• Title/Summary/Keyword: Plant Cell Culture

Search Result 622, Processing Time 0.034 seconds

Characterization of Anionic Peroxidase Induced by Low Host-Specific Elicitor in Suspension Cultures of Rose (Rosa sp.) (장미(Rosa sp.) 현탁배양세포에서 숙주 특이성이 낮은 Elicitor에 의해 유도되는 Anionic Peroxidase의 특성)

  • 신미선;양은진;이인철
    • Korean Journal of Plant Tissue Culture
    • /
    • v.25 no.4
    • /
    • pp.277-282
    • /
    • 1998
  • Whereas cationic extracellular peroxidases (PODs) were observed in the suspension cultures of rose (Rosa sp. L. cv Pual's scarlet) grown under normal conditions, new anionic isozymes were induced within 24 hr by the treatment of low host-specific elicitor (10 mg glucan/L media) prepared from yeast cell wall. Prominent anionic (pI 6.1) and cationic POD (pI 8.4) were purified and characterized to understand the physiological role of the enzymes. Both enzymes were purified (ca.200 fold) by the ammonium sulfate precipitation, ion exchange chromate-graphy and gel filtration chromatography. The Km values of the purified anionic POD for ferulic acid and $\textrm{H}_2\textrm{O}_2$ were 4.64 mM and 0.72 mM, whereas those of the cationic POD were 1.38 mM and 0.48 mM, respetively. The activity of the anionic POD as NADH oxidase was twice higher than that of cationic POD. The NADH oxidation in the anionic POD fraction was inhibited by 60% on the addition of 0.1 mM coniferyl alcohol, while that in the cationic fraction was inhibited by 15%.

  • PDF

Anti-inflammatory Effect of the Robinia pseudoacacia L. High Temperature Extract (아까시 나무 고온추출물의 항염증 효과)

  • Nho, Jong Hyun;Kang, Byoung Man;Jung, Won Seok
    • Korean Journal of Plant Resources
    • /
    • v.31 no.4
    • /
    • pp.294-302
    • /
    • 2018
  • This study was conducted to compare anti-inflammatory effect of Robinia pseudoacacia L. using different extraction methods (water extraction, ethanol extraction and high temperature extraction). We investigated anti-inflammatory effect of Robinia pseudoacacia L. extract (RP1, water extract; RP2, ethanol extract; RP3, high temperature extract) on lipopolysaccharide (LPS)-stimulated inflammation using Raw 264.7 cell. Cells were treated with various concentrations (12.5, 25, 50, 100 or $200{\mu}g/m{\ell}$) of water extract, ethanol extract and high temperature extract. Cytotoxicity was not observed on Raw 264.7 cells, LPS-stimulated production of NO (nitric oxide), $PGE_2$ (prostaglandin $E_2$) and cytokines ($TNF-{\alpha}$, IL-6 and $IL-1{\beta}$) was reduced by RP3 treatment more than RP1 and RP2. In conclusion, these results indicated that inflammation on Raw 264.7 cells was improved by RP3. Treatment of RP3 could be used to natural medicine for improving inflammatory response. However, further experiment is required to observe how the high temperature extraction at $500^{\circ}C$ for 48 h influences on alteration of active ingredient in Robinia pseudoacacia L., and conducts the inflammation signal pathway on Raw 264.7 cells.

Optimization of Solid State Fermentation of Mustard (Brassica campestris) Straw for Production of Animal Feed by White Rot Fungi (Ganoderma lucidum)

  • Misra, A.K.;Mishra, A.S.;Tripathi, M.K.;Prasad, R.;Vaithiyanathan, S.;Jakhmola, R.C.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.20 no.2
    • /
    • pp.208-213
    • /
    • 2007
  • The objective of the experiment was to determine the optimum cultural [moisture levels (55, 60 and 70%), days of fermentation (7, 14 and 21), temperature (25 and $35^{\circ}C$) of incubation)] and nutritional parameters (urea addition (0 and 2%) and variable levels of single super phosphate (0.25 and 0.50% SSP)) for bio-processing of the mustard (Brassica campestris) straw (MS) under solid-state fermentation (SSF) system. The performance of SSF was assessed in terms of favorable changes in cell wall constituents, protein content and in vitro DM digestibility of the MS. Sorghum based inoculum (seed culture) of Ganoderma lucidum to treat the MS was prepared. The 50 g DM of MS taken in autoclavable polypropylene bags was mixed with a pre-calculated amount of water and the particular nutrient in the straw to attained the desired levels of water and nutrient concentration in the substrate. A significant progressive increase in biodegradation of DM (p<0.001), NDF (p<0.01) and ADF (p<0.05) was observed with increasing levels of moisture. Among the cell wall constituents the loss of ADF fraction was greatest compared to that of NDF. The loss of DM increased progressively as the fermentation proceeded and maximum DM losses occurred at 28 days after incubation. The protein content of the treated MS samples increased linearly up to the day $21^{th}$ of the incubation and thereafter declined at day $28^{th}$, whereas the improvement in in vitro DM digestibility were apparent only up to the day $14^{th}$ of the incubation under SSF and there after it declined. The acid detergent lignin (ADL) degradation was slower during the first 7 days of SSF and thereafter increased progressively and maximum ADL losses were observed at the day $28^{th}$ of the SSF. The biodegradation of DM and ADL was not affected by the variation in incubation temperature. Addition of urea was found to have inhibitory effect on fungal growth. The effect of both the levels (0.25 and 0.50) of SSP addition in the substrate, on DM, NDF, ADF, cellulose and ADL biodegradation was similar. Similarly, the protein content and the in vitro DM digestibility remain unaffected affected due to variable levels of the SSP inclusion in the substrate. From the results it may be concluded that the incubation of MS with 60 percent moisture for 21 days at $35^{\circ}C$ with 0.25 percent SSP was most suitable for MS treatment with Ganoderma lucidum. Maximum delignification, enrichment in the protein content and improvement in in vitro DM digestibility were achieved by adopting this protocol of bioprocessing of MS.

