• Title/Summary/Keyword: shoot-tip culture

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Characterization of In Vitro Totipotency by Armoracia rusticana (서양고추냉이의 기내 전형성능에 관한 특성)

  • BAE, Chang-Hyu;MIN, Kyung-Soo;AHN, Chang-Soon;LIM, Yong Pyo;KAMEYA, Tosiaki;Lee, Hyo-Yeon
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.119-124
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    • 1997
  • Plantlets were regenerated from various explants (shoot tip, leaf blade, petiole and root segments) via organogenesis and/or somatic embryogenesis from Armoracia rusticana(Lam) Gaerth., Mey, et Scherb.. Shoot regeneration rate from callus was highest on the MS mediums supplemented with 0.5 ㎎/L IAA, 5.0㎎/L BA and 10.0㎎/L spermine. A Low frequency of regeneration occurred on hormone-free MS medium. Multiple shooks were regenerated at a pH of 4.0 to 8.0 on MS medium supplemented with 1.0 ㎎/L BA and 0.1 ㎎/L NAA. Polyamines promoted shoot- and root-formation by 2 to 4 times normal, Specific proteins associated with organogenesis were identified. Somatic embryogenesis occurred directly from the leaf blade, petiole and root segments cultured on MS medium with 2.0 ㎎/L BA and 2.0 ㎎/L BA and 2.0 ㎎/L NAA. Three types of regeneration in A, rusticana were clearly established, which could be applied to the study of morphogenesis and genetics at cell, tissue and organ levels.

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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.

In vitro Shoot Proliferation by Pulse Treatment from Shoot Cultures of Q. acutissima and Ex vitro Root Induction Using Peat Plug Systems in Quercus spp. (상수리나무 기내배양(器內培養)에서의 Pulse처리(處理)에 의한 줄기증식(增殖) 및 Peat Plug를 이용(利用)한 참나무류(類) 기내줄기의 기외삽목(器外揷木))

  • Moon, Heung Kyu;Youn, Yang;Son, Sung Ho;Lee, Suk Koo;Yi, Jae Seon
    • Journal of Korean Society of Forest Science
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    • v.82 no.3
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    • pp.221-226
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    • 1993
  • Methods for shoot proliferation via pulse treatment onto the microshoots of Quercus acutissima, and ex vitro root induction using peat plug systems of the microshoots of 4 oak trees were described. Pulsing solution was prepared by the addition of BA and/or BA plus zeatin onto the aqueous WPM and sterilized distilled water. Using the solution, pulsing time was adjusted at different levels(0. 1, 2, 5. 9, and 24 hours). Although the effect of pulsing solution prepared by the addition of cytokinins onto the sterilized distilled water was slightly lower in shoot proliferation rate, a little higher in shoot elongation was observed compared with that of aqueous WPM. One hour of pulse treatment revealed best in shoot proliferation and its elongation, whereas the increment of pulsing time slightly suppressed the response. In addition, prolonged pulse time resulted high frequency of hyperhydric shoot appearance. Single treatment of BA was better in shoot proliferation than that of BA combination with zeatin, whereas the latter treatment usually showed rapid and healthy shoot growth. For ex vitro root induction using peat plug systems, black oaks(Q. acutissima and Q. variabilis) revealed excellent rootability compared with white oaks(Q. serrata and Q. mongolica). Shoot-tip necrosis of white oaks eras one of the big problems for survival. In this study, we discribed the effect of pulse treatment, successful ex vitro rooting system by the incorporation of peat plug, and the possibilities for the overcoming the obstacles on micropropagation of oaks.

