• Journal of Plant Biotechnology
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• 제36권2호
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• pp.174-178
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• 2009

Presently, we report a simple, reproducible and high frequency plant regeneration in Hibiscus syriacus L. using axillary buds. H. syriacus was regenerated from axillary buds directly or through a callus phase. Regenerated shoots were directly induced from young and fresh axillary buds cultured on Murashige and Skoog medium (MS) supplemented with 0.01 mg/L of the growth regulator thidiazuron (TDZ) after 2 weeks of culture. Directly induced shoots were transferred to hormone-free MS medium and root development was observed after 6 weeks. On the other hand, old and stale axillary buds were regenerated to shoots via callus induction on MS medium containing 0.01–2 mg/L TDZ after 4 weeks. A TDZ concentration of 0.01 mg/L was most effective in callus formation. Green callus was transferred to MS medium containing 0.01 mg/L α-naphthalene acetic acid (NAA) and 0.5 mg/L benzylaminopurine (BA). After 4 weeks, callus had developed into multiple shoots. Plantlets were formed from 10 week cultures of single shoots on hormone-free MS medium. Regenerated plantlets were cultured on MS medium for one month and then transferred to pots containing garden soil. Potted plants were acclimatized for one month and grown to maturity under greenhouse conditions. The present study has shown that various concentrations of plant growth regulator can be effective for in vitro plant regeneration of H. syriacus. The direct and indirect regeneration protocol presented here will be useful for understanding the manipulation and propagation of H. syriacus.

• Journal of Plant Biotechnology
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• 제5권4호
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• pp.245-250
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• 2003

An efficient procedure was developed for adventitious shoot bud induction and plantlet regeneration from various explants of the ten genotypes of Pepper (Capsicum annuum L.) using Thidiazuron (TDZ). Among various treatments at 1.0-3.0 mg/L TDZ Induced maximum number of adventitious shoots depending upon the explant type and genotype compared to other treatments. Among the explants tested, leaf induced maximum number of adventitious shoots than the cotyledons. TDZ-mediated organo-genesis was possible in 10 pepper cultivars, the extent of the response being genotype-dependent. Of the ten genotypes tested, C. annuum cvs CA960, $G_4$ and X-235 were produced maximum number of adventitious shoots and Sell was the least, and all other genotypes gave moderate response. Elongation of multiple shoots was observed on medium supplemented with SA (0.05 mg/L) in combination of IAA (0.05 mg/L). Differences in ability for in vitro shoot regeneration and elongation depend upon the variety and explant type. The elongated shoots were success. Fully rooted on MS medium containing at 1.0 mG/L IAA. Plantlets regenerated from different explants of ten genotypes were found to be diploid (2n=24) and were devoid of any chromosomal aberrations. Regenerated plants were successfully established in soil where 85-90% of them developed into morphologically normal and fertile plants.

• 한국자원식물학회지
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• 제35권4호
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• pp.502-507
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• 2022

본 연구는 약용식물자원인 땅꽈리 유식물의 하배축 절편으로부터 부정아 형성을 통한 재분화를 조사하여 효율적인 재분화 조건을 확립하고자 하였다. 신초는 저농도 BAP를 함유하는 MS 배지에서 효과적으로 유도되었다. 신초 유도는 BAP 0.5~1.0 mg/L를 단독으로 또는 NAA 0.1~0.5 mg/L와 함께 처리한 MS 배지에서 활발히 이루어졌으며, 특히 BAP 1.0 mg/L가 포함된 MS 배지가 가장 효과적이어서 다발성 신초가 왕성하게 형성되었다. 유도된 신초를 뿌리 유도 배지로 옮겼을 때, 저농도의 NAA, IBA, IAA에서 뿌리가 잘형성되어 재분화 식물체를 만들어 내기에 적절하였다. 발근 수와 뿌리의 길이는 각각평균 2.0개, 8.0 cm 이상으로 높게 나타났다. 특히, 0.03 mg/L의 NAA, IBA, IAA를 포함한 MS 배지에서 뿌리가 더잘 성장하고, 뿌리의 전체적인 형태도 양호하였다. 그리고, 저농도의 NAA, IBA, IAA를 함유하는 MS 배지에서 새로운 신초의 형성도 잘 이루어져서, 줄기의 수가 2개 이상으로 많이, 그리고 길이는 6.0 cm 이상으로 길게 신장하였다. 배양토에 이식한 재분화 식물체는 100%의 생존율을 보였으며, 모두 정상적인 성체로 생장하였다. 따라서 땅꽈리의 부정아 형성을 이용한 재분화 식물체의 생산은 개체들을 대량 증식할 수 있어 균질한 조원료를 안정적으로 공급하는데 주요 수단이 될 것으로 보인다.

