• Journal of The Korean Society of Grassland and Forage Science
• /
• v.31 no.3
• /
• pp.223-228
• /
• 2011

The present study was conducted to determine the optimum in vitro culture condition for callus induction and plant regeneration from mature seeds of bermudagrass (Cynodon dactylon cv. Common). It was revealed that mature seeds cultured on MS medium supplemented with 2 mg/L 2,4-D, 0.5 g/L proline, 0.5 g/L casamino acid and 3 g/L Gelrite under light condition produced the highest percentage of callus formation (39.2%). The most suitable medium for plant regeneration from dehydrated calli was MS agar medium supplemented with 0.5 mg/L 2,4-D, 2 mg/L BA, 0.5 g/L proline, 0.5 g/L casamino acid 3 g/L Gelrite which induced the highest percentage of calli forming shoots (57.7%). The frequency of callus induction and plant regeneration were the highest on sucrose, followed by maltose. The shoots were rooted at the highest rate (100%) when transferred onto 1/2 MS medium. Regenerated plants were morphologically uniform with normal growth pattern.

• FLOWER RESEARCH JOURNAL
• /
• v.19 no.4
• /
• pp.231-237
• /
• 2011

In this study, we performed ovule culture after reciprocal crosses of two Alstroemeria accessions and investigated genetic contribution of parents by using RAPD markers. The best method was half-ovule culture on MS medium supplemented with $60g{\cdot}L^{-1}$ sucrose and $2.2g{\cdot}L^{-1}$ gelrite at 14 days after pollination. Embryos began to germinate after 6 weeks of culture. The complete plantlets were formed after 4 months of culture. In eight progenies and two parental cultivars, 59 polymorphic bands were obtained out of 89 total bands by RAPD analysis using 7 primers. Eight $F_1$ progenies from the crosses between two accessions using reciprocal crosses showed 1:1 contribution of maternal and paternal parents. It is confirmed that $F_1$ progenies were obtained from parental accessions by using RAPD markers. We conclude this cross combination showed pre-fertilization barriers with incompatibility between stigma or style, and pollen because progeny number was different in each cross combination. Thereby, it warrants overcoming pre-fertilization barrier together with post-fertilization barrier in order to broaden the heterozygosity within progeny populations in Alstroemeria breeding program.

• Journal of Life Science
• /
• v.12 no.1
• /
• pp.19-21
• /
• 2002

The pathway of embryos formed anther culture in herbaceous peony was influenced by addition of 2,4-D. MS medium with 2,4-Dichlorophenoxy acetic acid (2,4-D) alone did not arise direct embryogenesis, but was proliferate callus. Embryos through calli were produced on medium containing 0.2 mg/1 zeatin or without growth regulators. Direct embryogenesis was obtained from MS basal medium. However, after the anthers were cultured on medium with 0.1 mg/1 2,4-D, 3 g/1 AC, 30 g/1 sucrose, 2 g/1 gelrite for 40 days. Its efficiency (32.3 %) was markedly improved when anthers cultured on medium without 2,4-D. Embryo morphology was also affected by the 2,4-D used in medium. The induction of normal embryos with two cotyledons was higher in the embryos formed through direct embryogenesis than those formed callus. The embryos formed from calli were mainly showed abnormal embryo with one, three, four cotyledons or hors and bowling pin type.

• Plant Biotechnology Reports
• /
• v.2 no.3
• /
• pp.179-189
• /
• 2008

We established an efficient plant regeneration system for Catharanthus roseus L. (G.) Don through somatic embryogenesis. Embryogenic callus was induced from hypocotyl of seed germinated in vitro. Somatic embryogenesis in Catharanthus has been categorized into three distinct stages: (1) initiation and proliferation of embryo; (2) maturation, and; (3) germination or plantlet conversion. Beside plant growth regulators, various stages of embryogenesis were screened for their response to a wide variety of factors (pH, gelrite, light, sugar alcohols, polyethyleneglycol and amino acids), which affect embryogenesis. All of the tested factors had a small to marked influence on embryogeny and eventual conversion to plantlets. The plantlets were acclimatized successfully in a greenhouse. To our knowledge, this is the first report describing a detailed study of various cultural factors which regulate embryogenesis in C. roseus. The results discussed in this paper may be used in mass propagation to produce medicinal raw material, and the embryo precursor cells could be used in genetic modification programmes that aim to improve the alkaloid yield as well.

