• Journal of Plant Biotechnology
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• 제49권4호
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• pp.356-361
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• 2022

본 연구에서는 Bacillus amyloliquefaciens ISP-5 균주의 최적배지조성을 확인하기 위해 5종의 탄소원(glucose, fructose, sucrose, maltose, corn starch)과 5종의 질소원(yeast extract, peptone, tryptone, soytone, soy bean flour)을 이용하여 각각의 생육을 측정하였다. 그 결과 glucose와 soy bean flour를 각각 최적 탄소원과 질소원으로 확립하였으며 최적 배지를 이용하였을 때 상업용 배지(TSB)에 비해 약 2500배의 생육을 확인할 수 있었다. B. amyloliquefaciens ISP-5 배양액을 상추에 엽면 살포하였을 때 상추의 엽폭과 엽장은 대조군에 비해 8.6%, 12.9%의 증가하였으며, 생체중과 건물중은 대조군에 비해 각각 24.2%, 23.9% 증가하였다. 이러한 결과들을 종합해보았을 때 B. amyloloiquefaciens ISP-5 균주를 이용한 미생물 제제는 상추용 식물생장촉진제로써 활용할 수 있을 것이라 기대된다.

• The Plant Pathology Journal
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• 제39권5호
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• pp.430-448
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• 2023

Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1213-1222
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• 2009

The search for diverse plant growth-promoting (PGP) diazotrophic bacteria is gaining momentum as efforts are made to exploit them as biofertilizers for various economically important crops. In the present study, 17 diazotrophic strains belonging to eight different genera isolated from rice paddy fields were screened for multiple PGP traits and evaluated for their inoculation effects on canola and rice plants. All of the strains tested positive for 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity and production of indole 3-acetic acid (IAA) and ammonia ($NH_3$). Additionally, four of the strains were able to solubilize phosphorus (P), five tested positive for zinc (Zn) solubilization and sulfur (S) oxidation, and eight strains produced siderophores. Based on the presence of multiple PGP traits, 10 strains were selected for inoculation studies. Treatment with Herbaspirillum sp. RFNB26 resulted in maximum root length (54.3%), seedling vigor, and dry biomass in canola, whereas Paenibacillus sp. RFNB4 exhibited the lowest activity under gnotobiotic conditions. However, under pot culture conditions, Paenibacillus sp. RFNB4 significantly increased plant height and dry biomass production by 42.3% and 29.5%, respectively. Canola plants and rhizosphere soils inoculated with Bacillus sp. RFNB6 exhibited significantly higher nitrogenase activity. In greenhouse experiments, Serratia sp. RFNB18 increased rice plant height by 35.1%, Xanthomonas sp. RFNB24 enhanced biomass production by 84.6%, and rice rhizosphere soils inoculated with Herbaspirillum sp. RFNB26 exhibited the highest nitrogenase activity. Our findings indicate that most of the selected strains possess multiple PGP properties that significantly improve the growth parameters of the two plants when tested under controlled conditions.

• 한국미생물·생명공학회지
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• 제49권3호
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• pp.391-402
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• 2021

Earthworms play an important role in soil fertilization, interacting continually with microorganisms. This study aims to demonstrate the existence of beneficial microorganisms living in the earthworm's immune system, the coelomic fluid. To achieve this goal, a molecular identification technique was performed, using cytochrome c oxidase I (COI) barcoding to identify abundant endogenic earthworms inhabiting the temperate zone of Rabat, Morocco. Then, 16S rDNA and ITS sequencing techniques were adopted for bacteria and fungi, respectively. Biochemical analysis, showed the ability of bacteria to produce characteristic enzymes and utilize substrates. Qualitative screening of plant growth-promoting traits, including nitrogen fixation, phosphate and potassium solubilization, and indole acetic acid (IAA) production, was also performed. The result of mitochondrial COI barcoding allowed the identification of the earthworm species Aporrectodea molleri. Phenotypic and genotypic studies of the sixteen isolated bacteria and the two isolated fungi showed that they belong to the Pseudomonas, Aeromonas, Bacillus, Buttiauxella, Enterobacter, Pantoea, and Raoultella, and the Penicillium genera, respectively. Most of the isolated bacteria in the coelomic fluid showed the ability to produce β-glucosidase, β-glucosaminidase, Glutamyl-β-naphthylamidase, and aminopeptidase enzymes, utilizing substrates like aliphatic thiol, sorbitol, and fatty acid ester. Furthermore, three bacteria were able to fix nitrogen, solubilize phosphate and potassium, and produce IAA. This initial study demonstrated that despite the immune property of earthworms' coelomic fluid, it harbors beneficial microorganisms. Thus, the presence of resistant microorganisms in the earthworm's immune system highlights a possible selection process at the coelomic fluid level.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1119-1128
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• 2015

1-Aminocyclopropane-1-carboxylate (ACC) deaminase, which is encoded by some bacteria, can reduce the amount of ethylene, a root elongation inhibitor, and stimulate the growth of plants under various environmental stresses. The presence of ACC deaminase activity and the regulation of ACC in several rhizospheric bacteria have been reported. The nitrogen-fixing Pseudomonas stutzeri A1501 is capable of endophytic association with rice plants and promotes the growth of rice. However, the functional identification of ACC deaminase has not been performed. In this study, the proposed effect of ACC deaminase in P. stutzeri A1501 was investigated. Genome mining showed that P. stutzeri A1501 carries a single gene encoding ACC deaminase, designated acdS. The acdS mutant was devoid of ACC deaminase activity and was less resistant to NaCl and NiCl₂ compared with the wild-type. Furthermore, inactivation of acdS greatly impaired its nitrogenase activity under salt stress conditions. It was also observed that mutation of the acdS gene led to loss of the ability to promote the growth of rice under salt or heavy metal stress. Taken together, this study illustrates the essential role of ACC deaminase, not only in enhancing the salt or heavy metal tolerance of bacteria but also in improving the growth of plants, and provides a theoretical basis for studying the interaction between plant growth-promoting rhizobacteria and plants.