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In vitro micropropagation of two local taro cultivars for large-scale cultivation

  • Alam, Noor Camellia Noor;Kadir, Abdul Muhaimin Abdul
    • Journal of Plant Biotechnology
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    • v.49 no.2
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    • pp.124-130
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    • 2022
  • The application of traditional taro propagation methods for large-scale cultivation would be insufficient to meet the high demand for quality planting materials. Therefore, this study aimed to develop an in vitro micro-propagation technique for two local taro cultivars (cv.), Wangi and Putih. Taro cormels were collected from the Malaysian Agricultural Research and Development Institute (MARDI) germplasm (Serdang, Malaysia). Explants were taken from the shoot tip of cormels and initially cultured on Murashige and Skoog (MS) basal media for four weeks. The explants were then transferred to different multiplication media supplemented with different types and concentrations of cytokinins such as 6-benzylaminopurine (BAP ) and Thidiazuron (TDZ). Shoot production was quantified after six weeks of culture. The highest mean number of new shoots was produced by the Wangi cultivar on MS medium supplemented with 2.0 mg/l BAP (2.10 shoots), MS medium supplemented with 0.5 mg/l TDZ (2.18 shoots), and Gamborg B5 medium supplemented with 6.0 mg/l BAP (2.43 shoots). The maximum average number of the Putih cultivar shoots was obtained on MS supplemented with 2.0 mg/l BAP (3.57 shoots). MS basal media was used for root initiation, as it produced an average of 25 roots with an 11-cm length. Various types of substrate mixtures were used during acclimatization. The best acclimatization substrate for the Wangi cultivar was 100% peat soil, whereas the Putih cultivar grew optimally in a combination of peat and perlites at a 1:1 ratio. Taro plantlets require approximately 4 to 6 weeks to acclimatize before they can be transferred to the field.

Rosenvingea orientalis (Scytosiphonaceae, Phaeophyceae) from Chiapas, Mexico: life history in culture and molecular phylogeny

  • West, John A.;Zuccarello, Giuseppe C.;Pedroche, Francisco F.;De Goer, Susan Loiseaux
    • ALGAE
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    • v.25 no.4
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    • pp.187-195
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    • 2010
  • The genus Rosenvingea is well known in the tropics. Four species have been reported from Pacific Mexico: R. floridana, R. antillarum, R. intricata and R. sanctae-crucis. We collected a plant (Boca del Cielo, Chiapas) that we identified as Rosenvingea orientalis, a species not previously reported from Pacific Mexico. We were able to characterize the life cycle of this species for the first time in laboratory culture. It reproduced exclusively by plurilocular sporangia (plurangia). The mature plants were up to 6 cm long with cylindrical to compressed fronds (to 2 mm wide) with dichotomous branches in the upper half of the thallus. The medulla was hollow with 2-3 layers of large inflated colourless cells at the periphery. The cortex was comprised of 1 layer of small cells, each with a single chloroplast and pyrenoid. Linear plurangial sori with phaeophycean hairs formed along the mature fronds. Zoospore germlings developed into prostrate filamentous systems, each with a single phaeophycean hair that gave rise to a single erect shoot with multiple hairs arising near the tip. Molecular phylogeny using the psaA gene placed this isolate within the Scytosiphonaceae. It does not confirm the exact identification of R. orientalis, although its placement close to other Rosenvingea sequences was confirmed and morphological evidence supports its placement in R. orientalis. Our culture investigations indicated that it has an asexual life cycle. Further collections are needed to resolve the full generic and specific relationships of Rosenvingea and related taxa, and their reproductive patterns.

Micropropagation of Sweetpotato (Ipomoea batatas) in a novel $CO_2$-Enriched Vessel

