Comparison of Plant Growth Promoting Methylobacterium spp. and Exogenous Indole-3-Acetic Acid Application on Red Pepper and Tomato Seedling Development

Hari P. Deka Boruah,Puneet S. Chauhan,Woo-Jong Yim(임우종),Gwang-Hyun Han(한광현),Tong-Min Sa(사동민) 한국토양비료학회 2010 한국토양비료학회지 Vol.43 No.1

무균 및 온실조건에서 indole-3-acetic acid (IAA)의 처리와 1-aminocyclopropane-1-carboxylate deaminase (ACCD) 및 IAA 활성을 갖는 Methylobacterium 균주 접종 시 토마토와 고추의 생장을 비교 평가하였다. 무균조건에서 1.0 ㎍ mL?¹의 IAA는 고추와 토마토의 뿌리생장을 촉진시키는데 비해 10.0 ㎍ mL?¹ 이상의 높은 농도에서는 뿌리생장이 억제되었다. 그러나 높은 ACCD 활성을 갖고, IAA 활성은 낮거나 가지고 있지 않은 Methylobacterium 균주들을 접종하였을 때에는 고추와 토마토 모두 IAA 처리구 보다 뿌리생장이 증진되는 것을 확인하였다. 마찬가지로 온실조건에서 Methylobacterium 균주들을 접종했을 때, 마디길이와 잎의 크기 그리고 단위 면적당 잎의 무게(SLW)에서 유의성 있는 증진효과를 보였다. 전반적인 식물 생장에서 저농도의 IAA 처리 효과는 Methylobacterium의 효과와 비슷한 경향을 나타냈다. 유묘의 지상부 길이는 ACCD 활성과 IAA 생산능을 갖는 Methylobacterium 균주 처리구에서 유의성 있는 증가를 확인할 수 있었으며, 전체 건물중 또한 Methylobacterium 처리 시 유의성 있는 증진 효과를 확인 할 수 있었다. 하지만 고농도의 IAA는 고추와 토마토의 생물량을 억제시켰다. 이러한 결과는 접종 균주의 IAA와 ACCD가 고추와 토마토 유묘의 생장 증진에 영향을 끼친다는 것을 증명한다. A comparative study was performed in gnotobiotic and greenhouse conditions to evaluate the effect of exogenous application of indole-3-acetic acid (IAA) and inoculation of Methylobacterium spp. possessing 1-aminocyclopropane-1-carboxylate deaminase (ACCD) and IAA activity on red pepper and tomato seedling growth and development. Application of 1.0 ㎍ mL?¹ IAA positively influenced root growth while high concentrations (>10.0 ㎍ mL?¹) suppressed root growth of red pepper and tomato under gnotobiotic condition. On the other hand, inoculation of Methylobacterium strains with ACCD activity and IAA or without IAA enhanced root growth in both plants. Similarly, under greenhouse condition the inoculation of Methylobacterium sp. with ACCD activity and IAA enhanced plant fitness recorded as average nodal length and specific leaf weight (SLW) but the effect is comparable with the application of low concentrations of IAA. Seedling length was significantly increased by Methylobacterium strains while total biomass was enhanced by Methylobacterium spp. and exogenous applications of < 10.0 ㎍ mL?¹ IAA. High concentrations of IAA retard biomass accumulation in red pepper and tomato. These results confirm that bacterial strains with plant growth promoting characters such as IAA and ACCD have characteristic effects on different aspects of growth of red pepper and tomato seedlings which is comparable or better than exogenous applications of synthetic IAA.

Methylobacterium dankookense sp. nov., Isolated from Drinking Water

이시원,오현우,Kang-Hyun Lee,안태영 한국미생물학회 2009 The journal of microbiology Vol.47 No.6

A pink-pigmented bacterium, designated SW08-7T was isolated from the drinking water of a water purifier. Cells were Gram-negative, rod-shaped, strictly aerobic, and non-spore-forming. It grew optimally at 25°C, pH 6~7. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain SW08-7T belongs to the genus Methylobacterium. The highest 16S rRNA gene sequence similarities were found to Methylobacterium mesophilicum JCM 2829T (96.9%), Methylobacterium brachiatum B0021T (96.9%), Methylobacterium phyllosphaerae CBMB27T (96.6%), Methylobacterium radiotolerans JCM 2831T (96.6%), and Methylobacterium hispanicum GP34T (96.5%). DNA-DNA hybridization experiment revealed low-level (28.5%) of DNA- DNA relatedness between strain SW08-7T and Methylobacterium hispanicum. The genomic DNA G+C content was 68.9 mol% and the major isoprenoid quinone was Q-10. The major cellular fatty acid of strain SW08-7T was C18:1 ω7c (79.8±2.1%). Results of phylogenetic, phenotypic, and biochemical analyses revealed that strain SW08-7T could be classified as representing a novel species of genus Methylobacterium, for which the name Methylobacterium dankookense sp. nov. is proposed. The type strain is SW08-7T (=KCTC 22512T =DSM 22415T).

