• Title/Summary/Keyword: ex vitro rooting

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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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Effects of Excising In Vitro-Formed Roots on Acclimatization of Micropropagated Cassava Plantlets (카사바의 미세증식에서 기내 발생 부정근의 절단이 순화에 미치는 영향)

  • Yoon, Sil;Cho, Duck-Yee;Soh, Woong Young
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.103-108
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    • 2001
  • The in vitro plantlets of cassava (Manihot esculenta Crantz cv. MColl 22) could be regenerated from nodal explant cultures in a liquid MS basal medium containing 0.01 mg/L zeatin for 2 weeks. The plantlets of 1.5∼2.5 cm in shoot length were transplanted to a glass bottle containing fine sand and acclimated under non-sterile conditions after excising their intact roots by: 1) prune leaving roots base of 1∼1.5 cm; 2) complete removal of roots; and 3) cutting off the rooting zone. The majority of in vitro-formed intact roots continued growth after transferred to soil, and all of the damaged roots stopped further growth. The plantlets with excised roots began to develop new roots within 7∼10 days after being transferred to a glass bottle, and a few of the pruned roots developed lateral roots from the remaining portion. Pruning and removal of in vitro roots resulted in a high survival rate (>87%), and did not significantly affect ex vitro root regeneration and acclimation, but the plantlets in which the rooting zone had been cut-off showed 73% survival rate. Pruning or removal of in vitro roots before transfer of plantlets is recommended for useful method of commercial micropropagation because of easier handling and high survival rate of plantlets.

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In vitro regeneration of Lycaste aromatica (Graham ex Hook) Lindl. (Orchidaceae) from pseudobulb sections

  • Mata-Rosas, Martin;Baltazar-Garcia, Rosario J.;Moon, Pamela;Hietz, Peter;Luna-Monterrojo, Victor E.
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.157-163
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    • 2010
  • A protocol for in vitro propagation from pseudobulb sections of Lycaste armomatica (Graham ex Hook) Lindl., an ornamental and fragrant orchid, was developed. The effect of four cytokinins: kinetin (K), metatopolin (mT), $N^6$-benzyladenine (BA), and thidiazuron (TDZ), in equimolar concentrations, was investigated. Shoot formation from apical and basal pseudobulb sections was obtained in all treatments. A few medial sections cultured in media supplemented with BA formed protocorm-like bodies. Shoot formation was greater from the basal section than the apical, and mainly occurred in explants cultured in media containing TDZ. The highest average numbers of shoots per explant were achieved from basal sections cultured in media supplemented with TDZ at 4.4, 8.87 and 2.2 ${\mu}M$, forming on average 9.9, 8.6 and 7.3 shoots per explant, respectively. Since the medial pseudobulb section was the worst explant for propagation of L. aromatica, we recommend that pseudobulbs be divided into two sections; the basal half should be cultured in MS medium supplemented with TDZ at 4.4 ${\mu}M$ and the apical half with TDZ at 2.2 ${\mu}M$. Subculturing individual shoots in MS medium without plant growth regulators allows further development and rooting. A survival rate of more than 90% under greenhouse conditions was achieved. This research represents a direct contribution to the conservation and sustainable use of this valuable natural resource.

An Efficient Plant Regeneration System for Sorghum bicolor - a Valuable Major Cereal Crop

  • Baskaran P.;Jayabalan N.
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.247-257
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    • 2005
  • An efficient, rapid and large-scale in vitro clonal propagation of agronomically important Indian cereal crop genotypes (NSH27 & K5) of Sorghum bicolor (L.) Moench. by enhanced shoot proliferation in shoot tip segments was designed. MS medium fortified with plant growth regulators and coconut water markedly influenced in vitro propagation of Sorghum bicolor. In vitro plantlet production system has been investigated on Murashige and Skoog (MS) medium with the synergistic combination of 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), 5% coconut water and 3% sucrose which promoted the maximum number of shoots as well as beneficial shoot length. Subculturing of shoot tip segments on a similar medium enabled continuous production of more than 100 healthy shoots with similar frequency. When the healthy shoot clumps were cultured on MS medium fortified with 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), ${\alpha}$-naphthaleneacetic acid ($2.7\;{\mu}M$), ascorbic acid ($30.0\;{\mu}M$) and 5% coconut water, a rapid production of axillary and adventitious buds was developed after 8 wk culture. More than 300 shoots were produced 10 wk after culture. Rooting was highest (100%) on half strength MS medium containing 22.8 mM IAA. Micropropagated plants established in garden soil, farmyard soil and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics. These plants grew normally without showing any traits.

