KR20120108701A - The microbial agent with useful soil bacteria and a method for remediation of salt stressed soil using this agent - Google Patents

The microbial agent with useful soil bacteria and a method for remediation of salt stressed soil using this agent Download PDF

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KR20120108701A
KR20120108701A KR1020110026908A KR20110026908A KR20120108701A KR 20120108701 A KR20120108701 A KR 20120108701A KR 1020110026908 A KR1020110026908 A KR 1020110026908A KR 20110026908 A KR20110026908 A KR 20110026908A KR 20120108701 A KR20120108701 A KR 20120108701A
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salt
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강재곤
백상훈
최호근
박정찬
정종철
강훈석
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(주)한국바이오케미칼
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    • A01N25/00Biocides, 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
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • B09C1/10Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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Abstract

PURPOSE: A method for recovering salt-stressed soil using a microorganism formulation containing soil microorganism is provided to promote plant growth and to enhance yield. CONSTITUTION: A Pantoea sp. J11 strain(deposit number KCTC 11893BP) has an ability of decomposition or removing salt integrated in soil. The soil is salt-stressed soil. The strain is used in a greenhouse cultivation land or reclaimed land. A composition for removing salt contains the strain or culture thereof. A plant growth promoting agent contains the strain or culture liquid thereof. The plant growth promoting agent additionally contains carriers.

Description

유용한 토양미생물을 포함하는 미생물 제제 및 이를 이용한 염류 장해 토양의 복원 방법{The microbial agent with useful soil bacteria and a method for remediation of salt stressed soil using this agent}The microbial agent with useful soil bacteria and a method for remediation of salt stressed soil using this agent}

본 발명은 신규 토양 미생물, 이를 이용한 미생물 제제, 및 상기 토양 미생물을 이용한 염류 장해 토양 복원 방법에 관한 것이다.
The present invention relates to a novel soil microorganism, a microbial preparation using the same, and a method for restoring a salt obstacle soil using the soil microorganism.

염이란 화학적으로는 산과 염기가 결합된 것을 말하고, 비료로 사용되는 황산칼리는 황산(산)과 칼리(염기)가 결합한 염이며, 염기는 전기적으로 양성을 나타내는 칼리, 석회, 마그네슘, 나트륨, 철, 아연, 망간 등의 금속원소이고, 산은 전기적으로 음성을 띠는 질산, 황산 및 염소 이온 등이 있다. 따라서 염은 작물 생육에 꼭 필요로 하는 영양소이다. 그러나 이들 염류가 토양 중에 적정 수준 이상으로 존재할 때 우리는 염류 집적이라고 말하며, 과다한 염은 식물의 근권을 통해 흡수되어 식물에 치명적인 상해를 입히는 장해를 염류 장해라고 한다.
Salt refers chemically to the combination of acid and base, and the sulfate used as fertilizer is a salt of sulfuric acid (acid) and kali (base), and the base is electrically positively kali, lime, magnesium, sodium, iron , Metals such as zinc and manganese, and acids include electrically negative nitric acid, sulfuric acid and chlorine ions. Salt is therefore a vital nutrient for crop growth. However, when these salts are present in the soil above the appropriate level, we say salt accumulation, and the excess salt is absorbed through the root zone of the plant and the obstacle that causes fatal injury to the plant is called salt disorder.

염류 장해는 염류성 토양(saline soils) 및 나트륨 토양(sodic soils), 염류-나트륨 토양(saline-sodic)의 세 가지 형태로 발생한다. 염류성 토양(saline soils)은 과다한 용해성 염을 함유한 토양으로 심한 경우 토양표면이 잔류물로 인해 하얗게 되는데, 이는 식물의 수분과 양분 흡수를 어렵게 하고 독성을 가지고 있어 염의 여과와 배수가 필수적이다. 나트륨 토양(sodic soils)은 과다한 교환성 나트륨을 함유한 토양인데 그 자체는 직접적으로 식물에 해를 주지 않지만, 토양 구성 입자가 고운 토양으로 만들어 토양 간극을 좁게 하여 결국에는 수분투과 및 배수를 극히 어렵게 하고 식물의 뿌리가 성장하는 것을 방해한다. 염류-나트륨 토양(saline-sodic)은 용해성 염과 교환성 나트륨을 함유한 토양으로서 방치하면 토양 구조를 악화시켜 작물의 생육이 불가능한 상태로 되기 때문에 토양 개선이 필수적으로 이루어져야 한다.
Salt disturbances occur in three forms: saline soils, sodium soils, and saline-sodic. Saline soils are soils that contain excessive soluble salts and, in severe cases, the soil surface becomes white due to residues, which makes it difficult to absorb moisture and nutrients from plants and is toxic, requiring salt filtration and drainage. Sodic soils are soils that contain excessive amounts of exchangeable sodium, which themselves do not directly harm plants, but they make the soil constituent finer, narrowing the soil gap and ultimately making water penetration and drainage extremely difficult. And prevent the plant roots from growing. Saline-sodic soil is a soil containing soluble salts and exchangeable sodium, and the soil improvement must be made because the soil structure is deteriorated and crop growth is impossible.

국내외에서 수행되고 있는 염류 장해 극복 방안 중 하나는 염에 내성을 가진 식물(salt tolerant crops)을 재배하는 경작적 제어법에 의존하고 있다. 그러나 높은 내성을 가진 작물은 목화, 보리 등 몇 종에 국한될 뿐만 아니라, 종자가 발아하기 위해서는 토양의 염류농도가 한계수준 이하의 비교적 낮은 수준에서만 가능하다. 대부분의 식물은 이들보다 염류농도에 더 민감하다. 또한 물리적인 극복방안은 문제가 되는 포장에 토관 배수로(tile drainage)를 설치하여 문제를 해결하고 있는 실정이다. 대단히 광범위한 염류장해토양(시설재배지, 간척지)을 대상으로, 배수로 공사를 할 경우 천문학적인 비용이 소요되므로 경제적인 부담은 이루 헤아릴 수 없는 상황이다. 뿐만 아니라 대다수의 자연 배수로는 토양 염류를 잘 여과할 수 있는 능력이 결여되어 있다.
One of the measures to overcome salt disorders, which are carried out at home and abroad, relies on cultivation control methods for growing salt tolerant crops. However, high-tolerant crops are not only limited to a few species, such as cotton and barley, but also only at relatively low levels of salt concentrations below the limit for seed germination. Most plants are more sensitive to salt concentration than these. In addition, the physical overcoming situation is to solve the problem by installing a tile drainage (tile drainage) on the pavement in question. The economic burden is immeasurable because of the astronomical cost of drainage construction for a very wide range of salt disturbed soils (facility cultivation and reclaimed land). In addition, most natural drains lack the ability to filter soil salts well.

현재 대다수의 국내 포장 토양과 간척지 토양은 이미 염류 장해가 매우 심각한 상태에 놓여있다. 이러한 염류 장해는 농작물 생육에 큰 피해를 주어 농업 생산성에 가장 커다란 걸림돌이 되는 농업 환경 문제 중 하나이다. 그러나 현재까지 국내외적으로 염류 장해 토양의 복원에 대한 구체적이고도 성공적인 환경 친화형 기술은 전무한 실정이다. 따라서 염류 장해 토양 복원을 위한 환경 친화형 미생물 제제의 개발은 절실하다.
At present, the majority of domestic pavement and reclaimed soils are already very seriously salted. These salt disturbances are one of the agricultural environmental problems that cause great damage to the growth of crops, which is the biggest obstacle to agricultural productivity. However, there are no specific and successful environmentally friendly technologies for restoring salt impaired soils at home and abroad. Therefore, the development of environmentally friendly microbial preparations for the restoration of salt disturbed soils is urgent.

이에 본 발명자들은 토양으로부터 인산염 가용화 균주를 탐색 및 분리하여 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균주를 분리하고, 상기 균주를 포함하는 미생물 제제가 토양내 집적된 염류를 분해 또는 제거하고, 식물의 생장을 촉진하는 활성을 갖는다는 것을 확인함으로써 본 발명을 완성하였다.
Therefore, the present inventors search for and isolate phosphate solubilizing strains from the soil to isolate Pantoea sp. J11 KCTC 11893BP strain, and the microbial agent including the strain decomposes or removes salts accumulated in the soil. The present invention was completed by confirming that it has an activity of promoting plant growth.

