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KR20160128533A - Soil-microbial product containing new microorganism enterobacter sp. 04-p-1 having soluble phosphate activity and manufacturing method thereof - Google Patents

Soil-microbial product containing new microorganism enterobacter sp. 04-p-1 having soluble phosphate activity and manufacturing method thereof Download PDF

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KR20160128533A
KR20160128533A KR1020150059834A KR20150059834A KR20160128533A KR 20160128533 A KR20160128533 A KR 20160128533A KR 1020150059834 A KR1020150059834 A KR 1020150059834A KR 20150059834 A KR20150059834 A KR 20150059834A KR 20160128533 A KR20160128533 A KR 20160128533A
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김정선
원항연
박인철
유재홍
이관형
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대한민국(농촌진흥청장)
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Abstract

본 발명은 인산가용화 활성을 갖는 신규 미생물 엔테로박터 에스피 04-P-1 균주 및 이를 포함하는 토양미생물 비료에 관한 것이다.
본 발명의 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P)는 서열번호 1로 표현되는 염기서열로 이루어지며 인산 가용화 활성을 갖는 것이 특징이며, 내염성도 갖는 것이 특징이다.
본 발명에 의해, 신규 미생물 엔테로박터 에스피 04-P-1 균주 및 이를 포함하는 토양미생물 비료를 난용성인산염이 접적된 시설재배지에 제공됨에 따라 토양에서 쉽게 불용화되는 인산질 비료가 식물이 이용할 수 있는 형태로 가용화됨으로써, 토양에 집적되는 염류가 줄어들어 염류 장해로 인한 피해가 감소하게 된다.
The present invention relates to a novel microorganism Enterobacter sp. 04-P-1 strain having a phosphoric acid solubilizing activity and a soil microbial fertilizer containing the same.
The inventive Enterobacter The strain 04-P-1 (KACC 92041P) is composed of the nucleotide sequence shown in SEQ ID NO: 1 and is characterized by having a phosphoric acid solubilizing activity and also has salt resistance.
The present invention provides a new microbial Enterobacter sp. 04-P-1 strain and a soil microbial fertilizer containing the microbial enterobacter sp. 04-P-1 as provided in facility cultures with poorly soluble phosphate, , The amount of salts accumulated in the soil is reduced, and the damage caused by the salt obstacle is reduced.

Description

인산가용화 활성을 갖는 신규 미생물 엔테로박터 에스피 04-P-1 균주 및 이를 포함하는 토양미생물 비료{Soil-microbial product containing new microorganism enterobacter sp. 04-p-1 having soluble phosphate activity and manufacturing method thereof}TECHNICAL FIELD The present invention relates to a novel microorganism Enterobacter sp. 04-P-1 and a soil microorganism fertilizer containing the microorganism. 04-p-1 having soluble phosphate activity and manufacturing method thereof}

본 발명은 인산가용화 활성을 갖는 신규 미생물 엔테로박터 에스피 04-P-1 균주 및 이를 포함하는 토양미생물 비료에 관한 것으로서, 더욱 상세하게는 토양에서 쉽게 불용화되는 인산질 비료를 식물이 이용할 수 있는 형태로 가용화함으로써 토양에 집적되는 염류를 줄이고 토마토의 생육을 촉진시키는 신규 미생물 엔테로박터 에스피 04-P-1 균주 및 이를 포함하는 토양미생물 비료에 관한 것이다.The present invention relates to a novel microorganism Enterobacter sp. 04-P-1 having a solubilizing activity for phosphoric acid and a soil microorganism fertilizer containing the same. More particularly, the present invention relates to a novel microbial enterobacteria fertilizer P-1 strain and soil microorganism fertilizer containing the microorganism Enterobacter sp. 04-P-1 strain, which reduces salt accumulated in the soil by solubilization and promotes the growth of tomato.

인(phosphorus)은 식물의 생장과 발육을 위해 매우 중요한 필수적 영양원소로서 대량의 공급이 필요한 대량영양원소(macronutrient)이다. 따라서 농업에 있어서 주요 작물의 최대 수확을 위해서는 식물이 가용할 수 있는 인의 유무는 매우 중요한 조건이 된다. Phosphorus is a macronutrient that is a vital nutrient that is very important for the growth and development of plants and requires a large supply. Therefore, for the maximum harvesting of major crops in agriculture, the presence of phosphorus that is available to plants is a very important condition.

그러나, 토양내 대부분의 인은 불용성 상태이어서 식물은 사용할 수가 없다.However, most of the phosphorus in the soil is insoluble and plants can not be used.

즉, 토양내 인은 화학적으로 공고하게 결합된 불용성 복합체나 유기적으로 결합된 파이테이트(phytate)와 같은 상태로 존재하여 식물이 전혀 사용할 수 없는 불용성 인산염으로 존재한다.That is, phosphorus in soil exists as a chemically cohesively insoluble complex or an organically bound phytate and exists as an insoluble phosphate that plants can not use at all.

따라서, 자연상태의 토양에서 가용할 수 있는 인은 미량영양원소들 (micronurtients)의 농도보다도 적은 양으로 존재하고 있다. 결국, 가용성 인의 농도가 토양에서 식물의 생장과 발육의 증진에 있어 제한요소인 것이다. 더욱이, 토양에 가용성 인이 든 무기인산비료를 공급해도 이들 중 무려 75%이상이 매우 빠르게 불용성 인산염 상태로 변화되어, 식물은 공급된 가용성 인의 25% 이하만을 사용할 수 있다.Thus, phosphorus available in natural soils is present in an amount less than the concentration of micronurti- ties. As a result, the concentration of soluble phosphorus is a limiting factor in promoting plant growth and development in the soil. Moreover, even if inorganic phosphoric acid fertilizers soluble in soil are supplied, more than 75% of them are converted into insoluble phosphate state very quickly, and plants can use only less than 25% of the supplied soluble phosphorus.

따라서, 농부들은 가용성 인의 부족에 따른 식물의 생장 및 발육의 저하를 막고자 계속해서 과도한 무기인산비료를 농토에 공급하는 과도한 시비를 반복하게된다. 그러나, 이렇게 과도한 시비로 공급되는 대부분의 가용성 인은 농토에서 다량의 불용성 인산염으로 변화되고 이것이 축적되어 결국은 식물의 생장 및 발육이 정상적으로 이루어지지 않는 염류장해 토양이 된다.Therefore, the farmers repeated the excessive fertilizing of supplying the excessive inorganic phosphoric acid fertilizer to the farmland in order to prevent the decrease of the growth and development of the plant due to the lack of soluble phosphorus. However, most of the soluble phosphorus supplied in such excess fertilization is converted into a large amount of insoluble phosphate in the agricultural land, which accumulates and eventually becomes a salt obstacle soil in which plant growth and development are not normally performed.

더욱이 이런 염류장해토양은 고부가가치의 작물을 생산하는 시스템으로 널리 보급된 시설재배지에서 더욱 자주 발생하여 농업생산성 및 농업경제에 있어 매우 큰 문제가 되고 있다.In addition, these polluted soils become more and more frequent in plantation areas, which are widely used as a system for producing high value crops, which is a great problem in agriculture productivity and agricultural economy.

