• Plant Biotechnology Reports
• /
• v.2 no.2
• /
• pp.123-131
• /
• 2008

Vitrification methods are convenient for cryopreserving plant specimens, as the specimens are plunged directly into liquid nitrogen (LN) from ambient temperatures. However, tissues and species with poor survival are still not uncommon. The development of vitrification solutions with high survival that cover a range of materials is important. We attempted to develop new vitrification solutions using bromegrass cells and found that VSL, comprising 20% (w/v) glycerol, 30% (w/v) ethylene glycol, 5% (w/v) sucrose, 10% (w/v) DMSO and 10 mM $CaCl_2$, gave the highest survival following cryopreservation, as determined by fluorescein diacetate staining. However, the cryopreserved cells showed little regrowth, for unknown reasons. To check its applicability, VSL was used to cryopreserve gentian axillary buds and the performance was compared with those of conventional vitrification solutions. Excised gentian stem segments with axillary buds (shoot apices) were two-step precultured with sucrose to induce osmotic tolerance prior to cryopreservation. Gentian axillary buds cryopreserved using VSL following the appropriate preculturing approach exhibited 78% survival (determined by the regrowth capacity), which was comparable to PVS2 and PVS1 and far better than PVS3. VSL had a wider optimal incubation time (20-45 min) than PVS2 and was more suitable for cryopreserving gentian buds. The optimal duration of the first step of the preculture was 7-11 days, and preculturing with sucrose and glucose gave a much higher survival than fructose and maltose. VSL was able to vitrify during cooling to LN temperatures, as glass transition and devitrification points were detected in the warming profiles from differential scanning calorimetry. VSL and its derivative, VSL+, seem to have the potential to be good alternatives to PVS2 for the cryopreservation of some materials, as exemplified by gentian buds.

• Journal of Korean Society of Forest Science
• /
• v.76 no.4
• /
• pp.370-375
• /
• 1987

This study was conducted to examine the position effect of axillary buds on shoot multiplication and rooting with 1-year-old seedlings of Quercus acutissima. Shoot multiplication was greatly affected by axillary bud position: Productivity of multiple shoots were decreased in the order of top, mid and basal explants respectively. The best shoot multiplication (mean 6.1 shoots per explant) was obtained on WPM medium containing $1.0mg/{\ell}$ BAP and $0.1mg/{\ell}$ NAA using basal explant after 4-week-culture. Rooting was also greatly influenced by position. Its percentage was increased in the order of top, mid and basal explant respectively. Root initiation was better and more rapid on 1/2MS medium than GD medium. High rooting percentage (100%) was obtained on 1/2MS medium containing $0.2mg/{\ell}$ IBA after 15 days culture. Sucrose concentrations did not effect on rooting. However root development and shoot growth were greatly affected by them. Root was developed shortly on 1-2% levels and shoot growth was getting retarded, whereas both of them did not show significant difference at 3-6% levels. Rotting was decreased on 7-8% levels gradually, but shoot and leaf condition was better than any other concentrations. Survival rate of rooted explants in pot was varied according to the position of explants. Seedlings of top part were survived up to about 50% but most of mid and basal part seedlings did not survive over 4 weeks even in high humidity condition. Seedlings in pots showed normal growth over 10 months but most of them showed the condition of premature leaf shedding.

• Journal of Plant Biotechnology
• /
• v.29 no.2
• /
• pp.117-121
• /
• 2002

We have developed an in vitro micropropagation system via shoot formation from axillary buds using nodal segments of Corylopsis coreana. Explants from both juvenile tree (one-year-old greenhouse stock seedlings) and mature tree (ten-years-old tree in nursery) were compared with regard to propagation efficiency. Combined treatment of both BA and zeatin were effective on shoot proliferation since the best result was obtained on MS medium supplemented with 0.5∼3.0 mg/L zeatin and 0.2 mg/L BA. Generally, juvenile explants were better in both shoot proliferation and growth than mature explants. However, as the duration of in vitro culture was proceed to 6 months, explants from mature tree also produced three shoots per explant. Distinctive differences in rooting and adaptability to soil of shoots obtained from mother trees. Whereas shoots originated from juvenile explants rooted as high as 97%, those from adult explants showed 62% rooting. Similar result was also observed in soil acclimatization. The plantlets derived from juvenile plants survived 67%, while only 48% of those from adult trees survived. The results showed a possibility of the micropropagation of Corylopsis coreana through shoot formation from axillary buds. In addition, the advance of the research still remain to enhance the frequency of acclimatization of plantlets from mature trees for practical application.

• Korean Journal of Plant Resources
• /
• v.28 no.3
• /
• pp.357-362
• /
• 2015

Carbohydrate sources are one of important factors associated with macro- and micro nutrients and phytohormones in vitro culture medium for shoot growth. The optimal carbohydrates for eight species of the genus Prunus which are economically important fruit crop was evaluated at the initiation and elongation stages. All carbohydrate seemed utilized for the bud break and leaf growth at the early stage of culture. However, shoot elongation and fresh weight of species tested were superior in the medium containing 90 mM of fructose or glucose rather than sucrose. There was no difference between glucose and fructose. Adventitious shoots from the axillary buds were induced in most species but no significant differences were observed except for two species (P. salicina ‘Shiro’ and P. tomentosa). These result demonstrated that glucose and fructose were suitable carbohydrate sources for diverse Prunus species than sucrose, although the response to the carbohydrates in the medium were slightly different in the species.

