항진균 활성, 식물 생장촉진 활성, 미네랄 가용화능을 가진 생물학적 제제로서 잠재적 식물 생장 촉진 근권세균의 특성조사

이송민(Song Min Lee),김지윤(Ji-Youn Kim),김희숙(Hee Sook Kim),오가윤(Ka-Yoon Oh),이광희(Kwang Hui Lee),이상현(Sang-Hyeon Lee),장정수(Jeong Su Jang) 한국생명과학회 2021 생명과학회지 Vol.31 No.7

본 연구에서는 근권토양으로부터 18종의 세균을 순수분리하고, 이들의 식물 병원성 진균 생육억제 활성, 식물생장촉진 활성 및 미네랄 가용화능을 평가하였다. Bacillus 속과 Pseudomonas 속 분리균주의 항진균 활성을 통해 생물학적 방제제로서의 가능성을 확인할 수 있었으며, 이는 식물 병원성 진균의 세포벽에 작용하는 여러 가수분해효소 활성과 siderophore 생성능 등에 기인된 것으로 판단된다. 또한 대부분의 분리균주가 aminocyclopropane-1-carboxylate deaminase 생성능, indole-3-acetic acid 생성능 및 질소 고정능을 갖고 있는 것으로 나타났으며, 이러한 특성은 식물이 흔히 노출될 수 있는 환경 스트레스 조건 하에서 스트레스 에틸렌의 농도를 감소시킴으로써 뿌리 발달 및 성장, 그리고 작물의 생산성에 긍정적인 영향을 줄 것으로 판단된다. 그리고 분리균주의 인산, 규소, 탄산칼슘, 아연 가용화능 등을 확인한 결과, 일부 분리균주들의 미네랄 가용화능을 확인하였으며, 이들 분리균주를 식물 생장 시에 접종한다면 식물 생장에 필요한 영양분을 식물이 흡수할 수 있는 이용가능한 형태로 변환시켜 식물의 생장에 도움을 줄 수 있을 것으로 기대된다. 이러한 항진균 활성, 식물생장촉진 활성 및 미네랄 가용화능의 결과를 통해 분리균주 18종의 biocontrol agent로서의 이용가능성을 제시하고자 한다. The purpose of this study was to confirm the antifungal activity, plant growth-promoting activity, and mineral solubilizing activity of 18 types of bacteria isolated purely from rhizosphere soil. The potential of isolates of the genus Bacillus and Pseudomonas as biocontrol agents was confirmed through the antifungal activity of these isolates. This activity has been determined to be due to various hydrolytic enzymes on the cell wall of plant pathogenic fungi and the production of siderophores in isolates. In addition, most of the isolates have been found to have aminocyclopropane-1-carboxylate deaminase production activity, indole-3-acetic acid production activity, and nitrogen fixation activity. These characteristics are believed to have a positive effect on root development, growth, and the productivity of crops via a reduction in the concentration of ethylene under conditions of environmental stress, to which plants are commonly exposed. In addition, on testing for the solubilizing activity of the isolates for phosphoric acid, silicon, calcium carbonate, and zinc, some isolates were found to have mineral solubilizing activities. Inoculation of these isolates during plant growth is expected to assist plant growth by converting nutrients necessary for growth into usable forms that can be absorbed by plants. The 18 isolated strains can be used as biocontrol agents due to their antifungal activity, plant growth-promoting activity, and mineral solubilizing activity.

Assessment of Plant Growth Promoting Activities of Phosphorus Solubilizing Bacteria

Walpola, Buddhi Charana,Song, June-Seob,Yoon, Min-Ho Korean Society of Soil Science and Fertilizer 2012 한국토양비료학회지 Vol.45 No.1

Plant growth promoting traits like production of indoleacetic acid (IAA), ammonia, hydrogen cyanide (HCN), siderophore, and like the enzyme activities of catalase, ACC deaminase, cellulase, chitinase and protease were assayed in vitro for twenty one phosphorus solubilizing bacteria isolated from soil isolates. Except SPP-5 and SPP-15 strains, all the other isolated strains produced IAA in various amounts of 10 to $23{\mu}g\;ml^{-1}$. All strains showed positive response for ammonia production and ACC deaminase activity implying that they are capable of growing in a N-free basal medium. Catalase activity was found to be superior in SPP-2, SPP-7, SPP-12 and SPP-17 compared to the other strains tested. HCN production was detected by 15 strains and among them SPP-9, SPP-15, SAph-11, and SAph-24 were found to be strong HCN producers. Except the isolates SPP-10, SPP-12, SPP-13 and SPP-14, all the other isolates produced more than 80% siderophore units. None of the strains showed cellulose and chitinase activity. SAph-8, SAPh-11, SAPh-24 and SPP-15 strains showed 35.84, 50.33, 56.64 and 34.78 U/ml protease activities, respectively. SPP-1, SPP-2, SPP-3, SPP-11, SPP-17, SPP-18, SAph-11 and SAph-24 strains showed positive response for all the tested plant growth promotion traits except cell wall degrading enzyme activities. According to the results, all the tested phosphorus solubilizing isolates could exhibit more than three or four plant growth promoting traits, which may promote plant growth directly or indirectly or synergistically. Therefore, these phosphorus solubilizing strains could be employed as bio-inoculants for agriculture soils.

