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Euphorbiae Immifusae Sensitizes Apoptosis of TRAIL-resistant Human Gastric Adenocarcinoma AGS Cells (지금초 추출물에 의한 TRAIL 저항성 인체위암세포의 세포사멸 유도)

  • Lee, Jae-Jun; Shin, Dong-Hyuk;Park, Sang-Eun;Kim, Won-Il;Park, Dong-Il;Choi, Yung-Hyun;Hong, Sang-Hoon
    • Journal of Life Science
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    • v.18 no.1
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    • pp.120-128
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    • 2008
  • The death ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/ Apo1L is a cytokine that activates apoptosis through cell surface death receptors. TRAIL has sparked growing interest in oncology due to its reported ability to selectively trigger cancer cell death. Euphorbiae humifusae Wind has been used in traditional Oriental medicine as a folk remedy used for the treatment of cancer. However, the mechanism responsible for the anticancer effects of E. humifusae not clearly understood. Here, we show that treatment with subtoxic doses of water extract of E. humifusae (WEEH) in combination with TRAIL induces apoptosis in TRAIL-resistant human gastric carcinoma AGS cells. Combined treatment with WEEH and TRAIL induced chromatin condensation and sub-G1 phase DNA content. These indicators of apoptosis were correlated with the induction of caspase activity that resulted in the cleavage of poly (ADP-ribose) polymerase. Combined treatment also triggered the loss of mitochondrial membrane potential. Furthermore, co-treatment with WEEH and TRAIL down-regulated the protein levels of the anti-apoptotic proteins such as Bcl-2, Bcl-xL, XIAP and cIAP-1. Although more study will be needed to examine the detailed mechanisms, this combined treatment may offer an attractive strategy for safely treating gastric adenocarcinomas and the results provide important new insights into the possible molecular mechanisms of the anticancer activity of E. humifusae.

A Study on the Gene Expression in Shikonin-Induced Inhibition of Adipogenesis (Shikonin에 의한 지방세포형성 억제과정에서의 유전자 발현 연구)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Chung, Sang-In;Cho, Soo-Hyun;Oh, Dong-Jin;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1637-1643
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    • 2009
  • Shikonin, a component of Lithospermum erythrorhizon Sieb. et Zucc, exerts various characteristics such as anti-inflammatory, anti-cancer and anti-obesity functions. To elucidate the molecular mechanism of shikonin-induced inhibition of adipogenesis, we analyzed the mRNA expression level of various adipogenesis-related factors including C/EBPs (CCAAT/enhancerbinding proteins) and $PPAR{\gamma}$ (peroxisome proliferator-activated receptor $\gamma$). The data showed that mRNA expressions of C/$EBP{\beta}$ and C/$EPB{\delta}$ were only slightly changed by shikonin treatment, but mRNA expressions of $PPAR{\gamma}$ and C/$EPB{\alpha}$ were significantly down-regulated. Then, we tested whether upstream regulators of C/$EBP{\beta}$ and $PPAR{\gamma}$ were involved in anti-adipogenesis of shikonin. C/$EBP{\gamma}$ and CHOP (C/EBP homologous protein), which are upstream regulators of C/$EBP{\beta}$, were not affected by shikonin treatment. On the contrary, the mRNA level of KROX20 was markedly down-regulated by shikonin treatment. These results suggest that KROX20 might regulate downstream factors of adipogenesis through C/$EBP{\beta}$-independent pathway. The expression of KLF15 (Kruppel-like factor15), which is a member of KLF family and is a upstream regulator of $PPAR{\gamma}$, was dramatically decreased by shikonin treatment, but KLF2 was not changed. Shikonin had no impact on the expression of KLF5 in the early stage of adipogenesis, but shikonin increased expression of KLF5 in the late stage of adipogenesis. Even though mRNA expression of KLF5 was moderately changed by shikonin treatment, its effect may be small compared with the effect of KLF15, which was markedly inhibited. Taken together, these results suggest that shikonin inhibits adipogenesis through the down-regulation of $PPAR{\gamma}$ and C/$EPB{\alpha}$, which is mediated by the down-regulation of two pro-adipogenic factors, KROX20 and KLF15.

