• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.352-359
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• 2012

One hundred thirty-four putative Bacillus isolates were recovered from soybean rhizosphere soils of Nimar region to select effective zinc solubilizers for increased assimilation of zinc (Zn) in soybean seeds. These isolates were screened in vitro for zinc-solubilization ability on Tris-minimal agar medium supplemented separately with 0.1% zinc in the form of zinc oxide, zinc phosphate, and zinc carbonate. Of all, 9 isolates and a reference Bacillus cereus ATCC 13061 were characterized and identified as Bacillus species based on Gram-positive reaction, endospore-forming cells, and the presence of iso-$C_{15:0}$ and anteiso-$C_{15:0}$ as predominant fatty acids. On plate assay, two isolates KHBD-6 and KHBAR-1 showed a greater diameter of solubilization halo and colony diameter on all the three zinc compounds. The isolates KHBD-6, KHBAR-1, BDSD-2-2C, and KHTH-4-1 and the reference strain ATCC 13061 had higher soluble zinc concentration in liquid medium supplemented with zinc phosphate and zinc carbonate compounds as compared with the other isolates and uninoculated control. Evaluation under microcosm conditions showed that inoculation of isolates KHBD-6 (57.34 ${\mu}g/g$), KHBAR-1 (55.67 ${\mu}g/g$), and strain ATCC 13061 (53.10 ${\mu}g/g$) significantly increased the Zn concentration in soybean seeds as compared with the other isolates and uninoculated control (47.14 ${\mu}g/g$). This study suggests the occurrence of zinc-solubilizing Bacillus in soils of Nimar region and isolates KHBD-6 and KHBAR-1 were found to be promising zinc solubilizers for increased assimilation of Zn in soybean seeds.

• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1477-1482
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• 2007

Gluconacetobacter diazotrophicus strain PA15 exhibited a minimum inhibitory concentration value of 11 mM in an LGI medium amended with $ZnCl_2$. When an LGI medium was amended with Zn metal, solubilization halos were observed in a plate assay, and further solubilization was confirmed in a broth assay. The maximum solubilization was recorded after 120 h with a 0.1% Zn metal amendment. During solubilization, the culture growth and pH of the broth were indirectly correlated. Using a Fourier Transform Infrared Spectroscopy analysis, one of the agents solubilizing the Zn metal was identified as gluconic acid. When the Zn-amended broth was observed under a bright field microscope, long involution cells were observed, and further analysis with Atomic Force Microscopy revealed highly deformed, pleomorphic, aggregate-like cells.

• Journal of Microbiology and Biotechnology
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• v.32 no.11
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• pp.1435-1446
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• 2022

Zinc-solubilizing bacteria can convert the insoluble form of zinc into soluble forms available to plants. This study was conducted to isolate and screen zinc-solubilizing actinobacteria from rhizosphere soils and to assess their effect on vegetable soybean growth. In total, 200 actinobacteria strains belonging to 10 genera were isolated from rhizosphere soil samples. Among these isolates, four showed zinc solubilization with solubilizing index values ranging from 3.11 to 3.78 on Bunt and Rovira agar supplemented with 0.1% zinc oxide. For the quantitative assay, in broth culture, strains CME34 and EX51 solubilized maximum available zinc contents of 529.71 and 243.58 ㎍/ml. Furthermore, indole-3-acetic acid (IAA) and ammonia were produced by these two strains, the strain CME34 produced the highest amount of IAA 4.62 ㎍/ml and the strain EX51 produced the highest amount of ammonia 361.04 ㎍/ml. In addition, the phosphate-solubilizing abilities in Pikovskaya's medium of CME34 and EX51 were 64.67 and 115.67 ㎍/ml. Based on morphological and biochemical characterization and 16S rDNA sequencing, the strains CME34 and EX51 were closely related to the genus Streptomyces. In a greenhouse experiment, single-strain inoculation of Streptomyces sp. CME34 or EX51 significantly increased the shoot length, root length, plant dry weight, number of pods per plant and number of seeds per plant of vegetable soybean plants compared to the uninoculated control. These findings facilitated the conclusion that the two Streptomyces strains have potential as zinc solubilizers and can be suggested as bioinoculants to promote the growth and yield of soybean.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.480-491
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• 2017

