• Title/Summary/Keyword: plant regeneration.

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Development of an efficient protocol for high-frequency regeneration system in Hibiscus syriacus L.

  • Seo, Sang-Gyu;Ryu, Sun-Hee;Zhou, Yang;Kim, Sun-Hyung
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.164-170
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    • 2017
  • A rapid and efficient in vitro regeneration system was established for Hibiscus syriacus L. The successful regeneration protocol employs induction of shoot organogenesis on leaf, petiole, and root explants. Among the various plant growth regulators evaluated, thidiazuron (TDZ) was the most effective for inducing rapid shoot formation. Most efficient shoot regeneration frequency was obtained from Murashige and Skoog (MS) media containing 0.01 mg/L TDZ. Regeneration efficiency was highest in the roots, and lowest in the leaves. A combination of 0.01 mg/L TDZ with benzyladenine (BAP) markedly improved the frequency of shoot differentiation from the root (up to 98%) and petiole (up to 88%) explants. Furthermore, leaf and petiole explants showed the highest frequency of shoot induction in half-strength MS media containing 0.01 mg/L TDZ and 1.0 mg/L BAP, while root explants formed the greatest number of shoots when 0.01 mg/L TDZ and 0.1 mg/L BAP were added to half-strength MS media. Although the frequency of shoot differentiation from leaf explants was only 50%, the leaf is considered the most efficient plant organ for use in tissue culture because leaves are easier to obtain than roots and petioles. Our findings show that various organs of H. syriacus can be used for plant regeneration, and the protocol developed in this study may be applicable in the horticulture industry.

Effects of Medium , Plant Growth Regulators , and Explant Sources on Plant Regeneration of Rehmannia glutinosa (배지, 생장조절물질 및 치상조직이 지황 체세포조직으로부터 식물체분화에 미치는 영향)

    • Korean Journal of Plant Resources
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    • v.10 no.1
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    • pp.94-99
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    • 1997
  • When the leaf and stem tissues of Rehmannia glutinosa were cultured on MS medium with plant growth regulators, shoots were regenerated on MS medium with TDZ(0.01 to $2.0{\mu}M$), and root initiation was better on MS medium treated with NAA(0.01 to 2.0mg/$\iota$). On $B_5$ medium, shoot regeneration was better on medium with TDZ than those with Quincrac, NAA, and 2.4-D. The addition of Quincrac, NAA, and 2.4-D inhibited shoot regeneration on MS medium, but promoted shoot regeneration on $B_5$ medium. Shoot regeneration and growth was better on MS medium with the combination treatments of 2.4-D 0.01mg/$\iota$ and TDZ 0.01, 0.1 and $2{\mu}M$ compared with other treatments. Also shoot regeneration and growth on B_5 medium showed the similar results to that on MS medium.

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Effect of Embryogenic Callus Conditions on Plant Regeneration in Satsuma Mandarin (Citrus unshiu Marc.)

  • Han, Sang-Heon;Kang, Sung-Ku;An, Hyun-Joo;Kim, Han-Young
    • Journal of Plant Biotechnology
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    • v.4 no.1
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    • pp.29-32
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    • 2002
  • The ability to form embryiod from callus in satsuma mandarin is low and unstable. In this study, the conditions of embryogenic calli induced from nucellar tissue for promotion of plant regeneration in satsuma mandarin were investigated. The calli of I, II and III line were divided into two sizes of 0.5 mm and 1.0 mm in diameter and two weight gradients of percoll at 40% and 50% though the filter mesh. The frequency of embryo formation from $\phi$ 1.0mm-40% was slightly higher than callus that from others. Adventitious embryoids developed to a globular stage were transferred to regeneration medium. In 'Miyagawa Wase', the embryos from I and II line developed into a heart stage from most of $\phi$ 0.5 mm- 40% and $\phi$1.0 mm-40% calli, but it failed in 'Sugiyama Unshu'. In the cultivar of 'Miyagawa Wase', 63% of adventitious embryos transferred to the regeneration medium developed into the heart stage from the most $\phi$ 1.0 mm-40% calli of I line, but of 'Sugiyama Unshu' failed in some calli condition. The embryoids from two callus lines developed further to shoots and plantlets, while the embriods from III line abnormal failed to regenerate in the cultivar. From these results, it is suggested that the plant regeneration from embryogenic callus in satsuma mandarin could be affected by callus conditions.