Seasonal Changes of Chemical Compositions in Leaves, Shoot and Berries of ‘Delaware’Seedless Grapes induced by Gibberellic Acid (Delaware 포도의 무핵재배에서 엽${\cdot}$신초${\cdot}$과립의 생장 및 성숙에 따른 화학성분의 변화)

  • 최수주
    • Journal of Life Science
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    • v.7 no.2
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    • pp.142-148
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    • 1997
  • This study was carried out to clarify seasonal changes of chemical compositions and their interrelation in leaves, shoots and berries treated with gibberellin($GA$_{3}$)for seedless grapes in ‘Delaware’grapevines. the clusters were dipped twice with 100ppm $GA_{3}$: 10 days befor and after the full bloom. The reaults obtained as follows; 1. Cumulative growth curve of berry fresh weight showed a double sigmoid curve and the characteristics of three distinctive growth stages(I, II, III)were weekened with $GA_{3}$ treatment. 2. while the contents of ash, total carbon, total nitrogen and total carbohydratd had little reation with edvelopment and ripeness of berries, those of total sugar and starch jad close relation, viz., they decreased with enlargement and maturity fo berries, but increased rapidly after harvest in leaves and shoots. Especially, total sugars in leaves and shoots decreased coincidently with starch-increasing in shoots at November. 3. The contents of total soluble solid and reducing sugar in berries increased rapidly at growth stage III, but those of total titratable acidity and organic acid decreased coincidently with sugar-increasing. 4. The berry-hardness increased until growth stage I, and then stagnated until gtowth stage II, and then increased rapidly at growth stage III. pH of berry-juice decreased until growth stageII, afterwards increased at growth stage III. 5. By correlation and path coefficient analysis between qualitative characters and the ratio of total soluble solid to titratable acidity($^{\circ}$Brix/Acidity), total correlation coefficients were all highly significant. Of these characters, pH and viscosity of berry-juice were positive, but brightness and hardness of berry, negative. The direct effect of pH on $^{\circ}$Brix/Acidity ratio, p4y=0.9090, was large positively and those of berry-hardness and juice-viscosity, p1y=0, 5938, median and, p2y=0, 3550, small, respectively. Direct effect of brightness was negatively small.

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Expression of Yippee-Like 5 (YPEL5) Gene During Activation of Human Peripheral T Lymphocytes by Immobilized Anti-CD3 (인체 말초혈액의 활성화 과정 중 yippee-like 5 (YPEL5) 유전자의 발현 양상)

  • Jun, Do-Youn;Park, Hye-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1641-1648
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    • 2007
  • Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.

Antioxidant and Anticancer Activities of Euonymus porphyreus Extract in Human Lung Cancer Cells A549 (인체 폐암 세포주 A549에서 Euonymus porphyreus 추출물의 항산화 및 항암활성 분석)

  • Jin, Soojung;Oh, You Na;Son, Yu Ri;Bae, Soobin;Park, Jung-ha;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Life Science
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    • v.31 no.2
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    • pp.199-208
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    • 2021
  • Euonymus porphyreus, a species of plant in the Celastraceae family, is widely distributed in East Asia, especially in Southern China. The botanical characteristics of E. porphyreus have been reported, but its antioxidative and anticancer activities remain unclear. In this study, we evaluated the antioxidative and anticancer effects of ethanol extracts of E. porphyreus (EEEP) and the molecular mechanism of its anticancer activity in human lung adenocarcinoma A549 cells. The total polyphenol and flavonoid compound contents from EEEP were 115.42 mg/g and 23.07 mg/g, respectively. EEEP showed significant antioxidative effects with a concentration at 50% of the inhibition (IC50) value of 11.09 ㎍/ml, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. EEEP showed cytotoxic activity by increasing the SubG1 cell population of A549 cells in a dose-dependent manner. Apoptosis in A549 cells treated with EEEP was evident due to increased apoptotic cells and apoptotic bodies, as detected by Annexin V and 4,6-diamidino-2-phenylindole (DAPI) staining, respectively. EEEP-induced apoptosis resulted in increased expression of the First apoptosis signal (Fas), p53, and Bax, with decreased expression of Bcl-2 and subsequent activation of caspase-8, -9, and caspase-3, leading to cleavage of poly (ADP-ribose) polymerase (PARP). Collectively, these results suggest that EEEP may exert an anticancer effect by inducing apoptosis in A549 cells through both intrinsic and extrinsic pathways.

