• Title/Summary/Keyword: benzyladenine

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Use of Paromomycin as a Selectable Marker for the Transformation of Chinese Cabbage (배추의 형질전환용 선발항생제로서 Paromomycin의 이용)

  • Cho, Mi-Ae;Min, Sung-Ran;Ko, Suck-Min;Liu, Jang-Ryol;Lee, Jun-Haeng;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.271-276
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    • 2006
  • Hypocotyl explants of Chinese cabbage (us. 'Jeong Sang' and 'Seoul') produced adventitious shoots on Murashige and Skoog (MS) basal medium supplemented with 4mg/L $AgNO_3$, 5 mg/L acetosyringone, 4 mg/L 6-benzyladenine and 3mg/L alpha-naphthaleneacetic acid (SI) after cocoultivation with strains of Agrobacterium tumefaciens (LBA4404) harboring the pCAMBIA1301 and the $_PPTN290$ containing hygromycin-resistance gene and paromomycin-resistance gene as a selectable marker genes, respectively. There was a significant difference in the frequency of transgenic plants depending on antibiotics and cultivars used. Paromomycin was better than hygromycin, and cultivar 'Jeong-sang' was higher than 'c.v. Seoul' in the frequency of transgenic plants. In particular, the highest frequency (0.70%) of transgenic plants was obtained from selection medium (SI) containing 100mg/L paromomycin in c.v., 'Jeong-sang' GUS positive response were obtained 9 plants and 3 plants from the cultivars, 'Jeong-sang' and 'Seoul', respectively. They were grown to maturity in a greenhouse and normally produced $T_1$ seeds. GUS histochemical assay for progeny $(T_1)$ revealed that the transgenes were expressed in the plant genome.

Adventitious root induction in Ophiorrhiza prostrata: a tool for the production of camptothecin (an anticancer drug) and rapid propagation

  • Martin, Kottackal Poulose;Zhang, Chun-Lai;Hembrom, Manoj Emanuel;Slater, Adrian;Madassery, Joseph
    • Plant Biotechnology Reports
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    • v.2 no.2
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    • pp.163-169
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    • 2008
  • Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

A Bioassay Method with Radish Seedlings for Bioactive Substances (무우 유묘(幼苗)를 이용(利用)한 식물생리활성물질(植物生理活性物質)의 생물검정법(生物檢定法))

  • Hong, K.S.;Lim, H.K.;Cho, K.Y.
    • Korean Journal of Weed Science
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    • v.8 no.3
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    • pp.244-249
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    • 1988
  • Some experiments were conducted to utilize the raphanus test as a bioassay method for plant growth regulaters and hormonal herbicides. Among seven radish cultivars, the response of Jangbaegminong was the most sensitive and quantitative to ${\alpha}$-NAA and its coefficient of variation value was the lowest. The optimum application time of test compounds was estimated seven to nine days after seeding. After ${\alpha}$-NAA treatment the curvature angle of radish cotyledonary petiols was sharply increased to the maximum value at 24 hours after and was changed by light and temperature conditions. The curvature response was observed by application of ${\alpha}$-NAA, phenoxy and indole compounds but often plant growth regulators, $GA_3$ benzyladenine, ABA and homobrassinolide was not detected by this method. So this bioassay was useful to evaluate the activity of a compound as an auxin or a hormonal herbicide.

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Scopolamine Production in Suspension Cultures of Tumor Calli from Datura metel L. (흰독말풀(Datura metel L.)종양 캘러스의 현탁배양으로부터 Scopolamine 생성)

  • 이수경;윤길영;김용해;양덕조
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.203-211
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    • 2000
  • In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.

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Effect of Several Fungicides and Growth Regulators on Rice Seedling Growth and Damping-off in Seedling Boxes for Machine Transplanting (벼상자육묘(箱子育苗)에서 살균제(殺菌齊)와 생장조절제(生長調節劑) 처리(處理)가 묘생육(苗生育) 및 생리장해(生理障害)에 미치는 영향(影響))

  • Jeh, Sang Yull;Hwang, Chung Dong
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.1-11
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    • 1987
  • This study was conducted to evaluate the effect of several fungicides and growth regulators on rice seedling growth and damping-off in seedling boxes for machine transplanting. Fungicide treated plots were better seedling growth, shoot regrowth, rooting ability, change of moisture content than those of nontreated plot. Metalaxyl application of Samgangbyeo and SF8002 application of Nagdongdyeo apparently increased plant height, length of the third leaf and fourth leaf. And metaiaxyl application highly increased dry weight. Fungicide treated plots were highly effective in reducing the incidence of damping-off. Benzyladenine application of Samgangbye and $GA_3$ application of Nagdongbyeo apprently increased plant height. But ABA application highly decreased plant height. ABA application and aCE application resulted in highly increased rooting ability. Fungicide and $GA_3$ treated plots, Metalaxyl and growth regulator treated plots resulted in highly increased plant height. I soprothiolane and growth regulator treated plots resulted in decreased plant height. Dachigaren and lAA treated plot apprently increased dry weight and shoot dry weight/plant height. Fungicide and growth regulator treated plots were highly effective in reducing the incidence of damping-off.

