는 작용을 하며, 생장환경이 적절하지 않을 경우, 종자나 싹의 발아와 성장을 저해하여 식물체의 생존을 유지하는 역할을 하며, 특히 식물의 물공급이 충분하지 않으면 잎 안에 함량이 높아져 기공 개폐와 같은 기능을 조절하여 수분 스트레스의 영향을 낮게 하여 식물 생장에 도움을 주는 역할을 하는 것으로 알려져 있다( Seo, M. and Koshiba, T., Trends Plant Sci., 7: 41-48, 2002).
Plant hormones include IAA, indole acetic acid or auxin, indolebutyric acid (IBA) and abscisic acid (ABA), which are necessary for plants to germinate and grow in seeds, especially those involved in stem elongation. It is a kind of plant growth hormone, and the abscisic acid is a plant hormone associated with various physiological reactions such as seed development, dormancy or drought, response to environmental stress such as high salinity and low temperature, and is developing photosynthetic products. It acts to transport seeds and promote the synthesis of storage proteins, and when the environment is not appropriate, it inhibits germination and growth of seeds or shoots, and maintains the survival of the plant. If you do not do so, the content of the leaves will increase, and the functions such as pore opening and closing will be controlled to lower the effects of moisture stress. Role to help as it is known that (Seo, M. and Koshiba, T., Trends Plant Sci, 7:. 41-48, 2002).
억새는 C4 광합성 식물이고 영년생 작물이기 때문에 연간 마른 줄기 생산량이 스위치그래스 등 다른 비식량 에너지작물에 비해 많고 재식 이후 경운 및 파종작업을 하지 않고 15~20년간 수확할 수 있을 뿐만 아니라 줄기가 성숙하여 고사하기 시작하는 가을에 줄기의 양분이 땅속줄기로 이행한 후 수확하면 토양 비옥도가 대부분 유지되어 비료 요구도가 낮다. 자연상태에서 병해충 발생도 거의 없어 농약살포를 필요로 하지 않는 친환경 작물이기도 하며 최근에는 바이오 연료용 원료작물로 각광받고 있다.Pampas grass is a C 4 photosynthetic plant and is a perennial crop, so the annual dry stem production is higher than other non-food energy crops such as switchgrass, and it can be harvested for 15-20 years without tillage and sowing after planting. When the nutrients of the stalks are transferred to the substem in the autumn when they begin to die, the soil fertility is mostly maintained and the fertilizer demand is low. It is an eco-friendly crop that does not require pesticide spraying because it rarely causes pests in its natural state, and has recently been spotlighted as a raw material for biofuels.
억새 속에는 참억새(Miscanthus sinensis), 물억새(Miscanthus sacchariflorus) 등 17개 종(species)이 있는 것으로 알려져 있다. 주요 억새는 세 종류인데 참억새는 2 배체로서 염색체 수가 38개이며, 물억새는 주로 4 배체로 염색체 수는 76개이다. 바이오 매스 수량이 많아 주목받고 있는 3 배체인 Miscanthus X giganteus는 염색체 수가 57개이다.
It is known that there are 17 species, including Miscanthus sinensis and Miscanthus sacchariflorus. There are three main types of pampas antelope, which are doublets, 38 chromosomes, and pampas grasses are tetraploid and 76 chromosomes. Miscanthus X giganteus, a triplex that is attracting attention due to its high quantity of biomass, has 57 chromosomes.
한편, 대한민국 특허등록 제10-530885호에는 슈도모나스 플로레슨스 B16 균주 및 이를 이용한 작물의 생장촉진 방법 및 세균성 시들음병 방제 방법이 개시되어 있으며, 대한민국 특허등록 제10-755509호에는 질소고정력 및 작물 생장 촉진효과가 있는 신규한 균주 아조스피릴룸 브라실렌스 CW301, 이를 이용한 생물 비료 및 그 제조 방법이 개시되어 있으며, 대한민국 특허등록 제10-686596호에는 식물의 생장을 촉진하는 신균주 KLP1 및 이를 이용한 식물 생장 촉진 방법이 개시되어 있으나, 본 발명의 억새뿌리로부터 분리된 종자 발아촉진 및 식물 생장을 촉진하는 균주는 알려진바 없다.
Meanwhile, Korean Patent Registration No. 10-530885 discloses Pseudomonas florences B16 strain and a method for promoting growth and control of bacterial wilting disease, and Korean Patent Registration No. 10-755509 promotes nitrogen fixation and crop growth. A novel strain Azospirylbracilens CW301, a biological fertilizer using the same, and a method of preparing the same are disclosed, and Korean Patent Registration No. 10-686596 discloses a new strain KLP1 that promotes plant growth and a plant growth using the same. Although a method for promoting is disclosed, strains which promote seed germination and plant growth isolated from the silver grass root of the present invention are not known.
이에, 본 발명자들은 천연물에서 유래된 친환경적인 미생물 제제를 개발하기 위하여 노력한 결과, 억새 뿌리로부터 분리한 투메바실러스 속(Tumebacillus sp.) 균주, 바실러스 속(Bacillus sp.) 균주, 아그로박테리움 속(Agrobacterium sp.) 균주 및 라시니바실러스 속(Lysinibacillus sp.) 균주가 옥신(Auxin) 및 ACC 탈아미노화 효소(ACC deaminase)를 생산하고, 다양한 작물에 대하여 종아 발아 촉진효과 및 생장 촉진효과를 나타내므로, 식물 생장 촉진용 미생물 제제로 유용하게 사용될 수 있음을 확인함으로써 본 발명을 완성하였다.
Therefore, the present inventors endeavored to develop an eco-friendly microbial agent derived from natural products, the genus Tumebacillus isolated from Pampas roots sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp. Strains produce auxin and ACC deaminase and promote germination and growth of various crops, thus promoting plant growth. The present invention has been completed by confirming that it can be usefully used as a microbial preparation for.
본 발명의 목적은 투메바실러스 속(Tumebacillus sp.) 균주, 바실러스 속(Bacillus sp.) 균주, 아그로박테리움 속(Agrobacterium sp.) 균주 및 라시니바실러스 속(Lysinibacillus sp.) 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 식물 생장 촉진용 미생물 제제를 제공하는 것이다.The object of the present invention is the genus Tumebacillus sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp.) It is to provide a microbial agent for promoting plant growth containing at least one strain selected from the group consisting of strains, or a culture thereof as an active ingredient.
또한, 본 발명은 목적은 본 발명의 미생물 제제를 토양, 식물 또는 식물의 종자에 처리하는 단계를 포함하는 식물 생장 촉진 방법을 제공하는 것이다.It is also an object of the present invention to provide a method for promoting plant growth, which comprises treating the soil, plant or seed of a plant with the microbial agent of the invention.
또한, 본 발명은 목적은 투메바실러스 속 균주, 바실러스 속 균주, 아그로박테리움 속 균주 및 라시니바실러스 속 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 종자 발아 촉진용 미생물 제제를 제공하는 것이다.In addition, the present invention is an object of the genus Tumebacillus strains, Bacillus sp. Strains, Agrobacterium sp. And one or more strains selected from the group consisting of the strains of the genus Racinibacillus, or a microorganism for promoting seed germination containing the culture medium thereof as an active ingredient.
아울러, 본 발명은 목적은 투메바실러스 속 균주, 바실러스 속 균주, 아그로박테리움 속 균주 및 라시니바실러스 속 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 식물 생장 및 발아 촉진용 생물비료를 제공하는 것이다. In addition, the present invention is an object of the genus Tumebacillus strains, Bacillus strains, Agrobacterium strains And one or more strains selected from the group consisting of the genus Lacinibacillus, or to provide a biological fertilizer for promoting plant growth and germination containing the culture medium thereof as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 투메바실러스 속(Tumebacillus sp.) 균주, 바실러스 속(Bacillus sp.) 균주, 아그로박테리움 속(Agrobacterium sp.) 균주 및 라시니바실러스 속(Lysinibacillus sp.) 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 식물 생장 촉진용 미생물 제제를 제공한다.In order to achieve the above object, the present invention Tumebacillus genus ( Tumebacillus sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp.) Provides a microbial agent for promoting plant growth containing at least one strain selected from the group consisting of strains, or a culture thereof as an active ingredient.
또한, 본 발명은 본 발명의 미생물 제제를 토양, 식물 또는 식물의 종자에 처리하는 단계를 포함하는 식물 생장 촉진 방법을 제공한다.The present invention also provides a method for promoting plant growth, which comprises treating the soil, plant or seed of a plant with the microbial agent of the present invention.
또한, 본 발명은 투메바실러스 속 균주, 바실러스 속 균주, 아그로박테리움 속 균주 및 라시니바실러스 속 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 종자 발아 촉진용 미생물 제제를 제공한다.In addition, the present invention is Tumebacillus strain, Bacillus strain, Agrobacterium strain And one or more strains selected from the group consisting of the strains of the genus Racinebacillus, or microorganisms for promoting seed germination containing the culture medium thereof as an active ingredient.
아울러, 본 발명은 투메바실러스 속 균주, 바실러스 속 균주, 아그로박테리움 속 균주 및 라시니바실러스 속 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 식물 생장 및 발아 촉진용 생물비료.
In addition, the present invention is Tumebacillus strain, Bacillus strain, Agrobacterium strain And one or more strains selected from the group consisting of the genus Racinebacillus, or a fertilizer for promoting plant growth and germination, containing the culture medium thereof as an active ingredient.
본 발명의 억새 뿌리로부터 분리한 투메바실러스 속(Tumebacillus sp.) BE501, 바실러스 속(Bacillus sp.) BE506, 아그로박테리움 속(Agrobacterium sp.) BE516 및 라시니바실러스 속(Lysinibacillus sp.) BE533 균주는 식물 생장을 촉진하는 옥신(Auxin) 및 ACC 탈아미노화 효소(ACC deaminase)를 생산하고, 억새, 밀, 보리, 벼, 수단그라스, 배추, 오이 및 토마토에 대하여 현저한 종자 발아 촉진 효과 및 생장촉진 효과를 나타내므로 종자 발아 촉진용 미생물 제제 및 식물 생장 촉진용 미생물 제제의 유효성분으로 유용하게 사용될 수 있다.
Tumebacillus genus isolated from the pampas root of the present invention sp.) BE501, Bacillus sp.) BE506, Agrobacterium sp.) BE516 And Lysinibaci llus sp.) BE533 strains produce auxin and ACC deaminase, which promote plant growth, and marked seed germination for silver grass, wheat, barley, rice, sudangrass, cabbage, cucumber and tomato Since it exhibits a promoting effect and a growth promoting effect, it can be usefully used as an active ingredient of the microbial preparation for promoting seed germination and the microbial preparation for promoting plant growth.