Effects of Growth Regulators and Culture Environment on ex vitro Rooting and Acclimatization of Apple Rootstock in vitro Propagated (기내배양 사과 대목의 기외 삽목 시 발근과 순화에 미치는 배양조건 및 생장조절물질의 효과)

  • Kwon, Soon-Il;Kim, Jeong-Hee;Kang, In-Kyu;Kim, Mok-Jong
    • Journal of Plant Biotechnology
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    • v.31 no.2
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    • pp.133-138
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    • 2004
  • Growth of M.9 (Malus domestica Bark. cv) and M.26 (Malus domestica Bark. cv) of dwarf apple rootstock, cultured on MS agar medium in a vessel with ventilating stopper (VS) and then in vivo rooting and acclimatization under combined-treatment by some materials with IBA, were investigated. Concentration of $CO_2$ and ethylene in the vessel with VS was lower then in the vessel with non-VS. Change of temperature and humidity in the vessel with VS was repeated by light condition. Stomatal pares of tissue in the vessel with VS were immediately closed after plantlets were exposed to room humidity but those in the vessel with non-VS were opened after 20 minutes exposure to room humidity. Leaf area and chloroplast index of tissue in the vessel with VS was higher then in the vessel with non-VS. In vivo rooting ratio and acclimatization ratio of M.9 and M.26 was highest in 300mg/L IBA+3% sucrose dip-treatment among other combined- treatments.

In vitro plantlet regeneration of "dwarf" Indian olive (Elaeocarpus robustus Roxb.): a fruit plant of Bangladesh

  • Rahman, Md. Mahabubur;Amin, Muhammad Nurul;Ishiguri, Futoshi;Yokota, Shinso;Sultana, Rubaiyat Sharmin;Takashima, Yuya;Iizuka, Kazuya;Yoshizawa, Nobuo
    • Plant Biotechnology Reports
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    • v.3 no.3
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    • pp.259-266
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    • 2009
  • A plantlet regeneration protocol was developed on pot-grown mature plants of Elaeocarpus robustus Roxb. cv. Dwarf from nodal and leaf explants. The best yield of adventitious shoots was achieved from the leaf-derived calli in a modified MS ($MMS_1$, half strength of major salts, full strength of minor salts, and vitamins) medium containing $4.0{\mu}M$ BA + $4.0{\mu}M$ Kn + $0.5{\mu}M$ NAA + 15% coconut water (CW). The shoot multiplication rate was amplified about twofold per culture after the addition of 15% CW to the medium. The rate of shoot multiplication reached maximum at the 5th subculture, and it maintained this rate throughout the 3 subsequent subcultures. The best rooting in vitro was investigated by subculturing the microcuttings in an $MMS_2$ (half strength of both major salts and minor salts and full strength of vitamins) medium containing $1.0{\mu}M$ IBA in the dark for one initial week at $30^{\circ}C$, followed by subculturing them in a plant-growth regulator (PGR)-free medium in the light. The plantlets raised in vitro were successfully established under ex vitro conditions.

Effect of explant's position and culture method on shoot proliferation and micro-cuttings for a rare and endangered species, Abeliophyllum distichum Nakai (희귀 및 멸종위기 식물 미선나무(Abeliophyllum distichum Nakai)의 절편위치 및 치상방법에 따른 기내증식 및 미세삽목)

  • Lee, Na Nyum;Kim, Ji-Ah;Kim, Yong-Wook;Choi, Yong Eui;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.228-234
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    • 2015
  • Using either the apical or axillary bud of the endangered species Abeliophyllum distichum Nakai, we tested the effect of bud position and culture method on shoot proliferation and rooting. In shoot proliferation, the axillary bud explant was more effective than the apical bud and the effect was fostered by BA treatment, whereas no differences were observed in shoot elongation by the explant position. Spontaneous rooting was observed in the MS basal medium and resulted in conspicuous differences in the explant position : more than 80% in apical bud explant and 28% in axillary bud explant was achieved, respectively. The positional effects were also observed in BA pre-treatments: generally vertical culture method appeared to be better in shoot proliferation, growth, and rooting than that of the horizontal culture method regardless of the BA pre-treatment duration. The highest shoot multiplication was achieved through the vertical culture method with axillary bud explant, whereas the best shoot elongation and rooting was obtained using the vertical culture method with the apical bud explant. Apical bud explant was superior to axillary bud explant in ex vitro micro-cuttings and revealed a significant difference in shoot growth and root development. The above results suggest that explant position and culture method influence the efficiency of micropropagation for a rare and endangered plant Abeliophyllum distichum.