Fluorescent pseudomonads have been isolated from halophytes, mesophytes, and xerophytes of Pakistan. Among these, eight isolates, GS-1, GS-3, GS-4, GS-6, GS-7, FS-2 (cactus), ARS-38 (cotton), and RP-4 (para grass), showed antifungal activity and were selected for detailed study. Based on biochemical tests and 16S rRNA gene sequences, these were identified as strains of P. chlororaphis subsp. chlororaphis and aurantiaca. Secondary metabolites of these strains were analyzed by LC-MS. Phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine, Cyclic Lipopeptide (white line-inducing principle (WLIP)), and lahorenoic acid A were detected in variable amounts in these strains. P. aurantiaca PB-St2 was used as a reference as it is known for the production of these compounds. The phzO and PCA genes were amplified to assure that production of these compounds is not an artifact. Indole acetic acid production was confirmed and quantified by HPLC. HCN and siderophore production by all strains was observed by plate assays. These strains did not solubilize phosphate, but five strains were positive for zinc solubilization. Wheat seedlings were inoculated with these strains to observe their effect on plant growth. P. aurantiaca strains PB-St2 and GS-6 and P. chlororaphis RP-4 significantly increased both root and shoot dry weights, as compared with uninoculated plants. However, P. aurantiaca strains FS-2 and ARS-38 significantly increased root and shoot dry weights, respectively. All strains except PB-St2 and ARS-38 significantly increased the root length. This is the first report of the isolation of P. aurantiaca from cotton and cactus, P. chlororaphis from para grass, WLIP and lahorenoic acid A production by P. chlororaphis, and zinc solubilization by P. chlororaphis and P. aurantiaca.

• Journal of Microbiology and Biotechnology
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• v.21 no.6
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• pp.556-566
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• 2011

A total of 31 Acinetobacter isolates were obtained from the rhizosphere of Pennisetum glaucum and evaluated for their plant-growth-promoting traits. Two isolates, namely Acinetobacter sp. PUCM1007 and A. baumannii PUCM1029, produced indole acetic acid (10-13 ${\mu}g$/ml). A total of 26 and 27 isolates solubilized phosphates and zinc oxide, respectively. Among the mineral-solubilizing strains, A. calcoaceticus PUCM1006 solubilized phosphate most efficiently (84 mg/ml), whereas zinc oxide was solubilized by A. calcoaceticus PUCM1025 at the highest solubilization efficiency of 918%. All the Acinetobacter isolates, except PUCM1010, produced siderophores. The highest siderophore production (85.0 siderophore units) was exhibited by A. calcoaceticus PUCM1016. Strains PUCM1001 and PUCM1019 (both A. calcoaceticus) and PUCM1022 (Acinetobacter sp.) produced both hydroxamate-and catechol-type siderophores, whereas all the other strains only produced catechol-type siderophores. In vitro inhibition of Fusarium oxysporum under iron-limited conditions was demonstrated by the siderophore-producing Acinetobacter strains, where PUCM1018 was the most potent inhibitor of the fungal phytopathogen. Acinetobacter sp. PUCM1022 significantly enhanced the shoot height, root length, and root dry weights of pearl millet seedlings in pot experiments when compared with controls, underscoring the plant-growth-promoting potential of these isolates.