도 1은 본 발명 균주의 옥신(Auxin) 호르몬 생산능을 확인한 도이다.
도 2는 본 발명의 균주의 ACC 탈아미노화 효소(ACC deaminase)의 생성능을 확인한 도이다.
도 3은 본 발명의 균주의 셀룰라아제(Cellulase), 펙티나아제(Pectinase) 및 프로테아제(Protease) 생산능을 확인한 도이다.
도 4는 본 발명의 균주의 지하뿌리(rhizome) 이식 후 물억새 뿌리로 부터 새순 생장효과를 나타낸 도이다:
무처리: 대조군인 멸균수 처리군;
BE501: 본 발명의 투메바실러스 속(Tumebacillus sp.)BE501 처리군;
BE506: 본 발명의 바실러스 속(Bacillus sp.)BE506 처리군;
BE516: 본 발명의 아그로박테리움 속(Agrobacterium sp.)BE516 처리군; 및
BE533: 본 발명의 라시니바실러스 속(Lysinibacillus sp.) BE533 처리군.
도 5는 본 발명의 균주의 물억새 종자를 이용한 식물 생장 촉진효과를 나타낸 도이다:
무처리 : 대조군인 멸균수 처리군;
BE501: 본 발명의 투메바실러스 속 BE501 처리군;
BE506: 본 발명의 바실러스 속 BE506 처리군;
BE516: 본 발명의 아그로박테리움 속 BE516 처리군; 및
BE533: 본 발명의 라시니바실러스 속 BE533 처리군.
도 6은 본 발명의 균주의 수단그라스 종자발아 및 생장 촉진효과를 나타낸 도이다:
무처리: 대조군인 멸균수 처리군;
BE501: 본 발명의 투메바실러스 속 BE501 처리군;
BE506: 본 발명의 바실러스 속 BE506 처리군; 및
BE516: 본 발명의 아그로박테리움 속 BE516 처리군.
도 7은 본 발명의 균주의 보리 종자발아 및 생장 촉진효과를 나타낸 도이다:
무처리: 대조군인 멸균수 처리군;
BE501: 본 발명의 투메바실러스 속 BE501 처리군; 및
BE516: 본 발명의 아그로박테리움 속 BE516 처리군.
도 8은 본 발명의 균주의 벼 종자발아 및 생장 촉진효과를 나타낸 도이다:
무처리: 대조군인 멸균수 처리군; 및
BE501: 본 발명의 투메바실러스 속 BE501 처리군.
도 9는 본 발명의 균주의 배추 종자발아 및 생장 촉진효과를 나타낸 도이다:
무처리: 대조군인 멸균수 처리군;
BE501: 본 발명의 투메바실러스 속 BE501 처리군; 및
BE533: 본 발명의 라시니바실러스 속 BE533 처리군.
도 10은 본 발명의 균주의 토마토 종자발아 및 생장 촉진효과를 나타낸 도이다:
무처리: 대조군인 멸균수 처리군; 및
BE516: 본 발명의 아그로박테리움 속 BE516 처리군.1 is a diagram confirming the auxin (Auxin) hormone production capacity of the present invention.
Figure 2 is a diagram confirming the production capacity of ACC deaminase (ACC deaminase) of the strain of the present invention.
Figure 3 is a diagram confirming the cellulase (Cellulase), Pectinase (Pectinase) and protease (Protease) production capacity of the strain of the present invention.
Figure 4 is a diagram showing the growth effect of shoots from Pampas roots after transplantation of rhizome of the strain of the present invention:
No treatment: sterile water treatment group as control;
BE501: Tumebacillus sp. BE501 treatment group of the present invention;
BE506: Bacillus sp. BE506 treatment group of the present invention;
BE516: Agrobacterium sp. BE516 treatment group of the present invention; And
BE533: shinny la of the present invention, Bacillus (Lysinibaci llus sp.) BE533 treatment group.
Figure 5 is a diagram showing the effect of promoting plant growth using the pampasum seeds of the strain of the present invention:
No treatment: sterile water treatment group as control;
BE501: genus BE501 treatment group of the present invention;
BE506: the genus BE506 treatment group of the present invention;
BE516: Agrobacterium genus BE516 treatment group of the present invention; And
BE533: genus BE533 treatment group of the present invention.
Figure 6 is a diagram showing the effect of promoting seed germination and growth of the strain of the present invention:
No treatment: sterile water treatment group as control;
BE501: genus BE501 treatment group of the present invention;
BE506: the genus BE506 treatment group of the present invention; And
BE516: Agrobacterium genus BE516 treatment group of the present invention.
7 is a diagram showing the effect of promoting seed germination and growth of the strain of the present invention:
No treatment: sterile water treatment group as control;
BE501: genus BE501 treatment group of the present invention; And
BE516: Agrobacterium genus BE516 treatment group of the present invention.
Figure 8 is a diagram showing the effect of promoting rice seed germination and growth of the strain of the present invention:
No treatment: sterile water treatment group as control; And
BE501: A group of BE501 treatments of the genus Tomebacillus of the present invention.
9 is a diagram showing the effect of promoting the germination and growth of Chinese cabbage seed strain of the present invention:
No treatment: sterile water treatment group as control;
BE501: genus BE501 treatment group of the present invention; And
BE533: genus BE533 treatment group of the present invention.
10 is a diagram showing the tomato seed germination and growth promoting effect of the strain of the present invention:
No treatment: sterile water treatment group as control; And
BE516: Agrobacterium genus BE516 treatment group of the present invention.
이하, 본 발명을 상세히 설명한다.
Hereinafter, the present invention will be described in detail.
본 발명은 투메바실러스 속(Tumebacillus sp.) 균주, 바실러스 속(Bacillus sp.) 균주, 아그로박테리움 속(Agrobacterium sp.) 균주 및 라시니바실러스 속(Lysinibacillus sp.) 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 식물 생장 촉진용 미생물 제제를 제공한다.The present invention is the genus Tumebacillus sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp.) Provides a microbial agent for promoting plant growth containing at least one strain selected from the group consisting of strains, or a culture thereof as an active ingredient.
상기 균주는 억새로부터 분리한 것이 바람지하며, 물억새 뿌리로부터 분리한 것이 보다 바람직하나 이에 한정되지 않는다.The strain is preferably isolated from silver grass, and more preferably isolated from silver grass roots, but is not limited thereto.
상기 투메바실러스 속 균주는 수탁번호 KCTC12051BP로 기탁된 것이 바람직하나 이에 한정되지 않는다.The strain of the genus Tomebacillus is preferably deposited with accession number KCTC12051BP, but is not limited thereto.
상기 바실러스 속 균주는 KCTC12052BP로 기탁된 것이 바람직하나 이에 한정되지 않는다.The Bacillus strain is preferably deposited with KCTC12052BP, but is not limited thereto.
상기 아그로박테리움 속 균주는 KCTC12053BP로 기탁된 것이 바람직하나 이에 한정되지 않는다.The strain of the genus Agrobacterium is preferably deposited as KCTC12053BP, but is not limited thereto.
상기 라시니바실러스 속 균주는 KCTC12054BP로 기탁된 것이 바람직하나 이에 한정되지 않는다.The strain of the genus Racinebacillus is preferably deposited with KCTC12054BP, but is not limited thereto.
상기 식물은 억새, 밀, 보리, 수단그라스, 벼, 배추, 오이 또는 토마토인 것 바람직하나 이에 한정되지 않는다.The plant is preferably but not limited to silver grass, wheat, barley, sudangrass, rice, cabbage, cucumber or tomato.
상기 미생물 제제는 상기 균주 또는 이의 배양액을 유효성분으로 포함할 수 있다. 본 발명에 의한 미생물 제제는 통상적인 방법으로 식물 생장 촉진용으로 제형화할 수 있으며 건조분말 형태 또는 액상비료 형태로 제조할 수 있다. 구체적으로, 본 발명에 의한 미생물 제제는 액상 비료 형태로 제조될 수 있으며 이에 증량제를 첨가하여 가루분말의 형태로 이용하거나 이를 제형화하여 과립화시킬 수도 있다. 그러나 그 제형에 특별히 한정되지는 않는다. 바람직하게는 화학비료를 대체하기 위한 식물 생장 촉진 생물비료로 제형화 할 수 있고, 즉 화학비료 공급이 제한된 친환경 유기농업에서 이를 극복하기 위한 생물비료로 제형화가 가능하다.The microbial agent may include the strain or its culture as an active ingredient. Microbial preparations according to the invention can be formulated for promoting plant growth in a conventional manner and can be prepared in the form of dry powder or liquid fertilizer. Specifically, the microbial preparation according to the present invention may be prepared in the form of a liquid fertilizer and may be used in the form of powdered powder by adding an extender thereto or granulated by formulating it. However, the formulation is not particularly limited. Preferably it can be formulated as a plant growth promoting biofertilizer to replace the chemical fertilizer, that is, it can be formulated as a biofertilizer to overcome this in eco-friendly organic farming where the chemical fertilizer supply is limited.
상기 식물 생장 촉진제는 당업자에게 알려진 방법대로 제조되는 것이 바람직하며, 그 적용방법은 통상 일반적으로 행하고 있는 방법, 즉 살포(예를 들면 분무, 미스팅, 아토마이징, 분말 살포, 과립 살포, 수면시용, 상시용 등), 토양시용(예를 들면 혼입, 관주 등), 표면사용(예를 들면 도포, 도말법, 피복 등), 침지, 독이, 훈연 시용등에 의해 행할 수 있다. 그 사용량은, 그 체형, 피해상황, 적용방법, 적용장소 등에 따라 적절히 결정할 수 있다.The plant growth promoter is preferably prepared according to a method known to those skilled in the art, and the application method thereof is generally performed in general, that is, spraying (for example, spraying, misting, atomizing, powder spraying, granulating spraying, sleeping, Etc.), soil application (e.g. mixing, irrigation, etc.), surface use (e.g., coating, smearing, coating, etc.), dipping, poisoning, and smoking. The amount of use can be appropriately determined according to the body type, the damage situation, the application method, the application place, and the like.
