• 한국토양비료학회지
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• 제49권5호
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• pp.461-468
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• 2016

One of the important phytohormones produced by plant growth promoting bacteria is the auxin; indole-3-acetic acid (IAA), with L-tryptophan as the precursor. In this study, we focused on the investigation of optimal conditions for the production of IAA by Bacillus megaterium BM5. We investigated culturing conditions, such as incubation temperature, pH of the culture medium and incubation period, with varying media components such as inoculation volume, tryptophan concentration and carbon and nitrogen source. Besides, optimization study intended for high IAA production was carried out with fermentation parameters such as rpm and aeration. The initial yield of $42{\mu}g\;IAA\;ml^{-1}$ after 24 hr increased to $85{\mu}g\;ml^{-1}$ when 5% (v/v) of L-tryptophan was used in the culture broth. The maximum yield of $320{\mu}g\;IAA\;ml^{-1}$ was observed in trypticase soy broth (TSB) supplemented with starch and soybean meal as C and N sources with a C/N ratio of 3:1 (v/v) at $30^{\circ}C$, pH 8.0 for 48 hrs with 1.0 vvm and 250 rpm in 5 L working volume using 10 L scale fermenter. The bacterial auxin extracted from the culture broth was confirmed by thin layer chromatography and high-performance liquid chromatography and effect on plant growth was confirmed by root elongation test.

• 한국육종학회지
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• 제42권4호
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• pp.351-356
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• 2010

Brachypodium distachyon is rapidly emerged in biological study and has been currently used as a model system for genetics and functional studies for crop improvement and biofuel production. Phosphinothricin (PPT) has been widely used as a selectable agent, which raises ammonium content and induces toxicity in non-transformed plant cells. However PPT selection is not much effective on Brachypodium callus consequently reducing transformation efficiency. In order to identify the efficient conditions of PPT selection, calli obtained from mature seeds of Brachypodium (PI 254867) were cultured on the callus inducing medium (CIM) or regeneration medium (ReM) containing serial dilutions of the PPT (0, 2, 5, 10, and 15 mg/l) in dark or light condition. Callus growth and ammonium content of each treatment were measured 2 weeks after the treatment. Although callus growth and ammonium content did not show much difference in CIM, slow callus growth and increased ammonium accumulation were found in ReM. No significant difference of ammonium accumulation in response to PPT was found between dark and light conditions. In order to identify major factors affecting increased ammonium accumulation, callus was cultured on the media in combined with phytohormones (2,4-D or kinetin) and carbon sources (sucrose or maltose) containing with PPT (5 mg/l). The highest ammonium content in callus was found in the kinetin and maltose media.

• Journal of Plant Biotechnology
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• 제47권4호
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• pp.330-336
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• 2020

Ex-situ conservation of the ornamental and medicinal orchid, Coelogyne stricta, was performed by mass propagation using seed culture. Propagation stages were optimized using full- and half-strength solidified MS medium with different phytohormones. Maximum seed germination (88 ± 0.5% over 6 weeks of culture) was achieved on half-strength MS medium supplemented with 15% coconut water. Maximum shoot numbers were found on full-strength MS medium supplemented with 1 mg/L BAP, 2 mg/L Kinetin, and 10% coconut water, while the longest root was developed on full-strength MS medium with 1.5 mg/L IBA. A 2:1:1 combination of coco-peat, pine bark, and sphagnum moss was found to be a suitable potting mixture resulting in 80% seedling survivability. The cytotoxic activity of extracts of both wild plants and in vitro-developed protocorms was determined using an MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay on a cervical cancer cell line. The wild plant extract inhibited the growth of 41.99% of cells, showing that this extract has moderate cytotoxic activity toward cervical cancer cells.

• Plant Resources
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• 제1권2호
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• pp.98-104
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• 1998

The effects of basal media, sucrose and phytohormone concentrations, and gelling agent combinations on in vitro frond proliferation of Lemna gibba G3 and 24 additional Lemna gibba strains were examined. Frond proliferation was equivalent on Schenk and Hidebrand. Murashige and Skoog. Nitsch and Nitsch, and Gamborg's B5 media and poor on murashige and Skoog medium in the absence of benzyladenine. With the addition of benzyladenine, Schenk and Hildebrand and Gamborg's B5 Were superior and equivalent. The addition of benzuyladenine increased equally frond proliferation at either 1 or $10{\mu}M$, however at $10{\mu}M$ fronds were severely curled or fused. Benzyladenine and thidiazuron suppressed root growth but kinetin was found to greatly enhance root growth. Gibberellic acid inhibited frond proliferation. Frond proliferation was significantly different on the four sucrose concentrations of 0, 1, 3, and 5% Among them, 3% sucrose was found to be superior. The reduced frond size observed in cultures grown on 8% sucrose could be explained by showing medium osmotic potential in excess of frond water potential. Gell agents also varied significantly in their ability to promote frond proliferation with 0.25% Gelrite or a mixture of 0.15% Gelrite and 0.4% agar. Proliferation of 25 Lemna gibba strains on medium neat optimal for Lemna gibba G3 showed a six-fold variation across strains with Lemna gobba G3 placing in the top 5 fastest proliferating strains.

