• Title/Summary/Keyword: Shoot Regeneration

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Plant Regeneration from Mesophyll Protoplasts of Dianthus superbus (술패랭이꽃(Dianthus superbus)의 엽육원형질체로부터 식물체 재분화)

  • Lee, Eun-Ae;Kim, Joon-Chul;Kim, Won-Bae
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.1
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    • pp.41-46
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    • 1995
  • Leaf mesophyll protoplasts of Dianthus superbus were cultured in MSP1 liquid medium supplemented with 0.5 mg/L BAP, 2.0 mg/L NAA and 9% mannitol. Protoplast-derived colonies were formed after 3 to 4 weeks of culture in the dark at 27$^{\circ}C$. These colonies were kept under continuous illumination (21.5 $\mu$E. m-2 sec-1) for 2 weeks and finally most of the colonies became green microcalli, about 3 mm in diameter. When green microcalli were transferred to MS solidified medium with 2.0 mg/L 2,4-D, they formed embryogenic calli after 4 week of culture. These calli were then transferred onto $N_{6}$ medium containing 0.1mg/L 2,4-D, 0.1 mg/L NAA, 2.0 mg/L kinetin and 2.0 g/L casein hydrolysate and cultured under illumination. After 5 weeks of culture the calli gave rise to multiple shoots of 10 to 15 per callus. Upon transfer onto MS medium containing 2.0 mg/L NAA, they were noted. The regenerates were successfully transplanted into potting soil.

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In vitro regeneration of Lycaste aromatica (Graham ex Hook) Lindl. (Orchidaceae) from pseudobulb sections

  • Mata-Rosas, Martin;Baltazar-Garcia, Rosario J.;Moon, Pamela;Hietz, Peter;Luna-Monterrojo, Victor E.
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.157-163
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    • 2010
  • A protocol for in vitro propagation from pseudobulb sections of Lycaste armomatica (Graham ex Hook) Lindl., an ornamental and fragrant orchid, was developed. The effect of four cytokinins: kinetin (K), metatopolin (mT), $N^6$-benzyladenine (BA), and thidiazuron (TDZ), in equimolar concentrations, was investigated. Shoot formation from apical and basal pseudobulb sections was obtained in all treatments. A few medial sections cultured in media supplemented with BA formed protocorm-like bodies. Shoot formation was greater from the basal section than the apical, and mainly occurred in explants cultured in media containing TDZ. The highest average numbers of shoots per explant were achieved from basal sections cultured in media supplemented with TDZ at 4.4, 8.87 and 2.2 ${\mu}M$, forming on average 9.9, 8.6 and 7.3 shoots per explant, respectively. Since the medial pseudobulb section was the worst explant for propagation of L. aromatica, we recommend that pseudobulbs be divided into two sections; the basal half should be cultured in MS medium supplemented with TDZ at 4.4 ${\mu}M$ and the apical half with TDZ at 2.2 ${\mu}M$. Subculturing individual shoots in MS medium without plant growth regulators allows further development and rooting. A survival rate of more than 90% under greenhouse conditions was achieved. This research represents a direct contribution to the conservation and sustainable use of this valuable natural resource.

Shoot Regeneration of Mature Nodal Segments in Poplar (Populus deltoides) (미류나무 성숙 절간조직으로부터 식물체 증식)

  • Yang Hee-Sun;Kang Hoduck
    • Korean Journal of Plant Resources
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    • v.18 no.3
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    • pp.403-409
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    • 2005
  • Mature nodal segments of 2-year-old greenhouse stock plant were cultured on Murashige and Skoog basal medium supplemented with the different kinds and various concentrations of cytokinins to produce multiple shoots in in vitro condition. The most adventitious shoots were produced from excised ends of nodal segments. The highest average number $(24.6\;{\pm}\;4.6)$ of shoots was produced with the combination of BA 1.0mg/L and TDZ 0.1mg/L in MS medium. In addition, several shoots were formed from lenticels of bark cambium with the same treatment. These concentrations promoted high shooting capability upto $94.6\%$ and NAA was the best cytokinin among five different PGR sources.

