• Title/Summary/Keyword: plant cell culture

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Biocontrol of Rhizoctonia solani Damping-off of Cucumber by Bacillus cereus KJA-118 (Bacillus cereus KJA-118을 이용한 오이 모잘록병의 생물학적 방제)

  • An, Kyu-Nam;Jung, Woo-Jin;Chae, Dong-Hyun;Park, Ro-Dong;Kim, Tae-Hwan;Kim, Yong-Woong;Kim, Young-Cheol;Cha, Gyu-Suk;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.36 no.4
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    • pp.247-255
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    • 2003
  • A bacterium, KJA-118 showing a strong chitinase activity, was isolated and identified as Bacillus cereus. The strain produced maximum level of chitinase, when grown aerobically at $30^{\circ}C$ for 4 days in basal broth containing 1% colloidal chitin in the initial pH adjusted to 6.0. Among various carbon sources such as crab shell powder, chitin powder, colloidal chitin, and R. solani mycelium, maximum chitinase activity was found in culture broth supplemented with R. solani mycelium. When KJA-118 was incubated with R. solani, the cell wall of the fungus was found to be completely destroyed. SDS-PAGE and active staining results revealed that KJA-118 produced three isoforms of chitinase with molecular weights of 68 kDa, 47 kDa, and 37 kDa. When the suspension of KJA-118 was treated to cucumber seedlings, reducing rate of damping-off caused by R. solani was about 28.1%.

Antifungal Activity of Bacillus sp. KMU-1011 Against Gray Mold Causing Botrytis cinerea (잿빛 곰팡이병원균 Botrytis cinerea에 대한 Bacillus sp. KMU-1011의 항진균활성)

  • Park Sung-Min;Kim Hyun-Soo;Yu Tae-Shick
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.63-69
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    • 2006
  • We isolated a bacterium which produces antifungal substances from the Lake of Saimaa soils in Fin-land. The isolated strain was identified as Bacillus sp. and shown a strong antifungal activity on plant pathogenic fungi. Bacillus sp. KMU-1011 produced maximum level of antifungal substances under incubation aerobically at $24^{\circ}C$ for 48 hours in nutrient broth containing 1.0% glucose and 1.0% polypeptone at 180 rpm and initiated pH adjusted to 6.0. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against Botrytis cinerea KACC 40573 by dry cell weight. Chloroform extract of cultured broth also shown fungal growth inhibitory activity against C. gloeosporioides KACC 40804, D. bryoniae KACC 40669, F. oxysporum KACC 40037, F. oxysporum KACC 40052, F. oxysporum f. sp. radicis-lycopersici KACC 40537, F. oxysporum KACC 40902, M. cannonballus KACC 40940, P. cambivora KACC 40160, R. solani AG-1 KACC 40101, R. solani AG-4 KACC 40142, and S. scleotiorum KACC by agar diffusion method.

Anti-oxidant and Anti-inflammatory Effects of Ethanol Extracts from Aerial Part of Coriandrum sativum L. (고수(Coriandrum sativum L.) 지상부 추출물의 항산화 및 항염증 활성 효과)

  • Nan, Li;Lee, Chang-Hyun;Choi, You-Na;Choo, Byung-Kil
    • Korean Journal of Organic Agriculture
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    • v.27 no.4
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    • pp.513-528
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    • 2019
  • Coriandrum sativum L., an annual herbaceous plant of Apiaceae family. The present study evaluated the anti-oxidant activities and anti-inflammatory effects of ethanol extracts of C. sativum. The anti-oxidant activities of C. sativum were measured by total contents of polyphenol, flavonoid, DPPH and ABTS radical scavenging and reducing power activity. And anti-inflammatory effects of C. sativum were measured by LPS-induced RAW 264.7 cells. The results showed that the contents of total polyphenol and flavonoid were 76.03 ± 1.36 mg of gallic acid equivalents/g and 182.23 ± 4.32 mg of rutin equivalents/g at concentration 1 mg/mL of C. sativum. The DPPH radical scavenging activity was found to be 52.8% at 500 ㎍/mL. The ABTS radical scavenging activity was shown in 58.3% after exposure to 1,000 ㎍/mL. Reducing power activity was found to be 66.8% at 2,000 ㎍/mL. The inhibitory effect of NO production was found to be 65% concentration 500 ㎍/mL. In the generation quantity of inflammatory cytokines such as TNF-α and IL-1β in cell culture medium, the expression levels of inflammatory proteins in cells were showed decrease with the increase of concentration. Therefore, we suggest that the C. sativum should be a potential source of alternative anti-inflammatory drug with good anti-inflammatory effects.