A Study on Image Copyright Archive Model for Museums (미술관 이미지저작권 아카이브 모델 연구)

  • Nam, Hyun Woo;Jeong, Seong In
    • Korea Science and Art Forum
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    • v.23
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    • pp.111-122
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    • 2016
  • The purpose of this multi-disciplinary convergent study is to establish Image Copyright Archive Model for Museums to protect image copyright and vitalize the use of images out of necessity of research and development on copyright services over the life cycle of art contents created by the museums and out of the necessity to vitalize distribution market of image copyright contents in creative industry and to formulate management system of copyright services. This study made various suggestions for enhancement of transparency and efficiency of art contents ecosystem through vitalization of use and recycling of image copyright materials by proposing standard system for calculation, distribution, settlement and monitoring of copyright royalty of 1,000 domestic museums, galleries and exhibit halls. First, this study proposed contents and structure design of image copyright archive model and, by proposing art contents distribution service platform for prototype simulation, execution simulation and model operation simulation, established art contents copyright royalty process model. As billing system and technological development for image contents are still in incipient stage, this study used the existing contents billing framework as basic model for the development of billing technology for distribution of museum collections and artworks and automatic division and calculation engine for copyright royalty. Ultimately, study suggested image copyright archive model which can be used by artists, curators and distributors. In business strategy, study suggested niche market penetration of museum image copyright archive model. In sales expansion strategy, study established a business model in which effective process of image transaction can be conducted in the form of B2B, B2G, B2C and C2B through flexible connection of museum archive system and controllable management of image copyright materials can be possible. This study is expected to minimize disputes between copyright holder of artwork images and their owners and enhance manageability of copyrighted artworks through prevention of such disputes and provision of information on distribution and utilization of art contents (of collections and new creations) owned by the museums. In addition, by providing a guideline for archives of collections of museums and new creations, this study is expected to increase registration of image copyright and to make various convergent businesses possible such as billing, division and settlement of copyright royalty for image copyright distribution service.

Comparison of Fruit Characteristics of 'Fuji'/M.26 in Response to Ethephon Treatment and Combined Treatment of Ethephon and CaCl2 During Maturing Stages (Ethephon 단용처리와 Ethephon 및 염화칼슘 혼합처리에 따른 사과 'Fuji'/M.26의 성숙기 과실특성 비교)

  • Sewon Oh;Seong Ho Moon;Keum-Il Jang;Junsoo Lee;Daeil Kim
    • Korean Journal of Plant Resources
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    • v.36 no.5
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    • pp.517-526
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    • 2023
  • The harvest time of the late-ripening 'Fuji' apple (Malus × domestica) is variable, depending on the coloration of the fruit skin. Ethephon, a plant growth regulator, promotes the ethylene production and induces physiological responses associated with fruit maturation in climacteric fruit crops, such as apples. This study aimed to investigate the effect of ethephon treatment on fruit characteristics after fruit enlargement, with the objective of proposing an economical and stable harvest control method for 'Fuji'/M.26 apples. Fruit characteristics were assessed at 10-days intervals following the application of 100 mg/L ethephon and mixture of 100 mg/L ethephon and 0.5% CaCl2 at 145 days after full bloom (DAFB). Starch contents of ethephon-treated (ET) and ethephon with CaCl2-treated (EC) apples began to decrease from 155 DAFB, and the starch contents of ET and EC at 10 days before harvest were similar to those of control at harvest time. Red coloration of fruit skin in EC was lower compared to ET but higher than control. The average fruit firmness was low in ET, while the control and EC exhibited similar levels of firmness. Fruit sugar acid ratios did not show significant differences between treatments. However, the titratable acidity of EC was significantly lower than that of the control at 10 days before harvest. Moreover, fruit sugar acid ratio of ET and EC at 10 days before harvest in 2021 was similar to their sugar acid ratio at harvest time. Therefore, it was thought that fruit maturation and skin coloration could be accelerated by 10 days from the harvest time through the combined treatment of 100 mg/L ethephon and 0.5% CaCl2 at the end of fruit enlargement in 'Fuji'/M.26.

Attenuation of Oxidative Stress-Induced HepG2 Cellular Damage by Cirsiumjaponicum Root Extract (HepG2 세포에서 대계 추출물에 의한 산화적 스트레스 유발 세포 손상의 억제)

