• 제목/요약/키워드: Indirect regeneration

검색결과 26건 처리시간 0.079초

New protocol for the indirect regeneration of the Lilium ledebourii Bioss by using bulb explants

  • Ghanbari, Sina;Fakheri, Barat Ali;Naghavi, Mohammad Reza;Mahdinezhad, Nafiseh
    • Journal of Plant Biotechnology
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    • 제45권2호
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    • pp.146-153
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    • 2018
  • Lilium ledebourii Bioss is a wild species of Lilium, which grows naturally in some provinces of Iran. Previous studies on Lilium tissue culture have been linked to direct regeneration and a few studies have been conducted on indirect regeneration, which has been studied under bright conditions. In this study, for the first time in the world, all the stages of indirect regeneration (callus induction, shoot and root induction) have been studied under dark conditions. Callus formation and the regeneration levels of L. Ledebourii Bioss were examined for three replicates in an MS (Murashige and Skoog) medium with different hormonal compositions and by using a factorial experiment in the framework of a completely random plan. For callus initiation, 2,4-D and kinetin hormones were used in five and four levels, respectively, as auxin and cytokinin. Results showed that the highest percentage of the callus was found in $3{\mu}M$ of 2,4-D and $0.5{\mu}M$ of kinetin. In terms of callus wet weight, the highest amount was found in $3{\mu}M$ of 2,4-D and $0.5{\mu}M$ of kinetin. In addition, in terms of diameter, the highest amount was found in $3{\mu}M$ of 2,4-D, and $0.5{\mu}M$ of kinetin. In summary, the 2,4-D hormone had a major impact on the percentage of regeneration increase so that the best response was related to the composition of $3{\mu}M$ of 2,4-D, and $0.1{\mu}M$ of kinetin. This study contended that auxin and cytokinin can induce long shoots and roots through cell elongation in dark condition.

Plant regeneration via direct and indirect adventitious shoot formation and chromosome-doubled somaclonal variation in Titanotrichum oldhamii (Hemsl.) Solereder

  • Takagi, Hiroki;Sugawara, Shintaro;Saito, Tomoka;Tasaki, Haruka;Yuanxue, Lu;Kaiyun, Guan;Han, Dong-Sheng;Godo, Toshinari;Nakano, Masaru
    • 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.

An Interpretation of the Urban Folly in Gwangju, South Korea Through the Lens of Contextual Novelty

  • Lee, Min Jung;Lee, Dong-Eon
    • Architectural research
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    • 제18권4호
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    • pp.157-164
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    • 2016
  • Urban regeneration, which implies a certain type of change from what has previously existed, requires fundamental understandings of previous problematic ways of thinking for a meaningful change, as well as a new philosophy that is thought to support that change. This means a change in philosophy is needed to discuss 'newness' in urban regeneration. From this perspective, this paper explores the philosophies which shaped traditional cities as well as a philosophy that may be able to suggest changes to contemporary cities. This paper further proposes contextual novelty as necessary to pursue. This contextual novelty does not deny modern cities' problems but embraces them and carries them into contemporary cities. Arguing for qualitative novelty, this paper argues that contextualism, which explores the complex relationships of direct and indirect contributors of a given event, can help us to understand contemporary cities' heterogenic characteristics. In this context, this paper discusses the concept of novelty in contemporary discourse of urban regeneration by using the philosophy of contextualism. Finally a recent example of urban regeneration in the city of Gwangju, South Korea is interpreted using the concept of contextual novelty.

Shoot Organogenesis and Plantlet Regeneration from Stem Explants of Cleome rosea Vahl (Capparaceae)

  • Claudia Simoes;Alessandra S. Santos;Norma Albarello;Solange Faria Lua Figueiredo
    • Journal of Plant Biotechnology
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    • 제6권3호
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    • pp.199-204
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    • 2004
  • The medicinal value of the genus Cleome justifies bio-technological studies of Cleome rosea, a Brazilian annual species from sandy coastal ecosystems (restinga), which have been submitted to an intense process of antropogenic degradation. In the present work, was analyzed the influence of cytokinins, 6-benzyladenine (BA) and 6-furfurylaminopurine (kinetin) added to the Murashige and Skoog medium (MS), on the proliferation capacity of explants from the stem axis (hypocotyl, node and internode) for a period of five monthly subcultures (150 days). Regardless of the explant sources, plantlet regeneration by direct and indirect organogenesis was observed. The largest number of shoots proliferated through direct organogenesis was obtained on medium with 4.4 $\mu{M}$ BA. Also, the highest proliferation capacity through indirect organogenesis was found on medium with 4.4 $\mu{M}$ BA + 4.6 $\mu{M}$ kinetin. The presence of kinetin alone was not effective for multiplication of the species. Elongation and rooting were obtained when shoots were transferred onto growth regulator-free medium, and acclimatization rates from 70% to 81% were achieved depending on explant sources used. Plants were then successfully established in soil and showed normal phenotypes.