• Journal of Plant Biotechnology
• /
• v.5 no.2
• /
• pp.101-105
• /
• 2003

To eradicate several viruses such as PVX, PVY, and PLRV which often cause considerable damages to the growth and yields of potatoes, several stems including shoot tips were excised from the potato plants grown for 50 days and electric shock was treated. Shoot tips excised from electric-shocked stems were transferred into the medium supplemented with antiviral compound, ribavirin to examine the combinatorial effect. When treated only with 20 mg/L ribavirin, PVX concentration in the regenerated plant-lets was slowly decreased as repeating sub-culture and finally, it took 32 weeks to reach completely PVX-free stock. With an electric shock treatment (10 mA electric current), all the replicates became free from PVY. However, PLRV was not completely eradicated from 94P70-4 and 93P29-3 lines even by treating with 10 mA electric shock. In this case, both electric shock and antiviral compound treatments in axillary buds from the stem segment were successful in eradicating viral contamination.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.6
• /
• pp.357-360
• /
• 1997

Multiple shoot formation was obtained from excised axillary buds of Achyranthes japonica NAKAI cultured on MS media containing various growth regulators such as auxin and cytokinin. The highest average number of shoots was obtained in 1 mg/L NAA and 2 mg/L BA after 6 weeks (25.8 adventitious shoots per node). Although the regeneration rate was less than the former condition, optimal combination for the production of more shoots with a suitable size was 0.5 mg/L NAA and 1 mg/L BA (19.7 adventitious shoots per node). Roots were induced from regenerated shoots after 3 weeks culture, transferred to 1/2 MS medium supplemented with 0.1 mg/L IBA. Micropropagated plants were successfully transferred to soil.

• Journal of Korean Society of Forest Science
• /
• v.86 no.4
• /
• pp.430-434
• /
• 1997

Axillary bud explants from 3-year-old seedlings of Forsythia saxatilis N., rare and endangered species in Korea, were cultured on Murashige and Skoog's medium. The effect of various cytokinins(BAP, kinetin, and zeatin) at the different concentration(0.2, 0.5 and 1.0mg/L) was tested. Although an apparent shoot proliferation was not observed, zeatin showed slight promotional effect on normal shoot and leaf development. Both shoots and adventive roots could be induced simultaneously when the explants were cultured on the medium with kinetin, but adventive rooting was gradually reduced according as BAP and zeatin concentrations increased. Axillary shoot growth was promoted by the etiolation treatment. Shoot proliferation has been maintained more than three years with consecutive subculture. Rooted plantlets were successfully acclimatized in the artificial soil mixture and showed normal growth after transplantation into field.

• Korean Journal of Medicinal Crop Science
• /
• v.14 no.5
• /
• pp.278-281
• /
• 2006

The effect of plant growth regulators was investigated on in vitro shoot proliferation from axillary bud explants of Hypericum erectum. To determine the optimal cytokinin for proliferation of axillay buds, we carried out screening four cytokinins (BA, kinetin, 2iP, TDZ). When nodal segments were cultured on MS medium supplemented with $4.5\;{\mu}M$ TDZ (thidiazuron), a number of shoots were induced. Our results indicated that the addition of TDZ to culture medium resulted in the induction of significantly more axillary buds than in the addition of other cytokinins. The optimal concentration of TDZ for proliferation of axillary buds was $10\;{\mu}M$. 92% of shoots spontaneously rooted without any plant growth regulator (PGR) and formed whole plantlets within one month. More than 95% of these regenerants survived and they did not show any detectable variation in morphology or growth characteristics compared to their donor plants.

• Journal of Plant Biotechnology
• /
• v.35 no.3
• /
• pp.229-234
• /
• 2008

An efficient micropropagation was established by using axillary bud explants from two-year-old tree(Echinosphorea koreensis Nakai), which has been known as a rare and endangered species. Among various basal media tested, DKW medium was shown to be the best for axillary shoot elongation. The addition of both BA and TDZ to the medium induced 6 to 10 shoots per explant during eight weeks of culture, without showing any abnormal morphology at the shoot proliferation stage. However, high concentration of TDZ(>0.05 mg/L) appeared to cause hyperhydration on either leaf or shoot at the later developmental stage. Approximately 20% of shoots produced roots by the addition of 1.0 mg/L NAA but not by IBA($0.2{\sim}1.0$ mg/L). Ex vitro micro-cuttings were better source for root induction; up to 58.6% of the micro-cuttings rooted when 100 mg/L IBA was applied to the soil(vermiculite). More than 90% of plantlets with roots were successfully acclimatized and grew normally in the field. Therefore, we suggest that this endangered tree species can be effectively micropropagated by axillary bud culture system developed in this study.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2020.12a
• /
• pp.60-60
• /
• 2020

To establish in vitro axillary bud culture conditions of Echinosophora koreensis Nakai, one of Korean endemic endangered species famous for beautiful flowers, we tested the influence of plant growth regulators (PGRs) in shooting and rooting stage from in vitro plants. In shoot multiplication, addition of 6-benzylaminopurine (BA) to the media induced 2.5 to 3 shoots per bud during 4 weeks of culture. And media including 0.5 mg L^-1 thidiazuron (TDZ) produced 3 to 4 shoots per bud. However, zeatin and isopentenyl adenine (2-ip) were not successful to increase shoot number, and the combination treatments of BA with other PGRs were also not effective. Shoots were smaller than 2 cm in length, in most of the treatments. In rooting, naphthalene acetic acid (NAA) treatments in the range of 0.5 to 4.0 mg L^-1 appeared to increase rooting rate by 10% to 60% approximately when compared with the control but roots developed with callus clusters. Indole butyric acid (IBA) addition had little effect on rooting (below 10%), while some roots were longer than in NAA treatments and some shoots were longer on high IBA concentrations (4.0 to 8.0 mg L^-1). It is suggested that micropropagation is a highly applicable and promising to multiplication and conservation of rare and endangered endemic species.