• Journal of Plant Biotechnology
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• v.6 no.3
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• pp.181-188
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• 2004

Zingiber officinale buds from the rhizomes were used to produce in vitro shoots. These explants produced the largest number of multiple shoots, 9.8 shoots per explant, when were cultured on MS (Murashige and Skoog 1962) medium supplemented with 2.0 mg/L benzyladenine (BA) and 2.0 mg/L indole butyric acid (IBA). This medium was also found to be suitable for in vitro propagation of other Zingiberaceae species: Alpinia conchigera, Alpinia galanga, Curcuma domestica, C. zedoaria and Kaempferia galanga. Both C. domestica and C. zedoaria produced more multiple shoots when were cultured in the liquid proliferation medium, MS medium containing 2.0 mg/L BA and 2.0 mg/L IBA. To maintain the in vitro plantlets of Zingiberaceae species, they were required to subculture every four weeks. After executing proper acclimatization protocol, in vitro plantlets of Alpinia galanga, A. conchigera, Curcuma domestica, C. zedoaria, Kaempferia galanga and Zingiber officinale could be successfully planted in the field with high percentage of survival.

• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.111-118
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• 2009

Multiple shoot induction and plant regeneration using apical bud and nodal explants of 100 year old tree of Anthocephalus cadamba, an important sacred and medicinal tree in India was achieved for the first time. Aseptic explants cultured in Murashige and Skoog (MS) medium augmented with different concentrations of BAP (0.1, 0.5, 1.0, 2.5, 5.0 and 10 mg/l), when maintained for 60 days, healthy shoots were induced in presence of BAP (1 mg/l). Lower concentrations of BAP (0.1 - 0.5 mg/l) induced only one shoot per explant. Increase in number of shoots per explant was observed in presence of higher concentrations of BAP (2.5, 5.0 and 10 mg/l). However, elongation of shoots was completely inhibited. Bud break and shoot regeneration was largely associated with seasonal factors. Apical buds cultured during June to August exhibited early bud break within two weeks of initial culture. In rest of the months, bud break and shoot regeneration was very slow irrespective of the various concentrations of BAP used in the medium. Explants sourced from three different maturity levels of shoots indicated that actively growing shoots from the mother plant with 1 - 2 nodal segments was more suitable for culture initiation than the explants collected from mature shoots at dormant stage. Regenerated shoots with 2 - 3 pairs of leaves when transferred to half strength MS medium fortified with IBA (1 mg/l), 60% of the shoots induced healthy roots, indicating the possibility of large scale micropropagation.

• Journal of Crop Science and Biotechnology
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• v.10 no.3
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• pp.141-146
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• 2007

The roots of Aristolochia elegans Mast.(Aristolochiaceae) are widely used in Mexican traditional medicine as a remedy for scorpion venom. Current experimental evidence supports its purported antidote properties. However, collection from the wilderness has lead to local extinction of natural populations. In order to contribute to species preservation, cultivation, and standardization of morphological and pharmacological properties, a micropropagation method was developed. This includes in-vitro germination of seeds to produce aseptic plantlets, induction of multiple budding, and acclimatization. The treatment with benzylamino purine(10 ${\mu}M$) induced the highest number of buds(3.1 on average) in both types of explants. On the other hand, indolebutyric acid(1.5 ${\mu}M$) caused the highest root index(11.8) per explant. One hundred percent of the micropropagated plantlets developed vigorously after the acclimatization process.