그러나, 다행스럽게도 토양에는 이러한 불용성 인산염을 가용화 상태로 전환 할 수 있는 일부의 미생물들이 있음이 알려졌다. 이들 불용성 인산염 가용화 미생물(phosphate solubilizing bacteria, PSB)은 식물의 근권부에서 유기산을 분비하거나 특정 효소를 분비하여 토양의 불용성 인산염을 가용화 인으로 전환시켜 식물이 흡수하여 사용할 수 있도록 하는 일종의 공생관계를 유지할 것으로 추측되고 있다.Fortunately, however, it is known that there are some microorganisms in the soil that can convert such insoluble phosphate into a solubilized state. These insoluble phosphate solubilizing bacteria (PSBs) maintain a kind of symbiotic relationship that allows the plants to absorb and use insoluble phosphate by converting the insoluble phosphate of the soil into soluble phosphorus by secreting organic acids or releasing specific enzymes from the rhizosphere of the plant .

PSB 미생물은 슈도모나스(Pseudomonas), 바실러스(Bacillus), 라이조비움 (Rhizobium), 부르크홀데리아(Burkholderia), 아크로모박터(Achromobacter), 아그로박테리움(Agrobacterium), 마이크로코코스(Microccocus), 아에레오박터 (Aereobacter), 플라보박테리움(Flavobacterium)과 에위니아(Erwinia)에 속하는 몇몇 종들에서만 발견되었다.PSB microorganism Pseudomonas (Pseudomonas), Bacillus (Bacillus), rayijo Away (Rhizobium), Burkholderia (Burkholderia), Achromobacter (Achromobacter), Agrobacterium (Agrobacterium), micro Cocos (Microccocus), oh Leo bakteo ( Aleobacter ), Flavobacterium ( Flavobacterium ) and some species belonging to Erwinia .

그러나, 상기 균주들의 불용성 인산염 가용성 스펙트럼을 조사한 결과, 특정 불용성 인산염에 대해서만 가용성이 있거나 혹은 그 가용성이 광범위한 경우 전반적으로 낮은 활성을 보였다.However, the insoluble phosphate-soluble spectrum of the above-mentioned strains was examined. As a result, it was found that the insoluble phosphate was soluble only for a specific insoluble phosphate or exhibited low overall activity when its solubility was wide.

PSB 미생물은 토양 내에 존재하기는 하지만 항상 그 세포수가 식물 근권을 형성하는 다른 미생물들과 경쟁하여 우점할 정도로 충분히 높지 않다. 따라서, 토양내 PSB 미생물들이 불용성 인산염을 분해하여 만드는 가용성 인의 양은 식물의 생장과 발육을 증가시킬 정도로 높지 않다. 그러므로 인위적으로 다량의 PSB 미생물을 토양에 접종할 경우 식물의 생장과 발육을 증가시킬 수 있는 유용성이 있다.Although PSB microorganisms are present in soil, they are not always high enough to dominate by competing with other microorganisms that form plant rhizome. Therefore, the amount of soluble phosphorus produced by degradation of insoluble phosphate by PSB microorganisms in soil is not high enough to increase plant growth and development. Therefore, inoculation of a large amount of PSB microorganism into the soil is useful for increasing the growth and development of the plant.

이처럼 불용성 인산염 가용화 미생물을 사용하여 식물 생장을 증진시키고자하는 연구가 수행되어 효과가 있다고 보고되었다.It has been reported that the insoluble phosphate solubilizing microorganisms are used to enhance plant growth.

즉, 기존에 알려진 PSB 미생물외에 신규한 PSB 미생물을 발굴해내는 노력이 경주 되어 라넬라 아쿠아틸리스(Rahnella aquatilis)가 하이드록시아파타이트 (hydroxyapatite)를 가용화 할 수 있음이 보고된 바있다.In other words, it has been reported that Rahnella aquatilis can solubilize hydroxyapatite in an effort to discover new PSB microorganisms in addition to PSB microorganisms already known.

또한, 엔테로박터 아글로메란스(Enterobacter agglomerans)도 하이드록시아파타이트(hydroxyapatite)를 분해하여 가용성 인을 생성할 수 있음을 확인한 바 있다.In addition, it has been confirmed that Enterobacter agglomerans can also decompose hydroxyapatite to generate soluble phosphorus.

한편, 한국공개특허공보 특2002-0017516(화산회토양에 고정된 인산염을 가용화하는 새로운 미생물)에는 화산회 토양에 고정된 인산염을 가용화시키는 바실러스 스패리쿠스 PSB-13에 관한 것이 공개되어 있다.On the other hand, Korean Patent Laid-Open Publication No. 2002-0017516 (a new microorganism that solubilizes phosphate fixed on volcanic rock soils) discloses Bacillus sp. Picus PSB-13 which solubilizes phosphate fixed on volcanic soils.

그러나, 여전히 작물재배 토양내의 불용성 인산염을 가용화 하는 새로운 균주의 개발이 요구되고 있다.However, there is still a need to develop new strains that solubilize insoluble phosphate in crop growing soils.

본 발명의 목적은 토양에서 쉽게 불용화되는 인산질 비료를 식물이 이용할 수 있는 형태로 가용화함으로써 토양에 집적되는 염류를 줄이고 토마토의 생육을 촉진시키는 신규 미생물 엔테로박터 에스피 04-P-1 균주 및 이를 포함하는 토양미생물 비료를 제공하는데 있다.The object of the present invention is to provide a novel microorganism Enterobacter spi 04-P-1 strain which reduces the salts accumulated in the soil and promotes the growth of tomato by solubilizing the phosphoric acid fertilizer which is easily insolubilized in the soil in a form that plants can use, To provide soil microbial fertilizers.

본 발명이 이루고자 하는 기술적 과제들은 이상에서 언급한 기술적 과제들로 제한되지 않는다.The technical objects to be achieved by the present invention are not limited to the above-mentioned technical problems.

상기 목적을 달성하기 위한, 본 발명의 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P)는 서열번호 1로 표현되는 염기서열로 이루어지며 인산 가용화 활성을 갖는 것이 특징이며, 내염성도 갖는 것이 특징이다.In order to achieve the above object, the Enterobacter sp. 04-P-1 strain (KACC 92041P) of the present invention is characterized by having a base sequence represented by SEQ ID NO: 1 and having a phosphate solubilizing activity, .

특히, 상기 균주는 토양에서 불용화되는 인산질 비료를 가용화하여 토마토의 생육을 촉진하는 것이 특징이며, 미생물 농약, 종자코팅제, 토양개량제, 퇴비부숙제, 엽면살포제 또는 관주살포제 중 어느 하나의 유효성분으로 이용된다.In particular, the strain is characterized by solubilizing phosphoric acid fertilizer insolubilized in the soil to promote the growth of tomatoes. The strain is effective as a microbial pesticide, a seed coating agent, a soil improving agent, a composting homework, a foliar spraying agent, .

본 발명의 토양미생물 제제는 서열번호 1로 표현되는 염기서열로 이루어지며 인산 가용화 활성을 갖는 신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P)를 유효성분으로 포함하는 것이 특징이다. The soil microbial agent of the present invention comprises a nucleotide sequence represented by SEQ ID NO: 1 and is a novel microorganism Enterobacter sp.) 04-P-1 strain (KACC 92041P) as an active ingredient.