• Plant Biotechnology Reports
• /
• v.5 no.1
• /
• pp.35-43
• /
• 2011

This communication describes for the first time an efficient and reproducible protocol for large-scale multiplication of Bambusa nutans. Nodal segments collected from field-grown clumps and cultured on Murashige and Skoog (MS) medium supplemented with $4.4{\mu}M$ benzylaminopurine (BA) and $2.32{\mu}M$ kinetin (Kin) gelled with 0.2% gelrite yielded 80% aseptic cultures with 100% bud-break. The in vitro-formed shoots obtained after bud-break were successfully multiplied in MS liquid medium supplemented with $13.2{\mu}M$ BA, $2.32{\mu}M$ Kin, and $0.98{\mu}M$ indole-3-butyric acid (IBA). Sub-culturing of shoots every 3 weeks on fresh multiplication medium yielded a consistent proliferation rate of 3.5-fold. Shoot clusters containing three to five shoots were successfully rooted with 100% success on half-strength MS liquid medium supplemented with $9.8{\mu}M$ IBA, $2.85{\mu}M$ indole-3-acetic acid (IAA), $2.68{\mu}M$ naphthaleneacetic acid (NAA), and 3% sucrose. Plantlets grown in vitro were acclimatized and subsequently transferred to the field. Inter-simple sequence repeat analysis has confirmed the genetic uniformity of the tissue-cultured plants up to 27 passages.

• Korean Journal of Plant Tissue Culture
• /
• v.28 no.4
• /
• pp.215-219
• /
• 2001

This study was conducted to investigate the effect of culture media and environment on PLB proliferation by using PLBs produced from leaf segments excised from shoot of Phalanopsis flower stalk. The fresh weight of PLBs propagated was higher in MS medium than in NDM (New Dogashima medium) or VW, but the condition of PLB was better in NDM medium. Natural additives of Coconut water, potato and apple were absolutely required for the PLB propagation. PLB propagation was better in solid medium than in liquid medium including cotton as support. Optimal sucrose concentration for proliferation was 10 g/L. PLB proliferation was very effective condition 14.3 $\mu$mol.s$^{-1}$ m$^{-2}$ in PPFD and $25^{\circ}C$.

• Journal of Plant Biotechnology
• /
• v.6 no.2
• /
• pp.97-102
• /
• 2004

This study was carried out to establish The regeneration of healthy seedlings from the nodal segment culture of Chinese yam (Dioscorea opposita cv. Danma), cultivated in Korea. Different explants such as leaves, petioles, roots and nodal pieces, excised from the in vitro grown seedlings of Chinese yam, were cultured on MS medium supplemented with various combinations of growth regulators. All the growth regulators used induced plantlet regeneration from the nodal segments at a high frequency, while there was no induction of shoot or callus from leaf, petiole or root tissues. The medium supplemented with 0.01mg/L NAA, 0.5mg/L BA, 0.5-1.0mg/L kinetin and without plant growth regulator was effective for shoot development of buds from the nodal segment culture. The concentration of BA and NAA was an important factor in the bud induction of buds from the nodal segments of Chinese yam. Nodal segments cultured on the medium containing 1.0mg/L NAA and 0.5-1.0mg/L BA gave the best response to bud formation. The addition of GA$_3$ to the culture medium suppressed shoot induction and growth, while it increased microtuber formation. The shoot growth and microtuber formation were also affected by medium strength and solidity. The MS basal medium containing 1 g/L gelrite was suitable for microtuber formation from the nodal segment of Chinese yam.

