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Isolation of Agrobacterium sp. BE516 from the Root of Miscanthus sacchariflorus and Its Plant Growth Promoting Activity (물억새 뿌리로부터 Agrobacterium sp. BE516 균주의 분리 및 식물생육촉진활성)

  • Kang, Hye-Young;Park, Dong-Jin;Lee, Jae-Chan;Kwon, Mi-Kyung;Kim, Seung-Bum;Kim, Chang-Jin
    • Journal of Applied Biological Chemistry
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    • v.55 no.2
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    • pp.129-133
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    • 2012
  • To exploit plant growth promoting bacteria in the roots of Miscanthus sacchariflorus, a biomass energy crop, total 64 bacteria were isolated. For the investigation of plant growth promoting effects from the isolated bacteria, production of indole acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activities were tested and other cultural conditions were examined. As results, 8 isolates showed plant growth promoting effects on the M. sacchariflorus and an isolate designated Agrobacterium sp. BE516 has the highest activity by enhancing the shoot elongation over 2-fold than the control. Agrobacterium sp. BE516 produced 64 ${\mu}g$ IAA per mL and showed ACC deaminase activity which is involved in the resistance to environmental stress such as high salt and drought. It could grow at low temperature in the range from 4 to $15^{\circ}C$, at pH 4.0 and at 4% NaCl. These results indicate that the Agrobacterium sp. BE516 can be useful as a bio-fertilizer for M. sacchariflorus under the stressed conditions.

Biochemical and Molecular Characterization of High Population Density Bacteria Isolated from Sunflower

  • Goes, Kelly Campos Guerra Pinheiro De;Fisher, Maria Luisa De Castro;Cattelan, Alexandre Jose;Nogueira, Marco Antonio;Carvalho, Claudio Guilherme Portela De;Oliveira, Andre Luiz Martinez De
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.437-447
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    • 2012
  • Natural and beneficial associations between plants and bacteria have demonstrated potential commercial application for several agricultural crops. The sunflower has acquired increasing importance in Brazilian agribusiness owing to its agronomic characteristics such as the tolerance to edaphoclimatic variations, resistance to pests and diseases, and adaptation to the implements commonly used for maize and soybean, as well as the versatility of the products and by-products obtained from its cultivation. A study of the cultivable bacteria associated with two sunflower cultivars, using classical microbiological methods, successfully obtained isolates from different plant tissues (roots, stems, florets, and rhizosphere). Out of 57 plant-growth-promoting isolates obtained, 45 were identified at the genus level and phylogenetically positioned based on 16S rRNA gene sequencing: 42 Bacillus (B. subtilis, B. cereus, B. thuringiensis, B. pumilus, B. megaterium, and Bacillus sp.) and 3 Methylobacterium komagatae. Random amplified polymorphic DNA (RAPD) analysis showed a broad diversity among the Bacillus isolates, which clustered into 2 groups with 75% similarity and 13 subgroups with 85% similarity, suggesting that the genetic distance correlated with the source of isolation. The isolates were also analyzed for certain growth-promoting activities. Auxin synthesis was widely distributed among the isolates, with values ranging from 93.34 to 1653.37 ${\mu}M$ auxin per ${\mu}g$ of protein. The phosphate solubilization index ranged from 1.25 to 3.89, and siderophore index varied from 1.15 to 5.25. From a total of 57 isolates, 3 showed an ability to biologically fix atmospheric nitrogen, and 7 showed antagonism against the pathogen Sclerotinia sclerotiorum. The results of biochemical characterization allowed identification of potential candidates for the development of biofertilizers targeted to the sunflower crop.

Cloning and mutational analysis of pyrroquinoline quinone(PQQ) genes from a phosphate - solubilizing biocontrol bacterium Enterobacter intermedium.