본 발명의 토양미생물 제제의 제조방법은, 서열번호 1로 표현되는 염기서열로 이루어지며 인산 가용화 활성을 갖는 신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P)를 준비하는 단계 및, 상기 균주를 TSB배지에 접종한 후 20~35℃, pH 6~7의 조건하에서 배양하는 단계를 포함하는 것이 특징이다.The method for producing a soil microorganism preparation according to the present invention comprises a step of culturing a microorganism Enterobacter ( Enterobacter sp.) Comprising a nucleotide sequence represented by SEQ ID NO: 1 and having a phosphate solubilizing activity (KACC 92041P), and inoculating the strain into a TSB medium, followed by culturing under the conditions of 20 to 35 ° C and a pH of 6 to 7.

본 발명의 토양미생물 비료는 상기 서열번호 1로 표현되는 염기서열로 이루어지며 인산 가용화 활성을 갖는 신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P)의 균체 또는 상기 토양미생물 제제 중 어느 하나 이상을 유효성분으로 포함하는 것이 특징이다.The soil microorganism fertilizer of the present invention comprises a nucleotide sequence represented by SEQ ID NO: 1 and is a novel microorganism Enterobacter 04-P-1 strain (KACC 92041P) or the soil microorganism preparation as an active ingredient.

본 발명은 상기 토양미생물 비료를 토양에 관주처리하여 토양내 인산가용화능을 높여주는 방법을 제공한다.The present invention provides a method for enhancing the solubilization capacity of the soil in soil by treating the soil microbial fertilizer with the soil.

또는, 상기 토양미생물 비료를 3~4배 희석한 후 토마토 재배 토양에 관주처리하여 토양내 불용성 인산염을 가용화하여 토마토의 생육을 촉진하는 방법을 제공한다.Alternatively, there is provided a method for promoting the growth of tomato by solubilizing insoluble phosphate in the soil by treating the soil microorganism fertilizer 3 to 4 times and then cultivating the tomato growing soil.

본 발명에 의해, 신규 미생물 엔테로박터 에스피 04-P-1 균주 및 이를 포함하는 토양미생물 비료를 난용성 인산염이 접적된 시설재배지에 제공됨에 따라 토양에서 쉽게 불용화되는 인산질 비료가 식물이 이용할 수 있는 형태로 가용화됨으로써, 토양에 집적되는 염류가 줄어들어 염류 장해로 인한 피해가 감소하게 된다. The present invention provides a new microbial Enterobacter sp. 04-P-1 strain and a soil microbial fertilizer containing the microbial enterobacter sp. 04-P-1 as provided in facility cultures with poorly soluble phosphate, , The amount of salts accumulated in the soil is reduced, and the damage caused by the salt obstacle is reduced.

또한, 상기 균주를 토양에 관주처리 시 토마토의 생육이 촉진되어 기존 토마토 재배율보다 더 효율적인 토마토 재배율을 갖게 된다.In addition, when the strain is cultivated in the soil, the growth of tomato is promoted, and the tomato growth rate is more efficient than the existing tomato growth rate.

아울러, 인산질 비료 처리량 감소로 농가 경제여건에 도움이 된다.In addition, the reduction in phosphorus fertilizer throughput is beneficial to farmers' economic conditions.

도 1은 본 발명의 선발균주인 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주의 투명대를 나타낸 도면이다.
도 2는 도 1의 선발균주에 대하여 세균 16S rDNA 염기서열 상동성 및 유연관계 분석으로 인산가용화 미생물 Enterobacter sp. 04-P-1 동정한 결과를 나타낸 그래프이다.
도 3은 도 1의 선발균주에 대하여 인산 가용화능 정량 검정 및 pH 측정결과를 나타낸 그래프이다.
도 4는 도 1의 선발균주에 대하여 적정생육온도의 결과를 나타낸 그래프이다.
도 5는 도 1의 선발균주에 대하여 적정생육pH의 결과를 나타낸 그래프이다.
도 6은 도 1의 선발균주에 대하여 내염성을 확인한 그래프이다.
도 7은 도 1의 선발균주의 토양에 처리시 토양 내 유효인산량을 측정한 그래프이다.
FIG. 1 is a graph showing the effect of the Enterobacter sp.) 04-P-1 strain.
Fig. 2 is a graph showing the bacterial 16S rDNA nucleotide sequence homology and cross- linking analysis of the selected strain of Fig. 1, showing that the soluble bacterial Enterobacter sp. 04-P-1.
FIG. 3 is a graph showing the results of quantitative determination of pH-soluble solubility and pH measurement of the selected strain of FIG. 1;
FIG. 4 is a graph showing the results of the optimum growth temperature for the selected strain of FIG. 1; FIG.
FIG. 5 is a graph showing the results of the optimum growth pH for the selected strain of FIG. 1; FIG.
FIG. 6 is a graph showing the salt tolerance of the selected strain of FIG. 1; FIG.
FIG. 7 is a graph showing an effective amount of acid in the soil during the treatment of the soil of the selected strain of FIG. 1; FIG.

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

본 발명은 난용성인산(Ca3(PO4)2)이 첨가된 Pikovskaya 배지에 산림 토양에서 채집한 미생물을 배양하여 투명대를 형성하는 인산 가용화 균주를 선발하고, 이를 대상으로 세균 16S rDNA 염기서열 상동성 및 유연관계를 분석한 바, 상기 선발균주는 엔테로박터 에스피(Enterobacter sp.)의 신규한 균주로 밝혀졌다.The present invention relates to a method for isolating a bacterial strain capable of solubilizing phosphoric acid, which forms a zona pellucida, by culturing a microorganism collected from forest soil in a Pikovskaya medium supplemented with hardly soluble acid (Ca 3 (PO 4 ) 2 ) As a result of the analysis of homology and flexibility, the selected strain was found to be a novel strain of Enterobacter sp.

이에, 본 발명은 상기 선발균주를 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주로 명명하고, 이 균주를 국립농업과학원 농업유전자원센터에 기탁하여 2015년 2월 27일 수탁번호 KACC 92041P를 부여받았으며, 이는 서열번호 1로 기재되는 염기서열 전체 또는 그것의 일부로 이루어지며, 1,484bp의 오픈 리딩 프레임(ORF)으로 구성됨을 확인하였다. Therefore, the present invention named this strain as Enterobacter sp. 04-P-1 strain, which was deposited with the National Institute of Agricultural Science and Technology, National Institute of Agricultural Science and Technology on Feb. 27, 2015 under accession number KACC 92041P And it was confirmed that it consists of the entire nucleotide sequence shown in SEQ ID NO: 1 or a part thereof and is composed of an open reading frame (ORF) of 1,484 bp.

이렇게 구성된 상기 신규균주의 인산 가용화능을 정량 검정하고 pH를 측정한 바, 배양 기간이 지남에 따라 점차 유효 인산량이 증가하여 미생물의 인산가용화 효과가 확인되었으며, 배양 7일차에 597.6ppm까지 증가됨이 확인되었다. The phytic acid solubilizing ability of the new strain thus constructed was quantitatively assayed and its pH was measured. As a result, the effective amount of phosphoric acid was gradually increased over the period of incubation, and the effect of solubilizing the microorganism was confirmed, and it was confirmed to increase to 597.6 ppm on the 7th day of culture .

