• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.139.1-139
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• 2003

Tissues from woody plant contain higher amount of phenolic compounds and polysaccharides, which give inhibitory effects on reverse transcriptase and/or Taq ploymerase. The common multiple-step protocols using several additives to inhibit polyphenoic compounds during nucleic acid extraction are time consuming and laborious. Sodium sulfite (Na$_2$SO$_3$) was used as inhibitor of polyphenolic oxidases in extraction buffer and compare it's effect between commercial RNA extraction kit and small-scale double-stranded RNA (dsRNA) extraction by RT-PCR. During nucleic acid extraction procedure, addition of 0.5％-1.5％ (w/v) sodium sulfite to Iysis buffer or STE buffer resulted in lighter color change than extracts without sodium sulfite and improve the RT-PCR detection. When commercial RNA extraction kit used, optimal concentration of sodium sulfite were variable according to the host plant. However, using dsRNA as RT-PCR template, 1.5％ sodium sulfite in STE buffer improves the detection of both viruses and unspecific amplifications were reduced significantly, Furthermore, when viruses existed at low titers in host plant, small-scale dsRNA extractions were very reliable.

• The Plant Pathology Journal
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• 제20권2호
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• pp.142-146
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• 2004

Woody plant tissues contain great amounts of phenolic compounds and polysaccharides. These substances inhibit the activation of reverse transcriptase and/or Taq polymerase in RT-PCR. The commonly used multiple-step protocols using several additives to diminish polyphenolic compounds during nucleic acid extraction are time consuming and laborious. In this study, sodium sulfite was evaluated as an additive for nucleic acid extraction from woody plants and the efficiency of RT-PCR assay of commercial nucleic acid extraction kits and small-scale dsRNA extraction was compared. Sodium sulfite was used as an inhibitor against polyphenolic oxidases and its effects were compared in RNA extraction by commercial extraction kit and small-scale double-stranded RNA (dsRNA) extraction method for RT-PCR. During nucleic acid extraction, addition of 0.5%-1.5%(w/v) of sodium sulfite to lysis buffer or STE buffer resulted in lighter browning by oxidation than extracts without sodium sulfite and improved the RT-PCR detection. When commercial RNA extraction kit was used, optimal concentrations of sodium sulfite were variable according to the tested plant. However, with dsRNA as RT-PCR template, sodium sulfite 1.5% in STE buffer improved the detection efficiency of Apple chlorotic leaf spot virus (ACLSV) and Apple stem grooving virus (ASGV) in fruit trees, and reduced the unspecific amplifications signi-ficantly. Furthermore, when viruses existed at low titers in host plant, small-scale dsRNA extractions were very reliable.

• Plant Biotechnology Reports
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• 제4권1호
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• pp.49-52
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• 2010

A DNA extraction method using Chelex 100 is widely used for bacteria, Chlamydomonas, and animal cell lines, but only rarely for plant materials due to the need for additional time-consuming and tedious steps. We have modified the Chelex 100 protocol and successfully developed a rapid and simple method of DNA extraction for efficient PCR-based detection of transgenes from a variety of transgenic plant and algal species. Our protocol consists of homogenizing plant tissue with a pestle, boiling the homogenized tissue in a microfuge tube with 5% Chelex 100 for 5 min, and centrifuging the boiled mixture. The supernatant, which is used for PCR analysis, was able to successfully amplify transgenes in transgenic tobacco, tomato, potato, Arabidopsis, rice, strawberry, Spirodela polyrhiza, Chlamydomonas, and Porphyra tenera. The entire DNA extraction procedure requires <15 min and is therefore comparable to that used for bacteria, Chlamydomonas, and animal cell lines.

• Journal of Plant Biology
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• 제19권4호
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• pp.95-99
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• 1976

This work deals with different extraction media and reaction mixtures on photosystem II activity of Spinach chloroplasts. The photoreduction rate of ferricyanide and DPIP by intact chloroplasts which extracted with four kinds of extraction media; S-Tris-N pH 7.2, 8.0, S-Tricine-N pH 7.2, 8.0, was measured in five kinds of reaction mixtures; S-Tris-N pH 7.2, 8.0, S-Tricine-N pH 7.2, 8.0, 0.05 M-Tris pH 7.2. The extraction medium which shows the highest photoreduction rate was S-Tris-N at pH 7.2 and S-Tricine-N at pH 8.0. As to the reaciton mixture, S-Tricine-N pH 8.0 showed the highest rate. On the complex effects of extraction media and reaction mixtures, the highest photordeuction rate of Hill oxidant by intact chloroplasts was obtained by S-Tris-N pH 7.2 extraction medium and S-Tricine-N pH 8.0 reaction mixture. The second highest activity was obtained by S-Tricine-N pH 8.0 extraction medium and reaction mixture.

• 한국의류학회지
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• 제27권11호
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• pp.1350-1357
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• 2003

This research was aimed to establish the standard extraction and analytical procedures for examining the chromophoric substance in madder root with the ultimate goal of identifying the dyes in badly faded textiles of archaeological origin. The separation temperature of gas chromatography, pH and other extraction conditions were tested. The results were as follows: The suitable separation temperature for the GC cappillary column was 50∼305$^{\circ}C$, and methanol was a good GC solvent for both standard alizarin and madder extraction. The best extraction of madder was achieved by 90 min soaking in room temperature followed by filtration and the actual heat extraction procedure. The best pH for extracting alizarin was pH 3 and above pH 5 alizarin was not detectible. Only alizarin and no purpurin was found in the extraction of the currently used madder plant.