Changes and characteristics of the biochemical components on the differentiation of soybean cell tissue cultures: (1) Changes and characteristics of the proteins, amino acids and peroxidase isozymes on differentiation of soybean cell tissue cultures (대두 기내 배양체의 분화에 대한 생화학적 성분의 변화와 특성 : (I) 대두 기내 배양체의 분화에 대한 단백질, 아미노산 및 peroxidase 동위효소의 변화와 특성)

  • Nam, Sang-Hae;Choi, Sang-Uk;Yang, Min-Suk
    • Applied Biological Chemistry
    • /
    • v.34 no.2
    • /
    • pp.134-141
    • /
    • 1991
  • In order to investigate the changes and characteristics of biochemical metabolic substances of soybean tissue culture during the cultural period, immature cotyledons were detached form the plant on 15th days after flowering and cultured in vitro for 3 weeks. The cultures were classified into embryogenic(EC) and non-embryogenic callus(NEC). A part of the EC lines were subcultured for another 3 weeks and classified into root forming(RFC), and shoot forming cultures(SFC). Another part of the EC lines were used for isolation of protoplasts, which were subsequently cultured in vitro for 4 weeks. The cultures were classified into embryogenic(PEC) and non-embryogenic callus(PNEC) derived from the protoplasts. The cultures of EC and PEC lines showed higher phenylalanine content and lower methionine content than those of NEC and PNEC. At organ differentiation stage, both cultures showed the content of aspartic acid decreased, while the other amino acids increased as a whole. The protein pattern analysis of the cultures revealed that EC and NEC lines contained distinctive polypeptides, with mass of ca. 18KD for EC and ca. 22KD for NEC respectively. The EC and PEC lines also showed high activity of peroxidase isozyme A(piA), while the RFC and SFC lines showed that of peroxidase isozyme B(piB).

  • PDF

Enhanced Production of Oleanolic Acid by the Elicitation in Oldenlandia diffusa Suspension Cell Cultures (백화사설초의 현탁세포배양에서 Elicitation에 의한 Oleanolic acid 생산성 증대)

  • Lee Yong-Il;Kim Dong-Il
    • KSBB Journal
    • /
    • v.19 no.6 s.89
    • /
    • pp.471-477
    • /
    • 2004
  • Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.

Morphological and Cultural Characteristics of Fungi Causing Rice Sclerotial Diseases (수도균핵병(水稻菌核病)을 일으키는 진균(眞菌)의 형태적(形態的) 및 배양적(培養的) 특성(特性))