본 발명의 목적은 토양으로부터 유래된 신규한 인산염 가용화 균주를 제공하는 것이다.It is an object of the present invention to provide novel phosphate solubilizing strains derived from soil.

본 발명의 다른 목적은 상기 균주를 포함하는 토양 염류 집적 해소용 조성물을 제공하는 것이다.Another object of the present invention is to provide a composition for eliminating soil salt accumulation comprising the strain.

본 발명의 다른 목적은 상기 균주를 포함하는 식물 생장 촉진제를 제공하는 것이다.Another object of the present invention to provide a plant growth promoter comprising the strain.

본 발명의 다른 목적은 상기 균주를 포함하는 조성물을 토양에 처리하는 단계를 포함하는 염류 집적 해소 방법을 제공하는 것이다.Another object of the present invention is to provide a salt accumulation eliminating method comprising the step of treating the composition comprising the strain to the soil.

본 발명의 다른 목적은 상기 균주를 포함하는 조성물을 식물 또는 식물을 재배하는 토양에 처리하는 단계를 포함하는 식물 생육 촉진 방법을 제공하는 것이다.
Another object of the present invention is to provide a method for promoting plant growth, comprising the step of treating the composition comprising the strain to a plant or soil to grow plants.

상기 목적을 달성하기 위하여, 본 발명은 수탁번호 KCTC 11893BP로 기탁된 판토에아 속 J11(Pantoea sp. J11) 균주를 제공한다.In order to achieve the above object, the present invention provides a Pantoea sp. J11 strain deposited with accession number KCTC 11893BP.

또한, 본 발명은 상기 균주 또는 이의 배양액을 포함하는 토양 염류 집적 해소용 조성물을 제공한다.In addition, the present invention provides a composition for eliminating soil salt accumulation comprising the strain or culture thereof.

또한, 본 발명은 상기 균주 또는 이의 배양액을 포함하는 식물 생장 촉진제를 제공한다.The present invention also provides a plant growth promoter comprising the strain or its culture.

또한, 본 발명은 상기 균주, 이의 배양액 또는 이를 포함하는 조성물을 토양에 처리하는 단계를 포함하는 염류 집적 해소 방법을 제공한다.In addition, the present invention provides a salt accumulation elimination method comprising the step of treating the soil, a culture thereof or a composition comprising the same to the soil.

아울러, 본 발명은 상기 균주, 이의 배양액 또는 이를 포함하는 조성물을 식물 또는 식물을 재배하는 토양에 처리하는 단계를 포함하는 식물 생육 촉진 방법을 제공한다.
In addition, the present invention provides a method for promoting plant growth, comprising the step of treating the strain, its culture solution or a composition comprising the same to the plant or the soil for cultivating the plant.

본 발명의 판토에아 속(Pantoea sp.) J11 균주 또는 상기 균주의 배양액이 염류집적 토양에 대해 우수한 염류 분해 활성을 나타내고 식물 생육촉진 및 수확량에 대해 증수 효과를 나타내기 때문에, 상기 균주 또는 이의 배양액은 토양 염류 장해 극복 또는 식물 생육 촉진에 유용하게 이용될 수 있다.
Since the Pantoea sp. J11 strain of the present invention or the culture medium of the strain exhibits excellent hydrolysis activity against salt-integrated soils and shows a water-producing effect on plant growth promotion and yield, the strain or culture medium thereof. May be usefully used to overcome soil salt disorders or promote plant growth.

도 1은 트리칼슘포스페이트에 대한 가용화 활성을 나타내는 그림이다.
도 2는 판토에아 속 J11(Pantoea sp. J11)균주를 영양 한천 배지에서 계대배양하여 순수 배양된 균주를 나타내는 그림이다.
도 3은 판토에아 속 J11 균주의 16S rRNA 유전자 계통학적 위치를 나타내는 그림이다.
16S rRNA 서열에 기초하여 판토에아 속 J11 균주와 관련된 박테리아 분류군의 위치를 나타내는 계통수(Phylogenetic tree)를 작성하였다. 가지에 있는 수는 50 % 이상 지지되는 가지들로만 비롯된, 부트스트랩(bootstrap) 수치이다.
선은 위치당 0.002 치환이다.
T: 스트레인 유형
도 4는 J11 균주의 균체 지방산 조성 분석 결과를 나타내는 그래프와 그림이다.
1 is a diagram showing solubilizing activity for tricalcium phosphate.
Figure 2 is a diagram showing a strain cultured pure by subcultured Pantoea sp. J11 ( Pantoea sp. J11) strain in nutrient agar medium.
Figure 3 is a diagram showing the 16S rRNA gene phylogenetic position of Pantoea genus J11 strain.
Based on the 16S rRNA sequence, a Phylogenetic tree was created that indicates the location of the bacterial taxon associated with the J11 strain of genus Pantoea. The number on a branch is a bootstrap number that only comes from branches that are supported by 50% or more.
The line is 0.002 substitutions per position.
T: strain type
Figure 4 is a graph and a figure showing the results of analysis of the cell fatty acid composition of the J11 strain.

이하, 본 발명을 상세히 설명한다.
Hereinafter, the present invention will be described in detail.

본 발명은 수탁번호 KCTC 11893BP로 기탁된 판토에아 속 J11(Pantoea sp. J11) 균주를 제공한다.The present invention provides a Pantoea sp. J11 strain deposited with accession number KCTC 11893BP.

상기 균주는 토양 내 집적된 염류를 분해 또는 제거하는 활성을 갖는 것이 바람직하나, 이에 한정되지 않는다.The strain preferably has an activity of degrading or removing salts accumulated in the soil, but is not limited thereto.

상기 토양은 염류 장해 토양인 것이 바람직하나, 이에 한정되지 않는다.The soil is preferably a salt obstacle soil, but is not limited thereto.

상기 균주는 시설 재배지 또는 간척지의 토양에 사용하는 것이 바람직하나, 이에 한정되지 않는다.The strain is preferably used in the soil of plantation or reclaimed land, but is not limited thereto.

본 발명의 구체적인 실시예에서, 토양으로부터 인산 가용화 능력이 있는 균을 분리하기 위해서 전국 각지로부터 많은 토양시료를 수집하였다. 채취한 토양 시료 1 g을 멸균 증류수에 현탁한 후, 트리칼슘포스페이트(Ca3(PO4)2)의 최종 농도가 0.5 % 되도록 조제한 고형 배지에 평판희석법을 이용하여 페트리 디쉬에 도말, 접종하였다. 접종된 배지를 30 ℃ 항온기에서 2 내지 3일 배양하여 생육이 빠르고 투명환이 큰 균주를 인산 가용화능이 있는 균주로 선별하였다(도 1).In a specific embodiment of the present invention, a large number of soil samples were collected from all over the country in order to isolate bacteria having phosphate solubility. 1 g of the collected soil sample was suspended in sterile distilled water, and then plated and inoculated in a petri dish using a plate dilution method in a solid medium prepared so that the final concentration of tricalcium phosphate (Ca 3 (PO 4) 2 ) was 0.5%. Inoculated medium was incubated for 2 to 3 days at 30 ℃ thermostat was selected as a strain capable of phosphate solubilizing fast growth and large clear ring (Fig. 1).