• 농업과학연구
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• 제50권4호
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• pp.759-771
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• 2023

Plant growth-promoting rhizobacteria (PGPR) are naturally occurring bacteria that intensively colonize plant roots and are crucial in promoting the crop growth. These beneficial microorganisms have garnered considerable attention as potential bio-inoculants for sustainable agriculture. PGPR directly interacts with plants by providing essential nutrients through nitrogen fixation and phosphate solubilization and accelerating the accessibility of other trace elements such as Cu, Zn, and Fe. Additionally, they produce plant growth-promoting phytohormones, such as indole acetic acids (IAA), indole butyric acids (IBA), gibberellins, and cytokinins.PGPR interacts with plants indirectly by protecting them from diseases and infections by producing antibiotics, siderophores, hydrogen cyanide, and fungal cell wall-degrading enzymes such as glucanases, chitinases, and proteases. Furthermore, PGPR protects plants against abiotic stresses such as drought and salinity by producing 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase and modulating plant stress markers. Bacteria belonging to genera such as Bacillus, Pseudomonas, Burkholderia, Pantoa, and Enterobacter exhibit multiple plant growth-promoting traits, that can enhance plant growth directly, indirectly, or through synergetic effects. This comprehensive review emphasizes how PGPR influences plant growth promotion and presents promising prospects for its application in sustainable agriculture.

• Journal of Plant Biotechnology
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• 제2권2호
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• pp.79-82
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• 2000

We have isolated a cDNA encoding a calcium-dependent protein kinase (CDPK) in Nicotiana tabacum, which was designated NtCDPK1. Accumulation of the NtCDPK1 mRNA was stimulated by various stimuli, including phytohormones, CaCl$_2$ wounding, fungal elicitors, chitin and methyl jasmonate. The NtCDPK1 gene encodes a functional Ser/Thr protein kinase of which phosphorylation activity is strongly induced by calcium. By analyzing expression of the NtCDPK1-GFP fusion protein and by immunoblotting with antibody which reacts with NtCDPK1, we found that NtCDPK1 is localized in membrane and nucleus in plant cells. Silencing expression of the NtCDPK1 transgene resulted in marked decrease of lateral root development in the transgenic tobacco plants. Yeast two hybrid screening using NtCDPK1 as a bait identified a tobacco homologue of proteasome regulatory subunit 21D7, designated Nt21D7. The 21D7 mRNA has been shown to be predominantly expressed in proliferating tissues in the cell cycledependent manner in carrot. The recombinant NtCDPK1 protein associated with Nt21D7 in vitro, and could phosphorylate the Nt21D7 protein in vitro in the presence of calcium, suggesting that Nt21D7 protein is a natural substrate of NtCDPK1 in tobacco. These results suggest that NtCDPK1 may regulate tell proliferation processes, such as lateral root formation, by regulating specificity and/or activity of proteasome-mediated protein degradation pathway.

• 미생물학회지
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• 제46권1호
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• pp.33-37
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• 2010

근권토양에서 분리한 세균 Bacillus subtilis RFO41은 인산 가용화능이 높고 auxin 등의 식물호르몬을 생산한다. 이 균주를 이용하여 가시도꼬마리(Xanthium italicum Moore)의 생장 촉진 실험을 화천군 파로호의 호안나대지에서 수행하였다. 19주 경과 후 자라난 가시도꼬마리를 채취하여 줄기와 뿌리 길이를 측정하고 식물을 건조시켜 중량을 측정하였다. 균주를 처리한 접종군이 미접종 대조군보다 가시도꼬마리의 건조중량이 67.7%나 높았으며 또한 가시도꼬마리의 줄기 길이도 균주를 처리한 접종군이 미접종 대조군보다 21.1% 길었는데 모두 통계적 유의성을 가졌다. 접종균주의 동태를 denaturing gradient gel electrophoresis 방법으로 조사하였는데 나대지 토양에서 접종균주가 유지되고 있으며 토양의 고유 세균군집에는 큰 영향을 미치지 않는 것을 확인하였다. 따라서 척박한 호안나대지 토양에서 B. subtilis RFO41의 인산가용화능과 식물호르몬의 생성능 등의 식물생장촉진 효과가 가시도꼬마리의 생장에 직접적인 영향을 미치는 것으로 판단할 수 있다. 이러한 식물 생장 촉진효과는 친환경적 미생물비료, 특히 민감한 호안나대지 등의 미생물학적 식생복원방법으로서의 가치를 뒷받침하는 것이라 할 수 있다.