근권에 존재하는 Bacillus 속 균주들의 식물 생장 촉진 활성 특성

오가윤 ( Ka-yoon Oh ),김지윤 ( Ji-youn Kim ),이송민 ( Song Min Lee ),김희숙 ( Hee Sook Kim ),이광희 ( Kwang Hui Lee ),이상현 ( Sang-hyeon Lee ),장정수 ( Jeong Su Jang ) 한국미생물 · 생명공학회 2021 한국미생물·생명공학회지 Vol.49 No.3

본 연구에서는 토양 및 근권에 존재하는 Bacillus 속의 식물 생장 촉진 활성, 식물 병원성 곰팡이의 생장 억제활성, 미네랄 가용화능 및 세포 외 효소활성을 확인해 보고자 하였다. 식물 병원성 곰팡이에 대한 항진균 활성에서 DDP257은10종의 병원성 곰팡이에서 항진균 활성이 모두 나타났다. 식물 생장 촉진 인자인 indole-3-acetic acid 생성능에서는ANG20이 70.97 μg/ml로 가장 높게 나타났다. 추가적으로 1- aminocyclopropane-1-carboxylate deaminase 생성능 조사에서는 총 10종에서 생성능을 확인하였고, 질소 고정능과 siderophore 생성능 조사에서는 대부분의 분리균주에서 활성을 확인하였다. 이후 분리된 균주에 대하여 phosphate, calcite, zinc과 같은 미네랄 가용화능을 확인하였으며 세포외 효소활성에서도 대부분의 효소에서 활성이 나타났다. 특히 alkaline phosphatase, esterase (C4), acid phosphatase, naphtol-AS-BI-phosphohydrolase에서 선별된 균주 모두 유사한 활성을 보였다. 이는 Bacillus 속이 다양한 유기물과 항생물질 및 세포 외 효소를 분비함으로써 이러한 결과가 나타난 것으로 판단된다. 따라서, 본 연구 결과를 통해 토양의 환경 개선에 기여하는 균주를 활용하여 미생물 제제로써의 가능성을 제시한다. This study aimed to identify plant growth-promoting activity, phytopathogenic fungi growth inhibitory activity, mineral solubilization ability, and extracellular enzyme activity of the genus Bacillus in soil and the rhizosphere. With regards to antifungal activity against phytopathogenic fungi, DDP257 showed antifungal activity against all 10 pathogenic fungi tested. ANG20 showed the highest ability to produce indole- 3-acetic acid, a plant growth-promoting factor (70.97 μg/ml). In addition, 10 species were identified to have 1-aminocyclopropane-1-carboxylate deaminase production ability, and most isolates showed nitrogen fixation and siderophore production abilities. Thereafter, the isolated strains’ ability to solubilize minerals such as phosphate, calcite, and zinc was identified. With extracellular enzyme activity, the activity appeared in most enzymes. In particular, all the strains showed similar abilities for alkaline phosphatase, esterase (C4), acid phosphatase, and naphtol-AS-BI-phosphohydrolase production. This result was observed because the genus Bacillus secreted various organic substances, antibiotics, and extracellular enzymes. Therefore, through the results of this study, we suggest the possibility of using strains contributing to the improvement of the soil environment as microbial agents.

동해안 자생식물로부터 분리된 내생균류의 식물생장촉진활성 및 동정

유영현,진용주,강상모,오세종,이명철,김종국,You, Young-Hyun,Jin, Yong Ju,Kang, Sang-Mo,Oh, Sejong,Lee, Myung-Chul,Kim, Jong-Guk 한국미생물학회 2015 미생물학회지 Vol.51 No.1