Plant Regeneration of Iris koreana Nakai through Organogenesis for Ex-situ Conservation

  • Bae, Kee-Hwa;Yun, I-Seul;Jung, Ji-Sun;Kim, Chan-Beom;Kim, Hye-Won;Hong, Yong-Sik;Oak, Min-Kyeong;Kim, Hak-Koo;Lee, Ju-Hui
    • Journal of Forest and Environmental Science
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    • v.37 no.4
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    • pp.304-308
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    • 2021
  • Iris koreana (Iridaceae) is an endangered plant native to Korea. In order to develop an in vitro propagation method, we investigated the effect of 2,4-dichlorophenoxy acetic acid (2,4-D) and a-naphthalene acetic acid (NAA) on callus induction in different I. koreana tissues. In addition, we also investigated the effect of 2,4-D and Benzyl aminopurine (BA) treatments on adventitious shoot induction in viable calli and the effect of indole-3-butyric acid (IBA) on root formation in viable shoots. We found that callus production was highest with 1.0 mg/L NAA (94.4% cultured rhizome explants), and adding low concentrations of 2,4-D to BA containing media significantly increased the frequency of shoot primordial formation. The best rooting results were obtained with 1.0 mg/L IBA, on which 98% of regenerated shoots developed roots and produced an average of 7.4 roots within 45 days. This in vitro propagation protocol will be useful for conservation, as well as for mass propagation.

In vitro propagation and multiple shoot induction of Rhodiola rosea L. by axillary bud culture (홍경천(Rhodiola rosea L.)의 액아배양을 통한 다신초 유도 및 기내 대량증식)

  • Bae, Kee-Hwa;Ko, Myung-Suk;Kim, Nam-Young;Song, Jae-Mo;Song, Gwan-Pil
    • Journal of Plant Biotechnology
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    • v.39 no.2
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    • pp.114-120
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    • 2012
  • An efficient in vitro propagation was established by using axillary bud explants of roseroot (Rhodiola rosea L.), which has been known as a medicinal plant in East Asia. Among various media tested, MS medium supplemented with 1.0 mg/L BA and 1.0 mg/L $GA_3$ was found to be the best for multiple shoot formation (15 axillary shoots per axillary bud). In addition 1/2MS medium containing 50 g/L sucrose was best for shoot elongation (7.8 cm) and increasing total chlorophyll contents (8.64 mg/g) best. Maximum number of roots (17.7 roots per explant) was observed on the medium without plant growth regulators. Propagated plants were successfully acclimatized to ex vitro conditions, with a survival frequency of 97% after 12 weeks. Most rooted shoots grew well and produced viable seeds when grown in vitro culture conditions. Therefore, R. rosea can be effectively propagated in vitro by the system we developed in this study.

In Vitro Micropropagation of Polygonatum odoratum cv. Gungangbeaksea (둥굴레 '건강백세'의 기내 대량번식)

  • Kim, Man Bae;Park, Chun Geun;Yi, Sookyi;Han, Bong Hee
    • Korean Journal of Plant Resources
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    • v.27 no.5
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    • pp.540-545
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    • 2014
  • The Polygonatum odoratum cv. Gungangbeaksea, bred in Gyeongsangnam-Do Agricutural Research & Extension Service, was cultured in vitro for micropropagate rapidly through the culture of rhizome explants ($5{\times}5mm$). The $7{\times}7mm$ explants of adventitious multi-bud clusters (AMC), obtained through the culture of rhizome explants (MS + 3.0 mg/L BA) were cultured on MS media with BA and TDZ. The shoot multiplication was favorable on the MS medium containing 3.0 mg/L TDZ with 2.8 in shoot number. But the formation of AMC was low in all media tested. The explants of AMC were cultured on MS media containing 1.0~5.0 mg/L TDZ and NAA to multiplicate AMC more. The formation of AMC was a little more stimulated on combined MS media of TDZ and NAA, than that with TDZ alone. The multiplication of shoots and AMC was favorable on MS media with 3.0 mg/L TDZ and 5.0 mg/L NAA, and 5.0 mg/L TDZ and 3.0 mg/L NAA. As the concentration of MS salts increased, the formation of AMC was decreased. But the formation of AMC was more stimulated, as the concentration of sucrose increased to 7%. Therefore, the multiplication of shoots and AMC was suitable on media containing 3.0~5.0 mg/L TDZ and NAA, and 7% of sucrose. The explants of AMC were rooted on media with 3.0 mg/L IBA, or 2.0 mg/L NAA with more than 80% in rooting ratio. The plantlets were treated at $5^{\circ}C$ for 8 weeks, and cultured ex vitro for 8 weeks. The survival ratio of plantlets were 100% in vermiculite, and the mixed soil with perlite 1 volumn and vermiculite 1 volumn.