본 발명의 구체적인 실시예에서, 상기 균주의 게놈은 벤질 클로라이드(Benzyl chloride)법을 변형한 방법을 이용하여 상기 균주의 게놈 DNA를 추출하는 것이 바람직하나, 이에 한정되지 않는다. 상기 추출된 게놈 DNA에서 상기 균주의 16S rRNA 유전자는 서열번호: 1 및 2로 기재되는 프라이머(primer) 쌍을 이용하여 PCR 방법으로 증폭하는 것이 바람직하나, 이에 한정되지 않는다. 상기 균주의 16S rRNA 유전자 염기서열을 분석하여(서열번호: 3), DDBJ/NCBI/Genebank와 Ribosomal Database Project Ⅱ(RDP Ⅱ)의 데이터베이스에서 상동성 검색을 수행하는 것이 바람직하나, 이에 한정되지 않는다. 분자계통학적 분석결과에서 보는 바와 같이 선별된 균주는 판토에아 속(Pantoea sp.)에 속하는 종으로 판명되었다. 특히 상기 선발된 균주는 판토에아 속 CRLS069b(FJ593734)와 100 %, 그리고 표준 균주인 펙토박테리움 시프리페디(Pectobacterium cypripedii) DSM3873(AJ233413)의 염기서열과 98.3%의 유연관계를 나타내는 것으로 확인되어 상기 선별된 균주를 '판토에아 속 J11(Pantoea sp. J11)'로 명명하고 상기 균주를 2011년 3월 14일자로 국제기탁기관인 한국생명공학연구원 내 생물자원센터에 기탁하였다(기탁번호; KCTC 11893BP). In a specific embodiment of the present invention, it is preferable that the genome of the strain is extracted from the genomic DNA of the strain using a method modified from the benzyl chloride method, but is not limited thereto. In the extracted genomic DNA, the 16S rRNA gene of the strain is preferably amplified by PCR using a primer (primer) pairs set forth in SEQ ID NOs: 1 and 2, but is not limited thereto. By analyzing the 16S rRNA gene sequence of the strain (SEQ ID NO: 3), it is preferable to perform homology search in the database of DDBJ / NCBI / Genebank and Ribosomal Database Project II (RDP II), but is not limited thereto. As shown in the molecular analysis results, the selected strain was found to belong to Pantoea sp. In particular, the selected strain will remain on the panto in CRLS069b (FJ593734) and 100%, and the type strain of Peg tobak Te Solarium shifted Li Phedi (Pectobacterium cypripedii ) It was found to show a flexible relationship of 98.3% with the nucleotide sequence of DSM3873 (AJ233413), and thus the selected strain was named ' Pantoea sp. J11' and the strain was named March 14, 2011. The fund was deposited at the Korea Institute of Bioscience and Biotechnology, an international depository institution (KCTC 11893BP).

본 발명의 구체적인 실시예에서, 상기 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균주의 지방산 조성을 분석하여 주요 지방산으로 C16 :1 iso(36.99 %)를 나타내었으며, C12 :0, C14 :0, C16 :0 그리고 그것의 썸드피쳐 3(Summed feature 3)와 같은 다양한 지방산을 함유하는 특징을 나타내었다. 또한 상기 균주는 판토에아 속의 표준 균주들과도 주요 균체 지방산 조성과도 유사한 균체 지방산 조성을 갖는 것으로 확인되었다(표 1 및 도 3). In a specific embodiment of the present invention, the genus Pantoea J11 ( Pantoea sp. J11) Analysis of fatty acid composition of KCTC 11893BP strain showed C 16 : 1 iso (36.99%) as a major fatty acid, C 12 : 0 , C 14 : 0 , C 16 : 0 and its thumb feature 3 (Summed feature 3) It is characterized by containing a variety of fatty acids, such as). In addition, the strain was confirmed to have a cell fatty acid composition similar to the main cell fatty acid composition with standard strains of the genus Pantoea (Table 1 and Figure 3).

본 발명의 구체적인 실시예에서, 본 발명의 판토에아(Pantoea sp.) J11은 영양배지(Nutrient broth)에서 배양하는 것이 바람직하나 이에 한정되지 않고, 통상의 세균이 사용할 수 있는 영양원을 함유하는 배지에서 모두 배양이 가능하다. 상기 배양은 호기적 조건에서 진탕 배양하는 것이 바람직하고, 배양온도, 배양 pH 및 배양 시간은 28 ℃ 내지 35 ℃, pH 6.0 내지 7.0에서 2 내지 5일간 배양하는 것이 바람직하나 이에 한정되지 않는다.
In a specific embodiment of the present invention, Pantoea sp. J11 of the present invention is preferably cultured in nutrient broth (Nutrient broth), but is not limited thereto, a medium containing a nutrient source that can be used by conventional bacteria Both cultures are possible. The culture is preferably shaken in aerobic conditions, the culture temperature, the culture pH and the incubation time is preferably incubated for 2 to 5 days at 28 ℃ to 35 ℃, pH 6.0 to 7.0, but is not limited thereto.

또한, 본 발명은 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균주 또는 이의 배양액을 포함하는 토양 염류 집적 해소용 조성물을 제공한다.In addition, the present invention provides a composition for eliminating soil salt accumulation comprising Pantoea sp. J11 ( Pantoea sp. J11) KCTC 11893BP strain or a culture thereof.

상기 토양은 염류 장해 토양인 것이 바람직하나, 이에 한정되지 않는다.The soil is preferably a salt obstacle soil, but is not limited thereto.

상기 조성물은 시설 재배지 또는 간척지의 토양에 사용하는 것이 바람직하나, 이에 한정되지 않는다.The composition is preferably used in the soil of plantation or reclaimed land, but is not limited thereto.

본 발명의 구체적인 실시예에서, 상기 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균체는 미생물 수가 109~1010 CFU(colony forming unit)/㎖가 되도록 멸균 증류수로 현탁하여 이를 미생물 제제 원액으로 사용하고, 상기 미생물 제제 원액을 500 ㎖에 균수가 107~108 CFU/㎖가 되도록 희석하여 미생물 제제를 제조하는 것이 바람직하나, 이에 한정되지 않는다.In a specific embodiment of the present invention, the genus Pantoea J11 ( Pantoea sp. J11) KCTC 11893BP cells are suspended in sterile distilled water such that the number of microorganisms is 10 9 ~ 10 10 CFU (colony forming unit) / ㎖ and used as a stock solution of the microbial preparation, the microbial preparation stock solution in 500 10 7 ~ 10 It is preferred to prepare a microbial agent by diluting to 8 CFU / ml, but is not limited thereto.

본 발명의 토양 염류 집적 해소용 조성물은 판토에아(Pantoea sp.) J11의 균체, 이의 배양액, 이의 배양액 추출물 또는 배양 여액을 포함하는 것이 바람직하나, 이에 한정되지 않는다. The composition for eliminating soil salts of the present invention preferably includes the cells of Pantoea sp. J11, its culture solution, its culture solution extract or its culture filtrate, but is not limited thereto.

아울러, 본 발명에 따른 유효성분은 전체 조성물에 대하여 상기 균체 또는 배양액을 0.0625 ~ 90 중량%를 사용하는 것이 바람직하며, 0.125 ~ 50 중량%를 사용하는 것이 더욱 바람직하나, 이에 한정되지 않는다.In addition, the active ingredient according to the present invention is preferably used in the cell or culture medium with respect to the total composition 0.0625 ~ 90% by weight, more preferably 0.125 ~ 50% by weight, but is not limited thereto.

본 발명의 구체적인 실시예에서, 미생물 제제에 염류장해의 척도인 전기전도도(electrical conductivity, EC)의 수치가 약 20 내지 30 %이상 경감되었고, 또한 치환성 양이온인 칼륨(K), 칼슘(Ca), 마그네슘(Mg)도 30 내지 40 % 이상 경감시켰다(표 2).
In a specific embodiment of the present invention, the level of electrical conductivity (EC), which is a measure of salt damage in the microbial preparation, is reduced by about 20-30% or more, and the substitutional cations potassium (K) and calcium (Ca) , Magnesium (Mg) was also reduced by 30 to 40% or more (Table 2).

또한, 본 발명은 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균주 또는 이의 배양액을 포함하는 식물 생장 촉진제를 제공한다.The present invention also provides a plant growth promoter comprising Pantoea sp. J11 KCTC 11893BP strain or a culture thereof.

상기 식물 생장 촉진제는 미생물 농약, 종자 코팅제, 미생물 영양제, 토양 개량제, 퇴비 부숙제 및 엽면 살포제로 구성된 군으로부터 선택되는 것으로 사용하는 것이 바람직하나, 이에 한정되지 않는다.The plant growth promoter is preferably used as one selected from the group consisting of microbial pesticides, seed coatings, microbial nutrients, soil improvers, composting agents and foliar spreading agents, but is not limited thereto.

