• Title/Summary/Keyword: Regeneration frequency

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The Effect of agrobacterium Density on Transformation Efficiency in Apple (Agrobacterium 농도가 사과 형질전화 효율에 미치는 영향)

  • Seong, Eun-Soo;Cha, Ji-Eun;Kim, Jeong-Hee;Park, Seong-Whan;Yu, Chang-Yeon;Song, Kwan-Jeong
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.215-219
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    • 2003
  • This study was conducted to find optimum bacterial density for improving the efficiency of transformation mediated by Agrobacterium tumefaciens in apples. Regeneration(15%) and transformation frequency(10%) were increased in resuspension-culture density $A_{600}$ 1.3 from preculture density $A_{600}$ 0.7 of Agrobacterium tumefaciens in ′Fuji′. In ′Gala′, 20% regeneration and 16% transformation frequency were observed at optimum bacterial density $A_{600}$ 0.7 form preculture density $A_{600}$ 1.3. ′Mclntosh as well as "Gala" were 25%regeneration and 10% transformation frequency. Hence a frequency optimum condition of bacterial density for the efficient transformation of apple could be depend on apple genotypes.

Efficient Plant Regeneration from Petal Segment of Standard-Type Chrysanthemum

  • Chhetri, Mahesh;Jeon, Su-Min;Naing, Aung Htay;Kim, Chang Kil
    • Current Research on Agriculture and Life Sciences
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    • v.31 no.2
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    • pp.94-100
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    • 2013
  • An efficient plant regeneration protocol is developed for a standard-type chrysanthemum. When petal segments derived from flower buds (4 or 8cm in diameter) were used as the culture material, the highest shoot regeneration frequency (96%) was obtained on a Murashige and Skoog (MS) medium supplemented with 0.5 mg/L IAA, 2 mg/L BA, 3% sucrose, and a 0.8% agar. Pre-culturing the explants under dark conditions for 14 days produced better results for the shoot regeneration frequency than the explants cultured under a continuous 16 h photoperiod ($40{\mu}molm^{-2}s^{-1}$). The shoot regeneration frequency ranged from 19.0% for the Shinmato cultivar to 89.1% for the Baeksun cultivar. Activated charcoal (0.2%) enhanced the root formation of the regenerated shoots in a hormone-free MS medium. The rooted plantlets were acclimatized and successfully established in a greenhouse.

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High frequency plant regeneration from transverse thin cell layers in Indian mustard (Brassica juncea L.)

  • Bhuiyan, Mohammed Shafi Ullah;Lim, Yong-Pyo;Min, Sung-Ran;Choi, Kwan-Sam;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.81-86
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    • 2009
  • An efficient and reproducible plant regeneration system was established using transverse thin cell layers (tTCLs) in five cultivars of Brassjca juncea L. The effects of medium conditions, explant types (tTCLs of hypcotyl and cotyledonary petiole) on shoot regeneration were examined in this study. The maximum shoot regeneration frequency was obtained in Murashige and Skoog (MS) medium supplemented with 4 mg/L 6-benzylaminopurine (BA) and 0.2 mg/L 1-naphthaleneacetic acid (NAA). The hypocotyls derived tTCL explants had more shoot regeneration frequency (52%) than the cotyledonary petiole derived tTCL explants. Shoot induction was further improved by the addition of silver nitrate ($AgNO_3$) in the regeneration medium. A significant genotypic effect was also observed between the five cultivars; Rai-5 displayed higher capacities to produce shoots than other cultivars. Regenerated shoots were rooted on MS basal medium without PGRs which induced 90% of roots. The plantlets established in greenhouse conditions with 99% survival, flowered normally and set seeds. The regenerated plants were fertile and identical to source plants.

Plant Regeneration via Organogenesis from Leaf and Stipule Segments of Strawberry (Fragaria ananassa Duch.) (딸기의 잎과 탁엽 절편체로부터 기관형성을 통한 식물체 재생)

  • 최준영;김현정;형남인
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.347-351
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    • 1998
  • Plant regeneration via organogenesis from leaf and stipule explants of micropropagated shoots of strawberry (Fragaria $\times$ ananassa cv. Suhong) was achieved. Leaf and stipule explants were detached from shoot-tip cultured shoots and cultured on MS medium with various combinations of BA and NAA under light or dark condition. Shoot regeneration from leaf explant was observed after 3 weeks in culture and was good at the high ratio of BA and NAA among various combination treatments. The highest shoot regeneration frequency from leaf explants was obtained with 1.0 mg/L BA and 0.1 mg/L NAA, in which 31.1% shoot regeneration frequency(1.7 shoots per leaf explant) was yielded. In case of stipule explants, shoot regeneration was largely affected by plant growth regulators during incubation under dark condition for initial 4 weeks but not under continuous light condition. The combination treatment with 2.0 mg/L BA and 0.1 mg/L NAA showed the most excellent shoot regeneration from stipule explants, where 44.4% regeneration frequency(4.0 shoots per explants) obtained. Regenerated shoots were rooted on MS medium with 0.1 mg/L NAA after shoot elongation, and the plantlets regenerated were transferred to soil mixtures with vermiculite and perlite for acclimation.