An Efficient Plant Regeneration and Transformation System of Robinia pseudoacacia var. umbraculifera for Phytoremediation

  • Kwon, Hye-Jin;Woo, Seong-Min;Seul, Eun-Jun;Kim, Teh-Ryung;Shin, Dong-Un;Kim, Hag-Hyun
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.293-298
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    • 2007
  • Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

Plant regeneration from callus of Iris odaesanensis Y. N. Lee native to Korea via organogenesis

  • Bae, Kee-Hwa;Yoo, Kyoung-Hwa;Lee, Mi-Hyun;Jeong, Jae-Hun;Choi, Yong-Eui;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.40 no.3
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    • pp.163-168
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    • 2013
  • Iris odaesanensis Y. N. Lee. is an important endangered and native plant belonging to the family Iridaceae in Korea. This study describes a method for rapid micropropagation of this species via from leaf, rhizome and root explants derived calli. Leaf, rhizome and root explants were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) for callus induction. Rhizome explants yielded calli at a frequency of 72% when cultured at 1.0 mg/l 2,4-D. Calli were maintained at 1.0 mg/l 2,4-D. These calli were transferred to MS medium supplemented with 0, 0.5, 1.0, and 2.0 mg/l 2,4-D in combination with 0, 0.5, 1.0, and 3.0 mg/l BA for adventitious shoot induction. The highest number of adventitious shoot (228.9 per petri-dish) were formed at 1.0 mg/l 2,4-D and 1.0 mg/l BA. WPM medium was the best to convert calli into plantlets, where up to 98.2% of calli were regenerated into plantlets. This in vitro propagation protocol should be useful for conservation of this endangered plant.

Effects of Planting Density and Cutting Height on Production of Leaves for Processing Raw Materials in Goji Berry (구기자나무의 재식밀도 및 예취높이가 가공용 잎 생산에 미치는 영향)

  • Paik, Seung Woo;Lee, Jeong;Yun, Tug Sang;Park, Young Chun;Lee, Bo Hee;Son, Seung Wan;Ju, Jung Il
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.2
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    • pp.136-141
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    • 2020
  • Background: The leaves of the goji berry (Lycium chinense Mill.) are used as raw materials in processing and by replace fruits to some extent. The reason is that the leaves are cheaper, however, betaine content is higher than in the fruits. These experiments were conducted to determine the planting density and cutting height for producing a large number of leaves. Methods and Results: The cultivar 'Myeongan' with many branches was used. When the shoot height reached 50 cm - 70 cm, harvesting was possible four times a year. The time to next harvest was approximately 38 days after regeneration of new shoots. Leaf quantity was in the order of 1st > 2nd > 4th > 3rd harvest. Insect damage occurred during the third harvest in late July and early August, therefore, eco-friendly control was necessary. The total yield was higher at the planting density 60 cm × 30 cm than that of 60 cm × 20 cm or 60 cm × 40 cm. The yield at cutting for shoot height of 60 cm was increased by 6.3 percent compared to that of 50 cm, At the cutting height of 70 cm, harvest was difficult owing to hardening of stems and thorns. Betaine content, an indicator component of goji berry, was not significantly different according to planting densities and cutting height. Conclusions: The ideal cutting period to produce leaves of goji berry for processing is when the shoots grow to approximately 60 cm, and the leaves can be harvested 4 times a year. The dried-leaf yield was highest at the planting density of 60 cm × 30 cm.

Plant Regeneration from Zygotic Embryo-Derived Callus in Citrus junos SIEB. (유자(Citrus junos SIEB.)의 접합배로부터 캘러스 유도 및 식물체 재분화)

  • 박민희;정휘현;이숙영;김홍섭
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.189-194
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    • 1995
  • Calli were successfully induced from immature embryos of Citrus junos SIEB. cultured on 1/2 MS medium supplemented with 40.4 BA. Plant were regenerated from immature embryo derived callus on MS medium with 5 $\mu$M BA. The calli were morphologically characterized by two types: one was whitish and the other was yellowish. After 16 weeks of culture, shoots and root were formed on calli. Plantlets were transplanted to soil and successfully grown to a whole plant Also, the arrangement of the cells showed many differences according to developmental stages of callus and organogenesis. The small cells were compact in callus cultured for 6 weeks and the extended cells which divided actively appeared in it after 8 weeks of culture. The globular protrusion of compacted cells occurred in callus after 10 weeks of culture, and the neighboring cells were liquefied. Oil sac surrounded by the liquefied cell was observed in the leaf and was formed by rupture of liquefied cells.