• Journal of Plant Biotechnology
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• 제6권1호
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• pp.51-54
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• 2004

This study was conducted to examine differences in shoot regeneration among chrysanthemum cultivars. Leaf explants of chrysanthemum cultivars 'Sulhwa', 'Puma', 'Geummokseo' and 'Sulpoong' were used. Explants cultured on the medium for 2 weeks formed calli at the cut surfaces. Shoots regenerated on MS basal medium supplemented with various concentration combinations of NAA and BAP. Explants were cultured under cool-white fluorescent lamps with a light intensity of $40\mu{Mm}^{-2}$.$s^{-1}$ for 16 $hday^{-1}$, at $25^{\circ}c$ and 70-80% relative humidity. 'Geummokseo' and 'Sulpoong' were the most responsive cultivars in shoot regeneration. Most effective medium for 'Sulhwa' and 'Puma' was MS basal medium supplemented with 10.0 $\mu{M}$ NAA and 5.0 $\mu{M}$ BAP and for 'Geummokseo' MS supplemented with 10.0$\mu{M}$ NAA and 20.0$\mu{M}$ BAP. Regeneration of multiple shoots was observed on MS basal medium supplemented with 1.0$\mu{M}$ or 10.0 $\mu{M}$ NAA and 5.0$\mu{M}$ BAP. High frequency regeneration of adventitious shoots from leaf explants and efficient induction of root from these regenerated shoots were obtained.

• Journal of Plant Biotechnology
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• 제41권4호
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• pp.236-241
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• 2014

An efficient regeneration protocol for stem disc culture of Populus euramericana, which is important species for bioenergy resource in agroforestry, was established. The number of explants that were obtained and the number of explants that regenerated varied with the genotypes. However, in all the genotypes, stem disc culture produced more regenerated shoots than did in axillary bud culture. A comparison of the effects of cytokinin type and concentration on shoot regeneration in different explants (i.e., petiole, leaf, and root segments of P. euramericana) revealed that a concentration of $0.002mg\;l^{-1}$ thidiazuron (TDZ) used on petiole segments resulted in the greatest shoot regeneration (95.83%). The hormonal requirements for the greatest shoot regeneration in the three explant types varied. Different concentrations of $AgNO_3$ and $CoCl_2$ were added separately to the medium to stop the yellowing and subsequent necrosis of the regenerated shoots. Lower concentrations (3 and $5mg\;l^{-1}$) of these compounds improved shoot regeneration and elongation, compared with the control. The in vitro-regenerated shoots were transferred to rooting medium and subsequently acclimatized. The highly efficient regeneration system of P. euramericana reported here can be used for mass propagation of this recalcitrant for regeneration, economically important tree species.

• Plant Biotechnology Reports
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• 제5권2호
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• pp.187-195
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• 2011

The gesneriaceous perennial plant Titanotrichum oldhamii has beautiful foliage and attractive bright yellow flowers. However, breeding of T. oldhamii by conventional sexual hybridization may be difficult because sexual reproduction of this species is very rare. In the present study, plant regeneration systems via both direct and indirect formation of adventitious shoots from leaf explants were established as the first step toward breeding T. oldhamii by using biotechnological techniques. Adventitious shoots were formed efficiently on medium containing $0.1mg\;l^{-1}$ benzyladenine. Histological observation showed that shoot formation on this medium occurred directly from leaf epidermal cells without callus formation. On the other hand, leaf explants formed calluses on medium containing $0.1mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid. The calluses could be maintained by monthly subculturing to fresh medium of the same composition. When the calluses were transferred to plant growth regulator-free medium, they formed adventitious shoots. Directly and indirectly formed shoots rooted well on medium containing $0.1mg\;l^{-1}$ indole-3-butyric acid. Plantlets thus obtained were successfully acclimatized and grew vigorously in the greenhouse. Flow cytometry analysis indicated that no variation in the ploidy level was observed in plants regenerated via direct shoot formation, whereas chromosome doubling occurred in several plants regenerated via indirect shoot formation. Regenerated plants with the same ploidy level as the mother plants showed almost the same phenotype as the mother plants, whereas chromosome-doubled plants showed apparent morphological alterations: they had small and crispate flowers, and round and deep green leaves.