본 발명의 구체적인 실시예에서, 본 발명자들은 물억새(Miscanthus sacchariflorus) 뿌리로부터 내생하고 있는 미생물을 분리하기 위하여 물억새 군락지에서 물억새를 샘플링한 후, 물억새 뿌리로부터 내생하고 있는 4종의 균주를 분리하였다.In a specific embodiment of the present invention, the present inventors separated the four endogenous strains from the pampas root after sampling the pampas in the Pampas colony in order to separate the endogenous microorganisms from the Miscanthus sacchariflorus root.
본 발명의 또 다른 구체적인 실시예에서, 본 발명자들은 본 발명의 4종의 균주의 16s rRNA 시퀀싱(sequencing)을 수행한 결과, 본 발명의 BE501는 Tumebacillus permanentifrigoris strain Eur1 9.5와 99% 유사성을 나타내고, 본 발명의 BE506은 Bacillus thuringiensis strain 104XG46과 99% 유사성을 나타내며, 본 발명의 BE516은 Agrobacterium vitis strain K309와 99% 유사성을 나타내고, 본 발명의 BE533은 Lysinibacillus fusiformis isolate CCM1B와 99% 유사성을 나타내는 것을 확인하였다(표 1 참조). 또한, 본 발명의 BE501, BE506, BE516 및 BE533 균주를 각각 투메바실러스 속(Tumebacillus sp.) BE501, 바실러스 속(Bacillus sp.) BE506, 아그로박테리움 속(Agrobacterium sp.) BE516 및 라시니바실러스 속(Lysinibacillus sp.) BE533이라 명명하고, 한국생명공학연구원 유전자은행에 2011년 11월 08일자로 기탁하였다: 투메바실러스 속 BE501 수탁번호: KCTC12051BP; 바실러스 속 BE506 수탁번호: KCTC12052BP; 아그로박테리움 속 BE516 수탁번호: KCTC12053BP; 및 라시니바실러스 속 BE533 수탁번호: KCTC12054BP. In another specific embodiment of the present invention, the inventors performed 16s rRNA sequencing of four strains of the present invention, and as a result, BE501 of the present invention exhibited 99% similarity to Tumebacillus permanentifrigoris strain Eur1 9.5, BE506 of the invention is Bacillus thuringiensis 99% similarity with strain 104XG46, BE516 of the present invention is Agrobacterium 99% similarity to vitis strain K309, BE533 of the present invention is Lysinibacillus fusiformis It was confirmed that the isolate shows 99% similarity with CCM1B (see Table 1). In addition, BE501, BE506, BE516 and BE533 strains of the present invention, respectively ( Tumebacillus genus) sp.) BE501, Bacillus sp.) BE506, Agrobacterium sp.) BE516 And Lysinibaci llus sp.) was named BE533 and deposited with the Korea Institute of Biotechnology and Gene Bank on November 08, 2011: BE501 Accession No .: KCTC12051BP; Genus BE506 accession number: KCTC12052BP; BE516 accession number from Agrobacterium: KCTC12053BP; And BE533 Accession No .: KCTC12054BP.
본 발명의 또 다른 구체적인 실시예에서, 본 발명자들은 식물 생장 촉진효과를 확인하기 위하여 옥신(Auxin) 생성능을 확인한 결과, 본 발명의 균주는 식물 생장을 촉진하는 옥신을 유의적으로 생산하는 것을 확인하였다(도 1 및 표 4 참조).In another specific embodiment of the present invention, the inventors confirmed the ability to produce auxin (Auxin) in order to confirm the effect of promoting plant growth, it was confirmed that the strain of the present invention significantly produces auxin to promote plant growth. (See FIG. 1 and Table 4).
본 발명의 또 다른 구체적인 실시예에서, 본 발명자들은 식물 생장 촉진효과를 확인하기 위하여 ACC 탈아미노화 효소(ACC deaminase) 활성능을 확인한 결과, 본 발명의 균주는 식물 생장을 촉진하는 ACC 탈아미노화 효소를 유의적으로 생산하는 것을 확인하였다(도 2 및 표 4 참조).In another specific embodiment of the present invention, the present inventors confirmed the ACC deaminase activity in order to confirm the effect of promoting plant growth, the strain of the present invention ACC deamination to promote plant growth It was confirmed that the enzyme produced significantly (see FIG. 2 and Table 4).
본 발명의 또 다른 구체적인 실시예에서, 본 발명자들은 본 발명의 4종의 균주에 대한 식물 생장 촉진효과를 확인하기 위하여 가수분해 효소(Hydrolytic enzymes) 활성을 확인한 결과, 본 발명의 균주는 유의적인 가수분해 활성이 있는 것을 확인하였고, 특히 본 발명의 BE501 균주는 셀룰라아제, 펙티나아제 및 프로테아제에 현저한 활성을 나타내는 것을 확인하였다(도 3 및 표 4 참조).In another specific embodiment of the present invention, the present inventors confirmed the hydrolytic enzymes (Hydrolytic enzymes) activity to confirm the plant growth promoting effect on the four strains of the present invention, the strain of the present invention is a significant It was confirmed that the degradation activity, in particular the BE501 strain of the present invention was found to show significant activity on cellulase, pectinase and protease (see Fig. 3 and Table 4).
본 발명의 또 다른 구체적인 실시예에서, 본 발명자들은 본 발명의 균주의 억새 생장 촉진효과를 확인하기 위하여, 억새 지하경 뿌리에 본 발명의 균주를 접종한 결과, 본 발명의 균주를 접종한 실험군은 대조군인 무처리군과 비교하여 유의적인 생장촉진 효과를 나타내는 것을 확인하였다(도 4 참조).In another specific embodiment of the present invention, the present inventors inoculated the strain of the present invention to the locust root diameter roots to check the growth growth effect of the strain of the present invention, the experimental group inoculated strain of the present invention is a control group It was confirmed that it showed a significant growth promoting effect compared to the phosphorus treatment group (see Figure 4).
본 발명의 또 다른 구체적인 실시예에서, 본 발명자들은 본 발명의 균주의 억새 생장 촉진효과를 확인하기 위하여, 종자 발아된 유묘에 본 발명의 균주를 접종한 결과, 무처리군보다 본 발명의 균주를 처리하였을 때 줄기가 최대 2배까지 생장 촉진하는 것을 확인하였다(도 5 참조).In another specific embodiment of the present invention, the present inventors inoculated the strain of the present invention in seed germinated seedlings to confirm the effect of promoting growth of pampasum of the strain of the present invention, the strain of the present invention than the untreated group When treated, it was confirmed that the stem promotes growth up to two times (see FIG. 5).
본 발명의 또 다른 구체적인 실시예에서, 본 발명자들은 본 발명의 균주의 종자 발아촉진 및 생장 촉진효과를 확인하기 위하여, 밀, 보리, 벼, 수단그라스, 배추, 오이 및 토마토 종자를 대상으로 실험을 수행한 결과, 본 발명의 균주는 유의적인 종자 발아 촉진효과 및 식물 생장 촉진효과를 가지는 것을 확인하였다(도 6 내지 도 10, 및 표 5 참조).In another specific embodiment of the present invention, the inventors conducted experiments with wheat, barley, rice, Sudangrass, Chinese cabbage, cucumber and tomato seeds in order to confirm the seed germination and growth promoting effect of the strain of the present invention As a result, it was confirmed that the strain of the present invention has a significant seed germination promoting effect and plant growth promoting effect (see FIGS. 6 to 10, and Table 5).
따라서, 본 발명의 억새 뿌리로부터 분리한 투메바실러스 속(Tumebacillus sp.) BE501, 바실러스 속(Bacillus sp.) BE506, 아그로박테리움 속(Agrobacterium sp.) BE516 및 라시니바실러스 속(Lysinibacillus sp.) BE533 균주는 식물 생장을 촉진하는 옥신(Auxin) 및 ACC 탈아미노화 효소(ACC deaminase)를 생산하고, 억새, 밀, 보리, 벼, 수단그라스, 배추, 오이 및 토마토에 대하여 현저한 종자 발아 촉진 효과 및 생장촉진 효과를 나타내므로 식물 생장을 촉진하는 미생물 제제의 유효성분으로 유용하게 이용될 수 있다.
Therefore, the genus Tumebacillus isolated from the Pampas Root of the present invention sp.) BE501, Bacillus sp.) BE506, Agrobacterium sp.) BE516 And Lysinibaci llus sp.) BE533 strains produce auxin and ACC deaminase, which promote plant growth, and marked seed germination for silver grass, wheat, barley, rice, sudangrass, cabbage, cucumber and tomato Since it exhibits a promoting effect and a growth promoting effect, it can be usefully used as an active ingredient of a microbial agent for promoting plant growth.
또한, 본 발명은 본 발명의 미생물 제제를 토양, 식물 또는 식물의 종자에 처리하는 단계를 포함하는 식물 생장 촉진 방법을 제공한다.The present invention also provides a method for promoting plant growth, which comprises treating the soil, plant or seed of a plant with the microbial agent of the present invention.
상기 미생물 제제는 액체 상태로 식물에 관주, 작물의 종자에 침지 또는 분무하거나 종자에 코팅하여 이용하는 것이 바람직하나 이에 한정되지 않는다.The microbial agent is preferably used in irrigation in the liquid state, immersed or sprayed into the seed of the crop or coated on the seed, but is not limited thereto.
상기 식물은 억새, 밀, 보리, 벼, 수단그라스, 배추, 오이 또는 토마토인 것이 바람직하나 이에 한정되지 않는다. The plant is preferably silver grass, wheat, barley, rice, sudangrass, cabbage, cucumber or tomato, but is not limited thereto.
상기 침지하는 방법의 경우, 상기 미생물 제제를 식물체 주변의 토양에 붓거나 또는 종자를 제제에 담가둘 수 있고, 분무할 경우에는 당업계에 널리 공지된 기술로 식물체에 흐르도록 살포할 수 있으나 이에 한정하지 않는다.In the case of the immersion method, the microbial agent may be poured into the soil around the plant or the seeds may be immersed in the preparation, and when sprayed, the microbial agent may be sprayed to flow through the plant by techniques well known in the art, but is not limited thereto. I never do that.
본 발명의 억새 뿌리로부터 분리한 투메바실러스 속 BE501, 바실러스 속 BE506, 아그로박테리움 속 BE516 및 라시니바실러스 속 BE533 균주는 식물 생장을 촉진하는 옥신 및 ACC 탈아미노화 효소를 생산하고, 억새, 밀, 보리, 벼, 수단그라스, 배추, 오이 및 토마토에 대하여 현저한 종자 발아 촉진 효과 및 생장촉진 효과를 나타내므로 본 발명의 미생물 제제를 이용한 식물 생장을 촉진방법으로 유용하게 이용될 수 있다.