Effect of Ethylene Inhibitors on Plant Regeneration of Scrophularia buergeriana M. (에틸렌 억제 물질들이 현삼의 식물체재분화에 미치는 영향)

  • Kim, Young-Kyung;Park, Dong-Sik;Kim, Seong-Mu;Cho, Dong-Ha;Yu, Chang-Yeon;Park, Sang-Un
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.6
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    • pp.367-370
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    • 2006
  • The study was carried out to establish an improved protocol for shoot organogenesis and plant regeneration from leaf explant cultures of Scrophularia buergeriana M. with the treatment of ethylene inhibitors [silver nitrate (AgNO$_3$), aminoethox-yvinylglycine (AVG), Cobalt chloride (CoCl$_2$)]. The regenerated shoots obtained from leaf explant cultures on MS medium containing 2 mg/l BAP, The additions of AgNO$_3$. AVG and CoC1$_2$ substantially improved the shoot regeneration frequency, at the optimal concentration of 7 mg/L, 7 mg/L, and 3 mg/L respectively, The regenerated shoots could be easily rooted with 0.1 mg/L IBA treatment. The noted plants were hardened and transferred to vermiculite with a 85% survival rate where they grew normally.

Plant Regeneration Derived from Leaf Disk Cultures in Purple Sweetpotato (자색고구마의 잎 조직배양을 통한 식물체 재생)

  • Park, Hyae-Jeong;Ahn, Young-Sup;Jeong, Byeong-Choon;Park, Hyeon-Yong
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.245-249
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    • 2003
  • This study was carried out to establish a regeneration system from leaf explant of purple sweetpotato(Ipomoea batatas L.) The optimal concentrations of plant growth regulators for callus induction and shoot formation were determined. The optimal combination for callus formation was 1$\mu$M 2,4-D 5$\mu$M BM, and highest yield of embryogenic calli were observed on Murashige and Skoog basal medium containing 0.5$\mu$M 2,4-D under light condition after 4weeks of culture. Embryogenec callus was subcultured on medium supplemented with 5$\mu$M ABA for 4 days. Subsequently, regeneration of adventitious shoots occurred when these embryogenic calli were transferred onto medium with 3∼6$\mu$M gibberellic acid. Regenerated shoots were developed into normal plantlets.

Shoot regeneration via culture of leaf explants in pear (Pyrus pyrifolia cv. Niitaka) (잎 절편 배양을 이용한 배 '신고' (Pyrus pyrifolia cv. Niitaka)의 신초 재분화)

  • Kim, Se Hee;Shin, Il Sheob;Cho, Kang Hee;Kim, Dae Hyun;Kim, Hyun Ran;Kim, Ki Ok;Lee, Hyang Bun;Do, Kyung Ran;Chun, Jae An;Hwang, Hae Seong
    • Journal of Plant Biotechnology
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    • v.40 no.4
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    • pp.203-209
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    • 2013
  • Genetic manipulation of pear (Pyrus pyrifolia Nakai) breeding is still difficult due to lack of reliable regeneration system. The aim of this research is to establish shoot regeneration system from leaf explants for pear (P. pyrifolia cv. Niitaka) using various concentrations of plant growth regulators and carbon source supplemented to medium. The highest regeneration rate of about 20% was found on a medium containing 4.4 g/L of Murashige and Skoog (MS) without vitamins, Linsmaier and Skoog (LS) vitamins were added separately. Leaf explants of pear were cultured on MS medium containing 7 g/L of Daishin agar supplemented with various concentrations of NAA (0.01, 0.05, 0.1, 0.5 mg/L) in combination with BA(3, 5, 10 mg/L) for shoot regeneration. In medium with 5 mg/L of BA and 0.01 mg/L of NAA, adventitious shoot regeneration rate was higher than others treated. The optimal results were observed using MS medium supplemented with 30 g/L sorbitol as carbon source on regeneration system. Sorbitol is considered better carbon source than sucrose for shoot regeneration of pear (P. pyrifolia cv. Niitaka). In order to increase of shoot regeneration in pear (P. pyrifolia cv. Niitaka), plant agar and Daishin agar used as gelling agents, Daishin agar is more efficient in shoot regeneration.

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%.