  • Silva Jaime A. Teixeira da;Giang Dam Thi Thanh;Tanaka Michio
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.67-74
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    • 2005
  • To overcome various disadvantages of conventional cul-ture vessels for micropropagation, a novel disposable vessel, the 'Vitron', made of a multi-layered $OTP^{(R)}$ film and supported by a polypropylene frame, was developed. The film possesses superior properties such as: high light transmittance, low water vapor transmittance and thermal stability and in particular, high gas-permeability. Single nodal explants, which were excised from the multiple shoots derived from shoot-tip culture, were cultured in Vitron and polycarbonate vessels on $3\%$ sugar-containing agar on MS medium and placed at 3000 ppm $CO_2$-enrichment at a low photosynthetic photon flux density (PPFD) ($45{\mu}mol\;m^{-2}\;s^{-1}$). The in vitro and ex vitro growth, and the net photosynthetic rate of in vitro and ex vitro plantlets were significantly enhanced in the Vitron compared to those cultured in a polycarbonate vessel. Explants that were cultured on the same MS medium under low PPFD at various $CO_2$ concentrations were also cultured at 3000 ppm $CO_2$- enrichment at various PPFD: 30, 45, 60, 75 and $90{\mu}mol\;m^{-2}\;s^{-1}$. The best in vitro and ex vitro growth obtained for 3000 ppm $CO_2$-enrichment at $75{\mu}mol\;m^{-2}\;s^{-1}$ PPFD. The novel Vitron vessel, when placed under the two conditions, may replace conventional culture vessels for the successful micropropagation of sweetpotato.

Virus Free Stock Production by In vitro Stem Cutting of Shoot Tip Cultures of Grapes (포도 경정배양에서 얻은 유묘의 기내삽목에 의한 무병묘 생산)

  • 서정해;정재동;권오창
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.81-85
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    • 2001
  • The experiment was conducted to know the effect of plant growth regulators on axillary bud elongation from in vitro stem cutting and the possibility of virus-free stock production. Axillary buds were well elongated in 3/4 strength MS medium supplemented with 0.1 or 0.5 mg/1 BA and 0.05 mg/1 NAA. Transferred plantlets could be established well in vermiculite and peat moss mixture (3:1, v/v) compare to other mixtures. In virus indexing, all the varieties of mother plants were infected by GLRV Ⅲ. Infected percentages of the three varieties were ranged from 30% to 75%. But negative response was revealed against the other species of virus, GLRV Ⅰ, GFLV and ArMV. Plantlet of 'Schuyler' and 'Muscat of Alexandria', which were cultured in vitro, showed positive response against GLRV Ⅲ and infected percentage of the former was 37.5% but the latter, 12.5%. On the other hand, that of 'Campbell Early' negativiely responded against all the species of virus indexed.

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Optimization of apical tip culture condition for In Vitro propagation of 'Gisela 5' dwarf cherry rootstock (양앵두 왜성대목 'Gisela 5'의 기내번식을 위한 정단배양조건의 최적화)

  • Xu, Junping;Kang, In-Kyu;Kim, Chang Kil;Han, Jeung-Sul;Choi, Cheol
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.49-54
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    • 2015
  • Based on the results in this study, here we propose a systematic micropropagation process for 'Gisela 5' that is one of the important dwarfing cherry rootstocks. When the apical tips detached from newly developed shoot in spring season were cultured on the half strength MS media with 0.5 mg/L IBA and 0.5 ~ 1.0 mg/L BA, the cultures scored the highest acquisition rate at 90% for normal shoot with vigorous growth and without hyperhydricity. As next step, the young shoots maintained in vitro well multiplied on the full strength MS medium supplemented with 0.5 mg/L IBA and 0.5 mg/L BA, in which multiplication rate was approximately nine-fold. Given the half strength MS medium containing 2.0 mg/L IBA, each transplanted shoot further developed robust roots. Finally, the plantlets were easily acclimatized in the compost consisted of vermiculite, perlite, and peatmoss in the proportion of 1:1:1. We expect that the results are useful for cherry cultivation and its rootstock production.

Cloning and Characterization of Homeodomain-Zip Gene, Phc5 in Embryogenic Callus derived from Pimpinella brachycarpa Suspension Cultured Cells (참나물 현탁배양세포 유래 배발생캘러스에서 HD-Zip 유전자, Phc5의 클로닝과 특성)

  • 손수인;김준철
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.121-126
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    • 1999
  • Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

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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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