Tumebacillus BE501, Bacillus BE506, Agrobacterium BE516 Isolated from Pampas Root of the Present Invention And BE533 strains of the genus Racinibacillus produce auxin and ACC deamination enzymes that promote plant growth, and have marked seed germination and growth promoting effects on pampas grass, wheat, barley, rice, sudangrass, cabbage, cucumber and tomato. Since it shows an effect, it can be usefully used as a method for promoting plant growth using the microbial agent of the present invention.
또한, 본 발명은 투메바실러스 속(Tumebacillus sp.) 균주, 바실러스 속(Bacillus sp.) 균주, 아그로박테리움 속(Agrobacterium sp.) 균주 및 라시니바실러스 속(Lysinibacillus sp.) 균주로 구성된 군으로부터 선택되는 1개 이상의 균주, 또는 이의 배양액을 유효성분으로 함유하는 종자 발아 촉진용 미생물 제제를 제공한다.In addition, the present invention is the genus Tumebacillus sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp.) Provides a microbial agent for promoting seed germination containing at least one strain selected from the group consisting of strains, or a culture thereof as an active ingredient.
상기 균주는 물억새 뿌리로부터 분리한 것이 바람직하나 이에 한정되지 않는다.The strain is preferably separated from the pampas root, but is not limited thereto.
상기 투메바실러스 속 균주는 수탁번호 KCTC12051BP로 기탁된 것이 바람직하나 이에 한정되지 않는다.The strain of the genus Tomebacillus is preferably deposited with accession number KCTC12051BP, but is not limited thereto.
상기 바실러스 속 균주는 KCTC12052BP로 기탁된 것이 바람직하나 이에 한정되지 않는다.The Bacillus strain is preferably deposited with KCTC12052BP, but is not limited thereto.
상기 아그로박테리움 속 균주는 KCTC12053BP로 기탁된 것이 바람직하나 이에 한정되지 않는다.The strain of the genus Agrobacterium is preferably deposited as KCTC12053BP, but is not limited thereto.
상기 라시니바실러스 속 균주는 KCTC12054BP로 기탁된 것이 바람직하나 이에 한정되지 않는다.The strain of the genus Racinebacillus is preferably deposited with KCTC12054BP, but is not limited thereto.
상기 종자는 억새, 밀, 보리, 수단그라스, 벼, 배추, 오이 또는 토마토의 종자인 것이 바람직하나 이에 한정되지 않는다.The seed is preferably one of seeds of wheat grass, wheat, barley, sudangrass, rice, cabbage, cucumber or tomato, but is not limited thereto.
본 발명의 억새 뿌리로부터 분리한 투메바실러스 속(Tumebacillus sp.) BE501, 바실러스 속(Bacillus sp.) BE506, 아그로박테리움 속(Agrobacterium sp.) BE516 및 라시니바실러스 속(Lysinibacillus sp.) BE533 균주는 식물 생장을 촉진하는 옥신(Auxin) 및 ACC 탈아미노화 효소(ACC deaminase)를 생산하고, 억새, 밀, 보리, 벼, 수단그라스, 배추, 오이 및 토마토에 대하여 현저한 종자 발아 촉진 효과 및 생장촉진 효과를 나타내므로 종자 발아를 촉진하는 미생물 제제의 유효성분으로 유용하게 이용될 수 있다.
Tumebacillus genus isolated from the pampas root of the present invention sp.) BE501, Bacillus sp.) BE506, Agrobacterium sp.) BE516 And Lysinibaci llus sp.) BE533 strains produce auxin and ACC deaminase, which promote plant growth, and marked seed germination for silver grass, wheat, barley, rice, sudangrass, cabbage, cucumber and tomato Since it has a promoting effect and a growth promoting effect, it can be usefully used as an active ingredient of a microbial agent for promoting seed germination.
이하, 본 발명을 실시예 및 실험예에 의해 상세히 설명한다.Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.
단, 하기 실시예 및 실험예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예 및 실험예에 의해 한정되는 것은 아니다.
However, the following examples and experimental examples are illustrative of the present invention, and the content of the present invention is not limited by the following examples and experimental examples.
<< 실시예Example 1> 물억새 뿌리로부터 식물 1> Plants from pike roots 내생균Endogenous bacteria 분리 detach
물억새(Miscanthus sacchariflorus) 뿌리로부터 내생하고 있는 미생물을 분리하기 위하여 충청북도 청원군 문의면 대청호수주변에서 서식하고 있는 물억새를 수집한 후, 뿌리 표면에 있는 흙을 수돗물(tap water)을 이용하여 제거한 다음 상온에서 하루 동안 건조시켰다. 상기 건조된 물억새 뿌리를 1 cm 크기로 자른 후, 4% NaClO에 5분, 2.5% Na2S2O3에 10분간 침지시킨 다음 멸균수로 3 내지 4회 세척하였다. 그런 다음, 75% 에탄올(Ethanol)에 3분간 침지시킨 후, 멸균한 증류수로 3 내지 4회 세척한 다음 10% NaHCO3에 10분간 침지시킨 후 클린벤치에서 건조시켰다. 그런 다음, 표면 살균한 물억새 뿌리를 호모게나이저를 이용하여 5000 rpm으로 3분간 갈아서 분리용 시료로 TYA 및 R2A 배지에 도말한 후 28 배양기에서 7일간 배양한 후, 형성된 순수한 단일 콜로니(single colony)를 수득하였으며, 이를 BE501, BE506, BE516 및 BE533으로 명명하였다.
To isolate endogenous microorganisms from the roots of Miscanthus sacchariflorus, we collected water grasses in the vicinity of Daecheong Lake, Munmun-myeon, Cheongwon-gun, Chungcheongbuk-do, and then remove the soil on the root surface with tap water and Dried over. The dried Pampas Roots were cut to 1 cm size, immersed in 4% NaClO for 5 minutes, and 2.5% Na 2 S 2 O 3 for 10 minutes, and then washed 3 to 4 times with sterile water. Then, it was immersed in 75% ethanol (Ethanol) for 3 minutes, washed 3 to 4 times with sterile distilled water and then immersed in 10% NaHCO 3 for 10 minutes and dried in a clean bench. Then, using the homogenizer, the surface sterilized Pampas Roots were ground for 3 minutes at 5000 rpm and smeared on TYA and R2A medium as a separation sample, and then cultured in 28 incubators for 7 days, and then formed as a single colony. Were obtained, which were named BE501, BE506, BE516 and BE533.
<< 실시예Example 2> 16s 2> 16s rRNArRNA 시퀀싱( Sequencing sequencingsequencing )을 통한 균주의 동정Identification of strains through
상기 <실시예 1>에서 분리한 4종의 균주의 동정은 대상 균주에 대한 DB가 구축되어 안정된 분류동정 Key로 인정받고 있는 16s rRNA 부분 유전자 염기서열을 분석하여 실시하였다. 구체적으로 게놈(genomic) DNA 추출, PCR 및 염기서열 분석은 하기 실시예 <2-1> 및 <2-2>의 방법으로 수행하였으며, 분리된 균의 동정결과는 분리된 염기서열을 NCBI에서 블라스트(Blast) 검색을 통해 기 보고되어 있는 표준 균주와의 유사도를 고려하여 가장 유사도가 높은 표준 균주 명으로 동정결과를 반영하였다.
Identification of the four strains isolated in <Example 1> was carried out by analyzing the 16s rRNA partial gene nucleotide sequence recognized as a stable classification identification key DB was built for the target strain. Specifically, genomic DNA extraction, PCR, and sequencing were performed according to the methods of Examples <2-1> and <2-2>, and the result of identification of the isolated bacteria resulted in blasting the separated sequences from the NCBI. In consideration of the similarity with the previously reported standard strain through the (Blast) search, the identification result was reflected as the name of the standard strain with the highest similarity.
<2-1> 게놈(<2-1> Genome ( genomicgenomic ) ) DNADNA 추출 extraction
상기 <실시예 1>에서 수득한 균주의 게놈 DNA는 평판 배지에 자란 미량의 콜로니를 5%(w/v) Chelex-100(시그마) 100 ㎕가 든 E-tube에 넣고 100℃에서 5 내지 10분간 끓인 다음 상온에서 식혀 원심분리 한 후, 그 상등액을 콜로니 PCR에 이용하였다.
The genomic DNA of the strain obtained in Example 1 was placed in an E-tube containing 100 μl of 5% (w / v) Chelex-100 (Sigma) grown in a plate medium in 5-10 at 100 ° C. After boiling for a minute, cooled to room temperature, and centrifuged, the supernatant was used for colony PCR.
<2-2> 16s <2-2> 16s rRNArRNA 염기서열 분석 Sequencing
16s rRNA는 하기 두 개의 알려진 다범위 적용 프라이머(universal primer)를 사용하여 증폭하였다;16s rRNA was amplified using the following two known universal primers;
정방향 프라이머: 27F(서열번호: 1)(E. coli 16S rRNA positions 8-27;5'-AGAGTTTGATCCTGGCTCAG-3'); 및Forward primer: 27F (SEQ ID NO: 1) ( E. coli 16S rRNA positions 8-27; 5'-AGAGTTTGATCCTGGCTCAG-3 '); And
역방향 프라이머: 1492R(서열번호: 2)(E. coli 16S rRNA position 1492-1510; 5‘-GGTTACTTGTTACGACTT-3').Reverse primer: 1492R (SEQ ID NO: 2) ( E. coli 16S rRNA position 1492-1510; 5'-GGTTACTTGTTACGACTT-3 ').