Somatic Embryogenesis and Plant Regeneration from Immature Zygotic Embryo Culture in Pepper (Capsicum annuum L.)

  • Jo, Jeong-Yon;Choi, Eun-Young;Choi, Dong-Su;Lee, Kwang-Woong
    • Journal of Plant Biology
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    • 제39권2호
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    • pp.127-135
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    • 1996
  • An efficient system of somatic embryogenesis was established for the red pepper plant (Capsicum annuum L. cv. Nokkwang) usign immature zygotic embryos. The size of the immature zygotic embryos and the concentrations of 2, 4-D and sucrose were found to be critical. Somatic embryos were induced via callus or directly from explants and regenerated into plantlets successfully. When zygotic embryos 1~2 mm long were cultured on the modified Murashige-Skoong (MS) medium supplemented with 2 mg/L 2, 4-D for 3 weeks in the dark, somatic embryos were induced directly from the apical region of zygotic embryos with the highest frequency being approximately 90%. To mature the somatic embryos, ABA and an ethylene inhibitor AgNO3 were used. The highest frequency of shoot regeneration (25% in each) resulted at 2$\mu$M ABA or 20$\mu$M AgNO3 treatment at rates 3.7 and 1.6 times control, respectively. Shoots developed mainly from the cotyledonary node on CoCl2-containing medium, and from the upper side of cotyledon on medium containing AgNO3 while the embryos on the control medium produced shoots from both the cotyledonary node and the upper region of cotyledons both at frequencies of 50%. Indirect somatic embryogenesis via callus was induced at an efficiency of approximately 10% with zygotic embryos 3~4 mm long cultured on MS medium containing 5~10 mg/L, 2, 4-D for 5~7 weeks under a continuous light condition. The plants regenerated from the somatic embryos were morphologically normal. Using scanning electron microscopy, the direct and indirect somatic embryogeneses were observed to follow the globular, heart and torpedo stages, similar to zygotic embryogenesis. Also, suspensors appeared in the early globular and ovoid-shaped late globular embryos during indirect somatic embryogenesis.

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Preferences and flexibility in decision-making among dental clinicians regarding the treatment of multirooted teeth: an interactive communication device-based survey at two academic conferences

  • Lee, Jung-Seok;Lim, Hyun-Chang;Kim, Min-Soo;Choi, Seong-Ho;Jung, Ui-Won
    • Journal of Periodontal and Implant Science
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    • 제46권3호
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    • pp.166-175
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    • 2016
  • Purpose: Decision-making by dental and medical experts can be influenced by their biases, interests, and experiences, and academic arguments about controversial issues may additionally be considered indirect experiences capable of affecting decision-making. This study reports on the use of interactive communication devices to evaluate preferences and flexibility in decision-making among dental care providers who attended two distinct academic conferences. Methods: Two debates were presented by a team of two lecturers at two academic conferences (focusing on periodontology and implant dentistry, respectively) and the audience members of each session were surveyed. Before each lecture, two case modules about the diagnosis and treatment of multirooted molar lesions were provided, and interactive communication devices were used to collect responses about decision-making preferences in treatment planning immediately before and after a debate about treatment strategies. Results: In total, 81 and 84 completed answers from both conferences were obtained for the first and second case modules, respectively. The preferred treatment plan differed significantly according to the focus of the conference, and a tendency emerged for the clinicians participating in each conference to express uniform preferences. However, attending the debates resulted in significant changes in decision-making preferences regardless of the conference focus or the characteristics of the participants. Conclusions: Our findings suggest that providing continuing education via debates on controversial issues may be effective in widening conceptual knowledge and reducing biases among experts in the dental and medical fields.

In vitro plant regeneration from axillary buds of Hibiscus syriacus L.

  • Jeon, Seo-Bum;Kang, Seung-Won;Kim, Wan-Soon;Lee, Gung-Pyo;Kim, Sun-Hyung;Seo, Sang-Gyu
    • 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.

사과 'McIntosh Wijcik' 형질전환체 선발을 위한 Kanamycin 농도 (Kanamycin Concentration for Selection of 'Mcintosh Wijcik' Transgenic Apple)