또한, 상기 신규균주의 인산 가용화 균주로써의 생리적 특성을 검정하여 최적 생육 범위를 조사한 바, 적합생육온도는 20~35℃, 적합생육pH는 6~7, 내염성 범위는 염(NaCl)함유량이 배지전체조성 대비 0~10중량%이하인 것으로 확인되었다. The optimum growth range was examined by examining the physiological characteristics of the novel strain as a phosphate-solubilizing strain. The optimum growth temperature was 20 to 35 ° C, the optimum growth pH was 6 to 7, and the salt tolerance range was such that the salt (NaCl) It was confirmed that the content was 0 to 10% by weight or less based on the total composition.

또한, 토양 내에서의 상기 신규균주의 인산 가용화능 정량을 분석한 바, 761ppm으로 대조구에 비해 91 % 증가함을 확인하였다.In addition, the amount of phosphoric acid solubilization of the new strain in the soil was analyzed to be 761 ppm, which was 91% higher than that of the control.

또한, 상기 인산가용화 신규균주에 의한 시설재배지 내 식물 생장효과 분석하기 위해 난용성인산염이 집적된 시설재배지 토양에 상기 신규균주 Enterobacter sp. 04-P-1 배양액을 4배 희석하여 1주에 한 번씩 총 4회 관주 처리한 결과, 상기 신규균주 처리하지 않은 대조군 대비 토마토 과실수량 34.0%, 과중 14.8 %, 초장 17.1%가 증가됨을 확인하였다.In order to analyze the plant growth effect in the planting area by the above-mentioned new strain for solubilizing phosphoric acid, the new strain Enterobacter sp. 04-P-1 cultures were diluted 4-fold and treated once a week for 4 times. As a result, it was confirmed that tomato fruit yield 34.0%, fruit weight 14.8% and plant height 17.1% were increased compared with the untreated control strain .

이상 상기 결과를 토대로, 본 발명에서 선발된 신규균주 Enterobacter sp. 04-P-1는 토양에서 쉽게 불용화되는 인산질 비료를 식물이 이용할 수 있는 형태로 가용화함으로써 토양에 집적되는 염류를 줄이고 토마토의 생육을 촉진시키는 효과를 갖는 것을 알아내었다.Based on the above results, the novel strain Enterobacter sp. 04-P-1 has found that phosphate fertilizer, which is easily insolubilized in soil, is solubilized in a form that can be used by plants, thereby reducing salts accumulated in the soil and promoting the growth of tomatoes.

본 발명의 토양미생물 제제(배양액)는 상기 신규 균주를 포함하는 것으로, 상기 균주의 생장온도 및 pH는 어느 조건이든 균주의 인산가용화 활성을 나타내므로 조건을 한정하지 않으나, 상기 실험결과를 토대로 특히 20~35℃에서 pH 6~7일때 균주활성이 잘 일어남을 알 수 있다. 또한, 상기 균주는 내염성을 갖으며 그 범위가 염(NaCl)함유량이 배지전체조성 대비 0~10중량%이하인 것을 확인하였다.Since the soil microorganism preparation (culture solution) of the present invention includes the novel strain, the growth temperature and the pH of the strain exhibit the phosphate solubilizing activity of the strain under any condition, and thus the conditions are not limited. However, It can be seen that when the pH is 6 to 7 at 35 ° C, the strain activity is well established. In addition, it was confirmed that the strain had salt resistance and its salt (NaCl) content was 0 to 10% by weight or less with respect to the whole composition of the culture medium.

본 발명의 토양미생물 비료는 상기와 같은 신규 균주의 균체 또는 상기 토양미생물 제제 중 어느 하나이상을 포함하는 것이 특징이다.The soil microbial fertilizer of the present invention is characterized in that it comprises any one or more of the microbial cells of the novel strain or the soil microbial agent.

이러한 본 발명의 토양미생물 비료에는 미생물 보호제로서 적합한 글리세롤, 분유, 우유, 탈지분유 또는 동식물유 등을 추가로 첨가할 수 있으며, 영양 공급원으로서 섬유소 분말, 대두박 분말, 면실박 분말, 각종 동식물성 펩톤, 밀기울 분말, 옥수수 추출물, 효모 추출액, 당밀, 포도당, 설탕, 옥수수시럽 등과 같은 탄소원 및 질소원과 무기염류 등을 추가로 통상의 양으로 포함하여 상기 균주의 효능을 더 촉진하도록 한다.The soil microorganism fertilizer of the present invention may further contain glycerol, milk powder, milk, skim milk powder, animal or vegetable oil, etc. suitable as a microorganism protecting agent and may be added as a nutrient source such as fibrinogen powder, soybean meal powder, cottonseed powder, The present invention further includes a carbon source and nitrogen source such as bran powder, corn extract, yeast extract, molasses, glucose, sugar, corn syrup and the like and inorganic salts in an ordinary amount to further promote the efficacy of the strain.

상기와 같은 방법에 의해 수득된 토양미생물 비료는 일반적인 제형기를 사용하여 적당한 크기의 분말, 펠렛 또는 과립 등의 다양한 형태로 제형화 될 수 있다.The soil microorganism fertilizer obtained by the above method can be formulated into various forms such as powders, pellets or granules of appropriate size using a general formulator.

상기에서 제형화된 본 발명의 미생물 비료는 그대로 사용되거나 또는 실온에서 풍건하거나 동결 건조 또는 고온 건조 방법으로 건조시켜 사용할 수 있다. 또한, 쌀겨와 혼합하고 이를 상온에서 확대 배양한 후 토양에 적용시킬 수도 있다.The microbial fertilizer of the present invention formulated in the above may be used as it is or may be dried by air drying at room temperature or by lyophilization or high temperature drying method. It may also be applied to soil after mixing with rice bran and expanding it at room temperature.

이러한 본 발명의 토양미생물 비료를 토양에 적용하는 경우 토양에 관주하는 방법을 적용하여 토양내 인산가용화 능을 높여주며, 더욱 상세하게는 토양미생물 비료를 3~4배 희석한 후 토마토 재배 토양에 관주처리함으로써 토양내 불용성 인산염을 가용성 인산염으로 전환시켜줌으로서 토마토의 인산 흡수 및 생육을 촉진할 수 있는 효능이 장기간 동안 유지하게 된다.When the soil microbial fertilizer of the present invention is applied to the soil, the method of soil-related application improves the solubilization ability of the soil in the soil, more specifically, the soil microbial fertilizer is diluted 3 to 4 times, By treating the insoluble phosphate in the soil to soluble phosphate, the effect of promoting phosphoric acid uptake and growth of the tomato is maintained for a long period of time.

이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

<< 실시예1Example 1 > 인산 > Phosphoric acid 가용화Solubilization 균주의 선발 Selection of strain

1. 실험방법1. Experimental Method

난용성인산(Ca3(PO4)2)이 0.5% 첨가된 Pikovskaya 배지(1% Glucose, 0.05% (NH4)2SO4, 0.01% MgSO4·H2O, 0.02% KCl, 0.001% MnSO4, 0.5% Ca3(PO4)2, 1.8% Agar, pH 7.0)에 산림 토양에서 채집한 미생물을 배양하여 투명대를 형성하는 인산가용화 균주 선발하였다.Sparingly soluble adult acid (Ca 3 (PO 4) 2 ) is 0.5% of Pikovskaya medium (1% Glucose, 0.05% ( NH 4) 2 SO 4, 0.01% MgSO 4 · H 2 O, 0.02% KCl, 0.001% MnSO 4 , 0.5% Ca 3 (PO 4 ) 2 , 1.8% Agar, pH 7.0) were cultivated in the forest soil and selected for solubilization of phosphoric acid to form zona pellucida.