또한, 상기 식물 생장 촉진제는 농약 제제로서 사용가능한 담체를 추가로 포함하는 것이 바람직하나, 이에 한정되지 않는다. 상기 담체는 통상적으로 사용되는 농약 제형으로 허용 가능한 담체이면 모두 사용할 수 있다. 특히 동결보존제 스킴밀크(skim milk), 효모 추출물(yeast extract), PG와 전착제 SOF70, 계면활성제 TD20A, AS65D 등을 사용하는 것이 바람직하나, 이에 한정되지 않는다.In addition, the plant growth promoter preferably includes a carrier usable as a pesticide preparation, but is not limited thereto. The carrier may be used as long as it is an acceptable carrier in a commonly used pesticide formulation. In particular, cryopreservatives such as skim milk (skim milk), yeast extract (yeast extract), PG and electrodeposition agent SOF70, surfactant TD20A, AS65D and the like is preferably used, but not limited thereto.

본 발명의 구체적인 실시예에서, 상기 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균체는 미생물 수가 109~1010 CFU(colony forming unit)/㎖가 되도록 멸균 증류수로 현탁하여 이를 미생물 제제 원액으로 사용하고, 상기 미생물 제제 원액을 500 ㎖에 균수가 107~108 CFU/㎖가 되도록 희석하여 미생물 제제를 제조하는 것이 바람직하나, 이에 한정되지 않는다.In a specific embodiment of the present invention, the genus Pantoea J11 ( Pantoea sp. J11) KCTC 11893BP cells are suspended in sterile distilled water such that the number of microorganisms is 10 9 ~ 10 10 CFU (colony forming unit) / ㎖ and used as a stock solution of the microbial preparation, the microbial preparation stock solution in 500 10 7 ~ 10 It is preferred to prepare a microbial agent by diluting to 8 CFU / ml, but is not limited thereto.

본 발명의 토양 염류 집적 해소용 조성물은 판토에아(Pantoea sp.) J11의 균체, 이의 배양액, 이의 배양액 추출물 또는 배양 여액을 포함하는 것이 바람직하나, 이에 한정되지 않는다. The composition for eliminating soil salts of the present invention preferably includes the cells of Pantoea sp. J11, its culture solution, its culture solution extract or its culture filtrate, but is not limited thereto.

아울러, 본 발명에 따른 유효성분은 전체 조성물에 대하여 상기 균체 또는 배양액을 0.0625 ~ 90 중량%를 사용하는 것이 바람직하며, 0.125 ~ 50 중량%를 사용하는 것이 더욱 바람직하나, 이에 한정되지 않는다.In addition, the active ingredient according to the present invention is preferably used in the cell or culture medium with respect to the total composition 0.0625 ~ 90% by weight, more preferably 0.125 ~ 50% by weight, but is not limited thereto.

본 발명의 구체적인 실시예에서, 미생물 제제가 작물의 생육에 미치는 영향을 조사하였다. 작물은 상추인 것이 바람직하나, 이에 한정되지 않는다. In a specific embodiment of the present invention, the effect of the microbial agent on the growth of the crop was investigated. The crop is preferably lettuce, but not always limited thereto.

상추 비닐하우스에 대조구(관행), 반량 처리(1600배 희석), 적량 처리(800배 희석), 배양 처리(400배 희석)로 관주 처리하였다. 정식 후 8주 후 생육 및 수량 조사를 실시하여 평균값으로 계산하였다(표 3). 상추 초장은 일반관행에 비해 미생물 제제 시용구에서 다소 길었고, 엽수는 시용량이 증가함으로서 비례적으로 많았는데 대조구에 비해 적량처리 시용구가 평균 2 ~ 3 배(5 ㎝ 이상의 엽장)가 더 많은 경향을 나타내었다. 엽폭은 처리 구간 유의적인 차이가 없었고 생체중은 적량처리 이상 시용함으로 증가하였다. 뿌리길이는 시용량이 증가할수록 길었으며 측근 및 세근은 적량처리이상 시용구에서 잘 발달하였다.The lettuce green house was irrigated with control (practice), half volume (1600-fold dilution), titration (800-fold dilution), and culture treatment (400-fold dilution). Eight weeks after the establishment, growth and yield surveys were performed and calculated as average values (Table 3). The lettuce length was slightly longer in microbial application than in general practice, and the number of leaves was proportionately increased with increasing application volume. Indicated. Leaf width was not significantly different between treatments and fresh weight was increased by applying more than appropriate treatment. Root length was longer with increasing dose, and aides and roots were well developed at the time of proper treatment.

엽록체의 발달은 관행재배에 비해 미생물 제제 시용량에 따라 엽록체 함량이 증가하는 경향을 나타내었다.
The development of chloroplasts tended to increase chloroplast content according to the amount of microbial preparation compared to conventional cultivation.

또한, 본 발명은 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균주, 이의 배양액 또는 이를 포함하는 조성물을 토양에 처리하는 단계를 포함하는 염류 집적 해소 방법을 제공한다.In addition, the present invention provides a method for eliminating salt accumulation, comprising treating the soil with Pantoea sp. J11 KCTC 11893BP strain, a culture solution thereof, or a composition comprising the same.

상기 토양은 염류 장해 토양인 것이 바람직하나, 이에 한정되지 않는다.The soil is preferably a salt obstacle soil, but is not limited thereto.

상기 균주, 이의 배양액 또는 이를 포함하는 조성물을 시설 재배지 또는 간척지의 토양에 사용하는 것이 바람직하나, 이에 한정되지 않는다.It is preferable to use the strain, its culture solution or a composition comprising the same on the soil of facility plantation or reclaimed land, but is not limited thereto.

본 발명의 토양 염류 집적 해소 방법은 판토에아(Pantoea sp.) J11의 균체, 이의 배양액, 이의 배양액 추출물 또는 배양 여액을 포함하는 조성물을 토양에 처리하는 것이 바람직하나, 이에 한정되지 않는다. In the method for eliminating the accumulation of soil salt of the present invention, it is preferable to treat the composition including the cells of Pantoea sp. J11, its culture solution, its culture extract or its culture filtrate to the soil, but is not limited thereto.

아울러, 본 발명에 따른 토양에 처리하는 용량은 0.0625 ~ 90 중량%인 판토에아(Pantoea sp.) J11의 균체, 이의 배양액, 이의 배양액 추출물 또는 배양 여액을 포함하는 조성물을 토양에 처리하는 것이 바람직하며, 조성물을 800배 희석하여 관주하는 것이 더욱 바람직하나, 이에 한정되지 않는다.In addition, it is preferable to treat the soil with a composition comprising the cells of Pantoea sp. J11, a culture solution thereof, a culture extract thereof, or a culture filtrate of 0.0625 to 90% by weight. And, it is more preferable to irrigate the composition 800 times dilution, but is not limited thereto.

본 발명의 구체적인 실시예에서, 상기 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균체는 미생물 수가 109~1010 CFU(colony forming unit)/㎖가 되도록 멸균 증류수로 현탁하여 이를 미생물 제제 원액으로 사용하고, 상기 미생물 제제 원액을 500 ㎖에 균수가 107~108 CFU/㎖가 되도록 희석하여 미생물 제제를 제조하여 사용하는 방법으로 수행하는 것이 바람직하나, 이에 한정되지 않는다.
In a specific embodiment of the present invention, the genus Pantoea J11 ( Pantoea sp. J11) KCTC 11893BP cells are suspended in sterile distilled water such that the number of microorganisms is 10 9 ~ 10 10 CFU (colony forming unit) / ㎖ and used as a stock solution of the microbial preparation, the microbial preparation stock solution in 500 10 7 ~ 10 Dilution to 8 CFU / ㎖ is preferably carried out by the method of preparing and using a microbial preparation, but is not limited thereto.

아울러, 본 발명은 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균주, 이의 배양액 또는 이를 포함하는 조성물을 식물 또는 식물을 재배하는 토양에 처리하는 단계를 포함하는 식물 생육 촉진 방법을 제공한다.In addition, the present invention provides a method for promoting plant growth, comprising treating Pantoea sp. J11 (C Pantoea sp. J11) KCTC 11893BP strain, a culture solution thereof, or a composition comprising the same to a plant or a soil growing plant.