Toxicity and Characteristics of Antifungal Substances Produced by Bacillus amyloliquefaciens IUB158-03 (Bacillus amyloliquefaciens IUB158-03이 생산하는 항진균물질의 생화학적 특성 및 독성)

  • Kim, Hye-Young;Lee, Tae-Soo
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1672-1678
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    • 2009
  • The purified antifungal substances produced by Bacillus amyloliquefaciens IUB158-03 was positive to ninhydrin but negative to aniline, suggesting that the antifungal substance could be a peptide. FAB-MS, UV adsorption spectrum, and amino acid composition analysis revealed that the molecular weight of the antifungal substance was 1042 and that maximal adsorption was at 220 nm and 277 nm. The antifungal substance was composed of $Asn_3$, $Gln_2$, $Ser_1$, $Gly_1$, and $Tyr_1$. The composition and structural characteristics of antifungal substance were analysed by $^1H$-NMR spectrum, $^1H$-COSY, HMQC, which revealed that the compound belongs to the iturin A family. Temperature and pH had little effect on the stability of the antifungal substance in the ranges of $-70{\sim}121^{\circ}C$ and pH 6.0~10.0, respectively. It showed strong antibiotic activity against fungi. An in vitro cytotoxicity test using NIH3T3 cell showed that the antifungal substance does not have cytotoxicity. The number of circulating leukocytes and the hematobiological analysis of the mice administered with the antifungal substances was similar to those of the control group, indicating no cytotoxicity in vivo. Therefore, the antifungal substances extracted from culture broth of Bacillus amyloliquefaciens IUB158-03 have future potential as biocontrol agents against plant diseases caused by fungi.

Molecular Cloning and Expression of the Metallothionein Gene under Environmental Stresses in Sweet Potato (고구마 metallothionein 유전자의 클로닝 및 환경 스트레스 하에서 발현 분석)

  • Kim, Young-Hwa;Yu, Eun Jeong;Huh, Gyung-Hye
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1415-1420
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    • 2017
  • The metallothionein (MT) gene (IbMT3) was selected from an EST library of suspension-cultured sweet potato cells. The MT gene, which is one of abundant ESTs in the library, is involved in stress regulation of cells and tissues. A full-length IbMT3 cDNA was obtained and analysis of its nucleotide sequence revealed that IbMT3 encoded a type 3 MT protein, based on its structural characteristics. The function of type 3 MT in plants is not yet known. Northern blot analysis showed stronger expression of IbMT3 in suspension-cultured cells than in sweet potato plant leaves. Since cell culture is known to impose a state of oxidative stress on cells, sweet potato plants were subjected to oxidative stress to investigate the transcriptional regulation of IbMT3. When the herbicide methyl viologen (MV) was administered for 6, 12, and 24 hr, IbMT3 transcription rapidly increased at 6 hr and then decreased. A cold treatment at $15^{\circ}C$ for 24 and 48 hr resulted in a gradual increase in IbMT3 expression. These findings indicate that IbMT3 expression is regulated in response to environmental and oxidative stress. IbMT3 isoform is expected to have antioxidant effects in sweet potato plants and may play an important role in cellular adaptation to oxidative stress.