• The Plant Pathology Journal
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• 제31권2호
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• pp.152-164
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• 2015

Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.

• 한국토양비료학회지
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• 제42권4호
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• pp.266-273
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• 2009

화학비료와 유기질비료를 시비 수준 및 Methylobacterium suomiense CBMB120의 접종이 고추 생육에 미치는 영향을 확인하기 위하여 유기질비료와 화학비료의 시비량을 권장시용수준의 100%, 75% 및 50%로 처리하고 Methylobacterium suomiense CBMB120을 접종 한 후 정식 후 19, 36, 및 166일에 작물의 높이를 측정하였으며 이식 후 166일에 건물량 측정하였다. 균주를 접종한 처리구의 식물체 높이는 정식 후 19, 36일에 각각 14.17%, 10.03% 증가하였다. 건물 중 역시 10.30%, 6.84% 증가하여 유의성 있는 차이를 나타내었다. 또한 균주 접종은 유기질 비료 100% 시용구는 36일 후 고도의 유의성(p<0.01)을 갖는 차이를 보였으며 화학 비료 100% 시용구는 19일 후 고도의 유의성(p<0.01)을 갖는 차이를 나타내었다. 화학비료 100% 시용구에 균주를 접종하지 않은 처리 구와 화학비료 75% 시용구에 균주를 접종한 처리구의 생육을 분석한 결과 유의성 있는 차이가 없는 것으로 나타났다. 이는 향후 다양한 처리량 및 처리방법의 연구를 통하여 Methylobacterium suomiense CBMB120 균주의 접종이 화학비료를 일정 부분 감비 할 수 있다는 가능성을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제20권12호
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• pp.1614-1623
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• 2010

Bacteria were isolated from roots of sugarcane varieties grown in the fields of Punjab. They were identified by using API20E/NE bacterial identification kits and from sequences of 16S rRNA and amplicons of the cpn60 gene. The majority of bacteria were found to belong to the genera of Enterobacter, Pseudomonas, and Klebsiella, but members of genera Azospirillum, Rhizobium, Rahnella, Delftia, Caulobacter, Pannonibacter, Xanthomonas, and Stenotrophomonas were also found. The community, however, was dominated by members of the Pseudomonadaceae and Enterobacteriaceae, as representatives of these genera were found in samples from every variety and location examined. All isolates were tested for the presence of five enzymes and seven factors known to be associated with plant growth promotion. Ten isolates showed lipase activity and eight were positive for protease activity. Cellulase, chitinase, and pectinase were not detected in any strain. Nine strains showed nitrogen fixing ability (acetylene reduction assay) and 26 were capable of solubilizing phosphate. In the presence of 100 mg/l tryptophan, all strains except one produced indole acetic acid in the growth medium. All isolates were positive for ACC deaminase activity. Six strains produced homoserine lactones and three produced HCN and hexamate type siderophores. One isolate was capable of inhibiting the growth of 24 pathogenic fungal strains of Colletotrichum, Fusarium, Pythium, and Rhizoctonia spp. In tests of their abilities to grow under a range of temperature, pH, and NaCl concentrations, all isolates grew well on plates with 3% NaCl and most of them grew well at 4 to $41^{\circ}C$ and at pH 11.

• Applied Biological Chemistry
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• 제48권3호
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• pp.296-300
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• 2005

Among the root colonizing and plant growth promoting bacteria isolated from the bacterial wilt suppressive soil, five strains were detected to produce siderophores by CAS agar assay. The most effective isolate, TS3-7 strain induced significant suppression of bacterial wilt disease in tomato and pepper plants. Seed treatment followed by soil drench application with this strain resulted in over 80% reduction of bacterial wilt disease compared with the control. Significant disease suppression by TS3-7 strain was related to the production of siderophore. Besides iron competition, induction of resistance of the host plant with siderophore was suggested to be another mode of action that suppress bacterial wilt, based on the lack of direct antibiosis against pathogen in vitro. According to Bergey's Manual of Systemic Bacteriology and 16S rDNA sequence data, TS3-7 stain was identified as Pseudomonas sp. TS3-7.

• 한국토양비료학회지
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• 제45권2호
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• pp.169-176
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• 2012

Among the major nutrients, phosphorus is by far the least mobile and available to plants in most soil conditions. A large portion of soluble inorganic phosphate applied to soil in the form of phosphate fertilizers is immobilized rapidly and becomes unavailable to plants. To improve the plant growth and yield and to minimize P loss from soils, the ability of a few soil microorganisms converting insoluble forms into soluble forms for phosphorus is an important trait in several plant growth-promoting microorganisms belonging to the genera Bacillus and Pseudomonas and the fungi belonging to the genera Penicillium and Aspergillus in managing soil phosphorus. The principal mechanism of solubilization of mineral phosphate by phosphate solubilizing bacteria (PSB) is the release of low molecular weight organic acids such as formic, acetic, propionic, lactic, glycolic, fumaric, and succinic acids and acidic phosphatases like phytase synthesized by soil microorganisms in soil. Hydroxyl and carboxyl groups from the organic acids can chelate the cations bound to phosphate, thereby converting it into soluble forms.