• Journal of Microbiology and Biotechnology
• /
• 제23권10호
• /
• pp.1372-1380
• /
• 2013

Different endophytic fungi isolated from Himalayan Yew plants were tested for their ability to produce taxol. The BAPT gene (C-13 phenylpropanoid side chain-CoA acetyl transferase) involved in the taxol biosynthetic pathway was used as a molecular marker to screen taxol-producing endophytic fungi. Taxol extracted from fungal strain TBPJ-B was identified by HPLC and MS analysis. Strain TBPJ-B was identified as Fusarium redolens based on the morphology and internal transcribed spacer region of nrDNA analysis. HPLC quantification of fungal taxol showed that F. redolens was capable of producing $66{\mu}g/l$ of taxol in fermentation broth. The antitumour activity of the fungal taxol was tested by potato disc tumor induction assay using Agrobacterium tumefaciens as the tumor induction agent. The present study results showed that PCR amplification of genes involved in taxol biosynthesis is an efficient and reliable method for prescreening taxol-producing fungi. We are reporting for the first time the production of taxol by F. redolens from Taxus baccata L. subsp. wallichiana (Zucc.) Pilger. This study offers important information and a new source for the production of the important anticancer drug taxol by endophytic fungus fermentation.

• Plant Resources
• /
• 제6권1호
• /
• pp.1-6
• /
• 2003

Ginseng(Panax ginseng C. A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng hairy roots induced by introducing Ri-plasmid of Agrobacterium rhizogenes into genomic DNA of plant cells show vigorous growth, and the hairy roots produce the same or more saponins than natural ginseng roots. Therefore, hairy roots can be used for commercial purposes. The present study was carried out to induce hairy roots with both active growth and high saponin contents. Numerous hairy roots of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes R1000 A4T in dark condition after one month of culture. About 3 hundred lines of hairy roots were selected according as morphological characters on medium with carbenicillin. After pre-selection of fifteen lines of hairy roots with active growth, KGHR-l and KGHR-8 lines were finally selected which had characters of high content of ginsenoside-Rd and ginsenoside-Re, respectively. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark (22 $^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5L Erlenmeyer flasks, lL roller drums, 10L jar-fermenters, and especially in 20L air-lift culture vessels.

• 대한의생명과학회지
• /
• 제15권4호
• /
• pp.265-272
• /
• 2009

Recently, mesenchymal stem cells (MSCs) began to be utilized as a vehicle for ex vivo gene therapy based on their plasticity. Effective and safe transfection of therapeutic genes is a critical step for genetic modification of MSCs. Therefore, optimization of in vitro gene delivery into MSCs is essential to provide genetically modified stem cells. In this study, various cationic liposomes, O,O'-dimyristyl-N-lysyl aspartate (DMKD), DMKD/cholesterol, O,O'-dimyristyl-N-lysyl glutamate (DMKE), DMKE/cholesterol, and N-[1-(2,3-dioleoyloxy)]-N,N,N-trimethylammonium propane methyl sulfate (DOTAP)/cholesterol, were mixed with plasmid DNA encoding luciferase (pAAV-CMV-Luc) at varied ratios, and then used for transfection to MSCs under varied conditions. The MSCs were also transfected by electroporation under varied conditions, such as voltage, pulse length, and pulse interval. According to the experimental results, electroporation-mediated transfection was more efficient than cationic liposome-mediated transfection. The best MSC transfection was induced by electroporation 3 times pulses for 2 ms at 200 V with 10 seconds of a pulse interval.