PCR 반응물의 조성은 10 × 반응버퍼(reaction buffer), 2.5 mM Mgcl2, 50 mM dNTPs, 0.5 ㎕의 5U Taq polymerase(Bioneer co Ltd., 한국), 20 pmol 프라이머이며 약 20∼80 ng의 정제된 DNA를 주형으로 첨가하였으며 PCR 을 위한 각 반응의 온도 조건은 다음과 같다; DNA의 변성(denaturation)을 위해 94℃에서 40 초, 프라이머의 어닐링(annealing)을 위해 55℃에서 40초, DNA 스트랜드(DNA strand)의 합성을 위해 72℃에서 1분 과정을 28회 반복하는 조건을 사용하였다. 반응이 끝난 PCR purification kit(NucleoGen사)를 사용하여 염기서열을 결정하였다. 염기서열 결정 반응 산물은 최종적으로 자동염기서열장치(ABI 3730XL; Applied Biosystems)를 사용하여 염기서열을 결정한 후, 결정된 염기서열은 미국 NCBI의 BLAST (http://www.ncbi.nim.nih.gov/BLAST/) 검색방법을 사용하여 기존에 등록된 다양한 균주들의 상응하는 염기서열과 비교하여 유사도를 결정하였다.The composition of the PCR reaction was 10 × reaction buffer, 2.5 mM Mgcl 2 , 50 mM dNTPs, 0.5 μl of 5U Taq polymerase (Bioneer co Ltd., Korea), 20 pmol primer and about 20-80 ng of purified DNA was added as a template and the temperature conditions of each reaction for PCR were as follows; 40 seconds at 94 ° C for denaturation of DNA, 40 seconds at 55 ° C for annealing primers, and 1 minute at 72 ° C for DNA strand synthesis. Was used. The base sequence was determined using the PCR purification kit (NucleoGen). The sequencing reaction product was finally determined by sequencing using an autobase sequencer (ABI 3730XL; Applied Biosystems), and the sequencing sequence was determined by BLAST ( http://www.ncbi.nim.nih.gov) of the US NCBI. / BLAST / ) search method was used to determine the similarity compared to the corresponding sequencing of various previously registered strains.
그 결과, [표 1]에 나타낸 바와 같이 BE501(서열번호:3)는 Tumebacillus permanentifrigoris strain Eur1 9.5와 99% 유사성을 나타내고, BE506(서열번호: 4)은 Bacillus thuringiensis strain 104XG46과 99% 유사성을 나타내며, BE516(서열번호: 5)은 Agrobacterium vitis strain K309와 99% 유사성을 나타내고, BE533(서열번호: 6)은 Lysinibacillus fusiformis isolate CCM1B와 99% 유사성을 나타내는 것을 확인하였다(표 1). 또한, 상기 BE501, BE506, BE516 및 BE533 균주를 각각 투메바실러스 속(Tumebacillus sp.) BE501, 바실러스 속(Bacillus sp.) BE506, 아그로박테리움 속(Agrobacterium sp.) BE516 및 라시니바실러스 속(Lysinibacillus sp.) BE533이라 명명하고, 한국생명공학연구원 유전자은행에 2011년 11월 08일자로 기탁하였다:As a result, BE501 (SEQ ID NO: 3) showed 99% similarity to Tumebacillus permanentifrigoris strain Eur1 9.5 as shown in [Table 1], and BE506 (SEQ ID NO: 4) was Bacillus thuringiensis 99% similarity to strain 104XG46, BE516 (SEQ ID NO: 5) is Agrobacterium 99% similarity to vitis strain K309, BE533 (SEQ ID NO: 6) is Lysinibacillus fusiformis It was confirmed that the isolate exhibits 99% similarity with CCM1B (Table 1). In addition, the strains BE501, BE506, BE516 and BE533, respectively, Tumebacillus genus ( Tumebacillus sp.) BE501, Bacillus sp.) BE506, Agrobacterium sp.) BE516 And Lysinibaci llus sp.) Named BE533 and deposited with the Korea Biotechnology Research Institute Gene Bank on November 08, 2011:
투메바실러스 속 BE501 수탁번호: KCTC12051BP;Genus BE501 accession number: KCTC12051BP;
바실러스 속 BE506 수탁번호: KCTC12052BP;Genus BE506 accession number: KCTC12052BP;
아그로박테리움 속 BE516 수탁번호: KCTC12053BP; 및BE516 accession number from Agrobacterium: KCTC12053BP; And
라시니바실러스 속 BE533 수탁번호: KCTC12054BP.
Genus BE533 accession number: KCTC12054BP.
<< 실험예Experimental Example 1> 식물 생장 촉진효과 확인 1> Confirmation of plant growth promoting effect
<1-1> <1-1> 옥신Auxin (( AuxinAuxin ) ) 생성능Generation 확인 Confirm
상기 <실시예 1>에서 분리한 4종의 균주에 대한 식물 생장 촉진효과를 확인하기 위하여 식물 생장을 촉진하는 식물 생장 호르몬인 옥신 생성능을 확인하였다.In order to confirm the plant growth promoting effect of the four strains isolated in <Example 1> was confirmed the ability to produce auxin, a plant growth hormone that promotes plant growth.
구체적으로 옥신 호르몬 생산은 하기 [표 2]로 이루어진 King‘s medium B(KB) 배지에 옥신 전구체인 L-트립토판(L-tryptophan)을 0.1% 첨가하고 본 발명의 4종의 분리균을 접종하여 28℃에 24시간 배양하였다. 배양액을 4℃에서 6000 rpm으로 15분간 원심분리하고 분리된 상등액과 살코프스키(salkowski) 용액(35% HClO4, 50 mL, 0.5M FeCl3 1 mL)을 1:2로 30분간 암실에서 반응시킨 후 옥신 생산능을 확인하였다.Specifically, the production of auxin hormone was added 0.1% of auxin precursor L-tryptophan (L-tryptophan) to King's medium B (KB) medium consisting of the following [Table 2] and inoculated with four isolates of the present invention. Incubated for 24 hours at 28 ℃. Centrifuge the culture solution at 6000 rpm for 15 minutes at 4 ° C, and react the separated supernatant and salkowski solution (35% HClO4, 50 mL, 0.5 mL FeCl3 1 mL) in the dark at 1: 2 for 30 minutes. Auxin production was confirmed.
그 결과, 도 1 및 [표 4]에 나타낸 바와 같이 본 발명의 균주는 대조군으로 멸균수를 처리한 군과 비교하여 식물 생장을 촉진하는 옥신을 유의적으로 생산하는 것을 확인하였다(도 1 및 표 4).
As a result, as shown in Figure 1 and Table 4, the strain of the present invention was confirmed to produce auxin significantly promoting plant growth compared to the group treated with sterile water as a control (Fig. 1 and Table 4).
<1-2> <1-2> ACCACC 탈아미노화Deaminoation 효소( enzyme( ACCACC deaminasedeaminase ) ) 활성능Bow performance 확인 Confirm
상기 <실시예 1>에서 분리한 4종의 균주에 대한 식물 생장 촉진효과를 확인하기 위하여 식물 생장을 촉진하는 ACC(1-aminocyclopropane-1-carboxylic acid) 탈아미노화 효소 생산능을 확인하였다.In order to confirm the plant growth promoting effect of the four strains isolated in <Example 1> was confirmed the ability to produce ACC (1-aminocyclopropane-1-carboxylic acid) deaminoase enzyme promoting plant growth.
구체적으로, 질소원으로서 3 mM ACC만 첨가된 하기 [표 3]으로 이루어진 DF salt minimal 배지에 본 발명의 균주를 접종하여 28℃에서 2일간 배양한 후, ACC를 분해하는 ACC 탈아미노화 효소를 생산 여부를 확인하였다.Specifically, inoculated with the strain of the present invention in a DF salt minimal medium consisting of the following [Table 3] to which only 3 mM ACC was added as a nitrogen source and incubated at 28 ° C. for 2 days to produce an ACC deamination enzyme that degrades ACC. It was confirmed.
그 결과, 도 2 및 표 4에 나타낸 바와 같이 본 발명의 4종의 균주는 식물 생장을 촉진하는 ACC 탈아미노화 효소를 유의적으로 생산하는 것을 확인하였다(도 2 및 표 4).
As a result, as shown in Figure 2 and Table 4, the four strains of the present invention was confirmed to produce a significant ACC deamination enzyme to promote plant growth (Fig. 2 and Table 4).
<1-3> 가수분해 효소(<1-3> hydrolase ( HydrolyticHydrolytic enzymesenzymes ) ) 활성능Bow performance 확인 Confirm
상기 <실시예 1>에서 분리한 4종의 균주에 대한 식물 생장 촉진효과를 확인하기 위하여 가수분해 효소(Hydrolytic enzymes) 활성을 확인하였다.In order to confirm the plant growth promoting effect on the four strains isolated in <Example 1> was confirmed the hydrolytic enzymes (Hydrolytic enzymes) activity.
구체적으로, LB 액체 배지에 본 발명의 균주를 접종하고 28℃ 배양기에서 24시간 배양하였다. 배양액을 6000rpm에 15분간 원심분리하고 시험시료로 이용하였다. Specifically, LB liquid medium was inoculated with the strain of the present invention and incubated for 24 hours in a 28 ℃ incubator. The culture was centrifuged at 6000 rpm for 15 minutes and used as a test sample.
셀룰라아제(Cellulase) 생산능을 보기 위해 BMM[(0.2%(NH4)2SO4, 0.11% Na2HPO4, 0.07% KH2PO4, 0.0001% MgSO4, 0.0001% MnSO4] 배지에 0.4% 카르복시메틸셀룰로오스(Carboxymethyl Cellulose)가 포함된 고체 배지에 시료를 분주하여 투명환 형성으로 셀룰라아제(cellulase) 활성을 확인하였다. 0.4% Carboxymethyl Cellulose was added to BMM [(0.2% (NH 4 ) 2 SO 4 , 0.11% Na2HPO4, 0.07% KH2PO4, 0.0001% MgSO4, 0.0001% MnSO4] Dispensing the sample in the solid medium included to check the cellulase activity by the formation of a transparent ring.
펙티나아제(Pectinase)는 펙테이트 리에이즈 배지(pectate lyase medium)[1% 폴리갈락투론산(polygalacturonic acid), 1% 효모 추출물(yeast extract), 0.38μM CaCl2, 100mM Tris HCl(pH 8.5), 0.8% 아가로오스(agarose), 0.8% 소듐아자이드(sodium azide)]에 미생물 배양 상등액을 분주하여 투명환 형성으로 펙티나아제(pectinase) 활성을 확인하였다. Pectinase is a Pectate lyase medium [1% polygalacturonic acid, 1% yeast extract, 0.38 μM CaCl2, 100 mM Tris HCl (pH 8.5), 0.8% agarose (agarose), 0.8% sodium azide (sodium azide)] was dispensed microbial culture supernatant to confirm the pectinase (pectinase) activity by forming a transparent ring.
프로테아제(Protease)는 프로테아제 배지(protease medium)[3% 젤라틴(gelatin) 또는 탈지유(skim milk), 0.4% 영양배지(nutrient broth), 0.8% 아가로오스(agarose), 0.2% 소듐아자이드(sodium azide)]에 배양 상등액을 분주하여 투명환 형성으로 프로테아제(Protease) 활성을 확인하였다.Protease is protease medium [3% gelatin or skim milk, 0.4% nutrient broth, 0.8% agarose, 0.2% sodium azide azide)] was dispensed into the culture supernatant and the protease activity was confirmed by the formation of a transparent ring.