• Mass Spectrometry Letters
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• 제14권1호
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• pp.9-23
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• 2023

Plants are able to produce a large number of diverse bioactive compounds. Solvent extraction is used for isolation of plant metabolites. The extract yield for plant metabolite extraction strongly depends on the nature of solvent. A review showed the methanol can yield more bioactive compounds. Drying of the sample material is also important for the extraction of plant material. The present study was carried out to analyze the phytocomponents of 5 different latex producing plants. The plants like Calotropis gigantea, Carica papaya, Nerium oleander, Ficus benghalensis and Plumeria alba leaves and latex. The GC-MS analysis of the metabolites revealed phytocomponents. Calotropis gigantea leaves showed 14 compounds and latex produced 5 compounds out of this 4,4,6A,6B,8A,11,11,14B-Octamethyl-1,4,4A,5,6,6A,6B,7,8,8A,9,10,11,12,12A,14,14A,14B-Octadeca-hydro-2 and 2R- Acetoxymethyl-1,3,3-trimethyl-4T-(3-Methyl-2-Buten-1-Yl)-1T-Cyclohexanol compound was present in both latex and leaf extraction. Beta. -carotene compound was present in both latex and leaf of Carica papaya. It was observed that Ficus benghalensis contained 2R-Acetoxymethyl-1,3,3-trimethyl-4T-(3-Methyl-2-Buten-1-Yl)-1T-Cyclohexanol was same in latex and leaf extraction.

• The Plant Pathology Journal
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• 제26권3호
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• pp.286-288
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• 2010

Plant tissue homogenization using a mortar or mechanical equipment has been the preferred method for obtaining high yields of total RNA; this method, however, is both time-consuming and expensive. Additionally, homogenization may generate excessive endogenous RNases, polyphenolics, and other substances that reduce the quality and quantity of RNA. In this study, we describe the microwave irradiation-assisted RNA extraction (MIRE) technique which, without tissue disruption and homogenization, allows for the cost-effective and rapid generation of intact RNA from apple cane shavings and the reliable detection of apple virus by RT-PCR.

• Natural Product Sciences
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• 제24권3호
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• pp.199-205
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• 2018

To determine the optimum extraction conditions that give the highest yield of isoquercitrin and caffeic acid from Aster scaber, the effects of four extraction variables (solvent concentrations, extraction time, number of repeated extraction, and solvent volumes) on isoquercitrin and caffeic acid yield was examined via HPLC-UV. Our results showed that the highest extract and isoquercitrin yield were observed when A. scaber was extracted with 450 mL distilled water for 8 hr repeatedly for three times. In case of caffeic acid, the content was higher in the two repeated extracts. Also, content analysis of isoquercitrin in Aster species was performed in which A. fastigiatus, A. ageratoides, and A. scaber exhibited the highest isoquercitrin content at 6.39, 5.68, and 2.79 mg/g extract, respectively. In case of caffeic acid, the highest content of A. scaber and A. glehni was 0.64 and 0.56 mg/g extract, respectively. This study reports an optimized method for extraction of isoquercitrin and caffeic acid from A. scaber and evaluates potential sources of the compounds.

• Journal of Applied Biological Chemistry
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• 제64권2호
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• pp.153-158
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• 2021

Cordyceps militaris (CM) is one of the most important medicinal mushrooms known to possess various biological activities. Cordycepin (CP) is a bioactive compound present in the fruiting bodies of CM and is known to have anti-tumor, anti-metastatic immunomodulatory and anti-inflammatory activities. In this study, we aim to analyze CP quantitatively under various CM extraction conditions. CP was measured using high-performance liquid chromatography, quantified using a reversed phase column using a gradient elution system of water and acetonitrile, and detected with a UV absorbance wavelength of 260 nm. The CP content of CM was the highest in 100% ethanol extract of the fruiting bodies and 60% ethanol extract of the mycelium. This study provides an efficient analysis method to determine the optimal extraction conditions for CP that can be used as a basis for developing functional foods and pharmaceutical products derived from CM.

• Journal of Applied Biological Chemistry
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• 제64권1호
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• pp.57-61
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• 2021

Saikosaponin derivatives such as saikosaponins A, B1, B2, B3, B4, C, and D present in Bupleurum falcatum were analyzed by a high performance liquid chromatograph equipped with an evaporative light scattering detector, using different extraction solvents (water and 70% ethanol). The samples were injected into a YMC Pack Pro C18 column and separated using a gradient elution system with a mobile phase composed of acetonitrile and water at a flow rate of 1.1 mL/min. The content of saikosaponin derivatives was higher in 70% ethanol extract than in water extract. This study provides an efficient analytical method for determining the optimal conditions for extraction of saikosaponin derivatives, which can be used as a basis for development of functional foods and pharmaceutical products from B. falcatum.