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Apoptosis of Kinetin Riboside in Colorectal Cancer Cells Occurs by Promoting β-Catenin Degradation

  • TaeKyung Nam;Wonku Kang;Sangtaek Oh
    • Journal of Microbiology and Biotechnology
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    • v.33 no.9
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    • pp.1206-1212
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    • 2023
  • The Wnt/β-catenin pathway plays essential roles in regulating various cellular behaviors, including proliferation, survival, and differentiation [1-3]. The intracellular β-catenin level, which is regulated by a proteasomal degradation pathway, is critical to Wnt/β-catenin pathway control [4]. Normally, casein kinase 1 (CK1) and glycogen synthase kinase-3β (GSK-3β), which form a complex with the scaffolding protein Axin and the tumor suppressor protein adenomatous polyposis coli (APC), phosphorylate β-catenin at Ser45, Thr41, Ser37, and Ser33 [5, 6]. Phosphorylated β-catenin is ubiquitinated by the β-transducin repeat-containing protein (β-TrCP), an F-box E3 ubiquitin ligase complex, and ubiquitinated β-catenin is degraded via a proteasome pathway [7, 8]. Colorectal cancer is a significant cause of cancer-related deaths worldwide. Abnormal up-regulation of the Wnt/β-catenin pathway is a major pathological event in intestinal epithelial cells during human colorectal cancer oncogenesis [9]. Genetic mutations in the APC gene are observed in familial adenomatous polyposis coli (FAP) and sporadic colorectal cancers [10]. In addition, mutations in the N-terminal phosphorylation motif of the β-catenin gene were found in patients with colorectal cancer [11]. These mutations cause β-catenin to accumulate in the nucleus, where it forms complexes with transcription factors of the T-cell factor/lymphocyte enhancer factor (TCF/LEF) family to stimulate the expression of β-catenin responsive genes, such as c-Myc and cyclin D1, which leads to colorectal tumorigenesis [12-14]. Therefore, downregulating β-catenin response transcription (CRT) is a potential strategy for preventing and treating colorectal cancer. Plant cytokinins are N6-substituted purine derivatives; they promote cell division in plants and regulate developmental pathways. Natural cytokinins are classified as isoprenoid (isopentenyladenine, zeatin, and dihydrozeatin), aromatic (benzyladenine, topolin, and methoxytopolin), or furfural (kinetin and kinetin riboside), depending on their structure [15, 16]. Kinetin riboside was identified in coconut water and is a naturally produced cytokinin that induces apoptosis and exhibits antiproliferative activity in several human cancer cell lines [17]. However, little attention has been paid to kinetin riboside's mode of action. In this study, we show that kinetin riboside exerts its cytotoxic activity against colon cancer cells by suppressing the Wnt/β-catenin pathway and promoting intracellular β-catenin degradation.

Optimal concentrations of plant growth regulators and AgNO3 for the improvement of regeneration efficiency in Chrysanthemum morifolium 'Ohblang' (국화 '오블랑'의 재생 효율 증진을 위한 식물생장조절제와 AgNO3 적정 농도 선별)

  • Yeo Jin Youn;Yong Joon Yang
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.169-175
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    • 2023
  • A plant regeneration system was developed through shoot organogenesis from in vitro leaf explants of Chrysanthemum morifolium 'Ohblang'. The effects of different concentrations of plant growth regulators and AgNO3 on efficient shoot regeneration and inhibition of browning were evaluated in chrysanthemum. The explants were cultured on MS shoot induction medium supplemented with 12 combination treatments of 6-benzyladenine (BA) 0.5, 1.0 and 2.0 mg/L, and α-naphthaleneacetic acid (NAA) 0.2, 0.5, 1.0 and 2.0 mg/L in darkness for 6 weeks and cultured under a 16/8 h photoperiod for 6 weeks. The highest shoot regeneration was obtained from the explants cultured on the medium with 1.0 mg/L BA and 1.0 mg/L NAA. Based on this result, AgNO3 was added to a shoot induction medium containing MS salts, vitamins, 1.0 mg/L BA, 1.0 mg/L NAA, 30 g/L sucrose, and 6 g/L agar to reduce browning of chrysanthemum leaf explants. In the control treatment without AgNO3, leaf explants turned brown at the cut edge; however, browning was not observed in AgNO3 treatments. Shoot organogenesis was higher at low concentrations of AgNO3 and decreased with an increase in AgNO3 concentration. The explants cultured on shoot induction medium (MS salts, vitamins, 1.0 mg/L BA, 1.0 mg/L NAA) with 1 mg/L of AgNO3 produced the highest shoot regeneration with 2.6 shoots per explants and a browning index of 0.7. When the regenerated shoots were detached from the explants and cultured on MS medium, the shoots were elongated and rooted successfully.