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 2017년도 9th Asian Crop Science Association conference
• /
• pp.152-152
• /
• 2017

Plants exposed to environmental stress for long durations often can adapt to stress conditions with improved tolerance. Moreover this acquired tolerance to stress can be retained even after reverting to destressed growth conditions, which is known to stress memory. In these adaptation and stress memory processes, epigenetic regulation, such as DNA methylation and histone modifications play a key role. Here, we showed that regenerated rice plants from embryogenic callus exposed to gradually increasing NaCl concentrations (up to 120 mM NaCl) acquired salt tolerance and their enhanced tolerance are inherited to subsequent generations. The rice plants (R0) regenerated from rice callus under saline conditions were transplanted into normal paddy field and R1 seeds were harvested. These R1 seeds displayed higher germination rate on MS medium containing 100mM NaCl than wild-type. The callus derived from R1 seeds showed better growth than control callus on high salinity medium. And the salt-adapted R1 plants exhibited higher chlorophyll contents and also higher $K^+/Na^+$ ratio than wild-type rice under saline conditions. The results indicated that rice plants successfully adapted to saline growth conditions during regeneration on high salt medium and moreover this acquired tolerance to salt stress was inherited subsequent generation.

• Journal of Plant Biotechnology
• /
• 제36권3호
• /
• pp.294-300
• /
• 2009

Activation tagging that uses T-DNA vectors containing multimerized transcriptional enhancers from the cauliflower mosaic virus (CaMV) 35S gene is a powerful tool to determine gene function in plants. This approach has been successfully applied in screening various types of mutations and cloning the corresponding genes. We generated an activation tagged hairy root pool of ginseng (Panax ginseng C.A. Meyer) in an attempt to isolate genes involved in the biosynthetic pathway of ginsenoside (triterpene saponin), which is known as the major active ingredient of the root. Quantitative and qualitative variation of ginsenoside in activation tagged hairy root lines were profiled using LC/MS. Metabolic profiling data enabled selection of a specific hairy root line which accumulated ginsenoside at a higher level than other lines. The relative expression level of several genes of triterpene biosynthetic pathway in the selected hairy root line was determined by real time RT-PCR. Overall results suggest that the activation tagged ginseng hairy root system described in this study would be useful in isolating genes involved in a complex metabolic pathway from genetically intractable plant species by metabolic profiling.

• Journal of Microbiology and Biotechnology
• /
• 제14권4호
• /
• pp.766-771
• /
• 2004

Julong 601, a Gram-positive anaerobic bacterium strain capable of cleaving the C-ring of isoflavones daidzein and genistein, was isolated from human feces. BLAST search revealed that its complete 16S rDNA gene sequence has 99% similarity to Eubacterium ramulus. Metabolites of daidzein and genistein were determined as O-desmethylangolensin (O-Dma) and 2-(4-hydroxyphenyl) propionic acid (2-HPPA), respectively, based on UV, EI-MS, and $^1H$ and ^{13}C$ NMR spectral analyses. Enantiomers of O-Dma and 2-HPPA were isolated by chiral stationary-phase HPLC (CSP HPLC). Cleavage of the C-ring of daidzein and genistein by strain Julong 601 was highly enantioselective. Specific rotation ([$\alpha]_D$) and circular dichroism (CD) spectra of the enantiomers are reported here for the first time. Biotransformation kinetics of daidzein and genistein indicated that the C-ring of genistein has a higher susceptibility to bacterial degradation than that of daidzein.

• Plant Resources
• /
• 제1권1호
• /
• pp.6-12
• /
• 1998

The Fp1 gene, originally isolated from red pepper seedlings, encode the iron storage protein, and have a high homology with ferritin genes at DNA and amino acid level. In order to determine ferritin protein expression in vegetative tissue. Fp1 gene was constructed in plant expression vector(PIG12IHm) and introduced in red pepper(var. Bukang, Chungyang and Kalag-Kimjang 2) via Agrobacterium tumefaciensmediated transformation. After selection on MS media containing Kanamycin(Km), putatively selected transformants were confirmed by amplification of selectable marker gene(Fp1 and NPII) by polymerase chain reaction. Northern blot showed that transcripts of Fp1 gene were detected in mature leaves of the plants. In A6, A7 and A8 and A14 of transgenic plants, transcript of Fp1 gene was increased seven-fold to eight-fold than other transgenic plants. Also the proteins obtained from leaves of transgenic plants were immunologically detected by Western blot using rabbit anti-ferritin polyclonal antibody. The expression protein appeared as strong band of apparent mass of 23.5kDa. suggesting the iron accumulation in transgenic red pepper plants.

