• 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.

• 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.

• Journal of Plant Biotechnology
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• v.44 no.3
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• pp.343-348
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• 2017

Cymbidium aloifolium (L). Sw. is an exquisite epiphytic orchid of the Kolli Hills (Eastern Ghats) of Tamil Nadu in Southern India. It is fast disappearing from its natural habitats due to deforestation and low germination rate in natural habitat. In the present study, an attempt was made to germinate the seeds from un-dehisced capsule of Cymbidium aloifolium (L). Sw under in vitro condition. The seed germination and protocorm development were recorded in three different well known media namely Knudson C (KC), Half strength Murashige & Skoog (1/2 MS) and Vacin & Went (VW) media. The highest seed germination of 90% was observed KC basal media after $30^{th}$ days whereas germination percentages were 40% and 30% on 1/2 MS and VW media respectively. The well-developed protocorm were transferred to KC media supplemented with 6-Benzyl Amino Purine (BAP) and Naphthalene acetic acid (NAA) where BAP (1.0 mg/l) and NAA (1.0 mg/l) together were found to be optimum for the highest shoot formation. About 90% of the shoots found to be well rooted after transfer to the KC medium differently supplemented with 1.5 mg/l Indole-3-acetic acid (IAA) and 1.0 mg/l Indole-3-butyric acid (IBA). Though rooting also took place in the two basic media but the duration was longer when compared with the hormone-supplemented media. The rooted plantlets were hardened and kept under greenhouse conditions which can be relocated in natural habitats.

• Journal of Plant Biotechnology
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• v.36 no.3
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• pp.236-243
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• 2009

The halophyte Leymus chinensis (Trin.) is a perennial rhizome grass (tribe Gramineae) that is widely distributed throughout China, Mongolia and Siberia. This study was conducted to investigate an optimal condition for plant regeneration from mature seeds, leaf base segments, and root segments in L. chinensis. Plant growth regulators affecting embryogenic callus induction and plant regeneration were investigated by four-factor-three-level [L9 (34)] orthogonal test in this study. The effects of explants types (mature seeds, leaf base segments and root segments), callus types, medium types were examined in this study. Wild type (WT) and Jisheng No. 1 plants (JS) were used for primary callus induction. A clear explants difference was seen during callus induction; mature seeds were considered as the preferred explants; and the highest frequency of callus induction was obtained in Medium 6 using mature seeds as explants in WT. Plant regeneration ability was evaluated by frequencies of green callus forming, shooting, rooting, and shooting with roots. Effect of α-naphthalene acetic acid (NAA) on shoot regeneration was remarkable with the highest frequency of 70.8% in WT after 2-month culture. The medium with 0.2- 0.5 mg/L NAA was found to have the highest shoot induction. All regenerated shoots were successfully rooted when transferred on half-strength Murashige and Skoog (MS) basal medium. The acclimatized plantlets were grown to mature with flowering and seeds setting in green house conditions.

• Journal of Korean Society of Forest Science
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• v.99 no.5
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• pp.750-754
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• 2010

Among the environmental conditions employed in micropropagation, light quality plays an important role in growth, specially morphogenesis and photosynthesis. The effect of radiation quality (350-740 nm) on the development and growth of zygotic embryos and in vitro plantlets of open-pollinated chestnut (Castanea crenata S. et Z.) were studied. Two types of explants were exposed for 4 weeks to cool white (W, as control), monochromatic red (R, peak emission 650 nm), monochromatic blue (B, peak emission 440 nm), red+blue (R+B, 1:1), or red+far-red (R+Fr, 1:1, far-red peak emission 720 nm) radiation from a light-emitting-diode (LED) system. While the zygotic embryos showed positive photoblastic behavior, their germination was inhibited by blue radiation. Hypocotyl elongation and root development were promoted by red radiation. The emergence of primary leaf and its expansion were faster under blue than under red radiation. In the plantlets, red and red+far-red radiation significantly increased the formation and growth of the root, whereas blue light reduced rooting. Therefore, radiation quality appears to influence some steps in the development of zygotic embryos and plantlets in the chestnut.

• Journal of Plant Biotechnology
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• v.1 no.2
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• pp.97-100
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• 1999

In order to establish in vitro propagation method of sundew, Drosera rotundifolia L., the effects of MS medium concentration, cytokinin type and concentration, pH, and auxin type and concentration on shoot proliferation and root formation were investigated using shoots at 3 month after seed germination. The highest shoot production was obtained with the half strength of MS ($\frac{1}{2}$ MS) medium than with any other strength of MS medium tested. Addition of kinetin or BA in $\frac{1}{2}$ MS medium was strongly suppressed shoot proliferation. The suppression of shoot proliferation was more effective in BA-supplemented $\frac{1}{2}$ MS medium than kinetin-supplemented. The optimum pH of the media for shoot proliferation was pH 5.7-6.7. Shoots were subcultured in $\frac{1}{2}$ MS medium supplemented with 0.5mg/L 2,4-D for rooting every 8 weeks. All subcultured shoots produced extensive root systems after 5 to 6 week culture. Plantlets after root development were planted in plastic pots filled with moss. The survival rate of plantlets was almost 100%. On subculturing every 8 weeks, hundreds of the plants were propagated from a single plant within a year.