Elimination of Grapevine fleck virus from infected grapevines 'Kyoho' through meristem-tip culture of dormant buds (휴면아 경정 배양법을 통한 포도 '거봉' 에서 Grapevine fleck virus의 제거)

  • Kim, Mi Young;Cho, Kang Hee;Chun, Jae An;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.401-408
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    • 2017
  • Herein, we report the meristem-tip culture from dormant buds of grape 'Kyoho' single-infected with Grapevine fleck virus (GFkV), which is phloem-limited and transmitted by graft inoculation. We produced GFkV-free shoots without thermo- or chemotherapy using meristem-tip explants approximately 0.3 mm (73 explants) and 0.8 mm long (five explants) including shoot apical meristem, 2-5 leaf primordia, and 1-4 uncommitted primordia from dormant buds of the infected woody cuttings (stored at $4^{\circ}C$). Explants were cultured on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA). After 16 weeks of culture, shoot (10-mm long) regeneration frequency achieved from 0.3-mm explants was 4.1% and that obtained from 0.8-mm explants was 40.0%. Virus-free efficiency (expressed as the percentage of RT-PCR negative shoots regenerated) from 0.3- and 0.8-mm explants was 100% and 50%, respectively. Following in vitro multiplication, RT-PCR assays revealed identical results to assays of the first regenerated shoots. Our new methodological approach could be applied for eliminating other viruses in grapevines, as well as for producing virus-free plants in many other deciduous tree species, including fruit trees.

Micropropagation of Oak Seedlings from 37 Plus Half-Sib Families (참나무류(類) 수형목(秀型木) 37가계(家系)의 기내증식(器內增殖))

  • Moon, Heung Kyu;Youn, Yang;Son, Sung Ho;Lee, Suk Koo;Yi, Jae Sun
    • Journal of Korean Society of Forest Science
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    • v.82 no.1
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    • pp.26-33
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    • 1993
  • In vitro shoot proliferation and rooting were tested for 2-0 seedlings of half-sib families of 4 plus oaks trees. Nodal segments having axillary buds from 37 families(16 of Quercus acutissima, 10 of Q. variabilis, 7 of Q. serrata, and 4 of Q. mongolica) were cultured on WPM(Woody Plant Medium) supplemented with 0.5 mg/l BA (6-benzyladenine) and 0.01 mg/l NAA(${\alpha}$-naphthalene acetic acid) and subcultured at 2-3 weeks of intervals fur 6 months. In vitro rooting was carried out on GD(Gresshoff and Doy) medium supplemented with 0.5mg/l IBA(indole butyric acid). The capacity for shoot proliferation and rooting was highly varied with families. Generally, white oaks(Q. serrata and Q. mongolica) showed poor response than black oaks(Q. acutissima and Q, variabilis) in shoot proliferation and rooting. Among the total of 37 families, 7 of Q. acutissima, each 2 of Q. variabilis, Q. serrata, and Q. mongolica revealed abilities for continuous shoot proliferation, and the others failed to proliferate. Rooting of the selected oak trees also greatly varied among the families. In Q. acutissima, rooting ratio ranged from 10.0%(CB 25. KG 4) to 89.8%(CB 18). Although 26.7% of KG 16 in Q. variabilis, 3.3% of JN 15 in Q. serrata were rooted, Q. mongolica was not rooted at all in this experimental conditions. No relationship between shoot growth and the rooting ability was observed. Present results suggest the possibility of large-scale micropropagation, but further studies on family differences, shoot-tip necrosis, and callusing of rooting junction are still required to develop reliable micropropagation systems.

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Effect of Flower and Fruit Thinner on Fruit Set and Fruit Quality of 'Gamhong' Apples (적화제 및 적과제가 '감홍' 사과의 착과와 과실 품질에 미치는 영향)

  • Yoo, Jingi;Kang, Bong Kook;Kim, Dae Hyun;Lee, Jinwook;Lee, Dong Hoon;Kweon, Hun-Joong;Choi, In Myung;Jung, Hee Young;Choung, Myoung-Gun;Choi, Dong Geun;Kang, In-Kyu
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.24-31
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    • 2016
  • This study investigated the effects of flower and fruit thinning agents on fruit set and harvested fruit quality attributes in 'Gamhong' apples. Lime sulfur, MaxCel (1.9% BA), and Fruitone (3.5% NAA) were applied either at post-bloom or fruitlet stages to mature 'Gamhong/M.9' trees. In 2011, the numbers of fruits per cluster in terminal flowers were 1.74, 0.82, and 1.15 for the control, lime sulfur, and Maxcel (applied at 10-mm fruit stage) treatments, respectively. The percentages of single fruit per cluster were 36.0, 47.9, and 48.7% for the control, lime sulfur, and Maxcel (10 mm) treatments, respectively, while the percentages of clusters with three fruits per cluster were 22.9, 1.4, and 5.8%. In lateral flowers, fruit numbers per cluster were 1.20, 0.36, and 0.50 for the control, lime sulfur, and Maxcel (10 mm) treatments, respectively. In 2012, all the thinning treatments showed a positive effect on flower and fruit thinning, compared with the control. Moreover, the treatment with thinning agents did not affect fruit quality. Overall, the results suggest that a single application of flower or fruit thinning agents would be sufficient, rather than the mixed application of thinning agents, based on the observed decrease in fruit setting.