  • Han, Song-Hee;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.94.2-95
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    • 2003
  • E. intermedium 60-2G possessing a strong ability to solubilize insoluble phosphate, has plant growth-promoting activity, induced systemic resistance activity against scab pathogen in cucumber, and antifungal activity against various phytopathogenic fungi. The phosphate solubilizing activity of 60-2G may be mainly accomplished by production of gluconic acid through a direct extracellular oxidation of glucose by glucose dehydrogenase that required a PQQ cofactor for its activation. A pqq gene cluster conferred Phosphate-solubilizing activity in E. coli DH5${\alpha}$ was cloned and sequenced. The 6,783 bP pqq sequence had six open reading frames (from A to F) and showed 50-95% homology to pqq genes from other bacteria. The E. coli strain expressing the pqq genes solubilized phosphate from hydroxyapatite after a pH drop to 4.0, which paralleled in time the secretion of gluconic acid. To study the role of PQQ in biocontrol traits of E. intermedium, PQQ mutants of 60-2G were constructed by marker exchangee mutagenesis. The PQQ mutants of E. intermedium were lost activities of solubilizing phosphate, growth inhibition of phytopathogenic fungi, and plant growth promotion. These findings suggest that PQQ plays an important role, possibly activation of certain enzymes, in several beneficial bacterial traits of E. intermedium by as yet an unknown mechanism.

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Selection of Plant Growth-Promoting Pseudomonas spp. That Enhanced Productivity of Soybean-Wheat Cropping System in Central India

  • Sharma, Sushil K.;Johri, Bhavdish Narayan;Ramesh, Aketi;Joshi, Om Prakash;Sai Prasad, S.V.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.11
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    • pp.1127-1142
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    • 2011
  • The aim of this investigation was to select effective Pseudomonas sp. strains that can enhance the productivity of soybean-wheat cropping systems in Vertisols of Central India. Out of 13 strains of Pseudomonas species tested in vitro, only five strains displayed plant growth-promoting (PGP) properties. All the strains significantly increased soil enzyme activities, except acid phosphatase, total system productivity, and nutrient uptake in field evaluation; soil nutrient status was not significantly influenced. Available data indicated that six strains were better than the others. Principal component analysis (PCA) coupled cluster analysis of yield and nutrient data separated these strains into five distinct clusters with only two effective strains, GRP3 and HHRE81 in cluster IV. In spite of single cluster formation by strains GRP3 and HHRE81, they were diverse owing to greater intracluster distance (4.42) between each other. These results suggest that the GRP3 and HHRE81 strains may be used to increase the productivity efficiency of soybean-wheat cropping systems in Vertisols of Central India. Moreover, the PCA coupled cluster analysis tool may help in the selection of other such strains.

In Vivo Expression Technology (IVET) and Its Application in Plant-Associated Bacteria

  • Lee, Seon-Woo
    • The Plant Pathology Journal
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    • v.18 no.2
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    • pp.57-62
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    • 2002
  • In vivo expression technology (IVET) has been developed to study bacterial gene expression in Salmonella typhimurium during host infection. The expression of selected genes by IVET has been elevated in vivo but not in vitro. The selected genes turned out to be important for bacterial virulence and/or pathogenicity. IVET depends on a synthetic operon with a promoterless transcriptional fusion between a selection marker gene and a reporter gene. The IVET approach has been successfully adapted in other bacterial pathogens and plant-associated bacteria using different selection markers. Pseudomonas putida suppresses citrus root rot caused by Phytophthora parasitica and enhances citrus seedling growth. The WET strategy was adapted based on a transcriptional fusion, pyrBC'-lacZ, in P. putida to study the bacterial traits important far biocontrol activities. Several genes appeared to be induced on P. parasitica hyphae and were found to be related with metabolism and regulation of gene expression. It is likely that the biocontrol strain took a metabolic advantage from the plant pathogenic fungus and then suppressed citrus root rot effectively. The result was parallel with those from the adaptation of IVET in P. fluorescens, a plant growth promoting rhizobacteria (PGPR). Interestingly, genes encoding components for type III secretion system have been identified as rhizosphere-induced genes in the PGPR strain. The type III secretion system may play a certain role during interaction with its counterpart plants. Application of IVET has been demonstrated in a wide range of bacteria. It is an important strategy to genetically understand complicated bacterial traits in the environment.