그 결과, 도 3 및 표 4에 나타낸 바와 같이, 본 발명의 균주는 유의적인 가수분해 활성을 나타내는 것을 확인하였고, 특히 본 발명의 BE501 균주는 셀룰라아제, 펙티나아제 및 프로테아제에 현저한 활성을 나타내는 것을 확인하였다(도 3 및 표 4).
As a result, as shown in Figure 3 and Table 4, it was confirmed that the strain of the present invention shows a significant hydrolytic activity, in particular, the BE501 strain of the present invention was shown to have a significant activity on cellulase, pectinase and protease (FIG. 3 and Table 4).
균주
Strain
옥신
(Auxin)
Auxin
(Auxin)
ACC 탈아미노화 효소
(ACC deaminase)
ACC deaminolase
(ACC deaminase)
(Hydrolytic enzymes)Hydrolase
(Hydrolytic enzymes)
셀룰라아제
(Cellulase)
Cellulase
(Cellulase)
펙티나아제
(Pectinase)
Pectinase
(Pectinase)
프로테아제(Protease)
Protease
BE501 균주The
BE501 strain
BE506 균주The
BE506 strain
BE516 균주The
BE516 strain
BE533 균주The
BE533 strain
상기 (+): 양성, (-): 음성.
(+): Positive, (-): negative.
<< 실험예Experimental Example 2> 억새 생장 촉진효과 확인 2> Confirmation of Pamper Growth Promotion Effect
<2-1> 물억새 <2-1> Pike 지하경(rhizome)을Rhizome 이용한 식물 생장 촉진효과 확인 Confirmation of plant growth promoting effect
상기 <실시예 1>에서 수득한 본 발명의 균주의 물억새 생장촉진 효과를 확인하기 위하여, 물억새 뿌리로부터 잘 형성된 지하경(rhizome)을 엄선하고 지하경을 3마디 간격으로 전지용 가위로 잘라서 억새 발아 유목으로 이용하였다. 상기 엄선된 지하경은 직경 10 cm 실험용 포트에 멸균된 상토를 채워서 이용하였다. 접종 시료는 본 발명의 균주 배양균체의 배지 성분을 멸균수로 3∼4회 세척한 균주 현탁액을 108 CFU/ml 농도로 각 지하경 주위에 10 ml씩 분주하였고 대조군으로 멸균수를 동일 양으로 각각 3개체씩 3회 반으로 처리한 후, 접종 30일 후 선발 균주에 대한 억새의 생육촉진 효과를 확인하였다.In order to confirm the effect of promoting the growth of pampas grasses of the present invention obtained in Example 1, a well-formed rhizome (rhizome) was carefully selected from the pike roots and cut into three pangular intervals with pruning shears and used as a pampas germination driftwood. It was. The carefully selected underground diameter was used by filling a sterilized top soil in a 10 cm diameter experimental port. The inoculated sample was dispensed with 10 ml of each strain around the basement at a concentration of 10 8 CFU / ml of the strain suspension washed three to four times with the sterile water of the culture medium of the strain culture cells of the present invention, and the same amount of sterilized water as the control. After three and three treatments of three individuals, 30 days after the inoculation, the effect of promoting the growth of silver grass on the selected strains was confirmed.
그 결과, 도 4에 나타낸 바와 같이, 지하경 뿌리에 접종한 본 발명의 4가지 균주는 대조군인 무처리군 100%보다 물억새 줄기 길이가 118% 내지 120%정도 식물 생장 촉진활성을 나타내었고, 뿌리생성도 무처리군 보다 생장촉진 효과를 나타내는 것을 확인하였다(도 4).
As a result, as shown in Figure 4, the four strains of the present invention inoculated into the root diameter of the sperm showed a 118% to 120% plant growth promoting activity than the
<2-2> 물억새 종자를 이용한 식물 생장 촉진효과 확인<2-2> Confirmation of plant growth promoting effect
상기 <실시예 1>에서 수득한 본 발명의 균주의 물억새 생장촉진 효과를 확인하기 위하여, 물억새 종자를 2% NaClO용액에 10분간 침지하여 표면살균하고 멸균수로 4차례 세척한 종자를 실험에 이용하였다. 멸균된 여지를 페트리디시에 깔고 멸균수를 5 ml씩 분주한 후 준비한 물억새 종자를 여지에 올려 30℃ 배양기에서 7일간 발아시켰다. 종자 발아된 유묘를 포트에 이식하고 본 발명의 균주의 현탁액(108 CFU/ml)을 물억새 뿌리 주위에 10 ml씩 분주하였다. 대조군으로 멸균수를 10ml씩 동일하게 처리하였다. 각 처리군는 1개체씩 8회 반복 실험하여 처리 30일 후 억새의 생장을 확인하였다.In order to confirm the effect of promoting the growth of the pampas grass of the present invention obtained in the above <Example 1>, the pampas grass seeds were immersed in 2% NaClO solution for 10 minutes, surface sterilized and washed four times with sterile water for the experiment. It was. The sterilized lychee was placed in Petri Dish, and 5 ml of sterile water was dispensed, and then, the prepared pampas seeds were placed on the lychee and germinated in a 30 ° C. incubator for 7 days. Seed germinated seedlings were transplanted into pots and a suspension (10 8 CFU / ml) of the strains of the present invention was dispensed 10 ml around the Pampas Root. As a control, sterilized water was treated in the same manner. Each treatment group was tested eight times, one by one to confirm the growth of silver grass 30 days after treatment.
그 결과, 도 5에 나타낸 바와 같이, 물억새 종자 발아를 이용한 식물 생장촉진효과는 무처리군보다 본 발명의 균주를 처리하였을 때 줄기가 최대 2배까지 생장 촉진하는 것을 확인하였다(도 5).
As a result, as shown in Figure 5, it was confirmed that the plant growth promoting effect using the pampasilla seed germination promotes the growth of the stem up to two times when treated with the strain of the present invention than the untreated group (Fig. 5).
<< 실험예Experimental Example 3> 다양한 작물에 대한 종자 발아 및 생장 촉진효과 확인 3> Identification of seed germination and growth promoting effects on various crops
상기 <실시예 1>에서 수득한 4종의 균주의 종자 발아 및 생장 촉진효과를 확인하기 위하여, 밀, 보리, 벼, 수단그라스, 배추, 오이 및 토마토 종자를 대상으로 실험을 수행하였다. In order to confirm the seed germination and growth promoting effect of the four strains obtained in the <Example 1>, experiments were conducted on wheat, barley, rice, Sudangrass, Chinese cabbage, cucumber and tomato seeds.
구체적으로, 밀, 보리, 벼, 수단그라스, 배추, 오이 및 토마토 종자를 2% NaClO용액에 10분간 침지하고 멸균수로 4회 세척하여 종자표면을 살균한 다음, 종자 발아를 위하여 페트리디시내에 멸균된 여지 위에 종자를 20 립씩 깔고 28℃ 그로스쳄버에서 발아를 유도하였다. 본 발명의 균주처리는 여지에 본 발명의 균주 현탁액을 각각 108 CFU/ml의 농도로 3 ml씩 분주하였다. 대조군으로 멸균수를 각각 3 ml씩 처리하였으며 각 처리군는 20개체씩 3회 반복 실험하였다. 각 작물의 발아 및 식물 생장촉진효과는 처리 후 7일간 관찰하였다.Specifically, wheat, barley, rice, sudangrass, cabbage, cucumber and tomato seeds are immersed in 2% NaClO solution for 10 minutes and washed four times with sterile water to sterilize the seed surface and then sterilized in Petri dish for seed germination. Twenty grains of seeds were placed on the ground and induced germination in a 28 ° C gross chamber. The strain treatment of the present invention was dispensed 3 ml each of the strain suspension of the present invention at a concentration of 10 8 CFU / ml each. 3 ml of sterilized water was treated as a control group, and each treatment group was repeated three times with 20 objects. Germination and plant growth promoting effects of each crop were observed for 7 days after treatment.
그 결과, 도 6 및 표 5에 나타낸 바와 같이 본 발명의 BE501, BE506 및 BE516 균주는 유의적인 수단그라스 종자발아 및 식물 생장 촉진효과를 나타내는 것을 확인하였다(도 6 및 표 5)As a result, as shown in Figure 6 and Table 5, the strain BE501, BE506 and BE516 of the present invention was found to exhibit a significant means for promoting seed germination and plant growth (Fig. 6 and Table 5).
또한, 도 7 및 표 5에 나타낸 바와 같이 본 발명의 BE501 및 BE516 균주는 유의적인 보리 종자발아 및 식물 생장 촉진효과를 나타내는 것을 확인하였다(도 7 및 표 5).In addition, as shown in Figure 7 and Table 5, the strain BE501 and BE516 of the present invention was found to show significant barley seed germination and plant growth promoting effect (Fig. 7 and Table 5).
또한, 도 8 및 표 5에 나타낸 바와 같이 본 발명의 BE501 균주는 유의적인 벼 종자발아 및 식물 생장 촉진효과를 나타내는 것을 확인하였다(도 8 및 표 5).In addition, as shown in Figure 8 and Table 5, the strain BE501 of the present invention was confirmed to show significant rice seed germination and plant growth promoting effect (Fig. 8 and Table 5).
또한, 도 9 및 표 5에 나타낸 바와 같이 본 발명의 BE501 및 BE533은 배추 발아 및 식물 생장 촉진효과를 나타내며, 특히 본 발명의 균주를 처리한 군은 무처리군과 비교하여 줄기 직경이 현저히 두껍게 발아하는 것을 확인하였다(도 9 및 표 5).In addition, as shown in Figure 9 and Table 5 BE501 and BE533 of the present invention exhibits the effect of cabbage germination and plant growth, in particular, the group treated with the strain of the present invention germinates significantly thicker stem diameter compared to the untreated group It was confirmed that (Fig. 9 and Table 5).
아울러, 도 10에 나타낸 바와 같이 본 발명의 BE516 균주는 유의적인 토마토 종자발아 및 식물 생장 촉진효과를 나타내는 것을 확인하였다(도 10 및 표 5).
In addition, as shown in Figure 10 BE516 strain of the present invention was confirmed to show a significant tomato seed germination and plant growth promoting effect (Fig. 10 and Table 5).
상기 (+): 양성, (-): 음성.
(+): Positive, (-): negative.
