• Research in Plant Disease
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• v.26 no.1
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• pp.8-18
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• 2020

Red pepper, one of the major economic crops in Korea, is being affected by anthracnose disease caused by Colletotrichum acutatum. To control this disease, an antagonistic bacterial strain, Bacillus subtilis YGB36 identified by 16S rDNA sequencing, physiological and biochemical analyses is used as a biological control agent. In vitro screening revealed that the strain YGB36 possess strong antifungal activity against the pathogen Cylindrocarpon destructans. The strain exhibited cellulase, protease, amylase, siderophore production and phosphate solubility. In vitro conidial germination of C. acutatum was most drastically inhibited by YGB36 cell suspensions (10⁶ cfu/ml) or culture filtrate. Development of anthracnose symptoms was reduced on detached immature green pepper fruits by treatment with cell suspensions, and its control value was recorded as 65.7%. The YGB36 bacterial suspension treatment enhanced the germination rate of red pepper seeds and promoted root development and growth under greenhouse conditions. The in vitro screening of fungicide and insecticide sensitivity test against YGB36 revealed that the bacterial growth was not affected by any of the insecticides, and 11 fungicides out of 21 used. Collectively, our results clearly suggest that the strain YGB36 is considered as one of the potential biocontrol agents against anthracnose disease in red pepper.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.105-111
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• 2013

Biofilm formation of multifunctional plant growth promoting rhizobacterium (PGPR), Pseudomonas fluorescens 2112 is necessary for P. fluorescens 2112 to have a positive impact on the rhizosphere of red-pepper. This study investigated whether signal molecules of the quorum sensing AHLs are produced in order to confirm biofilm formative ability. Through the use of Petri dish bioassays a blue circle formed evidence of AHLs. It was confirmed that P. fluorescens 2112 produced six-carbon-chain-long AHLs by TLC bioassay. The bacterial density of P. fluorescens 2112 on the top and bottom of pepper plant roots was estimated as $3{\times}10^5$ and $8{\times}10^3$ CFU/g root, respectively. P. fluorescens 2112 exist more with high-density of $3.5{\times}10^6$ CFU/g soil at a depth of 1 cm but at a low-density of $1.1{\times}10$ CFU/g soil at a depth of 5 cm, from the surface of rhizosphere soil. In addition, biofilm formation of P. fluorescens 2112 on the epidermises and the tips of the red-pepper roots were confirmed visually by SEM. Thus, the production of AHLs by P. fluorescens 2112 brings about quorum sensing signaling and the formation of biofilm on the roots which has a positive effect on economically important crops such as red-pepper by additionally producing a variety of antifungal substances and auxin.

• Journal of Microbiology and Biotechnology
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• v.30 no.5
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• pp.668-680
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• 2020

Bacillus velezensis is an important plant growth-promoting rhizobacterium with immense potential in agriculture development. In the present study, Bacillus velezensis Lle-9 was isolated from the bulbs of Lilium leucanthum. The isolated strain showed antifungal activities against plant pathogens like Botryosphaeria dothidea, Fusarium oxysporum, Botrytis cinerea and Fusarium fujikuroi. The highest percentage of growth inhibition i.e., 68.56±2.35% was observed against Fusarium oxysporum followed by 63.12 ± 2.83%, 61.67 ± 3.39% and 55.82 ± 2.76% against Botrytis cinerea, Botryosphaeria dothidea, and Fusarium fujikuroi, respectively. The ethyl acetate fraction revealed a number of bioactive compounds and several were identified as antimicrobial agents such as diketopiperazines, cyclo-peptides, linear peptides, latrunculin A, 5α-hydroxy-6-ketocholesterol, (R)-S-lactoylglutathione, triamterene, rubiadin, moxifloxacin, 9-hydroxy-5Z,7E,11Z,14Z-eicosatetraenoic acid, D-erythro-C18-Sphingosine, citrinin, and 2-arachidonoyllysophosphatidylcholine. The presence of these antimicrobial compounds in the bacterial culture might have contributed to the antifungal activities of the isolated B. velezensis Lle-9. The strain showed plant growth-promoting traits such as production of organic acids, ACC deaminase, indole-3-acetic acid (IAA), siderophores, and nitrogen fixation and phosphate solubilization. IAA production was accelerated with application of exogenous tryptophan concentrations in the medium. Further, the lily plants upon inoculation with Lle-9 exhibited improved vegetative growth, more flowering shoots and longer roots than control plants under greenhouse condition. The isolated B. velezensis strain Lle-9 possessed broad-spectrum antifungal activities and multiple plant growth-promoting traits and thus may play an important role in promoting sustainable agriculture. This strain could be developed and applied in field experiments in order to promote plant growth and control disease pathogens.

• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.106-112
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• 2014

Very few plant growth-promoting rhizobacteria (PGPR) are known to produce gibberellins (GAs). The current study aimed to isolate a phytohormone-producing PGP rhizobacterium from soil and assess its potential to enhance plant growth. The newly isolated bacterium was identified as Leifsonia soli sp. SE134 on the basis of partial 16S ribosomal RNA gene sequence. Application of L. soli culture filtrate significantly increased the biomass, hypocotyl, and root lengths of cucumber seeds as compared with non-inoculated sole medium and distilled water treated controls. Furthermore, the PGPR culture was applied to the GA-deficient mutant rice cultivar Waito-C. Treatment with L. soli SE134 significantly increased the growth of Waito-C rice seedlings as compared with controls. Upon chromatographic analysis of L. soli culture, we isolated, detected and quantified different GAs; namely, $GA_1$ ($0.61{\pm}0.15$), $GA_4$ ($1.58{\pm}0.26$), $GA_7$ ($0.54{\pm}0.18$), $GA_8$ ($0.98{\pm}0.15$), $GA_9$ ($0.45{\pm}0.17$), $GA_{12}$ ($0.64{\pm}0.21$), $GA_{19}$ ($0.18{\pm}0.09$), $GA_{20}$ ($0.78{\pm}0.15$), $GA_{24}$ ($0.38{\pm}0.09$), $GA_{34}$ ($0.35{\pm}0.10$), and $GA_{53}$ ($0.17{\pm}0.05$). Plant growth promotion in cucumber, tomato, and young radish plants further evidenced the potential of this strain as a PGP bacterium. The results suggest that GA secretion by L. soli SE134 might prove advantageous for its ameliorative role in crop growth. These findings can be extended for improving the productivity of different crops under diverse environmental conditions.

• KSBB Journal
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• v.24 no.4
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• pp.334-340
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• 2009

For the isolation of a plant growth-promoting rhizobacteria, strain YK-5 was selected from approximately 400 thermostable strains isolated from special soil samples. Strain YK-5 produced an antifungal compound, and optimum carbon and nitrogen sources for the production of the antifungal compound were investigated against Aspergillus flavus as a test strain. Modified LB medium containing 1% peptone, 1% yeast extract and 5% black sugar was determined to be the optimal medium for growth and antifungal compound production. Culture broth of strain YK-5 potently inhibited growth of the phytopathogenic fungus Fusarium oxysporum KACC 40052 for 7 days. The plant growth-promotion function of strain YK-5 was tested against radish and rice in pot trials. Leaf number, plant height and root length in YK-5-treated radish markedly exceeded (> 60%) those of untreated radish. Leaf length and white rootlet development were markedly more prominent than in commercially-treated rice plants. Strain YK-5 was determined to be Bacillus subtilis YK-5 by physiological, chemotaxonomical, and phylogenetical analyses.

• Applied Biological Chemistry
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• v.50 no.2
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• pp.95-100
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• 2007

The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

• Korean Journal of Soil Science and Fertilizer
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• v.36 no.3
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• pp.134-144
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• 2003

A rhizobacterium Pseudomonas cholororaphis O6 produced several secondary metabolites, such as phenazines, protease, and HCN that may be involved in inhibition of the growth of phytopathogenic fungi. In field study, P. chlororaphis O6 treatment on wheat seed suppressed root rot disease caused by Fusarium culmorum. The major organic acids of cucumber root exudates were fumaric acid, malic acid, benzoic acid, and succinic acid. Glucose and fructose were major monosaccharides in cucumber root exudates. The total amount of organic acids was ten times higher than that of the sugars. P. chlororaphis O6 grew well on cucumber root exudates. The dctA gene of P. chlororaphis O6 consisted of a 1,335 bp open reading frame with a deduced amino acid sequence of 444 residues, corresponding to a molecular size of about 47 kD and pI 8.2. The deduced dctA sequence has ten putative transmembrane domains, as expected of a membrane-embedded protein. Our results indicated that organic acids in cucumber root exudates may play an important role in providing nutrient source for root colonization of biological control bacteria, and the dctA gene of P. chlororaphis O6 may be an important bacterial trait that is involved in utilization of root exudates.

• Korean Journal of Organic Agriculture
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• v.26 no.4
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• pp.677-686
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• 2018

This study was carried out to determine the effect of Kluyvera sp. CL-2 (KACC 91283P) on the growth of watermelon (Citrullus vulgaris L.). The study consisted of three treatments, no treatment (NT), twice application of Kluyvera sp. CL-2 before transplanting (KC1), and five times application around transplanting (KC2). We determined the chemical properties of soil before and after the treatments, and compared the growth characteristics of watermelon among treatments. The treatment of Kluyvera sp. CL-2 at $1.0{\times}10^6cfu\;mL^{-1}$ significantly increased available $P_2O_5$. The organic matter showed to increase for all treatments, while soil pH, exchangeable Ca and Mg tended to decrease for all treatments. The leaf width was increased by 11.6% for KC1 and 26.2% for KC2 compared to NT. But there were no significant differences in yield, leaf length, fruit weight, fruit length, fruit width, and pericarp thickness among treatments. The contents of free sugars such as fructose and glucose were increased by microbial treatments but sucrose was not different from NT. The content of glucose in watermelon was increased by 13.8% in KC1 and 12.8% in KC2 compared to NT. The content of fructose increased by 14.6% in KC1 and by 39.8% in KC2 compared to NT. The results from the study imply that Kluyvera sp. CL-2 can be used to increase sugar content in watermelon.