상기 처리는 식물의 종자에 도포하는 처리, 식물의 종자를 침지시키는 처리, 식물의 재배 토양에 관주(灌注)하는 처리, 식물의 재배 토양 표면에 분무하는 처리, 식물의 경엽에 살포하는 처리, 및 식물의 부상부(付傷剖)에 접촉시키는 처리로 이루어지는 군으로부터 선택되는 어느 하나인 것이 바람직하나, 이에 한정되지 않는다.The treatment is a treatment applied to the seed of the plant, a treatment of immersing the seed of the plant, a treatment irrigation to the cultivated soil of the plant, a spray to the surface of the cultivated soil of the plant, a treatment to spray on the foliage of the plant, and It is preferable that it is any one selected from the group which consists of a process which makes contact with the floating part of a plant, but it is not limited to this.

본 발명의 식물 생육 촉진 방법은 판토에아(Pantoea sp.) J11의 균체, 이의 배양액, 이의 배양액 추출물 또는 배양 여액을 포함하는 조성물을 식물에 처리하는 것이 바람직하나, 이에 한정되지 않는다. Plant growth promoting method of the present invention is preferably, but not limited to treating the plant comprising a composition comprising the cells of Pantoea sp. J11, its culture, its culture extract or culture filtrate.

아울러, 본 발명에 따른 식물에 처리하는 용량은 0.0625 ~ 90 중량%인 판토에아(Pantoea sp.) J11의 균체, 이의 배양액, 이의 배양액 추출물 또는 배양 여액을 포함하는 조성물을 식물에 처리하는 것이 바람직하며, 조성물을 800배 희석하여 관주하는 것이 더욱 바람직하나, 이에 한정되지 않는다.In addition, it is preferable to treat the plant with a composition comprising the cells of Pantoea sp. J11, a culture solution thereof, a culture extract thereof, or a culture filtrate of 0.0625 to 90% by weight. And, it is more preferable to irrigate the composition 800 times dilution, but is not limited thereto.

본 발명의 구체적인 실시예에서, 상기 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균체는 미생물 수가 109~1010 CFU(colony forming unit)/㎖가 되도록 멸균 증류수로 현탁하여 이를 미생물 제제 원액으로 사용하고, 상기 미생물 제제 원액을 500 ㎖에 균수가 107~108 CFU/㎖가 되도록 희석하여 미생물 제제를 제조하는 것이 바람직하나, 이에 한정되지 않는다.
In a specific embodiment of the present invention, the genus Pantoea J11 ( Pantoea sp. J11) KCTC 11893BP cells are suspended in sterile distilled water such that the number of microorganisms is 10 9 ~ 10 10 CFU (colony forming unit) / ㎖ and used as a stock solution of the microbial preparation, the microbial preparation stock solution in 500 10 7 ~ 10 It is preferred to prepare a microbial agent by diluting to 8 CFU / ml, but is not limited thereto.

이하 실시예를 통해 본 발명의 내용을 보다 상세히 설명한다.Hereinafter, the contents of the present invention will be described in more detail with reference to the following examples.

단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.
However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

인산염 phosphate 가용화Solubilization 균주의 탐색 및 분리 Exploration and Isolation of Strains

인산 가용화 능력이 있는 균을 분리하기 위해서 전국 각지로부터 토양 시료를 수집하였다. 채취한 토양 시료 1 g을 멸균 증류수에 현탁한 후, 트리칼슘포스페이트(Ca3(PO4)2)의 최종 농도가 0.5 % 되도록 조제한 고형 배지에 평판희석법을 이용하여 페트리 디쉬에 도말, 접종하였다. 접종된 배지를 30 ℃ 항온기에서 2 내지 3일 배양하여 생육이 빠르고 투명환이 큰 균주를 인산 가용화능이 있는 균주로 선별하였다(도 1).
Soil samples were collected from all over the country to isolate bacteria with phosphate solubilizing ability. 1 g of the collected soil sample was suspended in sterile distilled water, and then plated and inoculated in a petri dish using a plate dilution method in a solid medium prepared so that the final concentration of tricalcium phosphate (Ca 3 (PO 4) 2 ) was 0.5%. Inoculated medium was incubated for 2 to 3 days at 30 ℃ thermostat was selected as a strain capable of phosphate solubilizing fast growth and large clear ring (Fig. 1).

유전학적 특성 및 지방산 조성을 통한 선별된 미생물 균주의 동정Identification of Selected Microbial Strains by Genetic Properties and Fatty Acid Composition

본 발명자들은 선별된 균주의 종 분석을 위해, 목원대학교 미생물생태자원연구소에 의뢰하여 16S rRNA 유전자 전염기서열 해석 및 세포 지방산 분석을 실시하였다.
The present inventors conducted a 16S rRNA gene infectious sequence analysis and cellular fatty acid analysis by requesting the microbial ecology resource research institute of Mokwon University for species analysis of selected strains.

<2-1> 16S <2-1> 16S rRNArRNA 유전자의  Gene 상동성Homology 분석을 통한 선별된 미생물 균주의 동정 Identification of Selected Microbial Strains Through Analysis

<실시예 1>에서 선별된 균주를 영양 한천 배지(Nutrient agar, Difco, USA)에 접종하고 28 ℃에서 3일간 배양한 후 단일 균주임을 확인하였다. 상기 균주를 영양 배지(Nutrient broth, Difco, USA)에 접종한 후 28 ℃에서 2일간 진탕 배양한 후 벤질 클로라이드(Benzyl chloride)법을 변형한 방법을 이용하여 상기 균주의 게놈 DNA를 추출하였다. 추출된 게놈 DNA에서 상기 균주의 16S rRNA 유전자를 서열번호: 1 및 2로 기재되는 프라이머(primer) 쌍을 이용하여 PCR(Polymerase Chain Reaction) 방법으로 증폭하였다. The strains selected in <Example 1> were inoculated in nutrient agar medium (Nutrient agar, Difco, USA) and incubated at 28 ° C. for 3 days to confirm that they were single strains. The strain was inoculated in a nutrient medium (Nutrient broth, Difco, USA), followed by shaking culture at 28 ° C. for 2 days, and the genomic DNA of the strain was extracted using a modified method of benzyl chloride. In the extracted genomic DNA, the 16S rRNA gene of the strain was amplified by PCR (Polymerase Chain Reaction) method using a primer pair described by SEQ ID NOs: 1 and 2.

프라이머 27F : 5'-AGAGTTTGATCCTGGCTCAG-3'(서열번호: 1),Primer 27F: 5'-AGAGTTTGATCCTGGCTCAG-3 '(SEQ ID NO: 1),

프라이머 1492R : 5'-GGTTACCTTGTTACGACTT-3' (서열번호: 2)
Primer 1492R: 5'-GGTTACCTTGTTACGACTT-3 '(SEQ ID NO: 2)

여기서 PCR 조건은 하기와 같다. PCR conditions are as follows.

95 ℃ 2분,95 minutes 2 minutes,

94 ℃ 30초, ┐94 ℃ 30 seconds, ┐

58 ℃ 30초, │ 30 사이클58 ℃ 30 seconds, │ 30 cycles

72 ℃ 45초, ┘72 ℃ 45 seconds, ┘

72 ℃ 5분, 및5 minutes at 72 ° C., and

4 ℃ 보관
4 ℃ storage

16S rRNA 유전자 PCR 증폭 산물은 PCR Product Purification Kit (Qiagen)를 사용하여 정제하였고, Genetic analyzer 310A(Applied Biosystems)를 사용하여 염기서열을 분석하여 하기의 16 rRNA cDNA 1,333bp의 염기서열을 결정하였다.The 16S rRNA gene PCR amplification product was purified using a PCR Product Purification Kit (Qiagen), and the base sequence of 16 rRNA cDNA 1,333bp was determined by analyzing the nucleotide sequence using Genetic analyzer 310A (Applied Biosystems).