• 한국미생물·생명공학회지
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• 제35권1호
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• pp.58-65
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• 2007

중금속으로 오염된 토양을 정화하기 위한 rhizoremediation 기법에서 식물이 중금속을 흡수하고 이동시키는 효율을 증가시키기 위하여 토양 미생물 특히, 근권세균의 역할이 중요하다. 이를 위하여 본 연구에서는 정유공장 주변의 유류 및 중금속으로 장기간 오염된 토양에서 서식하는 4가지 식물의 근권토양으로부터 Methylobacterium sp. SY-NiR1 균주를 분리하였다. 분리한 Methylobacterium sp. SY-NiR1는 분홍색 콜로니 형성, 막대모양 및 $\alpha-proteobacteria$에 속하는 특성으로 보아 pink-pigmented facultative methylotroph인 것으로 사료된다. 이 균주는 식물성호르몬인 indole acetic acid(IAA) 생산능을 가지고 있으며, 카드뮴, 크롬, 구리, 납, 니켈 그리고 아연 등과 같은 다양한 중금속에 대하여 내성을 가지고 있었으며, $EC_{50}$을 기준으로 한 SY-NiR1의 중금속에 대한 내성은 Zn > Ni > Cu > Pb > Cd > Cr 순이다. 따라서 본 연구에서 분리한 Methylobacterium sp. SY-NiR1 균주는 중금속으로 오염된 토양에서 식물의 발아, 생장 및 발달을 도와 식물의 중금속 흡수를 증가시켜 rhizorememdiation 효율을 증가시킬 수 있을 것으로 기대된다.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.934-944
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• 2007

Paenibacillus polymyxa E681 is known to be able to suppress plant diseases by producing antimicrobial compounds and to promote plant growth by producing phytohormones, and secreting diverse degrading enzymes. In spite of these capabilities, little is known regarding the flow of information from the bacterial strain to the barley roots. In an attempt to determine the flow of information from the bacterial strain to barley roots, the strain was grown in the presence and absence of barley, and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MALDI-TOF mass spectrometry were used. 2D-PAGE detected approximately 1,000 spots in the cell and 1,100 spots in the supernatant at a pH 4-10 gradient. Interestingly, about 80 spots from each sample showed quantitative variations. Fifty-three spots from these were analyzed by MALDI-TOF mass spectrometry and 28 proteins were identified. Most of the cytosolic proteins expressed at higher levels were found in P. polymyxa E681 cells grown in the presence of barley rather than in the absence of barley. Proteins detected at a lower level in the surpernatant of P. polymyxa E68l cells grown in the presence of barley were lipoprotein, glucose-6-phosphate 1-dehydrogenase, heat-shock protein HtpG, spermidine synthase, OrfZ, ribonuclease PH, and coenzyme PQQ synthesis protein, and flagellar hook-associated protein 2 whereas proteins detected at a higher level in the surpernatant of P. polymyxa E681 cells grown in the presence of barley included D-alanyl-D-alanine ligase A, isopentenyl-diphosphate delta-isomerase, ABC transporter ATP-binding protein Uup, lipase. Many of the proteins belonging to plant-induced stimulons are associated with biosynthetic metabolism and metabolites of proteins and transport. Some of these proteins would be expected to be induced by environmental changes resulting from the accumulation of plant-secreted substances.

• Journal of Microbiology and Biotechnology
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• 제30권8호
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• pp.1156-1168
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• 2020

Drought stress is threatening the growth and productivity of many economical crops. Therefore, it is necessary to establish innovative and efficient approaches for improving crop growth and productivity. Here we investigated the potentials of the cell-free extract of Actinobacteria (Ac) isolated from a semi-arid habitat (Al-Jouf region, Saudi Arabia) to recover the reduction in maize growth and improve the physiological stress tolerance induced by drought. Three Ac isolates were screened for production of secondary metabolites, antioxidant and antimicrobial activities. The isolate Ac3 revealed the highest levels of flavonoids, antioxidant and antimicrobial activities in addition to having abilities to produce siderophores and phytohormones. Based on seed germination experiment, the selected bioactive fraction of Ac3 cell-free extract (F2.7, containing mainly isoquercetin), increased the growth and photosynthesis rate under drought stress. Moreover, F2.7 application significantly alleviated drought stress-induced increases in H₂O₂, lipid peroxidation (MDA) and protein oxidation (protein carbonyls). It also increased total antioxidant power and molecular antioxidant levels (total ascorbate, glutathione and tocopherols). F2.7 improved the primary metabolism of stressed maize plants; for example, it increased in several individuals of soluble carbohydrates, organic acids, amino acids, and fatty acids. Interestingly, to reduce stress impact, F2.7 accumulated some compatible solutes including total soluble sugars, sucrose and proline. Hence, this comprehensive assessment recommends the potentials of actinobacterial cell-free extract as an alternative ecofriendly approach to improve crop growth and quality under water deficit conditions.