  • Da Jung Ha;Seohwi Kim;Byunwoo Son;Myungho Jin;Sungwoo Cho;Sang Hoon Hong;Yung Hyun Choi;Sang Eun Park
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1002-1014
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    • 2023
  • The root of Cirsium japonicum var. maackii (Maxim.) has long been used in traditional medicine to prevent the onset and progression of various diseases and has been reported to exert a wide range of physiological effects, including antioxidant activity. However, research on its effects on hepatocytes remains scarce. This study used the human hepatocellular carcinoma HepG2 cell line to investigate the antioxidant activity of ethanol extract of C. japonicum root (EECJ) on hepatocytes. Hydrogen peroxide (H2O2) was used to mimic oxidative stress. The results showed that EECJ significantly reverted the decrease in cell viability and suppressed the release of lactate dehydrogenase in HepG2 cells treated with H2O2. Moreover, an analysis of changes in cell morphology, flow cytometry, and microtubule-associated protein light chain 3 (LC3) expression showed that EECJ significantly inhibited HepG2 cell autophagy induced by H2O2. Furthermore, it attenuated H2O2-induced apoptosis and cell cycle disruption by blocking intracellular reactive oxygen species and mitochondrial superoxide production, indicating strong antioxidant activity. EECJ also restored the decreased levels of intracellular glutathione (GSH) and enhanced the expression and activity of superoxide dismutase and GSH peroxidase in H2O2-treated HepG2 cells. Although an analysis of the components contained in EECJ and in vivo validation using animal models are needed, these findings indicate that EECJ is a promising candidate for the prevention and treatment of oxidative stress-induced liver cell damage.

Studies on the physio-chemical properties and the cultivation of oyster mushroom(Pleurotus ostreatus) (느타리버섯의 생리화학적성질(生理化學的性質) 및 재배(栽培)에 관(關)한 연구(硏究))

  • Hong, Jai-Sik
    • Applied Biological Chemistry
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    • v.21 no.3
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    • pp.150-184
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    • 1978
  • Nutritional characteristics and physio-chemical properties of mycelial growth and fruitbody formation of oyster mushroom(Pleurotus ostreatus)in synthetic media, the curtural condition for the commerical production in the rice straw and poplar sawdust media, and the changes of the chemical components of the media and mushroom during the cultivation were investigated. The results can be summarized as follows: 1. Among the carbon sources mannitol and sucrose gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while lactose and rhamnose gave no mycelial growth. Also, citric acid, succinic acid, ethyl alcohol and glycerol gave poor fruit-body formation, and acetic acid, formic acid, fumaric acid, n-butyl alcohol, n-propyl alcohol and iso-butyl alcohol inhibited mycelial growth. 2. Among the nitrogen sources peptone gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while D,L-alanine, asparatic acid, glycine and serine gave very poor fruit-body formation, and nitrite nitrogens, L-tryptophan and L-tyrosine inhibited mycelial growth. Inorganic nitrogens and amino acids added to peptone were effective for fruit-body growth, and thus addition of ammonium sulfate, ammonium tartarate, D,L-alanine and L-leucine resulted in about 10% increase fruit-body yield. L-asparic acid about 15%, L-arginine about 20%, L-glutamic acid, and L-lysine about 25%. 3. At C/N ratio of 15.23 fruit-body formation was fast, but the yield decreased, and at C/N ratio of 11.42 fruit-body formation was slow, but the yield increased. Also, at the same C/N ratio the higher the concentration of mannitol and petone, the higher yield was produced. Thus, from the view point of both yield of fruit-body and time required for fruiting the optimum C/N ratio would be 30. 46. 4. Thiamine, potassium dihydrogen phosphate and magnecium sulfate at the concentration of $50{\mu}g%$. 0.2% and 0.02-0.03%, respectively, gave excellent mycelial and fruit-body growth. Among the micronutrients ferrous sulfate, zinc sulfate and manganese sulfate showed synergetic growth promoting effect but lack of manganese resulted in a little reduction in mycelial and fruit-body growth. The optimum concentrati on of each these nutrients was 0.02mg%. 5. Cytosine and indole acetic acid at 0.2-1mg% and 0.01mg%, respectively, increased amount of mycelia, but had no effect on yield of fruit-body. The other purine and pyrimidine bases and plant hormones also had no effect on mycelial and fruit-belly yield. 6. Illumination inhibited mycelial growth, but illumination during the latter part of vegetative growth induced primordia formation. The optimum light intensity and exposure time was 100 to 500 lux and 6-12 hours per day, respectively. Higher intensity of light was injurous, and in darkness only vegetative growth without primordia formation was continued. 7. The optimum temperature for mycelial growth was $25^{\circ}C$ and for fruit-body formation 10 to $15^{\circi}C$. The optimum pH range was from 5.0 to 6.5. The most excellent fry it-body formation were produced from the mycelium grown for 7 to 10 days. The lesser the volume of media, the more rapid the formation of fruit-body; and the lower the yield of fruit-body; and the more the volume of media, the slower the formation of fruit-body, and the higher the yield of fruit-body. The primordia formation was inhibited by $CO_2$. 8. The optimum moisture content for mycelial growth was over 70% in the bottle media of rice straw and poplar sawdust. 10% addition of rice bran to the media exhibited excellent mycelial growth and fruit-body formation, and the addition of calciumcarbonate alone was effective, but the addition of calcium carbonate was ineffective in the presence of rice bran. 9. In the cultivation experiments the total yield of mushroom from the rice straw media was $14.99kg/m^2$, and from the sawdust media $6.52kg/m^2$, 90% of which was produced from the first and second cropping period. The total yield from the rice straw media was about 2.3 times as high as that from the sawdust media. 10. Among the chemical components of the media little change was observed in the content of ash on the dry weight basis, and organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the period of mycelial propagation, after which no change was observed. 11. During the period from spawning to the fourth cropping about 16.7% of the dry matter, about 19.3% of organic matter, and about 40% of nitrogen were lost from the rice straw media; about 7.5% of dry mallet, about 7.6% of organic matter, and about 20% of nitrogen were lost from the sawdust media. For the production of 1kg of mushroom about 232g of organic matter and about 7.0g of nitrogen were consumed from the rice straw media; about 235g of organic matter and about 6.8g of nitrogen were consumed from the sawdust media, 1㎏ of mushroom from either of media contains 82.4 and 82.3g of organic matter and 5.6 and 5.4g of nitrogen, respectively. 12. Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Content of amino nitrogen continued to increase up to the third cropping time, after which it decreased. 13. In the rice straw media 28.0 and 13.8% of the total pentosan and ${\alpha}$-cellulose, respectively, lost during the whole cultivation period was lost during the period of mycelial growth; in the sawdust media 24.1 and 11.9% of the total pentosan and ${\alpha}$-cellulose, respectively, was lost during the period of mycelial growth. Lignin content in the media began to decrease slightly from the second cropping time, while the content of reduced sugar, trehalose and mannitol continued to increase. C/N ratio of the rice straw media decreased from 33.2 at spawining to 30.0 at ending; that of the sawdust media decreased from 61.3 to 60.0. 14. In both media phosphorus, potassium, manganese and zinc decreased, at magnesium, calcium and copper showed irregular changes, and iron had a tendency to be increased. 15. Enzyme activities are much higher in the rice straw media than in the sawdust media. CMC saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased rapidly in the rice straw media, which was not observed in the sawdust media. Protease activity was highest after mycelial propagation, after which it gradually decreased. The pH of the rice straw media decreased from 6.3 at spawning to 5.0 after fourth cropping; that of the sawdust media decreased from 5.7 to 4.9. 16. The contents of all the components except crude fibre of the mushroom from the rice straw media were higher than those from the sawdust media. Little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.