Biological Control of Tomato and Red Pepper Powdery Mildew using Paenibacillus polymyxa CW (Paenibacillus polymyxa CW를 이용한 고추 및 토마토 흰가루병 방제)

  • Kim, Yong-Ki;Choi, Eun-Jung;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Jee, Hyeong-Jin;Park, Jong-Ho;Han, Eun-Jung;Jang, Bo-Kyung;Yun, Jong-Cheul
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.379-387
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    • 2013
  • In order to improve practical utility of agro-microorganisms (AMs) which had been cultured and disseminated to promote plant growth and to control crop diseases, 51 isolates of AMs were collected from 18 agricultural extension centers in local government and screened for multi-functions such as antifungal activity, activities of phosphorus solubilization, IAA and siderophore production, nitrogen fixation, and hydrolytic enzyme activity. Finally we selected one isolate showing good antifungal activity and multi-functions related to plant growth and disease control. The selected isolate, Paenibacillus polymyxa CW, showed good inhibitory effect against plant pathogens, Pyricularia gresea, Colletotrichum acutatum, Fusarium oxysporum, Phomopsis sp., Aspergillus niger, Rhizoctonia solani and Phytophthora capsici. Suppressive effect of P. polymyxa CW against the used plant pathogens except for R. solani was much higher than that of P. polymyxa AC-1 storing in National Academy of Agricultural Science. We found P. polymyxa CW isolate showed good activity in siderophore and IAA formation, and nitrogen fixation. With P. polymyxa CW isolate, siderophore formation activity was similar to that of P. polymyxa AC-1, but IAA formation and nitrogen fixation activity was much higher than that of P. polymyxa AC-1. However neither P. polymyxa CW nor P. polymyxa AC-1 showed hydrolytic enzyme (chitinase, pectinase and cellulase) activity. The treatment of P. polymyxa CW with culture suspension of different cell density ($10^8$, $10^7$. $10^6$ cfu/ml) showed that the highest density reduced incidence of red pepper powdery mildew by 68.3% after 10 days of application. As application density of P. polymyxa CW was decreased, its control efficacy was proportionally decreased. In addition, when P. polymyxa CW was treated to control tomato powdery mildew at the same concentrations and their control effects were investigated after 7 days of inoculation, disease incidence was 0.03, 19.5, 45.7%, respectively, compared to 56.3% that of untreated check. Like red pepper powdery mildew, increase of application density of P. polymyxa CW resulted in increase of its control efficacy proportionally. P. polymyxa CW showed a density-dependent control efficacy against red pepper and tomato powdery mildews. Therefore we think that mode of action of the antagonist for suppressing two powdery mildew diseases might be antibiosis and density of more than $10^8cfu/ml$ was needed to control effectively the two diseases. On this basis, we think that P. polymyxa CW can be a promising control agent for suppressing powdery mildews of red pepper and tomato.

The Effect of Kaempferol, guercetin on Hyaluronan-Synthesis Stimulation in Human Keratinocytes (HaCaT) (인체 피부 세포주 (HaCaT)에서 Kaempferol, Quercetin의 Hyaluronan 합성 촉진 효과에 대한 연구)

  • Kim, Seung-Hun;Nam, Gae-Won;Kang, Byung-Young;Lee, Hae-Kwang;Moon, Seong-Joon;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.97-102
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    • 2005
  • One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.