• Asian-Australasian Journal of Animal Sciences
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• v.10 no.4
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• pp.378-384
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• 1997

The ruminal solubility of copper (Cu), manganese (Mn), molybdenum (Mo), and zinc (Zn) in eight Philippine forages was studied. These forages were: 1) grasses: paragrass (Brachiaria mutica (Forsk.) Stapf), stargrass (Cynodon plectostachyum Pilger), and napiergrass (Pennisetum purpureum Schumach.); 2) creeping legumes: calopogonium or calopo (Calopogonium muconoides Desv.) and centrocema (Centrocema pubescens Benth.); 3) tree lequmes: gliricidia or kakawate (Gliricidia sepium (Jacq.) Walp.), leucaena or ipil-ipil (Leucaena leucocephala (Lam.) de Wit.), and sesbania or katuray (Sesbania grandiflora (L.) Poir). Nylon bags with samples were incubated for 0, 3, 6, 12, 24, 48 and 72h in three rumen cannulated sheep fed with timothy hay (Phleum pratense L.) at 2% body weight/d. The 0-h bags were washed with deionized water. There were species differences (p < 0.05) on the different solubilities of trace elements. At O-h incubation, the trend of solubility was Mo (54%) > Zn (43%) > Cu (38.7%) > Mn (29.5%). At 3-h incubation, all the elements except Mn had an average solubility above 50%. Combining particulate passage rate (1.9%/h) and various trace elements disappearance rates, the effective ruminal solubilization (ERS) of trace elements were computed. The ERS across species ranged from 44.6 to 89.9% for Cu, 29.9 to 84% for Mn, 66 to 95.1% for Mo, and 30.1 to 82.3% for Zn resulting to a trend of Mo > Zn > Cu > Mn.

• Journal of Applied Biological Chemistry
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• v.64 no.4
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• pp.333-341
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• 2021

In this study, to select strains suitable as microbial agent from among rhizosphere microorganisms present in rhizosphere soil and roots, the mineral solubilization ability, antifungal activity against 10 types of plant pathogenic fungi, and plant growth-promoting activity of rhizosphere microorganisms were evaluated. As a result, DDP346 was selected because it has solubilization ability of phosphoric acid, calcium carbonate, silicon, and zinc; nitrogen fixing ability; production ability of siderophore, indole-3-acetic acid, and aminocyclopropane-1-carboxylate deaminase; and antifungal activity against seven types of plant pathogenic fungi. DDP346 showed a 99.9% homology with Acinetobacter pittii DSM 21653 (NR_117621.1); phylogenetic analysis also revealed a close relationship with Acinetobacter pittii based on the 16S rRNA base sequence. The growth conditions of DDP346 were identified as temperatures in the range of 10-40 ℃, pH in the range of 5-11, and salt concentrations in the range of 0-5%. In addition, a negative correlation coefficient (r² = -0.913, p <0.01) was shown between pH change and the solubilized phosphoric acid content of Acinetobacter sp. DDP346, and this is assumed to be due to the organic acid generated during culture. Consequently, through the evaluation of its mineral solubilization ability, antifungal activity against plant pathogenic fungi, and plant growth-promoting activity, the potential for the utilization of Acinetobacter sp. DDP346 as a multi-purpose microbial agent is presented.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.403-412
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• 2021

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 ㎍/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.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.576-586
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• 2021

The purpose of this study was to evaluate the antifungal activity, plant-growth-promoting activity, and mineral solubilization ability of 10 species of phytopathogenic fungi to select a Bacillus sp. from rhizosphere soils and roots that can be used as a microbial agent. The antifungal activity for phytopathogenic fungi varied based on the Bacillus sp. Among the selected strains, DDP4, DDP16, DDP148, SN56, and SN95 exhibited antifungal activity for nine or more species of phytopathogenic fungi. Regarding nitrogen-fixation ability, all Bacillus sp. showed similar levels of activity, and siderophore production ability was relatively high in ANG42 and DDP427. The indole-3-acetic acid production abilities were in the range of 1.83-67.91 ㎍/ml, with variations in activity based on the Bacillus sp. One strain with a high activity was selected from each species, and their mineral solubilization abilities were examined. Most Bacillus sp. could solubilize phosphoric acid and calcium carbonate, and DDP148 and SN56 could solubilize silicon and zinc, respectively. These results suggested that Bacillus sp. can be considered potential multi-purpose microbial agents for plant growth promotion and disease prevention.