2. 실험결과 2. Experimental results

상기 실험결과 도 1에 나타나있듯이, 난용성인산(Ca3(PO4)2)이 0.5% 첨가된 Pikovskaya 배지 상에 투명대를 형성하는 균주하나가 표시됨을 확인하였으며, 이는 인산 가용화 능을 갖는 것으로 인지한 바, 이를 선발해 내고 이를 '04-P-1'로 표기하였다. As shown in FIG. 1, it was confirmed that one strain forming a zona pellucida was displayed on the Pikovskaya medium supplemented with 0.5% of hardly soluble acid (Ca 3 (PO 4 ) 2 ) It was selected as '04-P-1 '.

<< 실시예Example 2> 인산  2> Phosphorus 가용화Solubilization 균주의 분자생물학적 동정 Molecular biology of the strain

1. 실험방법1. Experimental Method

상기 실시예 1에서 선발된 신규균주를 대상으로 세균 16S rDNA 영역 염기서열 분석 및 세균 type strain의 유연관계를 DNAman program으로 분석하여 동정하였다.Bacterial 16S rDNA region sequencing analysis and the relationship of bacterial type strain were analyzed with the DNAman program for the new strains selected in Example 1 above.

2. 실험결과 2. Experimental results

상기 실험결과 도 2에 나타나 있듯이, 상기 실시예 1에서 선발된 신규균주는 인산 가용화 신규미생물 Enterobacter sp. 균주로 동정됨을 확인하였다.As a result of the above-mentioned experiment, as shown in Fig. 2, the new strain selected in Example 1 was a novel microorganism Enterobacter sp. Was identified as a strain.

이에 첨부된 기탁증명서에 기재되어 있듯이, 상기 신규균주는 토양 신규미생물인 엔테로박터 에스피(Enterobacter sp .) 04-P-1로 명명하였으며, 이 균주를 국립농업과학원 농업유전자원센터에 기탁하여 2015년 2월 27일 수탁번호 KACC 92041P를 부여받았다.As described in the deposit document attached thereto, the new strain is a new soil microorganism, Enterobacter sp . ) 04-P-1. This strain was deposited with the National Institute of Agricultural Science and Technology, National Institute of Agricultural Science and Technology, and received the accession number KACC 92041P on February 27, 2015.

<< 실시예Example 3> 인산  3> Phosphoric acid 가용화능Solubility 정량검정 및  Quantitative assay and pHpH 측정 Measure

1. 실험방법1. Experimental Method

난용성인산염(Ca3(PO4)2)이 0.5% 첨가된 TCP 배지(1% Glucose, 0.05% (NH4)2SO4, 0.01% MgSO4·7H2O, 0.02% KCl, 0.001% MnSO4, 0.5% Ca3(PO4)2, pH 7.0) 100ml에 상기 실시예 1에서 선발한 균주 1㎖(약 6×109/㎖)을 접종하였다.0.05% (NH 4 ) 2 SO 4 , 0.01% MgSO 4揃 7H 2 O, 0.02% KCl, 0.001% MnSO 4 ) containing 0.5% of weakly soluble phosphate (Ca 3 (PO 4 ) 2 ) 4, was inoculated with 0.5% Ca 3 (PO 4) 2, pH 7.0) strains selected in example 1 in 100ml 1㎖ (approximately 6 × 10 9 / ㎖).

그 다음, 상기 접종배지를 28℃, 150rpm으로 진탕배양 후 원심 분리하여 상등액의 유효인산 농도와 pH를 7일간 측정하였다.Then, the inoculation medium was shaken at 28 DEG C and 150 rpm, centrifuged, and the effective phosphoric acid concentration and pH of the supernatant were measured for 7 days.

이때, 상기 유리인산 농도는 Phosphate Colorimetric Assay Kit(BioVision co.)를 이용하여 30분간 추출액과 반응시켜 650nm에서 흡광도를 측정하여 유효인산 농도를 계산하였다. At this time, the free phosphoric acid concentration was measured by using a Phosphate Colorimetric Assay Kit (BioVision co.) For 30 minutes, and the absorbance at 650 nm was measured to calculate the effective phosphoric acid concentration.

2. 실험결과2. Experimental results

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

시간(일)Time (days) 00 1One 22 33 44 55 66 77 88 유효인산(ppm)Effective phosphoric acid (ppm) 00 278.3278.3 418.5418.5 471.4471.4 503.4503.4 539.3539.3 533.2533.2 597.6597.6 556.4556.4 pHpH 6.396.39 4.664.66 4.524.52 4.504.50 4.494.49 4.394.39 4.414.41 4.444.44 4.444.44

상기 표 1 및 도 3에 나타나 있듯이, 배양 기간이 지남에 따라 점차 유효 인산량이 증가하여 미생물의 인산가용화 효과가 확인되었으며, 배양 7일차에 597.6ppm까지 증가함을 확인하였다. As shown in Table 1 and FIG. 3, the phosphoric acid solubilization effect of the microorganism was confirmed by increasing the effective phosphoric acid amount gradually over the incubation period, and it was confirmed that it increased to 597.6 ppm at the 7th day of cultivation.

pH의 변화는 미생물 처리 전 pH는 6.39에서 배양기간 동안 4.44까지 감소하였고, 유효인산 생성량이 많을수록 pH는 낮아져 역상관관계를 나타내었음을 확인하였다.The pH was decreased from 6.39 before the microbial treatment to 4.44 during the incubation period, and the pH was decreased as the amount of available phosphoric acid was increased.

이상의 결과를 토대로, 미생물에 의한 인산 가용화는 미생물이 분비하는 유기산에 의하여 일어나는 것으로 알려져 있는데 본 실험 결과에서도 인산 가용화능이 높을수록 pH 저하가 많은 것으로 미루어 유기산에 의해 인산 가용화가 일어나는 것으로 사료된다.Based on the above results, it is known that the solubilization of phosphoric acid by microorganisms is caused by the organic acid secreted by the microorganisms. In this test result, the higher the solubility of the phosphoric acid is, the more the pH is lowered so that the solubilization of phosphoric acid is caused by the organic acid.

<< 실시예Example 4> 인산  4> Phosphorus 가용화Solubilization 균주의 생리적 특성 검정 Physiological characterization of the strain

1. 실험방법1. Experimental Method

상기 실시예 1에서 선발한 인산가용화 균주의 생리적 특성을 확인하고자, 배양 온도별, 배양 pH별, 염류(NaCl) 농도별로 접종 2일 후 측정하여 최적 생육 범위 조사하였다.To determine the physiological characteristics of the solubilized phosphoric acid strains selected in Example 1, the optimal growth range was determined by measurement at 2 days after inoculation by culture temperature, culture pH, and salt concentration (NaCl).