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Plant Regeneration from Protoplasts of Indica Rice (Indica 벼의 원형질체들로부터 식물체 재분화)

  • Sung-Ho, Lee;Young Goel, Shon;Soo In, Lee;Zhoo Hyeon, Kim;Moo Je, Cho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.5
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    • pp.615-625
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    • 1997
  • An efficient protocol for plant regeneration from protoplasts of the indica rice variety IR43 has been developed. The procedure involved plating of embryogenic suspension-derived protoplasts on the surface of a filter membrane overlaying agarose-embedded feeder cells. Lolium multiflorum cell suspensions were preferable to these of Oryza ridleyi as feeder cells and Lolium suspensions supported colony formation from up to 0.68% of the protoplasts, depending on the age of cell suspensions. Plant regeneration frequency was significantly improved by using maltose alone or in a 1:1(w/w) combination with sucrose as carbohydrate source and a simple dehydration treatment using a high concentration of agarose in the regeneration medium. Medium containing maltose or maltose mixed with sucrose increased the plant regeneration frequency compared with medium containing sucrose alone. The plant regeneration frequency was increased to 30.7 to 70.7% following dehydration treatment, while the non-treated controls showed a regeneration frequency of 3.1 to 30.6%. Protoplast-derived plants were transferred to the glasshouse, flowered with morphologically normal.

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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.

High frequency plant regeneration from zygotic-embryo-derived embryogenic cell suspension cultures of watershield (Brasenia schreberi)

  • Oh, Myung Jin;Na, Hye Ryun;Choi, Hong-Keun;Liu, Jang Ryol;Kim, Suk Weon
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.87-92
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    • 2008
  • An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when cultured on halfstrength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly with increasing concentrations of 2,4-D up to $3mg\;l^{-1}$, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryoderived white friable callus were established using half-strength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of water-shield, which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could be useful for the mass propagation and transformation of selected elite lines.

Effects of Embryo Development Stage and Gelrite Concentration on Plant Regeneration in Seed Culture of Rice (벼의 종자배양에서 배의 성숙정도와 Celrie 농도가 캘러스 형성 및 식물체 재분화에 미치는 영향)

  • 권용삼;김경민;김도훈;손재근
    • Journal of Life Science
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    • v.11 no.4
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    • pp.311-315
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    • 2001
  • To identify the effects embryo developmental stage and gelrite concentration on plant regeneration in seed culture of rice, mature and immature seeds of rice were cultured on the $N_{6}$ medium supplemented with2 mg/$\ell$ 2.4-D and different levels of gelrite(0.2~1.0%). The calli formed immature embryos were produced more plants than those from mature embryos. The maximum frequency of plant regeneration was achieved in the culture of the calli of immature embryos which was harvested at the 21$^{th}$ day after pollination. The plant regeneration on the medium with gelrite was more accelerate than that on the medium with agar. The highest frequency(55%) of plant regeneration was obtained from the calli transferred to the medium with 6g/$\ell$ gelrite.

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Factors Affecting Plant Regeneration in the Culture of Different Explants of Stevia(Stebia rebaudiana Bertoni) (Stevia 절편체 종류의 배양에 따른 식물체 재분화)

  • Park, Gyu-Hwan;Kim, Kyung-Min
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.151-154
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    • 2003
  • This study was conducted to determine the optimal concentrations of plant hormones (2,4-D, picloram, dicamba, NAA, kinetin) and the suitable explants among seeds, hypocotyl, and cotyledons on calls formation and plant regeneration of stevia(Stevia rebaudiana Bertoni). The frequency of cellus formation was higher in the young leaf-explants then the older ones, and in the seeds then the hypocotyls and cotyledons on MS medium with 1mg/L 2,4-D. After transfer of seed-derived stevia callus producing embryogenic callus on plant-regeneration medium, the frequency of plant regeneration from callus was 23.8% in MS medium with 1mg/L NAA and 3mg/L kinetin.

Formation, Regeneration, and Fusion of Protoplast of Micromonospora spp. (Micromonospora속 균주들의 protoplast생성, 균사체로의 환원 및 융합에 대한 연구)

  • 김광수;이세영
    • Korean Journal of Microbiology
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    • v.21 no.3
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    • pp.156-162
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    • 1983
  • Conditions for effcient formation and regeneration of protoplasts of Micromonospora rosaria and Micromonospora purpurea were investigated. The state of inoculm, culture stage and growth in a medium containing partially growth-inhibiting concentration of glycing have significant effects on portoplasting. A high frequency of regeneration (up to 30%) was accomplished with a hypertonic regeneration agar medium defined by Okanishi for Strptomyces. Using the optimal conditions for protroplasting and regeneration, protoplast fusion of auxotrophic M.rosaria was carried out. Polyethylene glycol 1,000 was chosen for fusogenic agent. When signgle auxotrophs were used, the recombinant frequency of auxortrophic markers varied from 1.3 to 3.2%. Using two double auxotrophs, the recombinant frequencies of 0.7-4.3% were obtained. Much lower frequencies(three or more orders of magnitude) were observed by the conventional matings.

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