Detection of Methylobacterium spp. Possessing Soluble Methane Monooxygenase (sMMO) Gene Isolated from Rice

강연경 ( Yeongyeong Kang ),( Denver Walitang ),( Poulami Chatterjee ),표채은 ( Chaeeun Pyo ),사동민 ( Tongmin Sa ) 한국환경농학회 2017 한국환경농학회 학술대회집 Vol.2017 No.-

Methane (CH4) is currently receiving great attention due to its severe impact on the environment as a greenhouse gas. The atmospheric concentration of methane is continuously increasing and its average concentration is around 1.87 mg L^-1. Methane is produced in nature when methanogens digest plant material in the absence of oxygen. Methane emission from rice fields is one of the largest individual sources in the global budget of atmospheric CH₄. Methane-oxidizing bacteria (MOB) are phylogenetically diverse groups defined by their ability to use methane as a major carbon and energy source. Aerobic MOB, which are widely present in natural environments, can utilize methane as the sole carbon and energy source through a unique enzyme system of methane monooxygenase (MMO). MMO has a soluble cytoplasmic form (sMMO) and a particulate membrane associated form (pMMO). The objective of this study is to investigate the presence of sMMO in Methylobacterium spp. The selected strains for the current study, Methylobacterium spp. were isolated from rice. MOB are a subset of a physiological group of bacteria known as methylotrophs. The sMMO consists of three components, proteins A, B, and C. Protein A is coded for by the mmoX, mmoY, and mmoZ genes. Protein B is coded for by the mmoB gene. Protein C is the reductase component of the enzyme and is coded for by the mmoC gene. The presence of sMMO in Methylobacterium spp. was proven through polymerase chain reaction (PCR) using mmoC specific primer. This results indicate that there were 18 out of 19 strains of Methylobacterium. spp with sMMO detected through PCR-based amplification of an mmoC gene. PCR products of the correct size (314 bp) were obtained with all of the Methylobacterium spp. used in this study. In the current study confirming the oxidative activity of the strains that contain the sMMO. Future study the growth of these isolates using methane as the sole carbon source and then assess methane oxidation potential of by gas chromatography and their contribution in atmospheric methane oxidation.

식물생장촉진미생물인 Methylobacterium을 처리한 고추에서 병 저항성유전자의 발현량 검정 (초록)

유제민 ( Je Min Yoo ),이수영 ( Su Young Lee ),엄유리 ( Yu Rry Um ),임우종 ( Woo Jong Yim ),사동민 ( Tong Min Sa ),이이 ( Yi Lee ) 한국토양비료학회 2012 한국토양비료학회 학술발표회 Vol.2012 No.-

식물생장촉진미생물인 Methylobacterium을 처리한 고추에서 병 저항성유전자의 발현량 검정

유제민(Je-Min Yoo),이수영(Su-Young Lee),엄유리(Yurry Um),임우종(Woojong Yim),사동민(Tongmin Sa),이이(Yi Lee) 한국토양비료학회 2012 한국토양비료학회 학술발표회 초록집 Vol.2012 No.6

Ralstonia Solanacearum에 감염된 토마토에서 ACC Deaminase를 생산하는 Methylobacterium spp.의 스트레스 에틸렌 수준 조절

임우종 ( Woo Jong Yim ),김기윤 ( Ki Yoon Kim ),우성만 ( Sung Man Woo ),이선미 ( Seon Mi Lee ),홍보희 ( Bo Hui Hong ),사동민 ( Tong Min Sa ) 한국환경농학회 2012 한국환경농학회 학술대회집 Vol.2012 No.-

