• Journal of Plant Biotechnology
• /
• v.8 no.1
• /
• pp.15-19
• /
• 2006

An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

• Plant Biotechnology Reports
• /
• v.2 no.1
• /
• pp.7-11
• /
• 2008

A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings as well as mature field-grown plants cultured on Murashige and Skoog's (MS) medium supplemented with thidiazuron (TDZ) ($2.27{\mu}M$), 6-benzylaminopurine (BA) ($2.22{\mu}M$) and indole-3-butyric acid (IBA) ($0.49{\mu}M$). The presence of TDZ in the induction medium has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented with BA ($4.44{\mu}M$), kinetin (Kn) ($2.33{\mu}M$), indole-3-acetic acid (IAA) ($1.43{\mu}M$), and gibberellic acid ($GA_3$) ($0.72{\mu}M$). Well-developed shoots were rooted on MS medium supplemented with IBA ($0.5{\mu}M$) after 30 days. Regenerated plants after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation.

• Journal of Plant Biotechnology
• /
• v.47 no.1
• /
• pp.40-45
• /
• 2020

An efficient protocol for multiple shoot induction and plant regeneration from axillary bud culture of Magnolia 'Vulcan' was developed in the present study. Primary shoots were obtained from axillary bud explants cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BA). To induce multiple shoots effectively, primary shoot tips were cultured on MS medium supplemented with different concentrations of BA and zeatin at 0, 0.2, 0.5, and 1.0 mg/L. Of these treatments, the MS medium with 0.5 mg/L BA resulted in the highest number of shoots per explant with an average value of 5.9, and it produced the greatest shoot height at 4.8 cm after 12 weeks of culturing. In the rooting of in vitro produced shoots, the greatest percentage of explants forming roots (91.3%), number of roots per explant (9.7), and root length (2.8 cm) were obtained in half-strength MS medium supplemented with 6.0 mg/L indole-3-butyric acid (IBA). Regenerated plantlets were successfully acclimatized and hardened off inside the culture room with 87.5% survival rate. Plants were transferred to a greenhouse with a 97.2% survival rate. The highly efficient shoot multiplication and plant regeneration system reported herein can be used for large-scale clonal propagation of valuable Magnolia species or cultivars.

• Korean Journal of Plant Resources
• /
• v.19 no.2
• /
• pp.308-314
• /
• 2006

This experiment was conducted out to investigate the effect of plant growth regulators on callus and shoot formation. The callus formation was effective on 1/2 MS medium containing 2,4-D, while shoot formation was suppressed. Shoot formation and differentiation were the highest in combination 0.1 mg/L of IAA and 0.1 mg/L of BA. The polarity of explants was investigated from cotyledon, which excised 20% of each basal and terminal parts. Formation of shoot was induced from excised ends of the basal part. In excised ends of the basal part, callus was induced vigorously and shoots were produced lately. Root induction was easily achieved in 1/3 MS medium from the adventitious shoot and more than 90% of regenerated plantlets acclimatized successfully and flowered normally.

• Journal of Life Science
• /
• v.9 no.1
• /
• pp.50-53
• /
• 1999

Effects of growth regulators and medium components were tested for shoot multiplication and callus growth from shoot explants of black locust. During shoot multiplication, callus growth at the cut end of shoot explants proceeded shoot development. The basal callus growth seemed to be a function of both mineral components and cytokinin supplemented in the medium. Maximum callus growth was induced by 0.5${\mu}$M BAP and the callus growth decreased as the level of BAP increased. Positive correlations were found between basal callus growth, and shoot multiplication and growth. Shoot multiplication was greatest on BSM medium (black locust shoot culture medium) supplemented with 1 $\mu$M BAP. With medium containing high nitrogen content, both shoot multiplication and growth were significantly enhanced. A new BRM medium was the most effective for rooting of black locust among three rooting media tested.