• Journal of Plant Biotechnology
• /
• v.3 no.2
• /
• pp.95-100
• /
• 2001

To select the section with shoot formation ability, the calli and shoot formation from three sections (first leaf including cotyledonary node, hypocotyl and cotyledon explants) of 5-days-seedlings of soybean were induced on MS medium supplemented with 1.0 mg/L BAP, 3% sucrose, and 0.3% gelrite for one month. The first leaf section exhibited the highest shoot formation rate (51%), followed the hypocotyl section (10%) and the cotyledon section (0%). The shoot formation rates and shoot number of the four excised sections (whole first leaf, a half of the first leaf, a third of the first leaf and only node) of the first leaf were also investigated on the same medium. A half of the first leaf explant and the third of the first leaf explant had higher shoot formation rates (76-80%) and numbers (3-4 / explants) than those in other two explants. Effects of six cytokinins (kinetin, zeatin, BAP, 2iP, PBA, and TDZ) on shoot formation were determined, using the half of the first leaf explants. Zeatin (1.0 mg/L) exhibited the highest in shoot formation rate (94%) and numbers (8 / explant). In addition, the combined effects of three cytokinins (zeatin, BAP, and TDZ; 0.5, 1.0, 2.0 mg/L, respectively) and an auxin (IAA; 0.0, 0.5, 1.0, 2.0 mg/L) were determined. The combination (1:1, v/v) of zeatin (1.0 mg/L) and IAA (1.0 mg/L) exhibited the highest in shoot formation rate (96%) and numbers (16 / explant), twice more than zeatin (1.0 mg/L) alone. The shoot cuttings were transferred and cultivated on the rooting media supplemented with only auxin, IBA at various concentrations. The highest root formation (8 / shoot) was achieved on the medium supplemented with 1.5 mg/L. After 4 weeks of cultivation, the plantlets with an extensive root system were transplanted in pots with a soil mixture of vermiculite and fine sand. Transferred to field, about 75% of the plantlets survived.

• Journal of Korean Society of Forest Science
• /
• v.98 no.2
• /
• pp.142-147
• /
• 2009

Hyoscyamus species is sources of the hypnotic and sedative drugs hyoscyamine and scopolamine. Single cells of Hyoscyamus niger were dissociated from suspension cultures and adventitious roots obtained from single-cell clones which were cultured on B5 medium containing 3% (w/v) sucrose, 0.1 mg/L IBA and 0.4% (w/v) gelrite. H. niger adventitious root lines showed wide variation in tropane alkaloids production and growth. An effective selection of 200 root lines was made possible by the application of the 'Dragendorff's reagent' for qualitative detection of the alkaloids from root. A high correlation coefficient (r=0.9390) was observed between the values obtained with the two methods based on HPLC and Dragendorff's reagent analysis. Among the selected roots, the highest scopolamine content was 16.64 mg/g DW (Hn-59), which was 8.82-fold more productive than the lowest alkaloid producing line (Hn-25). Here, we established an efficient selection method on tropane alkaloids production and suggest that the Dragendorff's reagent is of great practical value in selection of invisible compounds.

• Plant Biotechnology Reports
• /
• v.4 no.4
• /
• pp.321-328
• /
• 2010

An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro (15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g $1^{-1}$ sucrose, 2.2 g $1^{-1}$ Gelrite, and 7.7 lM naphthalene acetic acid (NAA) with 2.2 ${\mu}M$ thidiazuron (TDZ). Regeneration of adventitious shoots from callus (30-40 shoots per explant) was achieved on MS medium containing 5.5 ${\mu}M$ TDZ, 2.2 ${\mu}M$ NAA, and 3.3 ${\mu}M$ silver nitrate ($AgNO_3$). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 ${\mu}M$ gibberellic acid ($GA_3$). The elongated shoots were rooted in MS medium supplemented with 4.0 ${\mu}M$ indole 3-butyric acid (IBA). Rooted plants were acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an average of 40 plants per leaf explant with a culture period of 98 days.