이때, 처리 온도 및 배양 조건으로는 5~45℃의 온도 범위에서 R2A broth에 각 균주를 2일간 배양하여 600nm에서의 흡광도로 균주의 생육 정도를 측정하였다.At this time, each strain was cultured in R2A broth for 2 days at a temperature range of 5 ~ 45 ℃ for treatment temperature and culture conditions, and the degree of growth of the strain was measured by absorbance at 600 nm.

처리 pH 및 배양 조건으로는 인산 완충액 등으로 pH를 2~10으로 조정한 R2A broth에 각 균주를 28℃에서 2일간 배양하여 600nm에서의 흡광도로 균주의 생육 정도를 측정하였다.As the treatment pH and culture conditions, each strain was cultured at 28 ° C for 2 days in R2A broth adjusted to pH 2 to 10 with phosphate buffer, and the degree of growth of the strain was measured at an absorbance at 600 nm.

NaCl 처리 농도 및 배양 조건으로는 NaCl 0~20%의 염농도에서 R2A broth에 각 균주를 28℃에서 2일간 배양하여 600nm에서의 흡광도로 균주의 생육 정도를 측정하였다.The concentration of NaCl and culture conditions were determined by incubation of each strain on R2A broth at 0 ~ 20% NaCl concentration for 2 days at 28 ℃ and the growth of the strain with absorbance at 600nm.

2. 실험결과 2. Experimental results

상기 실험결과 도 4에 나타나 있듯이, 상기 실시예 1에서 선발한 신규균주의 생육온도는 20~35℃부근에서 생장이 양호한 중온성 미생물인 것을 확인하였다. As shown in FIG. 4, as shown in FIG. 4, it was confirmed that the growth temperature of the new strain selected in Example 1 was a mesophilic microorganism with good growth around 20 to 35 ° C.

또한 도 5에 나타나 있듯이, 상기 실시예 1에서 선발한 신규균주의 생육 pH는 pH 6~7부근에서 생장이 양호한 미생물인 것을 확인하였다.Also, as shown in FIG. 5, it was confirmed that the growth pH of the new strain selected in Example 1 was a good microorganism with growth around pH 6 to 7.

또한 도 6에 나타나 있듯이, 실시예 1에서 선발한 신규균주의 내염성은 NaCl이 증가할수록 생존율이 감소하여 10%까지 생존함을 확인하였다.Also, as shown in FIG. 6, the survival rate of the new strain selected in Example 1 decreased to 10% as NaCl increased.

<< 실시예Example 5> 토양 내에서의 인산  5> Phosphorus in the soil 가용화능Solubility 정량 분석 Quantitative analysis

1. 실험방법1. Experimental Method

유효인산은 Lancaster(토양화학분석법, 2010)법을 사용하여 정량 측정하였다. 설명하면, 난용성인산염(Ca3(PO4)2) 1% 첨가한 멸균토양 100g에 인산가용화 균주 5ml을 혼합하고 28℃에 정치 8일 후 토양을 채취하였다. 풍건시킨 토양 1g을 50㎖ tube에 취하고 추출액 4㎖를 가하여 10분간 진탕 후 No.2 여지로 여과하였다.The available phosphorus was quantitatively determined using Lancaster (Soil Chemical Analysis, 2010) method. In this case, 5 ml of the phosphoric acid solubilization strain was mixed with 100 g of the sterilized soil to which 1% of weakly soluble phosphate (Ca 3 (PO 4 ) 2 ) was added and the soil was collected at 28 ° C after 8 days. 1 g of the air-dried soil was taken in a 50 ml tube, 4 ml of the extract was added, and the mixture was shaken for 10 minutes and filtered through a No. 2 filter.

발색은 1-amino-2-naphtol-4-sulfonic acid에 의한 몰리브덴청법(토양화학분석법, 2010)을 참고하여 실험 진행하였다. 몰리브덴산 암몬황산 희석혼합액(조작용액)과 발색용액을 각각 첨가하여 30℃에서 30분간 발색시킨 후 파장 720nm에서 흡광도를 측정하였다.The color development was carried out by reference to molybdenum method (Soil Chemical Analysis, 2010) by 1-amino-2-naphthol-4-sulfonic acid. Ammonium molybdate diluted sulfuric acid mixture solution (working solution) and a coloring solution were added, respectively. The mixture was developed at 30 ° C for 30 minutes, and absorbance was measured at a wavelength of 720 nm.

2. 실험결과2. Experimental results

상기 실험결과 도 7에 나타나 있듯이, 상기 실시예1에서 분리한 Enterobacter sp. 04-P-1 균주를 처리한 토양에서 처리한지 8일 후 유효인산량을 측정한 결과 761ppm으로 대조구에 비해 91 % 증가함을 확인하였다.As a result of the above-mentioned experiment, as shown in FIG. 7, the Enterobacter sp. In the soil treated with the strain 04-P-1, the effective amount of phosphorus was measured 76 days after the treatment, and it was increased by 91% compared to the control.

<< 실시예Example 6>  6> 인산가용화Phosphoric acid solubilization 미생물에 의한 시설재배지 내 식물 생장 효과분석 Analysis of Plant Growth Effect in Microbiological Facilities

1. 실험방법1. Experimental Method

난용성인산염이 집적된 토마토 시설재배지에 20주씩 3반복 난괴법을 사용하여 배치된 토마토에 상기 실시예1의 신규균주 배양액을 처리하였다.The fresh strain culture of Example 1 above was treated with tomato seedlings which were placed using a three-round recuperative method for 20 weeks in a tomato plantation field containing an insoluble phosphate.

이때, 28℃에서 17시간 TSB 배지(1.7% tryptone, 0.3% soy peptone, 0.5% sodium chloride, 0.25% dipotassium hydrogen phosphate, 0.25% dextrose)에서 배양한 균주 50㎖을 4배 희석하여 1주일에 한 번씩 총 4회 관주 처리하였다. 그 다음 첫 균주 접종 10~12주 후 토마토 초장, 과중, 과실수량 측정하였다.At this time, 50 ml of the strain cultured in TSB medium (1.7% tryptone, 0.3% soy peptone, 0.5% sodium chloride, 0.25% dipotassium hydrogen phosphate, 0.25% dextrose) for 17 hours at 28 ° C was diluted 4 times 4 times. After 10 ~ 12 weeks of the first inoculation, tomato yield, fruit weight and fruit yield were measured.

2. 실험결과2. Experimental results

상기 실험결과, 하기 표 2와 같이 나타났다.The results of the experiment are shown in Table 2 below.

처리구Treatment 과실수(개)Number of fruit () 증감(%)increase(%) 과중(g)Overweight (g) 증감(%)increase(%) 초장(cm)Length (cm) 증감(%)increase(%) 대조구Control 6.66.6 00 101.9101.9 00 149.6149.6 00 실시예 1의 신규균주 Enterobacter sp. 04-P-1The new strain Enterobacter sp. 04-P-1 8.98.9 34.034.0 117.0117.0 14.814.8 175.3175.3 17.117.1

상기 표 2에 나타나 있듯이, 난용성인산염이 집적된 시설재배지 토양에 상기 실시예 1에서 분리된 인산가용화 미생물 Enterobacter sp. 04-P-1 배양액을 4배 희석하여 1주에 한 번씩 총 4회 관주 처리하였고, 그 결과 토마토 과실수량 34.0%, 과중 14.8 %, 초장 17.1% 증가함을 확인하였다.As shown in Table 2, the phosphate-soluble microbes Enterobacter isolated in Example 1 were added to the soil of the cultivation facility where the insoluble phosphate was accumulated, sp. 04-P-1 cultures were diluted 4 times and treated 4 times a week for 1 week. As a result, the yields of tomato fruit 34.0%, 14.8% and 17.1% were increased.