<110> Korea Research Institute of Bioscience & Biotechnology
<120> Plant growth promotion by using bacterial strains isolated from
roots of Miscanthus sacchariflorus
<130> 11p-09-33
<160> 6
<170> KopatentIn 1.71
<210> 1
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> forward primer for 16s rRNA
<400> 1
agagtttgat cctggctcag 20
<210> 2
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> reverse primer for 16s rRNA
<400> 2
ggttacttgt tacgactt 18
<210> 3
<211> 1425
<212> DNA
<213> Tumebacillus sp. BE501
<400> 3
tttgntaacc tttttacgac ttcaccccag ttatcgatca ccttaggcgg ctggctctgc 60
aagcagttac ctcaccgact tcgggtgtta ccaactccca tggtgtgacg ggcggtgtgt 120
acaaggcccg ggaacgaatt caccgcggca tgctgatccg cgattactag caattccggc 180
ttcatgcagg cgagttgcag cctgcaatcc gaactacgaa cggctttctg ggattggctt 240
cacctcgcgg cttcgcaacc ctttgtaccg tccattgtag cacgtgtgta gcccaggaca 300
taaggggcat gatgatttga cgtcatcccc gccttcctcc ggtttgtcac cggcagtctg 360
ttgtaagtgc tcaactaaat ggtagcaaca caacataggg gttgcgctcg ttgcgggact 420
taacccaaca tctcacgaca cgagctgacg acaaccatgc accacctgtc accgctgccc 480
cgaagggaaa ctctatctct agagcggtca gcgggatgtc aagtcctggt aaggttcttc 540
gcgttgcttc gaattaaacc acatgctcca ctgcttgtgc gggcccccgt caattccttt 600
gagtttcagt cttgcgaccg tactccccag gcggagtgct taatgcgtta gcttcggcac 660
taaggggtgg gccccctaac acctagcact catcgtttac ggcgtggact accagggtat 720
ctaatcctgt ttgctcccca cgctttcgcg cctcagcgtc agaaatcggc cagcaaggcg 780
ccttcgccac aggtgttcct ccacatctct acgcatttca ccgctacacg tggaattccc 840
cttgcctctc cgatcctcaa gtctccccgt atccaaggca atcccagagt tgagctctgg 900
gctttcaccc cggacgtgag agaccgcctg cgcgcgcttt acgcccagtg attccggaca 960
acgcttgccc cctacgtatt accgcggctg ctggcacgta gttagccggg gcttcctcct 1020
ctgttacngt caggtcctga gctttctctg cacaggatgg ttcttcacag aaracagagt 1080
tttacaaccc gaaggccttc atcactcacg cggcgttgct ccgtcaggct tgcgcccatt 1140
gcggaagatt ccctactgct gcctcccgta ggagtctggg ccgtgtctca gtcccagtgt 1200
ggccggtcac cctctcaggt cggctacgca tcgtcgcctt ggtgagccgt tacctcacca 1260
actagctaat gcgccgcggg tccatctggc agtgaagcaa aagctccttt tactctcgaa 1320
ccatgcgaty caagagatta tccggtatta gcactcgttt ccaagcgtta tcccagtctg 1380
ccaggcaggt tacccacgtg ttactcaccc gtccgccgct gacat 1425
<210> 4
<211> 1459
<212> DNA
<213> Bacillus sp. BE506
<400> 4
tagagttttg gatcctggct caggatgaac gctggcggcg tgcctaatac atgcaagtcg 60
agcgaatgga ttgagagctt gctctcaaga agttagcggc ggacgggtga gtaacacgtg 120
ggtaacctgc ccataagact gggataactc cgggaaaccg gggctaatac cggataacat 180
tttgaaccgc atggttcgaa attgaaaggc ggcttcggct gtcacttatg gatggacccg 240
cgtcgcatta gctagttggt gaggtaacgg ctcaccaagg caacgatgcg tagccgacct 300
gagagggtga tcggccacac tgggactgag acacggccca gactcctacg ggaggcagca 360
gtagggaatc ttccgcaatg gacgaaagtc tgacggagca acgccgcgtg agtgatgaag 420
gctttcgggt cgtaaaactc tgttgttagg gaagaacaag tgctagttga ataagctggc 480
accttgacgg tacctaacca gaaagccacg gctaactacg tgccagcagc cgcggtaata 540
cgtaggtggc aagcgttatc cggaattatt gggcgtaaag cgcgcgcagg tggtttctta 600
agtctgatgt gaaagcccac ggctcaaccg tggagggtca ttggaaactg ggagacttga 660
gtgcagaaga ggaaagtgga attccatgtg tagcggtgaa atgcgtagag atatggagga 720
acaccagtgg cgaaggcgac tttctggtct gtaactgaca ctgaggcgcg aaagcgtggg 780
gagcaaacag gattagatac cctggtagtc cacgccgtaa acgatgagtg ctaagtgtta 840
gagggtttcc gcccctttag tgctgaagtt aacgcattaa gcactccgcc tggggagtac 900
ggccgcaagg ctgaaactca aaggaattga cgggggcccg cacaagcggt ggagcatgtg 960
gtttaattcg aagcaacgcg aagaacctta ccaggtcttg acatcctctg aaaaccctag 1020
agatagggct tctccttcgg gagcagagtg acaggtggtg catggttgtc gtcagctcgt 1080
gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgatctta gttgccatca 1140
ttaagttggg cactctaagg tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt 1200
caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggac ggtacaaaga 1260
gctgcaagac cgcgaggtgg agctaatctc ataaaaccgt tctcagttcg gattgtaggc 1320
tgcaactcgc ctacatgaag ctggaatcgc tagtaatcgc ggatcagcat gccgcggtga 1380
atacgttccc gggccttgta cacaccgccc gtcacaccac gagagtttgt aacacccgaa 1440
gtcggtgggg taacctttt 1459
<210> 5
<211> 1449
<212> DNA
<213> Agrobacterium sp. BE516
<400> 5
ttgggttanc ctttgtacga cttcacccca gtcgctgacc ctaccgtggt tcgctgcctc 60
cttgcggtta gcgcacgacc ttcgggtaaa accaactccc atggtgtgac gggcggtgtg 120
tacaaggccc gggaacgtat tcaccgcagc atgctgatct gcgattacta gcgattccaa 180
cttcatgcac tcgagttgca gagtgcaatc cgaactgaga tggcttttgg agattagctc 240
gacctcgcgg tctcgctgcc cactgtcacc accattgtag cacgtgtgta gcccagcccg 300
taagggccat gaggacttga cgtcatcccc accttcctct cggcttatca ccggcagtcc 360
ccttagagtg cccaactgaa tgctggcaac taagggcgag ggttgcgctc gttgcgggac 420
ttaacccaac atctcacgac acgagctgac gacagccatg cagcacctgt ctctgtgtcc 480
ccgaaaggaa agctacgtct ccgtagcggt cacaggatgt caagagctgg taaggttctg 540
cgcgttgctt cgaattaaac cacatgctcc accgcttgtg cgggcccccg tcaattcctt 600
tgagttttaa tcttgcgacc gtactcccca ggcggaatgt ttaatgcgtt agctgcgcca 660
ccgacaagtc aacttgccga cggctaacat tcatcgttta cggcgtggac taccagggta 720
tctaatcctg tttgctcccc acgctttcgc acctcagcgt cagtaatgga ccagtaagcc 780
gccttcgcca ctggtgttcc tgcgaatatc tacgaatttc acctctacac tcgcaattcc 840
acttacctct tccatactca agatacccag tatcaaaggc agttccgcag ttgagctgcg 900
ggatttcacc cctgacttaa atacccgcct acgtgcgctt tacgcccagt aattccgaac 960
aacgctagcc cccttcgtat taccgcggct gctggcacga agttagcmgg ggcttcttct 1020
ccgactaccg tcattatctt catcggtgaa agagctttac aatcctaaga ccttcatcac 1080
tcacgcggca tggctggatc aggcttgcgc ccattgtcca atattcccca ctgctgcctc 1140
ccgtaggagt ttgggccgtg tctcagtccc aatgtggctg atcatcctct cagaccagct 1200
atggatcgtc gccttggtag gcctttaccc caccaactag ctaatccaac gcgggctcat 1260
cataccccga taaatctttc ccccgaaggg cgtatacggt attacttcca gtttcccaga 1320
gctattccgt agggtacggt agattcccac gcgttacttc acccgtctgc cgctcctctt 1380
gcgaggcgct cgactgcatg gtaagcctgc cgccwgcgtt cgttctgagc caggatccaa 1440
actcnaaan 1449
<210> 6
<211> 1518
<212> DNA
<213> Lysinibacillus sp. BE533
<400> 6
ttggttaccc tttktacgac ttcaccccaa tcatctatcc caccttcggc ggctggctcc 60
aaaaggttac cccaccgact tcgggtgtta caaactctcg tggtgtgacg ggcggtgtgt 120
acaaggcccg ggaacgtatt caccgcggca tgctgatccg cgattactag cgattccggc 180
ttcatgtagg cgagttgcag cctacaatcc gaactgagaa cgactttatc ggattagctc 240
cctctcgcga gttggcaacc gtttgtatcg tccattgtag cacgtgtgta gcccaggtca 300
taaggggcat gatgatttga cgtcatcccc accttcctcc ggtttgtcac cggcagtcac 360
cttagagtgc ccaactaaat gatggcaact aagatcaagg gttgcgctcg ttgcgggact 420
taacccaaca tctcacgaca cgagctgacg acaaccatgc accacctgtc accgttgtcc 480
ccgaagggaa aaccatatct ctacagtggt caacgggatg tcaagacctg gtaaggttct 540
tcgcgttgct tcgaattaaa ccacatgctc caccgcttgt gcgggccccc gtcaattcct 600
ttgagtttca gtcttgcgac cgtactcccc aggcggagtg cttaatgcgt tagctgcagc 660
actaaggggc ggaaaccccc taacacttag cactcatcgt ttacggcgtg gactaccagg 720
gtatctaatc ctgtttgctc cccacgcttt cgcgcctcag cgtcagttac agaccagaaa 780
gtcgccttcg ccactggtgt tcctccaaat ctctacgcat ttcaccgcta cacttggaat 840
tccactttcc tcttctgcac tcaagtcccc cagtttccaa tgaccctcca cggttgagcc 900
gtgggctttc acatcagact taaaggaccg cctgcgcgcg ctttacgccc aataattccg 960
gacaacgctt gccacctacg tattaccgcg gctgctggca cgtagttagc cgtggctttc 1020
taataaggta ccgtcaaggt acagccagtt actactgtac ttgttcttcc cttacaacag 1080
agttttacga tccgaaaacc ttcttcactc acgcggcgtt gctccatcag gctttcgccc 1140
attgtggaag attccctact gctgcctccc gtaggagtct gggccgtgtc tcagtcccag 1200
tgtggccgat caccctctca ggtcggctac gcatcgtcgc cttggtgagc cgttacctca 1260
ccaactagct aatgcgccgc gggcccatct tatagcgaca gccgaaaccg tctttcagta 1320
tttcaccatg aagtaaaata gattattcgg tattagcccc ggtttcccgg agttatccca 1380
aactataagg taggttgccc acgtgttact cacccgtccg ccgctaacgt caaaggagca 1440
agctccttct ctgttcgctc gacttgcatg ataggcacgc cgccwgcgtt cgtcctgagc 1500
cagancnaaa actctaaa 1518
<110> Korea Research Institute of Bioscience & Biotechnology
<120> Plant growth promotion by using bacterial strains isolated from
roots of Miscanthus sacchariflorus
<130> 11p-09-33
<160> 6
<170> Kopatentin 1.71
<210> 1
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> forward primer for 16s rRNA
<400> 1
Claims (12)
Tumebacillus sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp.) Microbial agent for promoting plant growth, containing one or more strains selected from the group consisting of strains, or cultures thereof as an active ingredient.