The Hepatotprotective and Antioxidative Effects of Onion (Allium cepa) Extracts in Rat Hepatocyte Primary Culture (양파(Allium cepa) 추출물의 간보호 및 항산화 효과)

  • Rhim, Tae-Jin;Lim, Sang-Cheol
    • Proceedings of the Plant Resources Society of Korea Conference
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    • v.18 no.1
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    • pp.52-60
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    • 2005
  • The objectives of present study were to investigate the hepatoprotective and antioxidative effects of onion extracts. Primary cultures of rat hepatocytes were incubated with 1.5 mM tert-butyl hydroperoxide(t-BHP), potent oxidizing agent for liver injury for 1 hr in the presence or absence of various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring catalase, glutathione peroxidase(GSH-Px), glutathione reductase(GSH-Rd) activities as well as DNA strand breaking assay. Incubation with t-BHP alone increased GOT and LDH activities and TBARS concentration but decreased MTT reduction. Onion extracts at the concentration of 0.05 mg/ml began to decrease GOT and LDH activities induced by 1.5 mM t-BHP. Decreased MTT reduction began to be increased by onion extract at the concentration of 0.01 mg/ml. Onion extracts at the concentration of 0.01 mg/ml began to decrease TBARS concentration induced by t-BHP. Taken together, onion extracts prevented t-BHP-induced hepatocyte injury and lipid peroxidation. Catalase, GSH-Px and GSH-Rd activities of hepatocytes were significantly decreased by 1.5 mM t-BHP for 1 hr incubation. Onion extracts, on the other hand, at the concentration of 0.1 mg/ml began to prevent t-BHP-induced decrease in catalase, GSH-Px and GSH-Rd activities. Onion extracts prevented hydroxyl radical-induced single-strand breakage in dose-dependent manner when plasmid DNA was incubated with various concentrations of onion extracts in the presence of Fenton regents producing hydroxyl radical. These results demonstrate that onion extracts suppressed t-BHP-induced cytoctoxicity, decreased viability and lipid peroxidation and increased GSH-Px, GSH-Rd and catalase activities. Thus hepatoprotective and antioxidant effects of onion extract seem to be due to, at least in part, the increase in antioxidant enzyme activities as well as prevention from hydroxyl radical-induced oxidation, followed by inhibition in lipid peroxidation.

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Recent Advancement in the Differentiation of Tissues and Organs and Regulation of Gene Expression (조직.기관의 분화와 유전자 발현의 조절, 최근의 진보)

  • Harn, Chang-Yawl
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.1-35
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    • 1997
  • Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

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Studies on the Ginseng Plants(III) -Radioactive Sodium $Acetate-U-C^{14}$ Feeding Experiments- (인삼식물(人參植物)에 관한 연구(III) -동위원소화합물(洞位元素化合物) Sodium $Acetate-U-C^{14}$을 투여한 실험-)

  • Kim, Jung-Yun;Staba, E. John
    • Korean Journal of Pharmacognosy
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    • v.5 no.2
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    • pp.111-124
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    • 1974
  • The radioactive compound sodium $acetate-U-C^{14}\;(C^{14}-acetate)$ was administered to two- and four-year-old July and September American ginseng (Araliaceae, Panax quinquefolium L.) plants and cuttings. The $C^{14}-acetate$ uptake was approximately 99%. The autoradiochromatograms suggest that the saponins isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration (% plant dry weight) of semi-purified saponins were high in the leaves (13.8%), as compared to fruits (9.8%), stems (7.9%) and roots (6.3%). The average percentage of $C^{14}-acetate$ incorporation into panaquilins was 4.8%. The average percentage of $C^{14}-acetate$ incorporation into panaquilins B and C was higher (1.40% and 1.13%, respectively) than that into panaquilins C, (d), G-1 and G-2 (0.75%, 0.65%, 0.13% and 0.53%, respectively). Panaquilin synthesis may be depending upon the part, collection period and age of the plant. The average percentage of $C^{14}-acetate$ incorporation into panaquilin B is high in roots (0.58%) and stems (0.48%); that into panaquilins C and (d) high in leaves (0.40% and 0.45%, respectively); and that into panaquilin E high in roots and leaves (0.55% and 0.50%, respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-1). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-1 may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and cailus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that $C^{14}-acetate$ was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act. 0.56 mmcCi/mg) and four-year-old plants $(sp.\;act.\;0.54\;m{\mu}Ci/mg)$.

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