• Title/Summary/Keyword: Shoot Regeneration

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The identification of optimum condition for direct regeneration in black raspberry

  • Ran, Choi-Heh;Park, Pill-Jae;Lee, Hee-Kwon;Joong, Yun-Song;Lee, In-Sok
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.163-167
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    • 2008
  • Adventitious buds appeared within 2 weeks on the base of the petiole explants and increased for two months. A maximum of regeneration (15.6%) was obtained on the medium containing $1.5\;{\mu}M$ TDZ in combination with $1\;{\mu}M$ IBA. To know which explants are the best for the induction of regeneration, three explants such as leaf, petiole and leaf-petiole were used. Among the explant types, the leaf-petiole explant was significantly more effective than leaf and petiole for promoting adventitious shoots, with leaf-petiole inducing at the highest regeneration frequency (33.7%). The regeneration frequency of adventitious shoots in the leaf-petiole explants was significantly affected by leaf size and the position of explants. The leaf-petiole smaller than 5 mm leaf in width was induced at the highest regeneration frequency (68.9%). The smaller leaf sizes, the greater regeneration frequency. Also when the leaves are nearer to the shoot tip, the regeneration frequency is higher. When the rooted micro-shoots were transferred to the soil after growing for 6 weeks in the media, the survival rate was 90%.

Optimization of Regeneration Condition Under Agrobacterium-mediated Transformation in in vitro Cultured Korean Soybean

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.36-36
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    • 2018
  • Soybean is the important crop in Asian countries as protein source, oil production and animal feed. Improving soybean using genetic transformation is the principal tool in nowadays. Developing herbicide resistant transgenic soybean plants through Agrobacterium-mediated transformation has been worked in many previous studied. However, the transformation efficiency is still low. Many attempts try to find the optimum media condition for plant regeneration after infection. After transformation, the plant regeneration is very important condition to promote growth of transgenic plant. In this study, we optimized a regeneration condition for two Korean soybean cultivar, Dawonkong and Pungsannamulkong using cotyledon, cotyledonary nodes and hypocotyl as explant. The results showed that shoot regeneration of cotyledonary nodes on B5 medium containing 2 mg/L 6-benzylaminopurine showed the highest percentage of regeneration in Dawonkong (75.8%) while Pungsannamulkong presented high number of shoots 2.12 shoots per explant. For transformation condition, co-cultivation in 7 days showed a high number of GUS positive expression. Most of explants can survived under media including 5 mg/L of glufocinate which refers phosphinotricin for 2-week selection. Washing with 400 mg/L of cefotaxime in several times and selection in plant regeneration media with 400 mg/L of cefotaxime can prevent bacteria growth, effectively.

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In vitro micropropagation of radish (Raphanus sativus L.) using callus induction and plant regeneration (캘러스 유기와 식물체 재분화를 이용한 무의 기내 대량증식)

  • You Kyoung Kim;Sug Youn Mo;Su Bin Choi;Han Yong Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.155-162
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    • 2023
  • Radish (Raphanus sativus L.), a root vegetable grown worldwide, is consumed in several ways. In the cross between parental lines to produce F1 seeds of radish, the problem of low purity may arise because of pollen contamination. Therefore, we aimed to establish conditions for callus induction and regeneration so that in vitro cultured plants could be used for the propagation of stock seeds. The most effective hormone combination containing various concentrations of 2,4-D, TDZ, and kinetin was selected for callus induction using radish hypocotyl, and the induced calli were transferred to two types of hormone media to investigate the optimal conditions for shoot regeneration of the callus. The combination of 1 mg/L 2,4-D + 0.05 mg/L kin was the most effective for callus induction of RA2 and RA10, 1 mg/L 2,4-D + 0.1 mg/L kin + 0.025 mg/L TDZ of RA4, and 1 mg/L 2,4-D + 0.2 mg/L kin of RA30. Shoot regeneration of the RA4 callus occurred in both shoot regeneration media, but the frequency was much higher in the 5H+1B medium (1 mg/L NAA + 0.1 mg/L 2,4-D + 1 mg/L IPA + 0.02 mg/L GA3 + 2 mg/L zeatin + 1 mg/L BA). For the in vitro micropropagation of radish, the conditions selected in this study can assist in the propagation and maintenance of stock seeds to produce F1 seeds.