• Plant Biotechnology Reports
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• 제4권1호
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• pp.85-94
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• 2010

In vitro shoot regeneration of gladiolus in three different culture systems, viz., semi-solid agar (AS), membrane raft (MR), and duroplast foam liquid (DF) cultures was evaluated following the kinetics of shoot multiplication and hyperhydricity at optimized growth regulator combinations. Compared to the AS system, matrixsupported liquid cultures enhanced shoot multiplication. The peak of shoot multiplication rate was attained at 18 days of incubation in the MR and DF systems, whereas the maximum rate in the AS system was attained at 21 days. An early decline in acceleration trend was observed in liquid cultures than the AS culture. The hyperhydric status of the regenerated shoots in the different culture systems was assessed in terms of stomatal attributes and antioxidative status. Stomatal behavior appeared to be normal in the AS and MR systems. However, structural anomaly of stomata such as large, round shaped guard cells with damage in bordering regions of stomatal pores was pronounced in the DF system along with a relatively higher $K^+$ ion concentration than in the AS and MR systems. Antioxidative status of regenerated shoots was comparable in the AS and MR systems, while a higher incidence of oxidative damages of lipid membrane as evidenced from malondialdehyde and ascorbate content was observed in the DF system. Higher oxidative stress in the DF system was also apparent by elevated activities of superoxide dismutase, ascorbate peroxidase, and catalase. Among the three culture systems, liquid culture with MR resulted in maximum shoot multiplication with little or no symptoms of hyperhydricity. Shoots in the DF system were more prone to hyperhydricity than those in the AS and MR systems. The use of matrix support such as membrane raft as an interface between liquid medium and propagating tissue could be an effective means for rapid and efficient mass propagation with little or no symptoms of hyperhydricity.

• 식물조직배양학회지
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• 제26권2호
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• pp.85-91
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• 1999

고구마의 정단분열조직을 배양하여 생장조절제의 종류와 농도 및 정단분열조직의 크기에 따른 다아체 증식효과를 조사하였다. NAA와 BA 혼용처리의 경우 0.1 ㎎/L NAA에 2.0 ㎎/L BA혼용처리에서 '목포 29호'와 '산천자'의 경우 배양 30일 후에 100% shoot분화율을 보였고, 뿌리발생률은 '목포 29호'는 66.7%, '산천자'는 69.2%였으며 줄기의 기부 부분에서 형성되었다. Cytokinin류인 kinetin과 BA 0.5∼4.0 ㎎/L 단독처리에서 다아체 분화율이 좋았으며 발달된 shoot의 대부분은 배양 60일 이내에 뿌리발생과 더불어 정상적인 식물체로 재분화되었다. 반면에 '금시'에서는 cytokinin류 단용처리에서 분화된 shoot가 캘러스화되어 정상적인 shoot를 생산하지 못하였다. 캘러스로 덮인 다아체의 shoot가 1∼2마디로 신장된 shoot를 분리하여 4.0 ㎎/L BA 또는 kinetin 처리구에 계대배양하였을 때, 분할된 shoot의 대부분이 줄기의 기부에 캘러스가 형성되면서 계대배양 30일 후에는 정상적인 shoot와 뿌리를 가진 재분화된 식물체의 생산이 가능하였다.

• Plant Biotechnology Reports
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• 제4권2호
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• pp.173-178
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• 2010

An efficient protocol for shoot regeneration was developed for sesame (Sesamum indicum L.) internodes using the transverse thin cell layer (tTCL) culture method. The frequency of shoot regeneration and the number of adventitious buds produced from regenerated shoots depend significantly on explant age, thickness of the tTCL sections, and the phytohormones supplemented to the culture medium. A combination of 6-benzyladenine (2.0 $mg\;l^{-1}$) and a-naphthaleneacetic acid (0.5 $mg\;l^{-1}$) was found to be the best phytohormone combination for shoot bud induction, with the maximum number of shoots obtained when the tTCL sections were 0.5-1.0 mm thick and derived from 4- to 6-week-old seedlings of sesame. Well-developed shoots were rooted on MS medium without phytohormones, and 80% of the regenerated plantlets were successfully established in soil.

• 식물조직배양학회지
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• 제25권5호
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• pp.347-351
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• 1998

딸기 '수홍'의 기내배양된 잎과 탁엽 절편체로부터 기관형성을 통한 식물체 재생 방법을 확립하고자 실험을 수행하였다. 엽절편체 배양에서는 생장조절제 처리에 따라 NAA 첨가시에는 뿌리 재생, BA 첨가시에는 신초 재생이 이루어졌으며, BA 1.0㎎/L + NAA 0.2 ㎎/L 처리구에서 재생률 31.1%, 절편체당 신초수 1.7개로 가장 양호하였다. 탁엽절편체 배양에서는 생장조절제와 광조건의 처리를 실시하였는데, 명조건에서는 생장조절제 처리에 따른 뚜렷한 경향을 발견할 수 없었으나, 암조건에서는 고농도 BA와 저농도 NAA의 혼용처리가 신초 재생에 필요함을 알 수 있었다. 탁엽절편체는 BA 2.0 ㎎/L + NAA 0.1㎎/L 처리구에서 재생률 44.4%, 절편체당 신초수 4.0개로 가장 양호한 반응을 나타내었다. 재생된 신초는 NAA 0.1㎎/L 가 첨가된 MS배지에서 발근되었으며, 이어서 인공토양(vermiculite : perlite = 1 : 1)에서 성공적으로 활착을 유도할 수 있었다.