Biosynthesis of ethylene is usually enhanced with severity of pathogenic infection. Some ethylene synthesis inhibitors are known to significantly decrease the severity of pathogen infections in plants. Bacteria of genus Methylobacterium have been found to stimulate plant growth and regulate the level of ethylene in crop plants. A selective decrease of ethylene can be achieved by plant growth promoting bacteria, for instance Methylobacterium, containing ACC (1-aminocyclopropane-1- carboxylate) deaminase, which cleaves ACC into -ketobutyrate and ammonia in higher plants which leads to reduced ethylene level. This work aimed to test the induction of defense responses in tomato against bacterial wilt by stress ethylene level reduction mediated by the ACC deaminase activity of Methylobacterium strains. In greenhouse experiments, the disease index value observed in Methylobacterium strains treated tomato and red pepper plants were lower than that of uninoculated treatment. Plants treated with Methylobacterium strains challenge inoculated with Ralstonia solanacearum (RS) showed significantly reduced disease symptom and lowered ethylene emission under greenhouse condition. The ACC and ACO (1-aminocyclopropane-1- carboxylate oxidase) accumulated in the tomato leaves were significantly reduced with Methylobacterium strains inoculation compared to treatment with pathogen alone. Similar to ACC oxidase activity, ACC oxidase related gene expression also increased in tomato treated with Ralstonia solanacearum alone compared to Methylobacterium strains treatment. In addition pathogenesis-related (PR) proteins related to ISR (Induced systemic resistance) including -1,3-glucanase, phenylalanine ammonia-lyase (PAL), peroxidase (PO) and polyphenol oxidase (PPO) were increased in Methylobacterium strain inoculated tomato plants. A significant increase in -1,3-glucanase and PAL related genes expression was found in all of the Methylobacterium spp. treatments compared to the Ralstonia solanacearum alone treatment. This study supports that the Methylobacterium strains might increase the activity of defense enzymes by modulating the ethylene biosynthesis pathway, suggesting the possibility of using Methylotrophic bacteria as potential bio-control agents.

Ethylene emission and PR protein synthesis in ACC deaminase producing Methylobacterium spp. inoculated tomato plants (Lycopersicon esculentum Mill.) challenged with Ralstonia solanacearum under greenhouse conditions

Yim, W.,Seshadri, S.,Kim, K.,Lee, G.,Sa, T. Gauthier-Villars ; Elsevier Science Ltd 2013 Plant physiology and biochemistry Vol.67 No.-

Bacteria of genus Methylobacterium have been found to promote plant growth and regulate the level of ethylene in crop plants. This work is aimed to test the induction of defense responses in tomato against bacterial wilt by stress ethylene level reduction mediated by the ACC deaminase activity of Methylobacterium strains. Under greenhouse conditions, the disease index value in Methylobacterium sp. inoculated tomato plants was lower than control plants. Plants treated with Methylobacterium sp. challenge inoculated with Ralstonia solanacearum (RS) showed significantly reduced disease symptoms and lowered ethylene emission under greenhouse condition. The ACC and ACO (1-aminocyclopropane-1-carboxylate oxidase) accumulation in tomato leaves were significantly reduced with Methylobacterium strains inoculation. While ACC oxidase gene expression was found higher in plants treated with R. solanacearum than Methylobacterium sp. treatment, PR proteins related to induced systemic resistance like β-1,3-glucanase, PAL, PO and PPO were increased in Methylobacterium sp. inoculated plants. A significant increase in β-1,3-glucanase and PAL gene expression was found in all the Methylobacterium spp. treatments compared to the R. solanacearum treatment. This study confirms the activity of Methylobacterium sp. in increasing the defense enzymes by modulating the ethylene biosynthesis pathway and suggests the use of methylotrophic bacteria as potential biocontrol agents in tomato cultivation.

Methylobacterium terrae sp. nov., a radiation-resistant bacterium isolated from gamma ray-irradiated soil

Jiyoun Kim,Geeta Chhetri,Inhyup Kim,Hyungdong Kim,Myung Kyum Kim,Taegun Seo 한국미생물학회 2019 The journal of microbiology Vol.57 No.11

A Gram-stain-negative, asporogenous, aerobic rods, motile by means of a single polar flagellum, catalase- and oxidase-positive, methylotrophic bacterium, designated 17Sr1-28T, was isolated from gamma ray-irradiated soil. The 16S rRNA gene sequence analysis showed that strain 17Sr1-28T was phylogenetically related to Methylobacterium currus PR1016AT (96.8%), Methylobacterium platani PMB02T (96.2%), Methylobacterium aquaticum DSM 16371T (96.3%), Methylobacterium tarhaniae N4211T (96.4%), Methylobacterium frigidaeris IER25-16T (95.8%), and Methylobacterium organophilum JCM 2833T (92.7%). The G+C content calculated based on genome sequence was 71.6%. The average nucleotide identity and in silico DNA-DNA hybridization values between strain 17Sr1- 28T and M. currus, M. platani, M. aquaticum, M. tarhaniae, M. frigidaeris, and M. organophilum were 77.7–90.4% and 22–39.6%, respectively. The major fatty acids of strain 17Sr1- 28T were summed feature 8 (C18:1 ω7c and/or C18:1 ω6c), and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c). The predominant quinone was ubiquinone 10 and the major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol. On the basis of the data from phenotypic tests and genotypic differences between strain 17Sr1-28T and its close phylogenetic relatives, strain 17Sr1-28T represents a new species belonging to the genus Methylobacterium, for which the name Methylobacterium terrae sp. nov. (= KCTC 52904T = NBRC 112873T) is proposed.