In Vitro Propagation of Cindium officinale Makino Through Shoot Tip Culture (천궁의 경정배양을 통한 기내번식)

  • 이현숙;정재동;김창배;윤재태;최부술
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.221-225
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    • 1994
  • This experiment was conducted to identify the optimal in vitro propagation condition of Cnidii rhizoma (Cnidium officinale Makino). It was effective to reduce contamination and improve regeneration of shoot when shoot tips taken in July were cultured in 1/2 strength Murashige and Skoog medium supplemented with 500 mg/L carbenicillin disodium 1.0 mg/L BA and 1.0mg/L $GA_3$followed by surface sterilization of explant source in solution of 1% sodium hypochlorite for 20 minutes. When shoot tips were 쳐cultured in 1/2 strength MS medium with 0.5 mg/L BA and 60 g/L sucrose, shoot elogation and subsequent multiplication of the formed shoot were favorable than in other media. Regenerants were well rooted in 1/2 strength MS medium containing 3.0 mg/L NAA.

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Plant Regeneration through Leaf Explant Culture of Boxthorn (Lycium chinense Mill.)

  • Ham, In-Ki;Park, Sang-Kyu;Lee, Bong-Chun;Lee, Mi-Ae;Kwon, Kyeong-Hak;Lee, Eun-Mo;Jo, Man-Hyun
    • Korean Journal of Plant Resources
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    • v.20 no.3
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    • pp.251-254
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    • 2007
  • This study was conducted to investigate the optimal plant growth regulator level for the shoot formation of Lycium chinense Mill. In vitro plant propagation was developed for leaf explants of boxthorn. Leaf explants were cultured on MS medium supplemented with cytokinins (BA and 2-iP) alone. Plants were successfully regenerated through in vitro culture by using leaf explants of boxthorn grown in the field. After 4 weeks of culture, 58% of shoot formation had developed from the leaf explants. The shoot formation rate of 'Jangmyeong' was highest followed by 'Myeongan', ;Cheondae', and 'Bullo'. The use of 0.2mg/L BA was critical for enhanced production of shoot formation and resulted in 58% of the culture producing shoot formations. Regenerated plantlets transplanted to pots were developed and successfully acclimatized to greenhouse.

Improvement of Regeneration Efficiency from Mature Embryo and Leaf Base Segment in Korean Oat Genotypes

  • Kim Kyung-Hee;Moon Jung-Hun;Lee Sang-Kyu;Lee Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.349-353
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    • 2004
  • Mature embryo and leaf base segment of Korean oat were used as materials in an experiment to check plant regeneration efficiency. MS media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin, and picloram were used for callus induction from mature embryos and leaf base segments. Three mg/l of 2,4­D and 3 mg/l of picloram in callus induction medium showed high frequency for plant regeneration from mature embryos. Leaf base segments were transferred to callus induction medium and incubated at $25^{\circ}C$ in 16/8 hr light/dark cycle for 3 weeks. Callus induction from leaf base segments of Malgwiri showed high efficiency in medium containing 3 mg/l of 2,4-D and 1 mg/l of kinetin $(91.8\%)$. In case of Samhangwiri, the combinations of phytohormones did not show significant difference. Regeneration from leaf base segments showed high frequency in shoot medium containing 1 mg/l of antiauxin, tri-iodobenzoic acid (TIBA) and 1 mg/l of 6-benzyladenine (BA). Calli induced from leaf base segments of Samhangwiri and Malgwiri in media containing 3 mg/l of 2,4-D and 3 mg/l of picloram showed high regeneration frequency. It appears that the callus initiation medium may be an important factor for subsequent plant regeneration.

Plant Regeneration Via Organogenesis on Petiole of Centella asiatica (L.) Urban

  • Choi, Kyung-Mi;Hwang, Sung-Jin;Chung, Sang-Jin;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.2
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    • pp.87-91
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    • 2006
  • An efficient plant regeneration of C. asiatica was achieved from organogenesis using petiole explants of in vitro plantlet on MS basal medium controled with different plant growth regulators (NAA,2,4-D, IAA kinetin, and BA). Best results that 50%, efficiency of regeneration per explant for regeneration were obtained with IAA $17.13\;{\mu}M$ and BA $8.9\;{\mu}M$. Formation of adventitious shoots via organogenesis from the petiole explant was verified by histological sectioning of plantlets. Regenerated plants were transplanted into soil.