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Development of a marker system to discern the flowering type in Brassica rapa crops (배추 속 작물의 개화형 판별 마커 시스템 개발)

  • Kim, Jin A;Kim, Jung Sun;Hong, Joon Ki;Lee, Yeon-Hee;Lee, Soo In;Jeong, Mi-Jeong
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.438-447
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    • 2017
  • Flowering is one of the most important development traits related to the production of Brassica rapa crops. After planting, a sudden low temperature triggers premature flowering, which leads to a reduction in the yield and quality of harvested production. Therefore, understanding the mechanism of flowering control is important in the agricultural productivity for preventing Brassica rapa crops. Vernalization is generally known as the main factor of flowering in the Brassica plant. However, in the subspecies of Brassica rapa, some accession such as Yellow sarson and Komatsuna display the flowering phenotype without vernalization. Circadian genes, which diurnally regulate plant physiology, have a role for photoperiodic flowering but are related to the regulation of the vernalizarion mechanism. In this report, the 22 B. rapa accession were divided into two groups, vernalization and non-vernalization, and the sequenced circadian gene, BrPRR1s. Among them, the BrPRR1b gene was found to have deletion regions, which could classify the two groups. The PCR primer was designed to amplify a short band of 422bp in the vernalization type and a long band of 451bp in the non-vernalization type. This primer set was applied to distinguish the flowering types in the 43 B. rapa accession and 4 Brassica genus crop, Broccoli, cabbage, mustard, and rape. The PCR analysis results and flowering time information of each crop demonstrated that the primer set can be used as marker to discern the flowering type in Brassica crops. This marker system can be applied to the B. rapa breeding when selecting the flowering character of new progenies or introducing varieties at an early stage. In addition, these results displayed that the circadian clock genes can be a good strategy for the flowering control of B. rapa crops.