• 약학회지
• /
• 제53권3호
• /
• pp.145-150
• /
• 2009

We evaluated cytotoxic effect based on the MTT assay and identified altered proteins in 5-FU(fluorouracil) treated HT29 cells using two-dimensional gel electrophoresis and MALDI-TOF/TOF-MS. As proteins inducing apoptosis, siah binding protein 1 and p47 protein isoform a were up-regulated and tumor protein translationally-controlled 1 was down-regulated by 5-FU treatment. And mannose 6 phosphate receptor binding protein 1 controls DNA mismatch repair system was increased. We suggest 5-FU promotes a cytotoxicity under the action of these proteins in colon cancer cells.

• Journal of Microbiology and Biotechnology
• /
• 제20권12호
• /
• pp.1672-1676
• /
• 2010

The Actinomycete strain KH29 is antagonistic to the multidrug-resistant Acinetobacter baumannii. Based on the diaminopimelic acid (DAP) type, and the morphological and physiological characteristics observed through the use of scanning electron microscopy (SEM), KH29 was confirmed as belonging to the genus Streptomyces. By way of its noted 16S rDNA nucleotide sequences, KH29 was found to have a relationship with Streptomyces cinnamonensis. The production of an antibiotic from this strain was found to be most favorable when cultured with glucose, polypeptone, and yeast extract (PY) medium for 6 days at $27^{\circ}C$. The antibiotic produced was identified, through comparisons with reported spectral data including MS and NMR as a cyclo(L-tryptophanyl-L-tryptophanyl). Cyclo(L-Trp-L-Trp), from the PY cultures of KH29, was seen to be highly effective against 41 of 49 multidrug-resistant Acinetobacter baumannii. Furthermore, cyclo(L-Trp-L-Trp) had antimicrobial activity against Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus, Saccharomyces cerevisiae, Aspergillus niger, and Candida albicans, However, it was ineffective against Streptomyces murinus.

• Advances in environmental research
• /
• 제9권2호
• /
• pp.109-121
• /
• 2020

Phenol is frequently present as the hazardous pollutant in petrochemical and pesticide industry wastewater. Because of its high toxicity and carcinogenic potential, a proper treatment is needed to reduce the hazards of phenol carrying effluent before being discharged into the environment. Phenol biodegradation with microbial consortium offers a very promising approach now a day's. This study focused on the formulation of phenol degrading bacterial consortium with three bacterial isolates. The bacterial strains Bacillus cereus strain VCRC B540, Bacillus cereus strain BRL02-43 and Oxalobacteraceae strain CC11D were isolated from detergent contaminated soil by soil enrichment technique and was identified by 16s rDNA sequence analysis. Individual cultures were degrade 100 μl phenol in 72 hrs. The formulated bacterial consortium was very effective in degrading 250 μl of phenol at a pH 7 with in 48 hrs. The study further focused on the analysis of the products of biodegradation with Fourier Transform Infrared Spectroscopy (FT/IR) and Gas Chromatography-Mass Spectroscopy (GC-MS). The analysis showed the complete degradation of phenol and the production of Benzene di-carboxylic acid mono (2-ethylhexyl) ester and Ethane 1,2- Diethoxy- as metabolic intermediates. Biodegradation with the aid of microorganisms is a potential approach in terms of cost-effectiveness and elimination of secondary pollutions. The present study established the efficiency of bacterial consortium to degrade phenol. Optimization of biodegradation conditions and construction of a bioreactor can be further exploited for large scale industrial applications.