Characterization of In Vitro Totipotency by Armoracia rusticana (서양고추냉이의 기내 전형성능에 관한 특성)

  • BAE, Chang-Hyu;MIN, Kyung-Soo;AHN, Chang-Soon;LIM, Yong Pyo;KAMEYA, Tosiaki;Lee, Hyo-Yeon
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.119-124
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    • 1997
  • Plantlets were regenerated from various explants (shoot tip, leaf blade, petiole and root segments) via organogenesis and/or somatic embryogenesis from Armoracia rusticana(Lam) Gaerth., Mey, et Scherb.. Shoot regeneration rate from callus was highest on the MS mediums supplemented with 0.5 ㎎/L IAA, 5.0㎎/L BA and 10.0㎎/L spermine. A Low frequency of regeneration occurred on hormone-free MS medium. Multiple shooks were regenerated at a pH of 4.0 to 8.0 on MS medium supplemented with 1.0 ㎎/L BA and 0.1 ㎎/L NAA. Polyamines promoted shoot- and root-formation by 2 to 4 times normal, Specific proteins associated with organogenesis were identified. Somatic embryogenesis occurred directly from the leaf blade, petiole and root segments cultured on MS medium with 2.0 ㎎/L BA and 2.0 ㎎/L BA and 2.0 ㎎/L NAA. Three types of regeneration in A, rusticana were clearly established, which could be applied to the study of morphogenesis and genetics at cell, tissue and organ levels.

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Factors Affecting the Production of In Vitro Plants from the Nodal Pieces of Chinese Yam (Dioscorea Opposita Thunb)

  • Shin, Jong-Hee;Kim, Sang-Kuk;Kwon, Jung-Bae;Lee, Bong-Ho;Sohn, Jae-Keun
    • Journal of Plant Biotechnology
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    • v.6 no.2
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    • pp.97-102
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    • 2004
  • This study was carried out to establish The regeneration of healthy seedlings from the nodal segment culture of Chinese yam (Dioscorea opposita cv. Danma), cultivated in Korea. Different explants such as leaves, petioles, roots and nodal pieces, excised from the in vitro grown seedlings of Chinese yam, were cultured on MS medium supplemented with various combinations of growth regulators. All the growth regulators used induced plantlet regeneration from the nodal segments at a high frequency, while there was no induction of shoot or callus from leaf, petiole or root tissues. The medium supplemented with 0.01mg/L NAA, 0.5mg/L BA, 0.5-1.0mg/L kinetin and without plant growth regulator was effective for shoot development of buds from the nodal segment culture. The concentration of BA and NAA was an important factor in the bud induction of buds from the nodal segments of Chinese yam. Nodal segments cultured on the medium containing 1.0mg/L NAA and 0.5-1.0mg/L BA gave the best response to bud formation. The addition of GA$_3$ to the culture medium suppressed shoot induction and growth, while it increased microtuber formation. The shoot growth and microtuber formation were also affected by medium strength and solidity. The MS basal medium containing 1 g/L gelrite was suitable for microtuber formation from the nodal segment of Chinese yam.

In Vitro Plant Regeneration from Stolen Node Explant in Eremochloa Ophiuroides (Munro) Hack

  • Barampuram, Shyamkumar;Chung, Byung-Yeoup;Lee, Seung-Sik;An, Byung-Chull;Kim, Jae-Sung
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.161-166
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    • 2007
  • In vitro shoot regeneration and multiple shoot induction has been obtained from the stolen node explants in Eremochloa ophiuroides (Munro) Hack. The highest number of shoots ($10.66{\pm}0.21$) was observed from initial explants after one month culture duration on Murashige and Skoog (MS) medium containing 6-benzyladenine (BA: 0.5 mg/l). First generation shoot was excised and sub-cultured on the same fresh media for further multiplication of shoots. An enhanced number of second round shoots ($15.33{\pm}0.21$) was obtained compared to the initial culture media containing BA (0.5 mg/l). The number of shoots/stolon node was higher among all the concentrations of BA than kinetin (KN). In vitro regenerated shoots were successfully rooted in the phytohormone free MS medium. Plantlets generated with roots were transferred to pots containing compound mixture of soil and kept in green house conditions. Acclimatized plants showed 100% survival rate with normal morphology in green house conditions. The present study demonstrates the effect of explant and different plant growth regulators towards in vitro response in E. ophiuroides. Moreover, the study reveals the effect of cytokinin on induction of shoot number per stolen node explant in E. ophiuroides.