• Journal of Plant Biotechnology
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• v.7 no.3
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• pp.149-153
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• 2005

In the present study, in vitro multiple shoot induction was achieved from cotyledonary node and stem nodal explants of cv. NARI-6 of safflower (Carthamus tinctorius L.). Among various growth regulators tested, MS salts and B5 vitamins supplemented with BA (6-Benzy-laminopurine) $17.76\;{\mu}M$ and KN (Kinetin) $6.96\;{\mu}M$ phytohormonal combination was found to be the most effective in initiating numerous shoot buds after 30 days of culture than BA ($4.44-44.39\;{\mu}M$) or KN ($2.32-46.40\;{\mu}M$) alone in the medium. In addition, 0.8% (w/v) agar (Hi-media) and 3.0% sucrose (w/v) was the optimum level for the formation of adventitious shoots. Further results showed the maximum shoot elongation occurred on MS medium with BA ($8.88\;{\mu}M$) and $GA_3$($11.56\;{\mu}M$) combinations. Efficient rooting occurred on quarter strength MS medium with NAA $10.74\;{\mu}M$. The regenerated plantlets were acclimatized and successfully transferred to the field.

• Journal of Plant Biotechnology
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• v.5 no.4
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• pp.239-244
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• 2003

A protocol has been developed for rapid large scale clonal propagation of an aromatic endangered medicinal plant, Hemidesmus indicus R. Br. with the elimination of the problems such as premature leaf fall and callus formation during caulogenesis and rhizogenesis. Multiple shoots were induced from shoot tip and nodal explants on Murashige and Skoog (MS) medium supplemented with 1 mg/L Benzylaminopurine (BAP) and 0.5 mg/L Napthaleneaceticacid (NAA). Addition of 15 mg/L adenine sulphate to the above medium checked leaf abscission completely, reduced the time required for caulogenesis and restored morphogenetic potential after several subcultures. The in vitro grown propagules were rooted in 1/2 MS medium supplemented with 2 mg/L Indolebutyric acid (IBA) +1 mg/L NAA and sucrose 0.7% (w/v). Addition of charcoal at 100 mg/L to the rooting medium quickened root initiation with a complete check on callus formation. The effect of sucrose concentration on both caulogenesis and rhizogenesis was also studied. The resultant plantlets were acclimatized and grown in fields where ninety eight percent of the rooted shoots survived and grew normally. The estimation of the secondary metabolite content in the shoots of the regenerated plant and the mother plant indicated that the concentration of the three secondary metabolites lupeol, vanillin and rutin was similar.

• Journal of Plant Biotechnology
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• v.4 no.1
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• pp.33-37
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• 2002

Micropropagation and adventitious root production via the culture of Polygonatum odoratum were performed. Stem segments of seedlings of Polygonatum odoratum were the most efficient explants for adventitious shoot formation compared to leaf and root segments. Exogenous cytokinin treatment was required for adventitious shoot formation. Among the cytokinin (BA, Kinetin and Zeatin) tested, BA was most effective for shoot formation from stem segments. Auxin (NAA or IBA) in combination with cytokinin significantly enhanced adventitious shoot formation. Twenty five percent of explants produced adventitious shoots on medium with 2.0 mg/L BAP alone, while 83% of explants produced adventitious shoots on medium with the combination of 2.0 mg/L BAP and 0.1 mg/L IBA. Rooting of adventitious shoots was achieved after transferring to 112 MS medium supplemented with 0.1 mg/L IBA and 0.5 mg/L zeatin. When stem segments were cultured on MS medium with various kinds of auxin (IBA, NAA and 2,4-D), adventitious roots were formed from callus. frequency of adventitious root formation was highest in 2,4-D than NAA and IBA. When roots were in clusters together with parental stem segments, growth of roots actively occurred in hormone-free MS liquid medium. The above results represent that possible application for the mass production of roots and plantlets through in vitro culture system of Polygonatum odoratum.

• Korean Journal of Plant Resources
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• v.13 no.1
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• pp.48-53
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• 2000

Lacquer tree can be proliferated by root or stem cutting, and seed. In case of proliferation by seed, however, the germination rate is very low. Thus, the present study was carried out to obtain aspceptic lacquer plant in vitro from seed because natural tissue culture was highly defiled by unknown fungi and bacteria. First seed grading on distilled water was 50.7% and second seed grading was 20.8% after 98% sulfuric acid treatment for 2 hours. Removal of inner seed coat was higher with 32.4% than non-removal of outer seed coat and removal outer seed coat in rooting rate. In germination rate according to pre-treatment, growth regulators were not effective at all, but sulfuric acid was effective a little with 3%. Removal outer seed coat was increased about 4%, that germinated about 10% in MS medium supplemented with 1.0mg/L BA and 0.05mg/L NAA, 1.0mg/L BA. Lacquer tree seeds germinated after 10 days in MS medium, and aspceptic seedling of lacquer tree were obtained after 3 weeks in vitro. Germination rate, however, was lower about 10%.