  • Kim, Wan-Gyu
    • The Korean Journal of Mycology
    • /
    • v.21 no.1
    • /
    • pp.16-22
    • /
    • 1993
  • Morphological and cultural characteristics of fungi causing rice sclerotial diseases were examined. Hyphal widths of R. solani and R. oryzae were same and ranged $6.0-12.0\;{\mu}m$ with average $9.0\;{\mu}m$, the widest among those of the sclerotial fungi examined. Hyphal width of R. oryzae sativae ranged $6.0-9.0{\mu}m$ with average $7.4{\mu}m$. Hyphal width of R. cerealis was the narrowest among those of Rhizoctonia species examined, and the same was hyphal width of S. oryzae among those of Sclerotium species. Nuclear staining by HCL-Giemsa method showed that R. solani and R. oryzae had many nuclei within one hyphal cell, S. oryzae one nucleus, and the other sclerotial fungi mostly two nuclei. The nuclear number of R. solani was the largest, which ranged 2-17 with average 6.3. Average size of sclerotia of the sclerotial fungi except S. hydrophilum and S. oryzae produced in lesions ranged 1.0-2.0mm. Average size of sclerotia of S. hydrophilum and S. oryzae was 0.5mm and 0.24mm, respectively. Sclerotia of R. solani and R. oryzae produced in culture were more variable in size and larger than those produced in lesions. However, the sclerotial sizes of the other sclerotial fungi produced in culture were almost the same as those produced in lesions. Sclerotial colors of sclerotial fungi produced in lesions were similar to those produced in culture, but sclerotial shapes of some sclerotial fungi exhibited somewhat difference between the sclerotia produced in lesions and in culture. Optimum temperature for mycelial growth of R. cerealis was $23^{\circ}C$, and that of the other sclerotial fungi ranged from $27\;to\;33^{\circ}C$. Maximum temperature for mycelial growth of some sclerotial fungi was as high as $41^{\circ}C$, while that of R. cerealis was as low as $31^{\circ}C$. Minimum temperature for mycelial growth of R. cerealis was $2^{\circ}C$, and that of the other sclerotial fungi ranged from $6\;to\;10^{\circ}C$.

  • PDF

Production of doubled haploid population derived from the microspore culture of rapeseed (Brassica napus L.) F1 generation and analysis of fatty acid composition (유채 잡종 1세대의 소포자 배양에 의한 배가반수체 집단 선발 및 지방산 조성 분석)

  • Lee, Ji Eun;Park, Ju Hyun;Kim, Kwang Soo;An, Da Hee;Cha, Young Lok
    • Journal of Plant Biotechnology
    • /
    • v.49 no.1
    • /
    • pp.74-81
    • /
    • 2022
  • Brassica napus, an oil crop that produces rapeseed oil, is an allotetraploid (AACC, 2n = 38) produced by natural hybridization between B. rapa and B. oleracea. In this study, microspore was cultured using the F1 developed from a cross between 'EMS26' line with high oleic acid content and 'J8634-B-30' lines. The flower bud size showing the nuclear development at the late uninucleate and binucleate stage with high embryogenesis rate was 2.6 ~ 3.5 mm. Microspores were cultured using only this size and after then most microspore embryo developed into secondary embryos and then regeneration plants obtained from the developed multilobe. The analysis of the ploidy of the plants revealed that 66.7% and 27.8% of the total lines were tetraploids and octoploids, respectively. The sizes of stomatal cells in tetraploids, octoploids, and diploids were 25.5, 35.6, and 19.9 ㎛, respectively, indicating that ploidy level was positively correlated with cell size. Furthermore, 62 tetraploid doubled haploid (DH) lines were selected. The average oleic acid (C18:1) and linolenic acid (C18:3) concentrations of DH were 72.3% and 6.2%, respectively. Oleic acid and linolenic acid concentrations exceeded the two parental values in 5 and 14 DH lines, respectively, suggesting that these two fatty acids had transgressive segregation. Therefore, the DH population can be utilized for the biosynthesis of unsaturated fatty acids in rapeseed and related genes. It can also be used as a breeding material for varieties with high oleic acid concentrations.

Characterization of Salt Tolerant Rice Mutant Lines Derived from Azetidine-2-Carboxylic Acid Resistant Cell Lines Induced by Gamma Ray Irradiation (AZCA 저항성 돌연변이 세포주로부터 선발 육성만 내염성 벼 돌연변이 계통의 특성 검정)

  • Song, Jae-Young;Kim, Dong-Sub;Lee, Geung-Joo;Lee, In-Sok;Kang, Kwon-Kyoo;Yun, Song-Joong;Kang, Si-Yong
    • Journal of Plant Biotechnology
    • /
    • v.34 no.1
    • /
    • pp.61-68
    • /
    • 2007
  • To develop rice (Oryza sativa L.) cultivars to be planted on salt-affected sites, cell lines with enhanced proline content and resistance to growth inhibition by Azetidine-2-carboxylic acid (AZCA), a proline analogue, were screened out among calli irradiated with gamma ray of 50, 70, 90, and 120 Gy. The calli had been derived from embryo culture of the cultivar Donganbyeo. Selected AZCA resistant lines that had high proline accumulation were used as sources for selection of NaCl resistant lines. To determine an optimum concentration for selection of NaCl resistant lines, Donganbyeo seeds were initially cultured on the media containing various NaCl concentrations (0 to 2.5%) for 40 days, and 1.5% NaCl concentration was determined as the optimum concentration. One hundred sixteen salt-tolerant (ST) lines were selected from bulked 20,000 seeds of the AZCA resistant $M_{3}$ seeds in the medium containing 1.5% NaCl. The putative 33 lines ($M_{4}$ generation) considered with salt-tolerance were further analyzed for salt tolerance, amino acid and ion contents, and expression patterns of the salt tolerance-related genes. Out of the 33 lines, 7 lines were confirmed to have superior salt tolerance. Based on growth comparison of the entries, the selected mutant lines exhibited greater shoot length with average 1.5 times, root length with 1.3 times, root numbers with 1.1 times, and fresh weight with 1.5 times than control. Proline contents were increased maximum 20%, 100% and 20% in the leaf, seed and callus, respectively, of the selected lines. Compared to control, amino acid contents of the mutants were 24 to 29%, 49 to 143%, 32 to 60% higher in the leaf, seed and callus, respectively. The ratio of $Na^{+}/K^{+}$ for most of the ST-lines were lower than that of control, ranging from 1.0 to 3.8 for the leaf and 11.5 to 28.5 for the root, while the control had 3.5 and 32.9 in the leaf and root, respectively. The transcription patterns for the P5CS and NHXI genes observed by RT-PCR analysis indicated that these genes were actively expressed under salt stress. The selected mutants will be useful for the development of rice cultivar resistant to salt stress.