• Horticultural Science & Technology
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• v.32 no.5
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• pp.694-701
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• 2014

Allium hookeri L. (Alliaceae family) is an important ethnomedicinal plant native to the Himalayan region of Asia. The aim of this research was to establish a high-frequency plant regeneration system for in vitro propagation of A. hookeri. Among the tissue types examined, receptacle explants derived from immature flower buds showed the highest regeneration rate of shoots ($93.33{\pm}4.63%$), roots ($76.67{\pm}7.85%$), and calli ($80.00{\pm}7.43%$) when cultured on Gamborg B5 (B5) medium containing $10{\mu}M$ 6-benzylaminopurine (BA) + $1{\mu}M$ naphthalene acetic acid (NAA), $0.5{\mu}M$ BA + $5{\mu}M$ NAA, and $1{\mu}M$ BA + $10{\mu}M$ NAA, respectively. Shoot multiplication was superior when cultured in liquid rather than on solid medium and relatively high concentrations of BA, ranging from 5 to $10{\mu}M$. Efficient bulblet formation following root induction from shoot clumps was achieved with culture in liquid B5 medium containing 7% (w/v) sucrose. Regenerated bulblets were successfully acclimatized to ex vitro conditions with a greater than 95% survival rate. By this method, a maximum of 62 plantlets per receptacle could be propagated within 9 weeks of initial culture. The in vitro propagation system established in this study will promote A. hookeri biotechnology, including large-scale production of healthy and aseptic clones, preserving parental genotypes with desirable traits, and genetic manipulation to enhance medicinal value.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.175-182
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• 2009

Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments from mature plus trees on MS liquid medium supplemented with $0.25mg\;1^{-1}$ KIN and $0.25mg\;1^{-1}$ IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment, while $0.25mg\;1^{-1}$ KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength MS medium with $0.15mg\;1^{-1}$ IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing 1:1 mixture of vermicompost and soil at $27{\pm}2^{\circ}C$ for hardening and acclimatization. 14- to 15-week-old well-established hardened plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation, providing a value addition to Eastern Cottonwood propagation in future.

• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.271-274
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• 1999

This study was undertaken to develop an efficient propagation technique for mature Betula davurica. Using aseptic materials taken from in vitro culture, the effects of media and plant growth regulators on shoot proliferation and rooting were investigated. DKW medium turned out to be the best in shoot proliferation among the media tested. Whereas axillary buds were better culture material than apical buds in proliferation of shoots, apical buds were slightly better than axillary buds on shoot elongation. Neither 1 /2 MS nor WPM medium seemed to be suitable for shoot multiplication or elongation. When the explants were cultured on 1/2 MS medium, shoot elongation was retarded by forming big callus at the base. In the case of WPM, shoots could be formed normally, but they exhibited slow growing. NAA was so effective on in vitro rooting that more than 80% rooting could be achieved on half-strength DKW medium supplemented with 1.0 mg/L NAA after 4 weeks in cultures. Ex vitro rooting using elongated shoot was also applicable to rooting and acclimatization. Rooted plantlets were successfully acclimatized in an artificial soil mixture and grew normally. The results demonstrate that efficient mass propagation of mature B. davurica can be done through tissue culture.

• FLOWER RESEARCH JOURNAL
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• v.18 no.1
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• pp.9-14
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• 2010

This study was conducted to examine the effects of IAA and kinetin on induction of gemmae and gametophytes in Hypnum plumaeforme moss during tissue culture. Explants were obtained from sterilized gametophyte tips cultured on solid basal medium containing Knop's major salts and Nitsch and Nitsch's trace elements. After culture, inoculated gametophyte tip produced protonema firstly, changed to new gametophytes after four weeks. Aseptic gametophytes were chopped and inoculated on the same media containing 0.01, 0.1, 1 and $10{\mu}M$ of IAA and kinetin. As a result, secondary protonemata as well as protonemal gemmae were formed from gametophytes. But protonemal gemmae formation was varied according to the concentration of IAA and kinetin. Lower concentration of IAA and kinetin promoted gemma formation and bud induction. Especially, $0.01{\mu}M$ of kinetin showed the highest frequency of bud and gemma production. All the materials, obtained from $0.01{\mu}M$ kinetin medium, were subcultured to media supplemented with 0.01, 0.1, 1 and $10{\mu}M$ of IAA and kinetin again to induce gametophytes. After subculture, protonema and calli were developed from secondary protonema, and induced gametophytes finally. IAA regulated induction and growth of gametophytes, and kinetin influenced gemma formation and gametophyte induction also. All aspects of development of this moss species were governed by the external growth regulators.