• Journal of Plant Biology
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• 제26권4호
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• pp.191-196
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• 1983

The isolatin and culture of protoplasts from hypocotyl originated callus of Phaseolus vulgaris cv. Damyang were carried out. The maximum protoplast yield of 4.6$\times$105 per gram fresh callus, using the 13-day-old callus, was obtained by digeston for 6 hours in the enzyme solution. After 10 day-culture of the isolated callus protoplsts, plating efficiency was 50%. Thereafter, cell cluster medium, and followed by leading to callus formation on an agar medium after 3 weeks of the liquid culture.

• Journal of Plant Biology
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• 제38권2호
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• pp.143-151
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• 1995

Addition of plant growth hormones [0.01 mg/L NAA and 0.2mg/L benzyladenine (BA)] to a woody plant medium stimulated the adventitious bud formation of poplar explants during culture. Endogenous IAA content increased rapidly at the initial culture stage and then decreased, being followed by rapid increment again at the late culture. But the content of trans-zeatin riboside (t-ZR) increased continuously during the culture. Cytoplasmic soluble proteins were analyzed by one- and two-dimensional SDS-PAGE. Increased amount of 40 kD band was detected by one-dimensional electrophoresis using Coomassie Blue staining during the culture and two distinctive proteins whose mol wt is 40,000 were detected by two-dimensional electrophoresis using autoradiography and these proteins were synthesized continuously prior to the adventitious bud formation. When the midvein segments were transferred to the actinomycin D-containing medium, the spots of adventitious bud-related proteins(ABRPs) did not disappeared but weakened in intensity. So, it is concluded that genes coding for the ABRPs are regulated to some degree at the transcriptional level. Also, they were not observed in BA-free medium, suggesting that these proteins be regulated by cytokinin, which made then possible to form the adventitious bud.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.27-35
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• 2006

Plantlet regeneration in Asteracantha longifolia(L.) Nees (Acanthaceae), a medicinal herb has been achieved from seedling explants on basal MS medium. Three different seedling explants including node, internode and leaf segments on used. Of these three explant, leaf explants gave better response for both callus mediated organogenesis and direct multiple shoot induction. Number of explants showing differentiation of shout buds was higher on MS media supplemented with BA compared to kinetin. MS medium fortified with BA ($2.0mgl^{-1}$) and NAA ($0.5mgl^{-1}$) was found to be most suitable for both callus mediated organogenesis and elongation of shouts. The elongated shoots were successfully routed on MS medium fortified with NAA or IBA. Among them $0.1mgl^{-1}$ NAA or $0.2mgl^{-1}$ IBA provides better response for rhizogenesis. Regenerated plantlets were successfully established in soil where 85.4% or them developed into morphologically normal and fertile plants. RAPD profiling using four decamer primers confirmed the genetic uniformity of the regenerated plantlets and substantiated the efficacy and suitability of this protocol for in vitro propagation of A. longifolia.

• 한국자원식물학회지
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• 제36권6호
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• pp.588-596
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• 2023

Long-term culture of cell lines is an important issue in in vitro culture and in plant science. In this study, the regeneration ability and ex vitro acclimatization of regenerants were evaluated. The ploidy level of regenerants derived from long-term cultured cell lines was measured in Imperata cylindrica 'Rubra', Poaceae. Adventitious buds (shoots) were successfully induced from five-year-cultured calli on MS medium containing 0.1 mg/L BA or 2.0 mg/L TDZ, combined with 0.01 mg/L auxins (IAA, IBA, NAA and 2,4-D), respectively. Adventitious roots were also induced on MS medium containing 0.01 mg/L auxins (IBA, NAA and 2,4-D), respectively. Interestingly, regenerants with both red and green leaf were successfully obtained when regenerants were cultured on MS medium with 9% sucrose. Regenerants derived from long-term cultured calli were transferred to pots using an optimal acclimatization process and successfully adapted to both pot and soil conditions. Moreover, the ploidy level was measured using calli and regenerants that had been kept on MS medium containing various kinds of plant growth regulators (PGRs).

• Plant Resources
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• 제8권2호
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• pp.81-86
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• 2005

An efficient plant regeneration system of Gentiana scabra through somatic embryogenesis was established. Leaves and roots of seedlings of Gentiana scabra excised after germination were cultured on MS basal medium with 2,4-D, NAA or BA. Embryogenic callus was obtained on MS medium with 0.5 mg/L 2,4-D alone or 0.1 mg/L 2,4-D combimation with 1.0 mg/L BA after 45 days of culture. These embryogenic calli gave rise to somatic embryos, which subsequently developed into plantlets on MS medium without PGRs. Also, shoots were effectively differentiated from embryogenic callus when root segments were cultured on MS medium supplement with 0.1 mg/L 2,4-D and 1.0 mg/L BA. Shoots were effectively rooted on MS medium without PGRs. In vitro flowers were formed from plantlets cultured on MS medium with $5\%$ sucrose after 60 days of culture.