Influences of the Plant Growth under Beta-Rays Irradiation at Low Dose (저 선량 베타선의 조사에 의한 식물의 생장에 미치는 영향)

  • Lee, Byung-Koo;Im, In-Chul;Kim, Jong-Eon
    • Journal of radiological science and technology
    • /
    • v.33 no.2
    • /
    • pp.143-148
    • /
    • 2010
  • This study is to analyze effects of the growth of Chunhyang Young Radish (CYR) and Altari Radish (AR) according to the exposure for 31 days at low dose ${\beta}$-rays. This test has one contrast sample and eleven test samples each as to AR and CYR. The seeds from contrast and test sample were planted in the culture soil after 8 seeds were chosen from each with identical condition. The accumulated dose of test samples has been measured at consistent time on a daily basis for 31 days. The growing process and germination have been measured twice at consistent time in each week. The number of leaves, length of first leave and weight have been acquired average value by measuring for 20 and 25 days, respectively after being planted. The result of test sample in case of 25 days shows that 5% increase in length and 36% increase in weight for AR each at accumulated dose 0.01 Gy compared to the contrast sample. And the length of CYR has increased by 13~17% and 1% at accumulated dose 0.01~0.08 Gy and 0.3 Gy compared to the contrast sample. For the weight at accumulated dose 0.05 Gy and 0.23 Gy has increased by 36% and 2% compared to contrast sample. As to the number of leaves, AR has increased by 0~50% at accumulated dose 0.01-0.32 Gy compared to contrast sample. It also shows that the CYR has increased to 0~67% at accumulated dose 0.01-0.62 Gy compared to contrast sample. As a result of this study, it indicates that both AR and CYR has generally increased in their length, weight, and the number of leaves at low level accumulated dose part 0.01~0.2 Gy. The size of cell, area of nucleus and density of cell for test sample has been observed quite similar to the ones from contrast sample through microscope. In conclusion, AR and CYR irradiated by ${\beta}$-rays have estimated that they are achieved a rapid growth at low level accumulated dose region corresponding to its radiation hormesis theory. Further studies need to confirm the correlation between the radiation hormesis and the growth of the plants.

Anti-obesity Effect of Salsola collina Ethanol Extract (솔장다리 추출물의 항비만 효과)

  • Jin, Kyong-Suk;Lee, Su Hyeon;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
    • /
    • v.27 no.8
    • /
    • pp.888-895
    • /
    • 2017
  • Salsola collina (S. collina) is an annual plant widely distributed in drought and semi-drought areas, which has been used for a long time as a kind of folk remedy in traditional Chinese medicine for the treatment of hypertension. Previously, the anti-oxidative and anti-cancer activities of S. collina were elucidated in our research group. In this study, the anti-obesity activities of S. collina ethanol extract (SCEE) were evaluated using a pancreatic lipase enzyme inhibition assay and cell culture model. The results showed that SCEE effectively suppressed pancreatic lipase enzyme activity in a dose-dependent manner. Furthermore, SCEE significantly suppressed adipocyte differentiation, lipid accumulation, and triglyceride (TG) content, and triggered lipolysis on insulin, dexamethasone, and 3-isobutyl-l-methylxanthine-treated 3T3-L1 preadipocytes in a dose-dependent manner without cytotoxicity. Its anti-obesity effect was modulated by cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$, and the peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) gene, as well as protein expressions. Taken together, these results offer the important new insight that S. collina possesses anti-obesity properties, such as pancreatic lipase inhibition and anti-adipogenic and lipolysis effects through the modulation of their upstream signaling pathway. It could become a promising source in the field of nutraceuticals, and the identification of active compounds that confer the biological activities of SCEE may be needed.