• Korean Journal of Medicinal Crop Science
• /
• v.5 no.1
• /
• pp.49-55
• /
• 1997

This study was conducted to establish mass propagation system from the tissue culture using immature embryos in Eleutherococcus senticosus. Immature embryos from seeds were removed under the microscope and placed on MS, $MSB_5\;and\;B_5$ medium containing several plant growth regulators. While the calli were well formed on media containing 2 mg/l of 2, 4-D, 2 mg/l of 2, 4-D and 0.7 mg/l of TDZ, shoot regeneration was better on MS medium with combinations of high concentrations of TDZ and low concentrations of 2, 4-D. Treatment of 2, 4-D alone was better than treatment of TDZ alone in callus induction, but plant regeneration was reversed. The results of callus formation and shoot regeneration on $MSB_5\;and\;B_5$ media were similar to those of MS media. The rate of callus formation was nearly 100% when 2, 4-D was added to $B_5$, medium on concentration of 2 mg/l or 0.7 mg/l. TDZ showed very significant effect on the formation of multiple shoots.

• Korean Journal of Plant Tissue Culture
• /
• v.22 no.1
• /
• pp.7-11
• /
• 1995

Protoplasts were isolated from hypocotyl tissues of 5-day-old Brassica oleracea var capitata Green Challenger seedlings. Several media were used for protoplast culture and shoot regeneration. The shoot-regeneration rapacity of protoplast derived callus depended on the initial culture medium. Protoplasts were cultured in liquid medium (B5 medium supplemented with CaCl2, 2H2O 600mg/L, g1ucose 20g/L, D-mannito1 70g/L, NAA lmg/L, BA lmg/L, 2.4-D 0.25 mg/L)at 27$^{\circ}C$ under the dark After 5 to 10 days, cultlues were diluted with medium with a reduced osmotic stabilizer and then transferred to illuminated conditions. The culture medium was changed with the fresh medium at 7- to 10-day-intervals until the formation of microcallus. Hypocotyl protoplast-derived callus proliferated when transferred to MS medium supplemented with NAA lmg/L, BA 1mg/L and GA$_3$ 0.02mg/L. Upon transfer to MS basal medium without growth regulators, roots were produced. In an attempt to increase the regeneration frequency, 10g/L polyvinylpyrrolidone was added to the regeneration medium, but the shoot regeneration was mot improved. The regenerated whole plants were acclimated in a sterized soilless mixture(vermiculite 2;perlite 2;peat moss1) in a culture room.

• Journal of Plant Biology
• /
• v.39 no.2
• /
• pp.93-98
• /
• 1996

In order to enhance the system of potato transformation and further regeneration, potato was transformed using the Agrobacterium tumefaciens harboring $\beta$-glucuronidase (GUS) gene. We found that a series fo modified medium ttained 100% shoot regeneration within 5 weeks after the preincubated explants on stage I medium were infected with Agrobacterium. Callus appeared at the cut edges of stem segments on stage II medium, mainly at the basal parts. Some explants started to form shoots after two to three weeks on stage III medium containing kanamycin (50 mg/L). When transferred to MS medium containing 200 mg/L kanamycin, 81% of the transformed shoots formed roots at the cut edge of the plantlets. In contrast, untrasformed shoots never rooted and became yellowish after few weeks under the same conditions. Southern and northern analysis indicated in vitro shoot regeneration on the callus derived from the potato explants, which were incubated with Agrobacteria. The regeneration cycle was shortened after the transformatin and finally the transformation efficiency was highly enhanced.

• Journal of Plant Biotechnology
• /
• v.36 no.3
• /
• pp.236-243
• /
• 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 Plant Biotechnology
• /
• v.36 no.4
• /
• pp.403-406
• /
• 2009

This study was conducted to develop an effective production of callus induction and plant regeneration system for garlic transformation. The best callus production occurred on in vitro root segment initially cultured on MS medium with 1.0 mg/L 2,4-D and 0.2 mg/L IAA in both ‘Danyang' and ‘Euseong'. The frequency of callus formation were 81.2% ‘Danyang' and 76.1% ‘Euseong'. Eight weeks after callus induction, callus lines were transferred to regeneration medium during 7 weeks. The best shoot regeneration medium was MS supplemented with 5 mg/L Kinetin and 1 mg/L NAA for ‘Danyang' and MS supplemented with 10 mg/L BAP for ‘Euseong'. The frequency of shoot regeneration were 51.5% ‘Danyang' and 56.6% ‘Euseong' The plantlets were acclimatized and transferred to the greenhouse with almost survival. This in vitro regeneration system should be useful for garlic transformation.