5'-GCTCCTTGGGTGACGAGTGGCGGACGGGTGAGTAATGTCTGGGGATCTGCCCGATGGAGGGGGATAACTACTGGAAACGGTAGCTAATACCGCATAACGTCGCAAGACCAAAGTGGGGGACCTTCGGGCCTCACACCATCGGATGAACCCAGATGGGATTAGCTAGTAGGTGGGGTAACGGCTCACCTAGGCGACGATCCCTAGCTGGTCTGAGAGGATGACCAGCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCCATGCCGCGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGTGTTGAAGTTAATAACTTCAGCAATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTCTGTCAAGTCGGATGTGAAATCCCCGGGCTTAACCTGGGAACTGCATTCGAAACTGGCAGGCTAGAGTCTTGTAGAGGGGGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGAGATCTGGAGGAATACCGGTGGCGAAGGCGGCCCCCTGGACAGAGACTGACGCTCAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCTGTAAACGATGTCGACTTGGAGGTTGTGCCCTTGAGGCGTGGCTTCCGGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGGCCTTGACATCCACGGAATTTGGCAGAGATGCCTTAGTGCCTTCGGGAACCGTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCCTTTGTTGCCAGCGATTCGGTCGGGAACTCAAAGGAGACTGCCGGTGATAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCAGGGCTACACACGTGCTACAATGGCGCATACAAAGAGAAGCGACCTCGCGAGAGCAAGCGGACCTCATAAAGTGCGTCGTAGTCCGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGTAGATCAGAATGCTACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGA-3'(서열번호: 3)
5 '-3' (SEQ ID NO: 3)

상기 균주의 16S rRNA 염기서열은 DDBJ/NCBI/Genebank와 Ribosomal Database Project Ⅱ(RDP Ⅱ)의 데이터베이스에서 상동성 검색을 수행하였다. 분자계통학적 분석결과에서 보는 바와 같이 선별된 균주는 판토에아 속(Pantoea sp.)에 속하는 종으로 판명되었다(도 3). 특히 상기 선발된 균주는 판토에아 속 CRLS069b(FJ593734)와 100 %, 그리고 표준 균주인 펙토박테리움 시프리페디(Pectobacterium cypripedii) DSM3873(AJ233413)의 염기서열과 98.3 %의 유연관계를 나타내는 것으로 확인되어 상기 선별된 균주를 '판토에아 속 J11(Pantoea sp. J11)'로 명명하였다. 상기 균주를 2011년 3월 14일자로 국제기탁기관인 한국생명공학연구원 내 생물자원센터에 기탁하였다(기탁번호; KCTC 11893BP).
The 16S rRNA sequences of the strains were subjected to homology search in the databases of DDBJ / NCBI / Genebank and Ribosomal Database Project II (RDP II). As shown in the molecular analysis results, the selected strain was found to belong to the genus Pantoea sp. (FIG. 3). In particular, the selected strain will remain on the panto in CRLS069b (FJ593734) and 100%, and the type strain of Peg tobak Te Solarium shifted Li Phedi (Pectobacterium cypripedii) the selected strain is identified as representing the base sequence and Genetic Relationship of 98.3% of the DSM3873 (AJ233413) 'in the panto O J11 (Pantoea sp. J11) '. The strain was deposited on March 14, 2011 to the Biological Resource Center of Korea Research Institute of Bioscience and Biotechnology (Accession No .; KCTC 11893BP).

<2-2> 지방산 조성을 통한 선별된 미생물 균주의 동정<2-2> Identification of Selected Microbial Strains Through Fatty Acid Composition

상기 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균주의 지방산 조성을 알아보기 위하여, 상기 균주를 TSA(Trypticase Soy Broth Agar, Difco, USA) 배지에 접종 후, 28 ℃에서 3일간 배양한 후 균체 50 ㎎(건조 중량)을 Ikemoto & Miyagawa의 방법에 의하여 균체 지방산 메틸 에스터(methyl ester)화 및 추출을 수행하였다. 지상산 조성은 Microbial Identification System (MIDI; Microbial ID, Inc., Newark, Del., USA)을 이용하여 6890N Gas Chromatograph(Agilent Technologies)로 분석하였다. Pantoea genus J11 ( Pantoea sp. J11) In order to determine the fatty acid composition of KCTC 11893BP strain, the strain was inoculated in TSA (Trypticase Soy Broth Agar, Difco, USA) medium, and then cultured at 28 ° C. for 3 days, and then 50 mg (dry weight) of cells were obtained from Ikemoto & Miyagawa. Cell fatty acid methyl esterification and extraction were carried out by the method of. The terrestrial acid composition was analyzed by 6890N Gas Chromatograph (Agilent Technologies) using the Microbial Identification System (MIDI; Microbial ID, Inc., Newark, Del., USA).

그 결과 주요 지방산으로 C16 :1 iso(36.99 %)를 나타내었으며, C12 :0, C14 :0, C16:0 그리고 썸드피쳐 3(Summed feature 3)과 같은 다양한 지방산을 함유하는 특징을 나타내었다. 또한 상기 균주는 판토에아 속의 표준 균주들과도 주요 균체 지방산 조성과도 유사한 균체 지방산 조성을 갖는 것으로 확인되었다(표 1 및 도 3).
As a result, C 16 : 1 iso (36.99%) was shown as the main fatty acid, and it was characterized by containing various fatty acids such as C 12 : 0 , C 14 : 0 , C 16: 0 and Summed feature 3 Indicated. In addition, the strain was confirmed to have a cell fatty acid composition similar to the main cell fatty acid composition with standard strains of the genus Pantoea (Table 1 and Figure 3).

판토에아 속(Pantoea sp.) J11의 균체 지방산 특성 Pantoea sp.) Cell fatty acid characteristics of J11 지방산(%)fatty acid(%) 판토에아 속 J11Pantoea in J11 판토에아 디스펄사
(Pantoea
dispersa)LMG2603T
Pantoea Dispulsar
( Pantoea
dispersa ) LMG2603 T
판토에아 아미로보라(Pantoea amylovora)LMG2024T Pantoea Amirobora amylovora ) LMG2024 T
포화 지방산
(Saturated acids)
Saturated fatty acids
(Saturated acids)
C12 :0 C 12 : 0 20.6420.64 4.24.2 5.15.1
C14 :0 C 14 : 0 11.3511.35 3.03.0 4.94.9 C16 :0 C 16 : 0 9.729.72 30.830.8 34.034.0 분지사슬 지방산
(Branched-chain acids)
Branched Chain Fatty Acids
Branched-chain acids
C16 :1 isoC 16 : 1 iso
36.96

36.96

10.6

10.6

24.5

24.5

썸드피쳐 3
(Summed feature 3)

Thumb Feature 3
(Summed feature 3)

7.50

7.50

9.0

9.0

9.1

9.1

액상형 미생물 제제의 제조Preparation of Liquid Microbial Formulations

상기 판토에아 속 J11(Pantoea sp. J11) KCTC 11893BP 균주의 배양은 영양 배지(Nutrient broth, Difco, USA)를 기본으로 사용하였다. 균주는 영양 한천 배지(Nutrient agar, Difco, USA)에서 1 내지 2일간 배양한 후 영양 배지 100 ㎖를 함유하는 250 ㎖ 삼각플라스크에 접종하여 48시간 동안 종균 배양을 하였다. 본 배양을 위해서 액체 배지 1.5 L를 함유하는 3 L 삼각플라스크에 상기 종균 배양액을 접종하여 배양하였다. 배양조건은 30 ℃에서 150 rpm으로 48시간 동안 진탕 배양하였다. 배양된 균은 원심분리기를 이용하여 배양액을 제거하여 균체를 회수하였다. 회수된 균체는 미생물 수가 109~1010 CFU(colony forming unit)/㎖가 되도록 멸균 증류수로 현탁하여 4 ℃에서 보관하고 이를 미생물 제제 원액으로 사용하였다. Pantoea genus J11 ( Pantoea sp. J11) The culture of KCTC 11893BP strain was based on nutrient medium (Nutrient broth, Difco, USA). The strains were cultured in nutrient agar medium (Nutrient agar, Difco, USA) for 1 to 2 days, and then seeded in 250 ml Erlenmeyer flasks containing 100 ml of nutrient medium and seeded for 48 hours. For the incubation, the seed culture was inoculated into a 3 L Erlenmeyer flask containing 1.5 L of liquid medium and cultured. Culture conditions were incubated for 48 hours at 30 ℃ 150 rpm. The cultured cells were recovered by removing the culture solution using a centrifuge. The recovered cells were suspended in sterile distilled water such that the number of microorganisms was 10 9 to 10 10 CFU (colony forming unit) / ml and stored at 4 ° C. and used as a stock solution of microbial preparation.

상기 미생물 제제 원액을 500 ㎖에 균수가 107~108 CFU/㎖가 되도록 희석하여 미생물 제제를 제조하였다.
The microbial preparation was prepared by diluting the stock solution of the microbial preparation to 10 7 ~ 10 8 CFU / ml.