Studies on the effects of medicinal plant extracts on the hair growth stimulation (數種의 韓藥材가 毛髮成長에 미치는 影響)

  • Choi, Woong;Choi, Jung-Hwa;Kim, Jong-Han
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.15 no.2
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    • pp.80-103
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    • 2002
  • To screen the effective materials for hair loss treatment, several natural extracts were tested using in vivo and in vitro test models. Firstly, all test materials were applicated onto the back skin of C57BL/6 mouse and then hair growth pormoting effect were measured using hair growth index As a result, Polygonum muitifiorum Thunb and Terrninalia chebula Retz. showed potent hair growth promoting effect, ranking as 1.5-2.0 of hair growth index. However, there were no plant extracts, which have remarkable potential of growth promotion of human hair dermal papilla cells cultured in vitro. In the experiments of 5${\alpha}$-reductase type Ⅱ inhibition assay, Morus alba L., Chaenomelis Fructus, Saussureae Radix, Angelicae Gigantis Radix, Polygonum multifiorum Thunb, and Angelica dahurica (Fischer) Bentham et Hooker f. showed effective potential to inhibit the activity of 5${\alpha}$-reductase type Ⅱ. To investigate the possible involvement of effects of several plant extracts on the gene expression of growth factors in human hair dermal papilla cells, RT-PCR analyses were performed. As a consequences, Mentha haplocalyx Briq., Cimicifuga foetida L., Eclipta prostrata (L.) L., Pinus densiflora S. et. Z, and Polygonum muitifiorum Thunb revealed the regulatory roles on the expression of growth factors such as IGF-I, KGF, HGF and VEGF in the dermal papilla cells. Another test for inhibition of microbial such as P. acne and P. ovale were also carried out to find whether these plant extracts have anti-microbial activities. Morus alba L. and Chaenomelis Fructus showed anti-microbial effects on Propionibacterium acnes, which is believed as a pathogen of acne. Together, these results showed several plant extracts can be used for hair growth promotion.

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Evaluation of Soil Streptomyces spp. for the Biological Control of Fusarium Wilt Disease and Growth Promotion in Tomato and Banana

  • Praphat, Kawicha;Jariya, Nitayaros;Prakob, Saman;Sirikanya, Thaporn;Thanwanit, Thanyasiriwat;Khanitta, Somtrakoon;Kusavadee, Sangdee;Aphidech, Sangdee
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.108-122
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    • 2023
  • Fusarium oxysporum f. sp. lycopersici (Fol) and Fusarium oxysporum f. sp. cubense (Foc), are the causal agent of Fusarium wilt disease of tomato and banana, respectively, and cause significant yield losses worldwide. A cost-effective measure, such as biological control agents, was used as an alternative method to control these pathogens. Therefore, in this study, six isolates of the Streptomyces-like colony were isolated from soils and their antagonistic activity against phytopathogenic fungi and plant growth-promoting (PGP) activity were assessed. The results showed that these isolates could inhibit the mycelial growth of Fol and Foc. Among them, isolate STRM304 showed the highest percentage of mycelial growth reduction and broad-spectrum antagonistic activity against all tested fungi. In the pot experiment study, the culture filtrate of isolates STRM103 and STRM104 significantly decreased disease severity and symptoms in Fol inoculated plants. Similarly, the culture filtrate of the STRM304 isolate significantly reduced the severity of the disease and symptoms of the disease in Foc inoculated plants. The PGP activity test presents PGP activities, such as indole acetic acid production, phosphate solubilization, starch hydrolysis, lignin hydrolysis, and cellulase activity. Interestingly, the application of the culture filtrate from all isolates increased the percentage of tomato seed germination and stimulated the growth of tomato plants and banana seedlings, increasing the elongation of the shoot and the root and shoot and root weight compared to the control treatment. Therefore, the isolate STRM103 and STRM104, and STRM304 could be used as biocontrol and PGP agents for tomato and banana, respectively, in sustainable agriculture.