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Expression of $\beta$-Glucuronidase (GUS) Gene in Transgenic Lettuce (Lactuca sativa L.) and Its Progeny Analysis (형질전환된 상추내에서 GUS 유전자의 발현 및 후대검정)

  • CHUNG, Jae Dong;KIM, Chang Kil;KIM, Kyung Min
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.225-229
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    • 1998
  • Agrobacterium tumefaciens LBA 4404 harboring binary vector pBI 121 was used for genetic transformation of lettuce(Lactuca Sativa L.). Optimal shoot regeneration from cotyledon explants was obtained in MS medium supplemented with 0.1mg/L NAA and 1.0 mg/L 2ip. In this condition, cotyledon explants were cocultivated with A, tumefaciens for 2 days, and then transferred to selection medium supplemented with 50 mg/L kanamycin and 500 mg/L carbenicillin. These explants were subsequently subcultured every 2 weeks on shoot induction medium. PCR analysis indicated that the GUS gene was stably integrated into the nuclear genome of lettuce. Histochemical analysis based on the enzymatic activity of the CUS protein showed that GUS activity was associated with vascular tissue in leaves and roots. Progenies of Ro plants demonstrated a linked monogenic segregation for GUS gene.

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Micropropagation of the hybrids of Actinidia deliciosa$\times$A. arguta by tissue culture (참다래$\times$다래 교잡종의 액아배양 및 캘러스 배양에 의한 기내번식)

  • 문흥규;권영진;이병실
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.227-230
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    • 2001
  • Kiwi (Actinidia deliciosa) is exotic plant and thus susceptible to cold climate in the middle part of Korean peninsular. Several hybrids have recently been developed to enhance cold tolerance by crossing them with domestic species (A. arguta), We have developed an efficient micropropagation technique for the hybrids using both axillary bud and callus culture systems. Shoot proliferation from axillary buds was possible on St medium supplemented with 0.2 mg/L Bh and 3.0 mg/L GA$_3$. In vivo cuttings of the proliferated shoots were more effective for root induction and subsequent survival than in vitro rooting. More than 95% of the plantlets were successfully transferred to field. Effective callus induction was achieved on MS or B$_{5}$ medium with 2,4-D or NAA. Although callus induction could be made from any combinations of media and auxins, shoot regeneration was observed only from the callus induced on medium containing NAA.A.

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Initiation, proliferation, and improvement of a micropropagation system for mass clonal production of banana through shoot-tip culture

  • Khaskheli, Allah Jurio;Ali, Muharam;Shah, Syad Zakir Hussain;Memon, Zohra Fatima;Awan, Saleem;Khaskheli, Muhammad Ibrahim;Khaskheli, Mohsin Ali;Magsi, Bilqees;Qambrani, Zareen;Khaskheli, Asad Ali
    • Journal of Plant Biotechnology
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    • v.48 no.2
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    • pp.86-92
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    • 2021
  • The present study aimed to evaluate the initiation, proliferation potential, and mass clonal production ability of a micropropagation system for banana through tissue culture. A total of 60 explants were cultured on basal media supplemented with various concentrations of BAP and NAA. Banana plants regenerated on MS basal medium (control) without the addition of BAP + NAA showed a significantly (P < 0.05) lower survival rate with no signs of shoots up to the end of the experimental period. The results further revealed that the performance in MSS-XI medium was almost 89%, followed by MSS-IX and MSS-X media, both of which showed performance up to 88%. In contrast, the performance in the MSS-XVI medium was less than 60%, at the less duration of time and highly shoot induction detected at MSS-XIII medium. The maximum number of shoots (4.9) was observed in the medium supplemented with growth adjuster MSS-XI, followed by the MSS-XII medium (4.5). Surprisingly, the best performance was observed for the MSR-VII medium approximately 16 days after initiation, while the lowest performance was observed with MSR-XI (approximately 31 days). The maximum rooting percentage (98%) was observed in the MSR-V to MSR-VIII media (98%), while the minimum rooting percentage was observed in MSR-XI (approximately 45%).