상기 결과들과 같이, 본 발명에서 토양내 미생물이 분비하는 유기산에 의해 불용성 인산염을 가용화시키는 우수한 능력을 나타내는 신규 미생물인 엔테로박터 에스피(Enterobacter sp.) 04-P-1를 제공함으로써 좀 더 효율적인 토마토 작물재배를 할 수 있게 되어 토양의 개량을 위한 비료의 유효성분으로도 이용가능하게 된다.As shown in the above results, the novel microorganism showing superior ability for solubilizing the insoluble phosphate by the organic acid to the soil microbial secretion in the present invention Enterobacter sp (Enterobacter sp.) 04-P-1, it is possible to cultivate more efficient tomato crops, so that it can be used as an effective ingredient of fertilizer for soil improvement.

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

기탁기관명 : 국립농업과학원 농업유전자원센터Depositor Name: National Institute of Agricultural Science

수탁번호 : KACC 92041PAccession number: KACC 92041P

수탁일자 : 2015년 02월 27일Funding date: February 27, 2015

<110> REPUBLIC OF KOREA(MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Soil-microbial product containing new microorganism enterobacter sp. 04-p-1 having soluble phosphate activity and manufacturing method thereof <130> P2015-0028 <160> 1 <170> KopatentIn 2.0 <210> 1 <211> 1484 <212> DNA <213> Enterobacter sp. 04-P-1 <400> 1 ttagagtttg ttcctggctc agattgaacg ctggcggcag gcctaacaca tgcaagtcga 60 gcggtagcac agagagcttg ctctcgggtg acgagcggcg gacgggtgag taatgtctgg 120 gaaactgcct gatggagggg gataactact ggaaacggta gctaataccg cataacgtcg 180 caagaccaaa gagggggacc ttcgggcctc ttgccatcag atgtgcccag atgggattag 240 ctagtaggtg gggtaatggc tcacctaggc gacgatccct agctggtctg agaggatgac 300 cagccacact ggaactgaga cacggtccag actcctacgg gaggcagcag tggggaatat 360 tgcacaatgg gcgcaagcct gatgcagcca tgccgcgtgt atgaagaagg ccttcgggtt 420 gtaaagtact ttcagcgagg aggaaggcat tgaggttaat aacctcagtg attgacgtta 480 ctcgcagaag aagcaccggc taactccgtg ccagcagccg cggtaatacg gagggtgcaa 540 gcgttaatcg gaattactgg gcgtaaagcg cacgcaggcg gtctgtcaag tcggatgtga 600 aatccccggg ctcaacctgg gaactgcatt cgaaactggc aggctagagt cttgtagagg 660 ggggtagaat tccaggtgta gcggtgaaat gcgtagagat ctggaggaat accggtggcg 720 aaggcggccc cctggacaaa gactgacgct caggtgcgaa agcgtgggga gcaaacagga 780 ttagataccc tggtagtcca cgccgtaaac gatgtcgact tggaggttgt tcccttgagg 840 agtggcttcc ggagctaacg cgttaagtcg accgcctggg gagtacggcc gcaaggttaa 900 aactcaaatg aattgacggg ggcccgcaca agcggtggag catgtggttt aattcgatgc 960 aacgcgaaga accttaccta ctcttgacat ccacggaatt tagcagagat gctttagtgc 1020 cttcgggaac cgtgagacag gtgctgcatg gctgtcgtca gctcgtgttg tgaaatgttg 1080 ggttaagtcc cgcaacgagc gcaaccctta tcctttgttg ccagcggttc ggccgggaac 1140 tcaaaggaga ctgccagtga taaactggag gaaggtgggg atgacgtcaa gtcatcatgg 1200 cccttacgag tagggctaca cacgtgctac aatggcgtat acaaagagaa gcgacctcgc 1260 gagagcaagc ggacctcata aagtacgtcg tagtccggat tggagtctgc aactcgactc 1320 catgaagtcg gaatcgctag taatcgtaga tcagaatgct acggtgaata cgttcccggg 1380 ccttgtacac accgcccgtc acaccatggg agtgggttgc aaaagaagta ggtagcttaa 1440 ccttcgggag ggcgcttacc actttgtgat tcatgactgg ggtg 1484 <110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Soil-microbial product containing new microorganism enterobacter          sp. 04-p-1 having soluble phosphate activity and manufacturing          method thereof <130> P2015-0028 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1484 <212> DNA <213> Enterobacter sp. 04-P-1 <400> 1 ttagagtttg ttcctggctc agattgaacg ctggcggcag gcctaacaca tgcaagtcga 60 gcggtagcac agagagcttg ctctcgggtg acgagcggcg gacgggtgag taatgtctgg 120 gaaactgcct gataggaggg gataactact ggaaacggta gctaataccg cataacgtcg 180 caagaccaaa gagggggacc ttcgggcctc ttgccatcag atgtgcccag atgggattag 240 ctagtaggtg gggtaatggc tcacctaggc gacgatccct agctggtctg agaggatgac 300 cagccacact ggaactgaga cacggtccag actcctacgg gaggcagcag tggggaatat 360 tgcacaatgg gcgcaagcct gatgcagcca tgccgcgtgt atgaagaagg ccttcgggtt 420 gtaaagtact ttcagcgagg aggaaggcat tgaggttaat aacctcagtg attgacgtta 480 ctcgcagaag aagcaccggc taactccgtg ccagcagccg cggtaatacg gagggtgcaa 540 gcgttaatcg gaattactgg gcgtaaagcg cacgcaggcg gtctgtcaag tcggatgtga 600 aatccccggg ctcaacctgg gaactgcatt cgaaactggc aggctagagt cttgtagagg 660 ggggtagaat tccaggtgta gcggtgaaat gcgtagagat ctggaggaat accggtggcg 720 aaggcggccc cctggacaaa gactgacgct caggtgcgaa agcgtgggga gcaaacagga 780 ttagataccc tggtagtcca cgccgtaaac gatgtcgact tggaggttgt tcccttgagg 840 agtggcttcc ggagctaacg cgttaagtcg accgcctggg gagtacggcc gcaaggttaa 900 aactcaaatg aattgacggg ggcccgcaca agcggtggag catgtggttt aattcgatgc 960 aacgcgaaga accttaccta ctcttgacat ccacggaatt tagcagagat gctttagtgc 1020 cttcgggaac cgtgagacag gtgctgcatg gctgtcgtca gctcgtgttg tgaaatgttg 1080 ggttaagtcc cgcaacgagc gcaaccctta tcctttgttg ccagcggttc ggccgggaac 1140 tcaaaggaga ctgccagtga taaactggag gaaggtgggg atgacgtcaa gtcatcatgg 1200 cccttacgag tagggctaca cacgtgctac aatggcgtat acaaagagaa gcgacctcgc 1260 gagagcaagc ggacctcata aagtacgtcg tagtccggat tggagtctgc aactcgactc 1320 catgaagtcg gaatcgctag taatcgtaga tcagaatgct acggtgaata cgttcccggg 1380 ccttgtacac accgcccgtc acaccatggg agtgggttgc aaaagaagta ggtagcttaa 1440 ccttcgggag ggcgcttacc actttgtgat tcatgactgg ggtg 1484