• 식물조직배양학회지
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• 제22권1호
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• pp.7-11
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• 1995

종자발아 후 5일된 양배추 그린챌린져 배축으로부터 분리된 원형질체로부터 완전한 식물체를 재분화 시켰다. Kao배지와 B5 배지를 기본으로 한 기존의 몇가지 배지에서 원형질체를 배양한 결과 변형시킨 BS배지에서 Plating efficiency는 낮았으나 식물체 재분화율은 가장 높게 나타나 거의 원형질체 배양배지 조성, 생장조절제 농도 등이 재분화에 상당한 영향을 주는 것으로 나타났다. 또한 사용된25가지 종류의 재분화 배지중에서 MS배지에 l% sucrose zeatin 3 mg/L, 1.6% T.C. agar가 첨가된 배지에서 재분화율이 가장 좋았다. 식물체 재분화에 미치는 PVP (polyvinyl Polypyrrolidone)와 agar 농도의 영향으로는 PVP는 큰 효과가 없었고 agar농도를 1.6%로 2배 증가시켰을 때 재분화율이 다소 높아졌다. 현재까지 재분화된 식물체는 150여 개체로서 포장에서 재배중에 있다.

• 한국작물학회지
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• 제38권4호
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• pp.366-370
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• 1993

본 연구는 기내에서 두릅의 잎조직배양을 통한 효율적인 종묘대량증식체계를 확립하기 위한 기초연구로서 엽내으로부터 캘루스 유기 및 식물체 재분화를 시도한 결과이다. 1. 유연조직으로부터 캘루스 유기는 1.0mg/1 Thidiazuron 첨가배지에서 70%의 유기율을 보였다. 2. 0.1～0.2mg/1 Dicamba 및 1.0mg/1 Picloram 첨가가 캘루스의 증식에 효과가 있었다. 3. 호르몬 첨가배지에서는 식물체 재분화가 이루어지지 않은 반면 Hormon-free 배지에서 shoot 및 root의 분화가 이루어져 완전한 식물체를 얻었다.

• Journal of Plant Biotechnology
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• 제1권2호
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• pp.97-100
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• 1999

In order to establish in vitro propagation method of sundew, Drosera rotundifolia L., the effects of MS medium concentration, cytokinin type and concentration, pH, and auxin type and concentration on shoot proliferation and root formation were investigated using shoots at 3 month after seed germination. The highest shoot production was obtained with the half strength of MS ($\frac{1}{2}$ MS) medium than with any other strength of MS medium tested. Addition of kinetin or BA in $\frac{1}{2}$ MS medium was strongly suppressed shoot proliferation. The suppression of shoot proliferation was more effective in BA-supplemented $\frac{1}{2}$ MS medium than kinetin-supplemented. The optimum pH of the media for shoot proliferation was pH 5.7-6.7. Shoots were subcultured in $\frac{1}{2}$ MS medium supplemented with 0.5mg/L 2,4-D for rooting every 8 weeks. All subcultured shoots produced extensive root systems after 5 to 6 week culture. Plantlets after root development were planted in plastic pots filled with moss. The survival rate of plantlets was almost 100%. On subculturing every 8 weeks, hundreds of the plants were propagated from a single plant within a year.

• Journal of Plant Biology
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• 제36권2호
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• pp.183-192
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• 1993

Efficient plant regeneration system was established through the culture of rice (Oryza sativa L.) microspores. Microspores released by anther shedding culture developed into proembryos and calluses in B5 medium within two weeks of culture. The optimal hormone and carbon sources were dependent on the genotypes used. Microspore's viability decreased rapidly in culture time, therefore less than 3% of the total microspores were viable at the 9th day when the first microspore division was observed. Of two types of microspores (pollen dimorphism) observed in culture, only the P-grain, larger microspores than normal one was able to divide. Using 4',6-diamidino-2-phenylindole (DAPI) fluorescence staining, it was confirmed that the symmetrical division of uninucleate microspore was the first step leading to continuous microspore development. Microspore-derived proembryos and calluses were regenerated to plants in N6 medium containing 1 mg/L NAA and 5 mg/L kinetin, and 78.3% of the regenerated plants were haploids.

• Plant Biotechnology Reports
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• 제3권4호
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• pp.341-345
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• 2009

This study describes culture conditions for a plant regeneration system via a combined pathway of somatic embryogenesis and organogenesis in root explant cultures of the commercial rose cultivar 'Charming'. Root explants formed white calluses at a frequency of 30% after 6 weeks of culture on Schenk and Hildebrandt (SH) medium supplemented with $11mg\;1^{-1}$ 2,4-dichlorophenoxyacetic acid. After 6 weeks of transfer to SH medium without growth regulators, initial white calluses gave rise to globular somatic embryos at a frequency of 2.8%, which were subsequently dedifferentiated to embryonic tissues. Somatic embryos or embryonic tissues initially derived from root explants did not undergo development beyond cotyledonary stage. To produce adventitious shoots, embryonic tissues were sliced and cultured on SH medium with $0.5mg\;1^{-1}$ 6-benzyladenine. After 4 weeks of culture, 28% of embryonic tissue explants formed adventitious shoots. Regenerated shoots were rooted on half strength SH medium with $0.1mg\;1^{-1}$ ${\alpha}-naphthalaneacetic$ acid and subsequently grown to maturity. Root-derived embryonic tissues were proliferated by subculture, while retaining the capacity for shoot production for a few years.