The method of claim 1, wherein the strain is Miscanthus sacchariflorus ) Microbial agent for promoting plant growth, isolated from the root.
The microorganism preparation for promoting plant growth according to claim 1, wherein the strain of the genus Tomebacillus is deposited with accession number KCTC12051BP.
The microorganism preparation for promoting plant growth according to claim 1, wherein the strain of genus Bacillus is deposited with KCTC12052BP.
The microorganism preparation for promoting plant growth according to claim 1, wherein the Agrobacterium strain is deposited with KCTC12053BP.
The microorganism preparation for promoting plant growth according to claim 1, wherein the strain of the genus Lacinibacillus is deposited with KCTC12054BP.
The microorganism preparation for promoting plant growth according to claim 1, wherein the plant is silver grass, wheat, barley, sudangrass, rice, cabbage, cucumber or tomato.
A method for promoting plant growth, comprising treating the soil, plant or seed of a plant with the microbial agent of claim 1.
The method according to claim 8, wherein the microbial agent is irrigated or sprayed on a plant in a liquid state, immersed or sprayed on a seed of a crop, or coated on a seed.
Tumebacillus sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp.) Microbial agent for promoting seed germination containing at least one strain selected from the group consisting of strains, or a culture thereof as an active ingredient.
The microorganism preparation for promoting seed germination according to claim 10, wherein the seed is a seed of silver grass, wheat, barley, sudangrass, rice, cabbage, cucumber or tomato.
Tumebacillus sp.) strain, Bacillus sp.) strain, Agrobacterium sp.) strain And Lysinibaci llus sp.) Bio-fertilizer for promoting plant growth and germination containing at least one strain selected from the group consisting of strains, or cultures thereof as an active ingredient.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020110122257A KR101456171B1 (en) | 2011-11-22 | 2011-11-22 | Plant growth promotion by using bacterial strains isolated from roots of Miscanthus sacchariflorus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020110122257A KR101456171B1 (en) | 2011-11-22 | 2011-11-22 | Plant growth promotion by using bacterial strains isolated from roots of Miscanthus sacchariflorus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20130056585A true KR20130056585A (en) | 2013-05-30 |
| KR101456171B1 KR101456171B1 (en) | 2014-10-31 |
Family
ID=48664575
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020110122257A KR101456171B1 (en) | 2011-11-22 | 2011-11-22 | Plant growth promotion by using bacterial strains isolated from roots of Miscanthus sacchariflorus |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR101456171B1 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108056111A (en) * | 2017-12-14 | 2018-05-22 | 吉林师范大学 | A kind of microbial bacterial agent that wheat nutrient is promoted to absorb and preparation method thereof |
| WO2018129018A1 (en) * | 2017-01-05 | 2018-07-12 | Novozymes Bioag A/S | Lysinibacillus isolates and uses thereof |
| WO2020067555A1 (en) * | 2018-09-28 | 2020-04-02 | 住友化学株式会社 | Bacterium degrading microorganism, microbial preparation, and method and device for degrading microorganism |
| WO2021193750A1 (en) * | 2020-03-26 | 2021-09-30 | 住友化学株式会社 | Monoclonal antibody, bacteria detection method, bacteria detection kit, and hybridoma |
| CN115798588A (en) * | 2022-01-10 | 2023-03-14 | 北京理工大学 | Method for carrying out endophytic physiological screening capable of promoting accumulation of active products at root of Glycyrrhiza uralensis Fisch based on bioinformatics means |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102138585B1 (en) * | 2018-01-15 | 2020-07-28 | 경북대학교 산학협력단 | Erwinia gerundensis KUDC9201 strain having antifungal activity against pathogens, and uses thereof |
| KR20220126123A (en) | 2021-03-08 | 2022-09-15 | 주식회사 코스믹그린 | Ignatzschineria strain producing auxin and promoting plant growth and uses thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4635520B2 (en) | 2004-08-31 | 2011-02-23 | 不二製油株式会社 | Plant growth promoter |
| JP4630627B2 (en) | 2004-10-26 | 2011-02-09 | クミアイ化学工業株式会社 | Plant seed germination rate improver |
| KR100946633B1 (en) * | 2007-12-20 | 2010-03-09 | 한국생명공학연구원 | Methods for plant growth promotion and plant protection by bacterial metabolites |
| KR20100115006A (en) * | 2009-04-17 | 2010-10-27 | 충북대학교 산학협력단 | Bacterial strain cbcn5 promoting plant growth isolated from rhizosphere soils of chinese cabbage and uses thereof |
-
2011
- 2011-11-22 KR KR1020110122257A patent/KR101456171B1/en active IP Right Grant
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018129018A1 (en) * | 2017-01-05 | 2018-07-12 | Novozymes Bioag A/S | Lysinibacillus isolates and uses thereof |
| CN108056111A (en) * | 2017-12-14 | 2018-05-22 | 吉林师范大学 | A kind of microbial bacterial agent that wheat nutrient is promoted to absorb and preparation method thereof |
| WO2020067555A1 (en) * | 2018-09-28 | 2020-04-02 | 住友化学株式会社 | Bacterium degrading microorganism, microbial preparation, and method and device for degrading microorganism |
| JPWO2020067555A1 (en) * | 2018-09-28 | 2021-09-02 | 住友化学株式会社 | Microbial decomposition bacteria, microbial preparations, microbial decomposition methods and decomposition equipment |
| WO2021193750A1 (en) * | 2020-03-26 | 2021-09-30 | 住友化学株式会社 | Monoclonal antibody, bacteria detection method, bacteria detection kit, and hybridoma |
| CN115798588A (en) * | 2022-01-10 | 2023-03-14 | 北京理工大学 | Method for carrying out endophytic physiological screening capable of promoting accumulation of active products at root of Glycyrrhiza uralensis Fisch based on bioinformatics means |
| CN115798588B (en) * | 2022-01-10 | 2024-02-20 | 北京理工大学 | Endophyte for promoting accumulation of active product of roots of uralensis, screening method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| KR101456171B1 (en) | 2014-10-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2017319484B2 (en) | Defined microbial compositions | |
| CN104745483B (en) | A kind of Paecilonyces variotii strain SJ1 and its application | |
| KR101086367B1 (en) | Bacillus aryabhattai LS9 promoting the growth of plants and use thereof | |
| CN113151062B (en) | Bacillus belgii LJBV19 and application thereof | |
| CN110616171B (en) | Saline-alkali-resistant Pacific bacillus and viable bacteria preparation and application thereof | |
| EA009126B1 (en) | Micro-organisms for the treatment of soil and process for obtaining them | |
| KR101456171B1 (en) | Plant growth promotion by using bacterial strains isolated from roots of Miscanthus sacchariflorus | |
| CN111690578B (en) | Salt and alkali resistant Siamese bacillus and production method and application of viable bacteria preparation thereof | |
| US20220369648A1 (en) | Endophytic falciphora oryzae fo-r20 and its application | |
| CN111778173B (en) | Bacillus subtilis Pro1A2, microbial inoculum and preparation method thereof, and application of bacillus subtilis Pro1A2 in cultivation of melons | |
| KR102000472B1 (en) | Bacillus aryabhattai strain promoting resistance of plants against abiotic stress and use thereof | |
| KR101908729B1 (en) | Chryseobacterium soldanellicola strain having biocontrol efficacy against plant diseases and plant growth promoting activity and use thereof | |
| CN108913625B (en) | Salt-tolerant streptomycete, microbial inoculum thereof and application of microbial inoculum thereof in promoting plant growth | |
| US11674118B2 (en) | PGPR compositions and methods for improved cultivation of tomato and potato species | |
| CN109609403B (en) | Biocontrol bacterium and application thereof in prevention and control of downy mildew of crops | |
| CN117467557A (en) | Paenibacillus strain with growth promoting and biocontrol functions and application thereof | |
| CN116240126A (en) | Multifunctional bacillus belgium SB10 and application thereof | |
| CN111548951B (en) | Bacillus subtilis Pro6A5, microbial inoculum and preparation method thereof, and application of bacillus subtilis Pro6A5 in cultivation of melons | |
| CN113416679A (en) | Bacillus methylotrophicus, microbial inoculum comprising bacillus methylotrophicus and application of bacillus methylotrophicus | |
| CN110066755A (en) | One plant of fructus arctii endogenetic streptomycete, the microbial bacterial agent containing the endophyte and application | |
| CN114164126B (en) | Rhizopus oryzae XERF-1 and application thereof in relieving apple continuous cropping obstacle | |
| CN113278551B (en) | Burkholderia, bacterial agent comprising Burkholderia, bacterial fertilizer, preparation method and application | |
| KR102437555B1 (en) | ALLEVIATION OF DROUGHT STRESS IN PUMPKIN PLANTS BY Kushneria konosiri H05E-12 AND USES THEREOF | |
| US20230240304A1 (en) | Aspergillus terreus strain and composition for controlling bacterial plant diseases using strain or culture filtrate thereof | |
| CN116426445B (en) | Pseudomonas bacteria NJAU-T102 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant | ||
| FPAY | Annual fee payment |
Payment date: 20170824 Year of fee payment: 4 |
|
| FPAY | Annual fee payment |
Payment date: 20190909 Year of fee payment: 6 |