IAA-Producing Penicillium sp. NICS01 Triggers Plant Growth and Suppresses Fusarium sp.-Induced Oxidative Stress in Sesame (Sesamum indicum L.)

  • Radhakrishnan, Ramalingam;Shim, Kang-Bo;Lee, Byeong-Won;Hwang, Chung-Dong;Pae, Suk-Bok;Park, Chang-Hwan;Kim, Sung-Up;Lee, Choon-Ki;Baek, In-Youl
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.856-863
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    • 2013
  • Application of rhizospheric fungi is an effective and environmentally friendly method of improving plant growth and controlling many plant diseases. The current study was aimed to identify phytohormone-producing fungi from soil, to understand their roles in sesame plant growth, and to control Fusarium disease. Three predominant fungi (PNF1, PNF2, and PNF3) isolated from the rhizospheric soil of peanut plants were screened for their growth-promoting efficiency on sesame seedlings. Among these isolates, PNF2 significantly increased the shoot length and fresh weight of seedlings compared with controls. Analysis of the fungal culture filtrate showed a higher concentration of indole acetic acid in PNF2 than in the other isolates. PNF2 was identified as Penicillium sp. on the basis of phylogenetic analysis of ITS sequence similarity. The in vitro biocontrol activity of Penicillium sp. against Fusarium sp. was exhibited by a 49% inhibition of mycelial growth in a dual culture bioassay and by hyphal injuries as observed by scanning electron microscopy. In addition, greenhouse experiments revealed that Fusarium inhibited growth in sesame plants by damaging lipid membranes and reducing protein content. Co-cultivation with Penicillium sp. mitigated Fusarium-induced oxidative stress in sesame plants by limiting membrane lipid peroxidation, and by increasing the protein concentration, levels of antioxidants such as total polyphenols, and peroxidase and polyphenoloxidase activities. Thus, our findings suggest that Penicillium sp. is a potent plant growth-promoting fungus that has the ability to ameliorate damage caused by Fusarium infection in sesame cultivation.

Phosphate solubilization by phosphate solubilizing microorganisms: insight into the mechanisms

  • Buddhi Charana, Walpola;Kodithuwakku Kankanange Indika Upali, Arunakumara;Min Ho, Yoon
    • Korean Journal of Agricultural Science
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    • v.49 no.3
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    • pp.463-481
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    • 2022
  • Phosphorous (P) is considered to be one of the key essential elements demanded by crop plants. Approximately 70 - 90% of phosphatic fertilizers applied to crops are fixed in soil as Ca, Fe, and Al metal cations, which are insoluble and thus not readily available for plant uptake. Therefore, most soils are deficient in plant available P. This is usually rectified by applying phosphate fertilizers continuously, although this is not economically viable or environmentally acceptable. The present paper reviews the mechanisms involved with phosphate solubilization and mineralization by phosphate solubilizing microorganisms (PSMs) with the associated factors that determine the success. PSMs are effectively involved in mediating the bioavailability of soil P. Their contribution includes mineralization of organic P solubilization of inorganic P minerals, and storing sizable amounts of P in biomass through different mechanisms such as the production of organic and inorganic acids, H2S, siderophores, exopolysaccharides, and production of enzymes such as phosphatases, phytase, and phosphonatases/C-P lyases, which are capable of chelating the metal ions, forming complexes, and making plant available P. PSMs manifest a wide range of metabolic functions in different environments, resulting in significantly higher plant growth, enhanced soil properties, and increased biological activities. Therefore, development of bio-inoculants with efficient novel PSM strains and further investigations on exploring such strains from diverse ecological niches with multifunctional plant-growth-promoting traits are needed.

Evaluation and Genome Mining of Bacillus stercoris Isolate B.PNR1 as Potential Agent for Fusarium Wilt Control and Growth Promotion of Tomato

  • Rattana Pengproh;Thanwanit Thanyasiriwat;Kusavadee Sangdee;Juthaporn Saengprajak;Praphat Kawicha;Aphidech Sangdee
    • The Plant Pathology Journal
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    • v.39 no.5
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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.