제조된 미생물 제제의 활성 시험Activity test of the prepared microbial agent

상기 <실시예 3>에서 얻은 미생물 제제를 시설하우스 토양 300평에 800배 희석하여 관주 처리하고 8주 후 그 결과를 관찰하였다. The microbial preparations obtained in Example 3 were diluted 800-fold in 300 pyeong of the soil of the facility house and irrigated and observed after 8 weeks.

<4-1> 제조된 미생물 제제의 토양 염류 집적 해소 활성 시험<4-1> Soil Salt Reduction Activity Test of Prepared Microbial Preparation

미생물 제제를 토양에 처리한 결과, 염류장해의 척도인 전기전도도(electrical conductivity, EC)의 수치가 약 20 내지 30 %이상 경감되었고, 또한 치환성 양이온인 칼륨(K), 칼슘(Ca), 마그네슘(Mg)도 30 내지 40 %이상 경감되었다(표 2).As a result of treating the soil with microbial agents, the level of electrical conductivity (EC), which is a measure of salt disturbance, was reduced by about 20 to 30% or more, and the substitutional cations such as potassium (K), calcium (Ca) and magnesium (Mg) was also reduced by 30 to 40% or more (Table 2).

미생물 제제 처리에 따른 토양 분석Soil Analysis According to Microbial Preparation
구분

division

pH

pH
전기
전도도
(EC)
Electricity
conductivity
(EC)

유효인산
(ppm)

Effective Phosphoric Acid
(ppm)

치환성 양이온(me/100g)

Substituent Cation (me / 100g)

유기물
(%)

Organic matter
(%)
칼륨potassium 칼슘calcium 마그네슘magnesium
처리
1

process
One


I'm

7.2

7.2

3.85

3.85

385

385

4.92

4.92

20.1

20.1

4.9

4.9

4.1

4.1


after

7.4

7.4

2.96

2.96

617

617

2.14

2.14

9.35

9.35

3.6

3.6

4.2

4.2

처리
2

process
2


I'm

6.7

6.7

4.57

4.57

423

423

5.85

5.85

17.6

17.6

6.2

6.2

4.0

4.0


after

6.5

6.5

3.65

3.65

507

507

3.04

3.04

6.85

6.85

3.9

3.9

3.8

3.8

<4-2> 제조된 미생물 제제의 식물 생장 촉진 활성 시험<4-2> Plant growth promoting activity test of the prepared microbial agent

또한 미생물제제가 작물의 생육에 미치는 영향을 조사하기 위하여 상추 비닐하우스에 대조구(관행), 반량 처리(1600배 희석), 적량 처리(800배 희석), 배량 처리(400배 희석)로 관주 처리하였다. 정식 후 8주 후 생육 및 수량 조사를 실시하여 평균값으로 계산하였다(표 3). 처리간 평균 비교는 5 % 수준에서 DMRT에 의해 분석하였다.
In addition, in order to investigate the effect of microbial agents on the growth of crops, lettuce plants were irrigated with control (practice), half-treatment (1600-fold dilution), titration (800-fold dilution), and doubling (400-fold dilution). Eight weeks after the establishment, growth and yield surveys were performed and calculated as average values (Table 3). Average comparisons between treatments were analyzed by DMRT at the 5% level.

미생물 제제가 상추의 생육에 미치는 효과Effect of microbial preparations on the growth of lettuce
처 리

process

초 장
(㎝)

Candle sheet
(Cm)

엽 수
(매)

Conifer
(every)

엽 폭
(cm)

Lobe width
(cm)

근 장
(㎝)

Muscle
(Cm)

생체중(g)

Live weight (g)

건물중(g)

(G) in building

엽록체
(㎎/㎡)

Chloroplast
(Mg / ㎡)

엽중

Leaflet

근중

Near-term

엽중

Leaflet

근중

Near-term
대조구
(관행)
Control
(Practice)

27.48b

27.48b

9.74c

9.74c

16.33a

16.33a

13.65c

13.65c

106.0c

106.0c

4.06c

4.06c

5.31c

5.31c

0.42c

0.42c

20.4b

20.4b
반량
처리수
Half volume
Treated water

28.89a

28.89a

10.89c

10.89c

15.95a

15.95a

15.72b

15.72b

117.2c

117.2c

4.90b

4.90b

5.45c

5.45c

0.54bc

0.54bc

21.3b

21.3b
적량
처리구
Suitable amount
Treatment

29.53a

29.53a

12.40b

12.40b

16.78a

16.78a

17.60a

17.60a

151.9b

151.9b

5.46b

5.46b

7.78b

7.78b

0.60b

0.60b

24.6a

24.6a
배량
처리구
Displacement
Treatment

30.40a

30.40a

13.46a

13.46a

18.03a

18.03a

16.37ab

16.37ab

177.1a

177.1a

6.70a

6.70a

9.74a

9.74a

0.77a

0.77a

25.3a

25.3a

상추 초장은 일반 관행에 비해 미생물 제제 시용구에서 다소 길었고, 엽수는 시용량이 증가함으로써 비례적으로 많았는데 대조구에 비해 적량 처리 시용구가 평균 2 ~ 3배(5 ㎝이상의 엽장)가 더 많은 경향을 나타내었다. 엽폭은 처리 구간 유의적인 차이가 없었고 생체중은 적량처리 이상 시용함으로 증가하였다. 뿌리 길이는 시용량이 증가할수록 길었으며 측근 및 세근은 적량 처리 이상 시용구에서 잘 발달하였다.
Lettuce height was slightly longer in microbial application than in general practice, and the number of leaves was proportionately increased with increasing application volume, and the average number of treatments was 2 ~ 3 times higher than those of control. Indicated. Leaf width was not significantly different between treatments and fresh weight was increased by applying more than appropriate treatment. Root length was longer with increasing dose, and the aides and roots were well developed in the right group.

한국생명공학연구원Korea Biotechnology Research Institute KCTC11893BPKCTC11893BP 2011031420110314

