• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.115-119
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• 1999

The single node cuttings of sweet potato (cv. Mokpo #29) plantlets maintained in vitro were cultured with (MF+) or without membrane filter (MF-) under photomixotrophic (PM), hetrotrophic (HT) and autotrophic (AT) conditions. Shoot length was the greatest (11.9cm) in 3$0^{\circ}C$ (HT) treatment and it was the shortest (3.4 cm) in $25^{\circ}C$ (PM) treatment. Nodal explants cultured in 3$0^{\circ}C$ treatment looked more vigorous than those of $25^{\circ}C$ in appearance, and node number was the greatest (10.5 per plantlet) among the treatments. But plantlets grew in 3$0^{\circ}C$ (HT) treatment were observed all overgrown. The size in leaf area was about 2 times greater and shoot length was about 2 times shorter in PM than in HT condition. Percent dry matter of shoots was 5.9% (HT) and 7.4% (PM) in $25^{\circ}C$ treatment and 6.1% (HT) and 7.4% (PM) in 3$0^{\circ}C$ treatment. Plantlets cultured in the MF+ treatments were less succulent than those cultured in the MF- treatment. Vitrified plantlets were examinated 14.8% (both $25^{\circ}C$ and 3$0^{\circ}C$) in PM condition and 22.2% ($25^{\circ}C$) and 31.5% (3$0^{\circ}C$) in HT condition. Sucrose was necessary for the survival of in vitro plantlets. In the sucrose-free medium, explants cultured in the MF- had turned yellow and were dead after 30 days of culture. But explants cultured in the MF+ were alive and produced plantlets with shoot and root (AT). On the other hand, the survival of explants on the MS basal medium (sucrose-free and hormone-free) depended entirely upon the MF attachment.

• Korean Journal of Plant Resources
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• v.19 no.3
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• pp.385-391
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• 2006

In vitro-grown axillary buds of Melia aredarach were successfully cryopreserved by vitrification. On the MS medium supplemented with BA 1 mg/L, multiple shoots were developed within $4{\sim}5$ weeks. Plantlets of Melia azedarach were cold-hardened at $10^{\circ}C$ for a 16-hr photo-period for 6 weeks. Excised axillary shoot-tips from hardened plantlets were precultured on a solidified Murashige & Skoog agar medium (MS) supplemented with 0.7 M sucrose for 1 day at $25^{\circ}C$. Axillary shoot-tip meristems wert dehydrated using a highly concentrated vitrification solution (PVS2) for 60 min at $0^{\circ}C$ prior to a direct plunge into liquid nitrogen (LN). The PVS2 vitrification solution consisted of 30% glycerol (w/v), 15% ethylene glycol (w/v), 15% DMSO (w/v) in MS medium containing 0.4M sucrose. After short-term warming in a water bath at $40^{\circ}C$, the meristems were transferred into 2 ml of MS medium containing 1.2M sucrose for 15 min and then planted on solidified MS culture medium. Successfully vitrified and warmed meristems resumed growth within 2 weeks and directly developed shoots without intermediary callus formation. The survival rate of cold-hardened plantlets for 3 and 4 weeks was 90%. We did not find any difference in PCR-band patterns between control and cryopreserved plants. This method appears to be a promising technique for cryopreserving axillary shoot-tips from in vitro-grown plantlets of Medicinal plants.

• Korean Journal of Plant Tissue Culture
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• v.24 no.6
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• pp.323-327
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• 1997

Addition of growth inhibitors such as ancymidol, ABA, chloromequat, and pachlobutrazol into MS medium had no effect to preventing vitrification in cultures of Rehmania glutinosa. Anatomical investigation revealed that vitrified thick leaf tissue in vitro had larger intercellular space with poor development of sponge and pallisade tissue compared to those of in vitro grown glaucous and field grown plants. In vitro grown glaucous leaf had smaller and round type stomata showing distinguishable guard and subsidiary cell than those of reestablished plantlets into soil whereas abnormal stomata and poor development of epicuticular wax on the surface of leaf was observed in verified plantlet.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.257-261
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• 1998

The repeated subcultures of in vitro plant materials in wasabi became highly vitrified and the capacity for multiple shoot formation from the vitrified plant materials was very low. In order to improve the quality of in vitro propagated planting materials, the experiments were carried out using culture vessels capped with membrane filter(MF). When vitrified shoots were cultured on MS medium with 0.2mg/L BA in the vessels with MF or without MF for 60 days, the shoots in the vessels with MF did not vitrified. In contrast, the shoots grown in the vessels without MF vitrified at 65%. The stomates of vitrified leaves were circular and inflated, whereas those of normal leaves acclimatizated in the vessels with MF were ovate in shape. The hardened shoots were also cultured on MS media without sucrose containing 0.01mg/L IBA in vessels with(photoautotrophic culture) or without(control) MF. Sucrose was necessary for survival of the in vitro plantlets in the vessels without MF. After 20 days of culture, the shoots in the vessels without MF on the sucrose-free media turned yellow and died. But the shoots in the vessels with MF in the sucrose-free media produced a lot of roots. When shoots were cultured on MS medium with 2% sucrose containing 0.01mg/L IBA in the vessels with(photomixotrophic culture) or without(heterotrophic culture) MF, best growth occured in photomixotrophic culture.