Claims (10)

서열번호 1로 표현되는 염기서열로 이루어진,
신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P).
1 &lt; / RTI &gt;
New microbial Enterobacter sp.) 04-P-1 strain (KACC 92041P).
제1항에 있어서,
상기 균주는 인산 가용화 활성을 갖는 것이 특징인,
신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P).
The method according to claim 1,
Wherein the strain has a phosphate solubilizing activity.
New microbial Enterobacter sp.) 04-P-1 strain (KACC 92041P).
제1항에 있어서,
상기 균주는 토양에서 불용화되는 인산질 비료를 가용화하여 토마토의 생육을 촉진시키는,
신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P).
The method according to claim 1,
The above-mentioned strains are used for solubilizing phosphate fertilizer insolubilized in soil to promote the growth of tomatoes,
New microbial Enterobacter sp.) 04-P-1 strain (KACC 92041P).
제1항에 있어서,
상기 균주는 내염성을 갖는 것이 특징인,
신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P).
The method according to claim 1,
Wherein said strain is salt tolerant.
New microbial Enterobacter sp.) 04-P-1 strain (KACC 92041P).
제1항에 있어서,
미생물 농약, 종자코팅제, 토양개량제, 퇴비부숙제, 엽면살포제 또는 관주살포제 중 어느 하나의 유효성분으로 이용되는,
신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P).
The method according to claim 1,
A microbial pesticide, a seed coating agent, a soil improving agent, a compostant homework agent, a foliar spraying agent, or a root spraying agent,
New microbial Enterobacter sp.) 04-P-1 strain (KACC 92041P).
제1항에 따른 신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P)를 유효성분으로 포함하는,
토양미생물 제제.
A novel microorganism according to claim 1, which is Enterobacter sp.) 04-P-1 strain (KACC 92041P)
Soil microorganism preparation.
제1항에 따른 신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P)를 준비하는 단계 및,
상기 균주를 TSB배지에 접종한 후 20~35℃, pH 6~7의 조건하에서 배양하는 단계;를 포함하는,
토양미생물 제제의 제조방법.
A novel microorganism according to claim 1, which is Enterobacter sp.) 04-P-1 strain (KACC 92041P)
Culturing the strain at a temperature of 20 to 35 DEG C and a pH of 6 to 7 after inoculating the strain into a TSB medium;
A method for producing a soil microorganism preparation.
제1항에 따른 신규 미생물 엔테로박터 에스피(Enterobacter sp.) 04-P-1 균주 (KACC 92041P)의 균체 또는 제6항에 따른 토양미생물 제제 중 어느 하나 이상을 유효성분으로 포함하는, 토양미생물 비료.
A soil microbial fertilizer comprising at least one of the microorganism Enterobacter sp. 04-P-1 strain (KACC 92041P) according to claim 1 or the soil microorganism preparation according to claim 6 as an active ingredient. .
제8항에 따른 토양미생물 비료를 토양에 관주처리하여 토양내 인산가용화능을 높여주는 방법.
A method for enhancing the solubilization ability of the soil in the soil by treating the soil microbial fertilizer according to claim 8 with the soil.
제6항에 따른 토양미생물 제제를 3~4배 희석한 후 토마토 재배 토양에 관주처리하여 토양내 불용성 인산염이 가용화됨으로써 토마토의 생육을 촉진하는 방법.A method for promoting the growth of tomato by solubilizing the insoluble phosphate in the soil by treating the soil microorganism preparation according to claim 6 with 3 to 4 times of diluting and cultivating the tomato growing soil.
KR1020150059834A 2015-04-28 2015-04-28 Soil-microbial product containing new microorganism enterobacter sp. 04-p-1 having soluble phosphate activity and manufacturing method thereof Ceased KR20160128533A (en)

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114381405A (en) * 2022-01-22 2022-04-22 赣南师范大学 A strain of Enterobacter sp.Jin-002 and its prepared pesticide degradation preparation and degradation method
CN114592032A (en) * 2022-03-04 2022-06-07 武汉科技大学 Screening and identifying method of phosphorus-dissolving strain and application of phosphorus-dissolving strain in biofertilizer
CN114752538A (en) * 2022-05-25 2022-07-15 湖南大学 Camellia oleifera endophytic actinomycetes with soil improvement function and its application
CN114921362A (en) * 2022-04-19 2022-08-19 湖南大学 Camellia oleifera endophytic actinomycetes with insoluble phosphorus dissolving and growth promoting functions and application thereof
CN117327601A (en) * 2023-04-21 2024-01-02 中国科学院地理科学与资源研究所 Enterobacter and application thereof
KR102793586B1 (en) * 2023-12-28 2025-04-14 (주)현농 A soil conditioner for improving salt accumulation comprising Enterobacter intermedium 60-2G strain as an active ingredient
KR102793589B1 (en) * 2023-12-28 2025-04-14 (주)현농 A method of manufacturing soil conditioner for improving salt accumulation comprising Enterobacter intermedium 60-2G strain as an active ingredient
CN120097770A (en) * 2025-03-06 2025-06-06 广东工业大学 A method for converting insoluble phosphorus using microorganisms of the genus Enterobacter and a highly efficient phosphorus-dissolving bacterial agent and application thereof

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114381405A (en) * 2022-01-22 2022-04-22 赣南师范大学 A strain of Enterobacter sp.Jin-002 and its prepared pesticide degradation preparation and degradation method
CN114381405B (en) * 2022-01-22 2023-08-18 赣南师范大学 A strain of Enterobacter sp.Jin-002 and its prepared pesticide degradation preparation and degradation method
CN114592032A (en) * 2022-03-04 2022-06-07 武汉科技大学 Screening and identifying method of phosphorus-dissolving strain and application of phosphorus-dissolving strain in biofertilizer
CN114921362A (en) * 2022-04-19 2022-08-19 湖南大学 Camellia oleifera endophytic actinomycetes with insoluble phosphorus dissolving and growth promoting functions and application thereof
CN114752538A (en) * 2022-05-25 2022-07-15 湖南大学 Camellia oleifera endophytic actinomycetes with soil improvement function and its application
CN117327601A (en) * 2023-04-21 2024-01-02 中国科学院地理科学与资源研究所 Enterobacter and application thereof
CN117327601B (en) * 2023-04-21 2024-05-14 中国科学院地理科学与资源研究所 Enterobacter and application thereof
KR102793586B1 (en) * 2023-12-28 2025-04-14 (주)현농 A soil conditioner for improving salt accumulation comprising Enterobacter intermedium 60-2G strain as an active ingredient
KR102793589B1 (en) * 2023-12-28 2025-04-14 (주)현농 A method of manufacturing soil conditioner for improving salt accumulation comprising Enterobacter intermedium 60-2G strain as an active ingredient
CN120097770A (en) * 2025-03-06 2025-06-06 广东工业大学 A method for converting insoluble phosphorus using microorganisms of the genus Enterobacter and a highly efficient phosphorus-dissolving bacterial agent and application thereof

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