동해안에 자생하는 해안식물인 개질경이를 채집하였고, 뿌리로부터 형태적으로 다른 내생균류 20균주를 선발하였다. 개질경이 뿌리에 공생하는 내생균류의 ITS-rDNA 염기서열을 분석하였으며, 분리된 내생균류의 유연관계는 Bayesian 프로그램을 이용하여 계통분석을 하였다. 모든 내생균류의 농축배양여과액을 난장이벼에 처리하여 식물생장촉진활성을 스크리닝하였고, 분리된 균류 중에서 E/PC/10/1 균주가 생장촉진활성이 우수한 것으로 확인되었다. E/PC/10/1 균주의 배양여과액을 HPLC와 GC/MS-SIM을 이용하여 분석한 결과, 식물호르몬인 지베렐린 $GA_1$, $GA_3$, $GA_4$가 정량분석 되었다. 그리고 E/PC/10/1 균주의 명확한 동정을 위하여, beta-tubulin 유전자 염기서열을 이용한 분자적인 방법과 현미경을 이용한 형태적인 방법으로 동정하였다. 최종적으로 E/PC/10/1 균주는 GAs을 생산하는 새로운 P. spinulosum으로 동정되었다. Coastal plant species, Plantago camtschatica Cham. native to the coastal region of the East Sea were sampled and then morphologically different 20 endophytic fungal strains were purely isolated. Phylogenetic analysis of isolates was done by the Bayesian program based on sequenced internal transcribed spacer (ITS-rDNA) region. Culture filtrates of each of 20 isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity, respectively. As the results, E/PC/10/1 strain showed the highest plant growth-promoting activity among them. The culture filtrate of the strain E/PC/10/1 was revealed as containing gibberellins ($GA_1$, $GA_3$, $GA_4$) by using HPLC, and gas GC/MS with selected ion monitoring (SIM). Finally, this strain was identified as novel Penicillium spinulosum species that producing new GAs with microscopic observation and further molecular analysis with beta-tubulin gene sequence.

Plant Growth-promoting Activity of Acremonium strictum MJN1 Isolated from Roots of Panax ginseng

Lim, Hyung-Bum,Chung, Yang-Jo,Bae, Ju-Yun,Kim, Dong-Jin,Kwon, Hyung-Jin,Lee, In Hyung,Chung, Byung-Chul,Lee, Woong-Sang,Suh, Joo-Won 한국응용생명화학회 2000 Journal of Applied Biological Chemistry (J. Appl. Vol.43 No.2

The plant growth-promoting activity of Acremonium strictum MJN1 isolated from roots of Panox ginseng was explored. The myceliaI extract of A. strictum MJN1 enhanced the rice seedling growth by 14.5 and 9.0% in the dried weight of shoots and roots, and the growth of red pepper by 54 and 85% in the top length and the dried weight in pot experiments, respectively. The plant growth-promoting substances in the myceliaI extract of Acremonium strictum MJN1 were identified as D-adenosine and glycerol. Both commercial D-adenosine and glycerol also promoted significantly the rice seedling growth but, unlike the mycelial extract of A. strictum MJN1, hardly affected the yields of plants grown in pots or field. Therefore, it is possible that other plant growth-promoting substances are produced by A. strictum MJN1. However, this study shows that A. strictum MJN1 has a great potential as a biofertilizer.

Isolation and Characterization of Phosphate Solubilizing Pseudomonas Species and Their Effect on Plant Growth Promotion

조경서,김상윤,조주식,정용화 한국토양비료학회 2019 한국토양비료학회지 Vol.52 No.4

Plant growth-promoting rhizobacteria (PGPR) affect plant growth via various direct or indirect mechanisms. In this study, eight different Pseudomonas strains (PM1 - PM8) isolated from rhizospheric soil were characterized plant growth-promoting characteristics, including indole-3-acetic acid (IAA) and Gibberellin (GA3) production, phosphate solubilization, 1- aminocyclopropane-1-carboxylic acid (ACC) deaminase activity and effects of direct inoculation on plant growth parameters (fresh weight, shoot length). All of these strains have their ability for solubilization of insoluble phosphate depending on pH decrease at the range around pH4 - 5 at 2 days after inoculation. PM1, PM4, PM5 and PM7 species was capable of IAA production and PM4, PM5, PM7 and PM8 species have high activity for production of gibberellin (GA3) hormone. For ACC deaminase activity, PM1, PM3 and PM8 show high activity. Furthermore, all these species have significantly promoted the growth of the lettuce seedling plants for fresh weight and shoot length enhancement, so that these Pseudomonas species could be used as potential bio-fertilizer agents.

Assessment of Plant Growth Promoting Activities of Phosphorus Solubilizing Bacteria

Buddhi Charana Walpola,June-Seob Song,Min-Ho Yoon 한국토양비료학회 2012 한국토양비료학회지 Vol.45 No.1