Efficient Fertile Plant Regeneration from Protoplasts of Javanica Rice and Their Ploidy Determination by Flow Cytometry (Javanica 벼 원형질체로 부터 효율적인 식물체 재분화와 flow cytometry에 의한 ploidy 검정)

  • LEE, Sung-Ho;Lee, Soo In;SHON, Young Goel;GAL, Sang Wan;CHOI, Young Ju;CHO, Moo Je
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.81-88
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    • 1998
  • The Southeast Asian javanica rice variety Tinawen was investigated for efficient protoplast culture and plant regeneration from cell suspension-derived protoplasts using a feeder cell culture method. Feeder cells of both Lolium multiflorum and Oryza ridleyi, either alone, or in combination, were employed and plants were regenerated from protoplast-derived colonies on several plant regeneration media. Dehydration of protoplast-derived colonies was also investigated as a means of enhancing plant regeneration. In the presence of L. multiflorum or O. ridleyi feeder cells, the protoplast plating efficiency ranged from 0.09% to 1.48%, depending on the feeder cell type and the age of the cell suspension. L. multiflorum feeder cells induced approximately 6-fold higher plating efficiency compared with those of O. ridleyi. The plant regeneration frequencies were 19.3-31.7% with L. multiflorum, 13.0-18.0% with O. ridleyi and 18.0-22.0% with a mixture of both in various plant regeneration media when protoplast-derived colonies were dehydrated, while for the non-dehydrated colonies, the values were 2.0-7.0%, 3.0-5.0% and 0-4.0%, respectively. Flow cytometric analysis of 34 protoplast-derived plants showed that the majority of plants were diploids and only 2 plants were tetraploids. The plants which were transferred to glasshouse were fertile.

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Influence of plant growth regulators on adventitious shoot formation of Glehnia littoralis Fr. Schmid (갯방풍의 기내부정아 형성에 미치는 식물생정조절물질의 영향)

  • Choo, Byung-Kil;Ji, Yun-Ui;Moon, Byeong-Cheol;Choi, Go-Ya;Lee, Hye-Won;Lee, A-Yeong;Kim, Ho-Kyoung
    • Korean Journal of Oriental Medicine
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    • v.14 no.1
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    • pp.113-116
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    • 2008
  • Petiole explant of Glehnia littoralis Fr. Schmidt was in vitro cultured MS plant medium(DUCHEFA co.) supplemented with various plant growth regulators and examined to find out their optimum combination and concentration for plantlet regeneration. We investigated optimal condition for efficient plant regeneration through adventitious shoot formation on MS plant medium with various kinds of plant growth regulators. Embryogenic calli and adventitious shoot formation were greatly influenced by plant growth regulators. Embryogenic calli induction showed a good response on MS medium supplemented with NAA and BA than others. Especially, combination of 1.0 mg/L NAA and 0.5 mg/L BA on MS medium led to the greatest frequency in adventitious shoot. The results suggest that plant regulator selection be important factor to achieve an efficient regeneration.

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Effect of Medium Components and Culture Methods on Shoots Regeneration from Athyrium niponicum (개고사리의 기내 포자체 재생에 미치는 배지구성물질 및 배양방법의 영향)

  • Shin, So-Lim;Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.24 no.2
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    • pp.113-120
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    • 2011
  • Present studies are carried out to find media components and culture methods for in vitro propagation of Athyrium niponicum and to establish the optimal economic masspropagation systems. Among pinnae, petiole and rhizome segments only rhizome segments produced young plants. Rhizome segments showed vigorous plant regeneration on 1/2MS medium and supplement to 1% sucrose and 50 $mg{\cdot}L^{-1}$ $NaH_2PO_4$ were promoted the plant regeneration from rhizome segments. Kinetin was better than BA for plant regeneration and combination with 2 ${\mu}M$ kinetin and 5 ${\mu}M$ IBA was most efficient for plant regeneration. Solid or liquid medium with or without 0.1% qactivated charcoal in modified 1/2MS medium (1% sucrose, 50 $mg{\cdot}L^{-1}$ $NaH_2PO_4$, 2 ${\mu}M$ kinetin, 5 ${\mu}M$ IBA, pH 5.8) were used to find the optimal culture methods. The plant regeneration from rhizome segments were most vigorous on solid medium without activated charcoal. The addition of activated charcoal were inhibited the plant regeneration from rhizome segments not only on solid medium but also liquid stationary or suspension culture.