<110> Korea Bio Chemical Co., Ltd. <120> The microbial agent with useful soil bacteria and a method for remediation of salt stressed soil using this agent <130> 11p-03-10 <160> 3 <170> KopatentIn 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> An artificially synthesized primer for PCR <400> 1 agagtttgat cctggctcag 20 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> An artificially synthesized primer for PCR <400> 2 ggttaccttg ttacgactt 19 <210> 3 <211> 1333 <212> DNA <213> Pantoea sp. <220> <221> rRNA <222> (1)..(1333) <400> 3 gctccttggg tgacgagtgg cggacgggtg agtaatgtct ggggatctgc ccgatggagg 60 gggataacta ctggaaacgg tagctaatac cgcataacgt cgcaagacca aagtggggga 120 ccttcgggcc tcacaccatc ggatgaaccc agatgggatt agctagtagg tggggtaacg 180 gctcacctag gcgacgatcc ctagctggtc tgagaggatg accagccaca ctggaactga 240 gacacggtcc agactcctac gggaggcagc agtggggaat attgcacaat gggcgcaagc 300 ctgatgcagc catgccgcgt gtatgaagaa ggccttcggg ttgtaaagta ctttcagcgg 360 ggaggaaggt gttgaagtta ataacttcag caattgacgt tacccgcaga agaagcaccg 420 gctaactccg tgccagcagc cgcggtaata cggagggtgc aagcgttaat cggaattact 480 gggcgtaaag cgcacgcagg cggtctgtca agtcggatgt gaaatccccg ggcttaacct 540 gggaactgca ttcgaaactg gcaggctaga gtcttgtaga ggggggtaga attccaggtg 600 tagcggtgaa atgcgtagag atctggagga ataccggtgg cgaaggcggc cccctggaca 660 gagactgacg ctcaggtgcg aaagcgtggg gagcaaacag gattagatac cctggtagtc 720 cacgctgtaa acgatgtcga cttggaggtt gtgcccttga ggcgtggctt ccggagctaa 780 cgcgttaagt cgaccgcctg gggagtacgg ccgcaaggtt aaaactcaaa tgaattgacg 840 ggggcccgca caagcggtgg agcatgtggt ttaattcgat gcaacgcgaa gaaccttacc 900 tggccttgac atccacggaa tttggcagag atgccttagt gccttcggga accgtgagac 960 aggtgctgca tggctgtcgt cagctcgtgt tgtgaaatgt tgggttaagt cccgcaacga 1020 gcgcaaccct tatcctttgt tgccagcgat tcggtcggga actcaaagga gactgccggt 1080 gataaaccgg aggaaggtgg ggatgacgtc aagtcatcat ggcccttacg gccagggcta 1140 cacacgtgct acaatggcgc atacaaagag aagcgacctc gcgagagcaa gcggacctca 1200 taaagtgcgt cgtagtccgg attggagtct gcaactcgac tccatgaagt cggaatcgct 1260 agtaatcgta gatcagaatg ctacggtgaa tacgttcccg ggccttgtac acaccgcccg 1320 tcacaccatg gga 1333 <110> Korea Bio Chemical Co., Ltd. <120> The microbial agent with useful soil bacteria and a method for          remediation of salt stressed soil using this agent <130> 11p-03-10 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> An artificially synthesized primer for PCR <400> 1 agagtttgat cctggctcag 20 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> An artificially synthesized primer for PCR <400> 2 ggttaccttg ttacgactt 19 <210> 3 <211> 1333 <212> DNA <213> Pantoea sp. <220> <221> rRNA (222) (1) .. (1333) <400> 3 gctccttggg tgacgagtgg cggacgggtg agtaatgtct ggggatctgc ccgatggagg 60 gggataacta ctggaaacgg tagctaatac cgcataacgt cgcaagacca aagtggggga 120 ccttcgggcc tcacaccatc ggatgaaccc agatgggatt agctagtagg tggggtaacg 180 gctcacctag gcgacgatcc ctagctggtc tgagaggatg accagccaca ctggaactga 240 gacacggtcc agactcctac gggaggcagc agtggggaat attgcacaat gggcgcaagc 300 ctgatgcagc catgccgcgt gtatgaagaa ggccttcggg ttgtaaagta ctttcagcgg 360 ggaggaaggt gttgaagtta ataacttcag caattgacgt tacccgcaga agaagcaccg 420 gctaactccg tgccagcagc cgcggtaata cggagggtgc aagcgttaat cggaattact 480 gggcgtaaag cgcacgcagg cggtctgtca agtcggatgt gaaatccccg ggcttaacct 540 gggaactgca ttcgaaactg gcaggctaga gtcttgtaga ggggggtaga attccaggtg 600 tagcggtgaa atgcgtagag atctggagga ataccggtgg cgaaggcggc cccctggaca 660 gagactgacg ctcaggtgcg aaagcgtggg gagcaaacag gattagatac cctggtagtc 720 cacgctgtaa acgatgtcga cttggaggtt gtgcccttga ggcgtggctt ccggagctaa 780 cgcgttaagt cgaccgcctg gggagtacgg ccgcaaggtt aaaactcaaa tgaattgacg 840 ggggcccgca caagcggtgg agcatgtggt ttaattcgat gcaacgcgaa gaaccttacc 900 tggccttgac atccacggaa tttggcagag atgccttagt gccttcggga accgtgagac 960 aggtgctgca tggctgtcgt cagctcgtgt tgtgaaatgt tgggttaagt cccgcaacga 1020 gcgcaaccct tatcctttgt tgccagcgat tcggtcggga actcaaagga gactgccggt 1080 gataaaccgg aggaaggtgg ggatgacgtc aagtcatcat ggcccttacg gccagggcta 1140 cacacgtgct acaatggcgc atacaaagag aagcgacctc gcgagagcaa gcggacctca 1200 taaagtgcgt cgtagtccgg attggagtct gcaactcgac tccatgaagt cggaatcgct 1260 agtaatcgta gatcagaatg ctacggtgaa tacgttcccg ggccttgtac acaccgcccg 1320 tcacaccatg gga 1333

Claims (15)

수탁번호 KCTC 11893BP로 기탁된 판토에아 속 J11(Pantoea sp. J11) 균주.
Pantoea sp. J11 strain deposited with accession number KCTC 11893BP.
제 1항에 있어서, 상기 균주는 토양 내 집적된 염류를 분해 또는 제거하는 활성을 가지는 것을 특징으로 하는 균주.
According to claim 1, wherein the strain is characterized in that the strain has the activity to decompose or remove the salt accumulated in the soil.
제 2항에 있어서, 상기 토양은 염류 장해 토양인 것을 특징으로 하는 균주.
The strain according to claim 2, wherein the soil is a salt disturbed soil.
제 1항에 있어서, 상기 균주는 시설 재배지 또는 간척지의 토양에 사용하는 것을 특징으로 하는 균주.
According to claim 1, wherein the strain is characterized in that the strain is used in the soil of plantation or reclaimed land.
제 1항의 균주 또는 이의 배양액을 포함하는 토양 염류 집적 해소용 조성물.
Claim 1 strain or a composition for solving the salt accumulation of soil containing the culture medium thereof.
제 5항에 있어서, 상기 토양은 염류 장해 토양인 것을 특징으로 하는 조성물.
6. The composition of claim 5 wherein the soil is a salt impediment soil.
제 5항에 있어서, 상기 조성물은 시설 재배지 또는 간척지의 토양에 사용하는 것을 특징으로 하는 조성물.
6. The composition of claim 5, wherein the composition is used for soil in facility plantations or reclaimed land.
제 1항의 균주 또는 이의 배양액을 포함하는 식물 생장 촉진제.
Plant growth promoter comprising the strain of claim 1 or its culture.
제 8항에 있어서, 상기 식물 생장 촉진제는 미생물 농약, 종자 코팅제, 미생물 영양제, 토양 개량제, 퇴비 부숙제 및 엽면 살포제로 구성된 군으로부터 선택되는 것으로 사용되는 것을 특징으로 하는 식물 생장 촉진제.
9. The plant growth promoter according to claim 8, wherein the plant growth promoter is used selected from the group consisting of microbial pesticides, seed coating agents, microbial nutrients, soil improvers, composting nutrients and foliar spreading agents.
제 8항에 있어서, 상기 식물 생장 촉진제는 농약 제제로서 사용가능한 담체를 추가로 포함하는 것을 특징으로 하는 식물 생장 촉진제.
9. The plant growth promoter of claim 8, wherein the plant growth promoter further comprises a carrier usable as a pesticide preparation.
제 1항의 균주, 이의 배양액 또는 이를 포함하는 조성물을 토양에 처리하는 단계를 포함하는 염류 집적 해소 방법.
The salt accumulation resolution method comprising the step of treating the soil of claim 1, its culture or composition comprising the same.
제 11항에 있어서, 상기 토양은 염류 장해 토양인 것을 특징으로 하는 방법.
12. The method of claim 11 wherein the soil is a salt barrier soil.
제 11항에 있어서, 상기 균주, 이의 배양액 또는 이를 포함하는 조성물을 시설 재배지 또는 간척지의 토양에 사용하는 것을 특징으로 하는 방법.
12. The method of claim 11, wherein the strain, its culture, or a composition comprising the same is used for soil in a facility cultivation or reclaimed land.
제 1항의 균주, 이의 배양액 또는 이를 포함하는 조성물을 식물 또는 식물을 재배하는 토양에 처리하는 단계를 포함하는 식물 생육 촉진 방법.
Claim 1, a method for promoting plant growth comprising the step of treating the plant or a culture containing the same or a composition comprising the same to the plant or the soil for growing the plant.
제 14항에 있어서, 상기 처리는 식물의 종자에 도포하는 처리, 식물의 종자를 침지시키는 처리, 식물의 재배 토양에 관주(灌注)하는 처리, 식물의 재배 토양 표면에 분무하는 처리, 식물의 경엽에 살포하는 처리, 및 식물의 부상부(付傷剖)에 접촉시키는 처리로 이루어지는 군으로부터 선택되는 어느 하나인 것을 특징으로 하는 방법.The treatment according to claim 14, wherein the treatment is a treatment applied to the seed of the plant, a treatment of immersing the seed of the plant, a treatment irrigation to the cultivated soil of the plant, a treatment sprayed on the surface of the cultivated soil of the plant, the foliage of the plant It is any one selected from the group which consists of a process which sprays on and the process which makes contact with the floating part of a plant.
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