Plant growth promoting traits like production of indoleacetic acid (IAA), ammonia, hydrogen cyanide (HCN), siderophore, and like the enzyme activities of catalase, ACC deaminase, cellulase, chitinase and protease were assayed in vitro for twenty one phosphorus solubilizing bacteria isolated from soil isolates. Except SPP-5 and SPP-15 strains, all the other isolated strains produced IAA in various amounts of 10 to 23 ㎍ ml<SUP>-1</SUP>. All strains showed positive response for ammonia production and ACC deaminase activity implying that they are capable of growing in a N-free basal medium. Catalase activity was found to be superior in SPP-2, SPP-7, SPP-12 and SPP-17 compared to the other strains tested. HCN production was detected by 15 strains and among them SPP-9, SPP-15, SAph-11, and SAph-24 were found to be strong HCN producers. Except the isolates SPP-10, SPP-12, SPP-13 and SPP-14, all the other isolates produced more than 80% siderophore units. None of the strains showed cellulose and chitinase activity. SAph-8, SAPh-11, SAPh-24 and SPP-15 strains showed 35.84, 50.33, 56.64 and 34.78 U/ml protease activities, respectively. SPP-1, SPP-2, SPP-3, SPP-11, SPP-17, SPP-18, SAph-11 and SAph-24 strains showed positive response for all the tested plant growth promotion traits except cell wall degrading enzyme activities. According to the results, all the tested phosphorus solubilizing isolates could exhibit more than three or four plant growth promoting traits, which may promote plant growth directly or indirectly or synergistically. Therefore, these phosphorus solubilizing strains could be employed as bio-inoculants for agriculture soils.

단보 : 담수에 자생하는 수생식물에서 분리된 내생균류의 지베렐 린 생산과 동정

유영현 ( Young Hyun You ),강상모 ( Sang Mo Kang ),최유미 ( Yu Mi Choi ),이명철 ( Myung Chul Lee ),김종국 ( Jong Guk Kim ) 한국균학회 2015 Mycobiology Vol.43 No.1

Aquatic plant Hydrocharis dubia (Blume) Backer was collected from the Dalsung wetland in Daegu. Sixteen endophytic fungi with different colony morphologies were isolated from the roots of aquatic plants. Waito-c rice (WR) seedlings were treated with fungal culture filtrates (FCF) for screening plant growth-promoting activity. In the results, HD1008 strain isolated from aquatic plant showed highest plant growth-promoting activity. The FCF of HD1008 strain was analyzed using gas chromatography mass spectrometry (GC/MS) with selected ion monitoring (SIM). Analysis of the FCF of HD1008 strain found that it contained gibberellins (GA) (GA1, 1.2 ng/100 mL; GA4, 5 ng/100 mL). Phylogenetic tree of HD1008 strain was constructed by partial internal transcribed spacer (ITS) region and partial beta-tubulin gene sequences. Therefore, we describe HD1008 strain as a new gibberellin-producing Penicillium trzebinskii based on morphological and molecular characteristics.

Isolation and Characterization of an Antifungal and Plant Growth-Promoting Microbe

( Se Won Park ),( Hee-jong Yang ),( Ji Won Seo ),( Jinwon Kim ),( Su-ji Jeong ),( Gwangsu Ha ),( Myeong Seon Ryu ),( Hee Gun Yang ),( Do-youn Jeong ),( Hyang Burm Lee ) 한국균학회 2021 韓國菌學會誌 Vol.49 No.4

Fungal diseases including anthracnose, stem rot, blight, wilting, and root rot of crops are caused by phytopathogens such as Colletotrichum species, Sclerotinia sclerotiorum, Phytophthora species, and Fusarium oxysporum and F. solani which threaten the production of chili pepper. In this study, to identify biological control agents (BCAs) of phytopathogenic fungi, potentially useful Bacillus species were isolated from the field soils. We screened out five Bacillus strains with antagonistic capacity that are efficiently inhibiting the growth of phytopathogenic fungi. Bacillus species were characterized by the production of extracellular enzymes, siderophores, and indole-3-acetic acid (IAA). Furthermore, the influence of bacterial strains on the plant growth promoting activity and seedling vigor index were assessed using Brassica juncea as a model plant. Inoculation with Bacillus subtilis SRCM 121379 significantly increased the length of B. juncea shoots and roots by 45.6% and 52.0%, respectively. Among the bacterial isolates, Bacillus subtilis SRCM 121379 showed the superior enzyme activities, antagonistic capacity and plant growth promoting effects. Based on the experimental results, Bacillus subtilis SRCM 121379 (GenBank accession no. NR027552) was finally selected as a BCA candidate.

Biocontrol of Fusarium Crown and Root Rot and Promotion of Growth of Tomato by Paenibacillus Strains Isolated from Soil

( Sheng Jun Xu ),( Byung Sup Kim ) 한국균학회 2014 韓國菌學會誌 Vol.42 No.2

In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited the mycelial growth of Fusarium oxysporum f. sp. radicis-lycopersici in vitro. The seven Paenibacillus strains enhanced a range of growth parameters in tomato plants under greenhouse conditions, in comparison with non-inoculated control plants. Notably, treatment of tomato plants with one identified strain, P. polymyxa SC09-21, resulted in 80.0% suppression of fusarium crown and root rot under greenhouse conditions. The plant growth promoting and antifungal activity of P. polymyxa SC09-21 identified in this study highlight its potential suitability as a bioinoculant.