  • 송관정;성은수
    • 원예과학기술지
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    • 제18권6호
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    • pp.811-814
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    • 2000
  • 사과에서 효율적인 형질전환 체계를 마련코자 'Mclntosh Wijcik' 형질전환체 계통을 이용하여 재분화 및 발근 배지내 kanamycin 농도에 따른 반응을 조사하였다. Gelrite 고형 재분화 배지에서는 $150mg{\cdot}L^{-1}$의 고농도에서도 재분화율에 대한 저해 정도는 낮았으나, 정상 신초 재분화 면에서는 $100mg{\cdot}L^{-1}$이상의 농도에서 는 저해 정도가 높았다. Agar 고형 발근배지에서 kanamycin 농도에 따른 발근 반응은 계통간 차이를 보여, $30mg{\cdot}L^{-1}$이상의 농도에서 발근율이 크게 저해되는 계통과 거의 영향을 받지 않는 계통이 존재하였다. 또한 이들 형질전환체 계통간 발근율 차이는 전이 유전자 삽입수보다는 삽입 위치에 따른 효과로 보아졌다. 따라서 'McIntosh Wijcik' 품종의 형질전환시 gelrite 고형 재분화 배지내 적정 kanamycin 농도는 $100mg{\cdot}L^{-1}$정도로 판단되며, 형질전환체에 대한 일차적인 간접선발에 적당한 agar 고형 발근배지내 kanamycin 농도는 $30mg{\cdot}L^{-1}$정도인 것으로 판단되었다.

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간접구 시술이 골격근 Adiposity 유발 쥐의 근육조직에 미치는 영향 (Effects of Indirect Moxibustion on Skeletal Muscles in Mouse Model of Skeletal Muscle Adiposity)

  • 이기수;홍권의
    • Journal of Acupuncture Research
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    • 제31권1호
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    • pp.7-21
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    • 2014
  • Objectives : To observe the regenerative effects of indirect moxibustion, a traditional Korean medical treatment on skeletal muscles using mouse model of skeletal muscle adiposity. Methods : Twenty seven ICR male mice were randomly assigned into Intact control(n=3), glycerol treatment together without moxibustion(n=12), and glycerol treatment together with moxibustion (n=12) groups. Mice of glycerol treatment groups were injected with 50 ${\mu}l$ DW(distilled water) containing 50 % of glycerol into the two tibialis anterior. After injection, moxibustion was applied at 'Shenshu'($BL_{23}$) and 'Zusanli'($ST_{36}$) acupoints three times per each session, every days for twelve days(total 12 treatments). Phospho-Erk1/2, Myostatin protein levels were analyzed by western blotting and immunofluo-rescence staining techniques for tissues of the tibialis anterior muscle. Smad, phospho-Smad were analyzed by immunofluorescence staining. Results : 1. Histological analysis of sections from injected TA muscles showed that glycerol induced rapidly muscle necrosis, with a maximum at day 3. 6 days and 9 days after injection, muscle was regenerating. 2. According to western blotting and immunofluorescence staining, phospho-Erk1/2 protein signals in glycerol treatment with moxibustion group were stronger compared to Intact and glycerol treatment without moxibustion group. 3. According to western blotting and immunofluorescence staining, myostatin protein signals in glycerol treatment without moxibustion group were stronger compared to Intact and glycerol treatment with moxibustion group. 4. According to immunofluorescence staining, Smad protein signals in glycerol treatment without moxibustion group were stronger compared to Intact and glycerol treatment with moxibustion group. 5. According to immunofluorescence staining, phospho-Smad protein signals in glycerol treatment without moxibustion group were stronger compared to Intact and glycerol treatment with moxibustion group. Conclusions : These results confirm that indirect moxibustion of 'Shenshu'($BL_{23}$) and 'Zusanli'($ST_{36}$) influences muscle regeneration in mouse models of skeletal muscle adiposity. Further discussion, and the establishment of moxibustion mechanism will prompt clinical application of moxibustion.

Direct Somatic Embryogenesis of Curculigo orchioides Gaertn., an Endangered Medicinal Herb

  • Thomas, T.Dennis;Jacob, Alphonsa
    • Journal of Plant Biotechnology
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    • 제6권3호
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    • pp.193-197
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    • 2004
  • In vitro multiplication of Curculigo orchioides was achieved by direct somatic embryogenesis in young leaf segments. Immature leaf segments of about 0.5 cm in length were cultured on MS medium supplemented with different concentrations of BAP (2-10 $\mu{M}$) or Kin (2-10 $\mu{M}$). Optimum response in terms of per cent cultures responding (89%) and the number of embryos per explant (16) were observed on MS medium supplemented with 8 $\mu$M BAP. The emergence of several somatic embryos on the adaxial side of the leaf segments was observed one month after the culture. Germinated somatic embryos were grown up to about 1.5 cm length before transferring to maturation medium. For maturation, the individual embryos were isolated and transferred to MS medium supplemented with BAP (5 $\mu{M}$) and NAA (0.5 $\mu{M}$). The plantlets emerged from the embryos were transferred to soil containing 1 peat: 1 sand with 90% success. The embryos were formed directly on the leaf segments without any callus phase. Direct regeneration of somatic embryos is important for the conservation of this endangered species, as rare somaclonal variants are likely to arise than from indirect regeneration.