Effect of Methylotrophic Bacteria in Seedling Development of Some Crops under Gnotobiotic Condition

In-Soo Hong(홍인수),Jun-Seok Kim(김준석),Min-Kyong Lee(이민경),Woo-Jong Yim(임우종),Md. Rashedul Islam,Hari P. Deka Boruah,Puneet Singh Chauhan,Guang-Hyun Han(한광현),Tong-Min Sa(사동민) 한국토양비료학회 2009 한국토양비료학회지 Vol.42 No.4

농업유용미생물을 이용한 원예 및 채소작물의 건전 유묘 생산을 위하여 본 연구에서는 growth pouch 실험을 통하여 6가지 식물생육촉진 미생물을 접종하여 작물의 초기 뿌리 생장에 미치는 영향을 살펴 보았다. 본 실험에 사용한 6가지 균주는 Methylobacterium oryzae CBMB20, Methylobacterium phyllosphaerae CBMB27, Methylobacterium suomiense CBMB120, Methylobacterium strains CBMB12, CBMB15와 CBMB17이었다. 대조구와 비교했을 CBMB12, CBMB17, 및 CBMB20접종은 상추 초기 뿌리 생육에 유의성있는 효과를 보였고, CBMB102접종은 배추 초기 뿌리 생육에 유의성있는 효과를, CBMB27접종은 토마토의 초기 뿌리 생육에 유의성있는 효과를 보였다. 또한 Methylobacterium suomiense CBMB120접종은 오이, 토마토, 배추, 그리고 Methylobacterium strain CBMB12접종은 고추의 뿌리 초기 생장을 크게 촉진시켰음을 알 수 있었다. 위의 결과를 통하여 실험한 균주들을 각각의 작물 육묘 포트에 처리할 때에도 유묘의 생장속도를 촉진 시킷수 있음을 예상할 수 있다. Healthy seedling generation is the major concern in overcoming adverse effects of biotic and abiotic stresses during tender stage of development in vegetables and horticultural crops. Because of this, priority is given to research leading to the generation of healthy seedlings in crops subjected to transplanting and bedding. In this study, growth pouch experiments were conducted to determine the effect of inoculation of six different strains of Methylobacterium sp. namely, M. oryzae CBMB20, M. phyllosphaerae CBMB27, M. suomiense CBMB120, and Methylobacterium strains CBMB12, CBMB15 and CBMB17 on the seedling development of the vegetable crops cabbage, Chinese cabbage and cucumber; and horticultural crops tomato and red pepper. Crops treated with the test strains generally showed higher seedling dry matte accumulation compared to the control. Significantly higher accumulation was exhibited by CBMB12, CBMB17, and CBMB20 in cabbage, as well as for CBMB27 and CBMB120 on tomato and Chinese cabbage, respectively. Furthermore, all the strains promoted root elongation in cucumber and tomato seedlings while in Chinese cabbage and red pepper, root elongation was observed with CBMB120 and CBMB12 inoculation, respectively. Large scale nursery study is needed to develop a thorough protocol for healthy seedling development with the use of these strains.

Regulation of Ethylene Emission in Tomato (Lycopersicon esculentum Mill.) and Red Pepper (Capsicum annuum L.) Inoculated with ACC Deaminase Producing Methylobacterium spp.

Yim, Woo-Jong,Woo, Sung-Man,Kim, Ki-Yoon,Sa, Tong-Min Korean Society of Soil Science and Fertilizer 2012 한국토양비료학회지 Vol.45 No.1

Improvement of plant growth by Methylotrophic bacteria can be influenced through alterations in growth modulating enzymes or hormones, especially by decreasing ethylene levels enzymatically by 1-aminocyclopropane-1-carboxylate (ACC) deaminase or by production of indole-3-acetic acid (IAA). In this study, the effect of seven strains of Methylobacterium on seedling ethylene emission of tomato and red pepper plants was evaluated under greenhouse condition. Ethylene emission was lowest in Methylobacterium oryzae CBMB20 inoculated tomato plants and CBMB110 inoculated red pepper plants at 47 days after sowing (DAS). However, at 58 DAS all inoculated plants showed almost similar pattern of ethylene emission. Methylobacterium inoculated tomato and red pepper plants showed significantly less ethylene emission compared to control. Our results demonstrated that Methylobacterium spp. inoculation promotes plant growth due to the reduction of ethylene emission and therefore can be potentially used in sustainable agriculture production systems.