• Title/Summary/Keyword: shoots production

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High-frequency plant regeneration from leaf-disc cultures of Jatropha curcas L.: an important biodiesel plant

  • Deore, Ajay C.;Johnson, T. Sudhakar
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.7-11
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    • 2008
  • A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings as well as mature field-grown plants cultured on Murashige and Skoog's (MS) medium supplemented with thidiazuron (TDZ) ($2.27{\mu}M$), 6-benzylaminopurine (BA) ($2.22{\mu}M$) and indole-3-butyric acid (IBA) ($0.49{\mu}M$). The presence of TDZ in the induction medium has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented with BA ($4.44{\mu}M$), kinetin (Kn) ($2.33{\mu}M$), indole-3-acetic acid (IAA) ($1.43{\mu}M$), and gibberellic acid ($GA_3$) ($0.72{\mu}M$). Well-developed shoots were rooted on MS medium supplemented with IBA ($0.5{\mu}M$) after 30 days. Regenerated plants after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation.

Oxidative Stress in Rice (Oryza sativa L.) Seedlings Induced by Flooding

  • Lee, Keun Pyo;Jung, Jin
    • Journal of Applied Biological Chemistry
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    • v.44 no.4
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    • pp.159-162
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    • 2001
  • Plant stress incurred by flooding was studied in terms of oxidative stress, using greened rice seedlings subjected to a complete submergence followed by re-exposure to air under illumination ($30W/m^2$). It appeared that shoot tissues of the seedlings suffered oxygen deficiency during the flooding treatment, pertinent to the general concept. Interestingly enough, however, membrane peroxidation in shoots was enhanced by the submergence, as assessed by the content of 2-thiobarbituric acid-reactive substances (TBARS), and the re-aeration resulted in a rapid reduction of TBARS content. Such pattern of response was also seen in the change in the steady state level of $H_2O_2$. In contrast, superoxide dismutase and glutathione reductase that are involved in the detoxifying processes of superoxide in plant cells were significantly activated only during the re-aeration. These results allowed us to suggest the followings as a working hypothesis. Photorespiration-linked production of $H_2O_2$ may largely contribute to the increase in $H_2O_2$ level as well as TBARS production in shoots during the submergence. An abrupt re-supply of $CO_2$ by the re-aeration brings the photosynthetic apparatus back to full operation, suppressing photorespiration and probably causing a momentary, excess formation of superoxide and its dismutation product through side reaction, which gives rise to activating substrate-inducible antioxidative enzymes.

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Production of transgenic Alstroemeria plants containing virus resistance genes via particle bombardment

  • Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.164-171
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    • 2020
  • Transgenic Alstroemeria plants resistant to Alstroemeria mosaic virus (AlMV) were generated through RNA-mediated resistance. To this end, the friable embryogenic callus (FEC) of Alstroemeria was induced from the leaf axil tissue and transformed with a DNA fragment containing the coat protein gene and 3'-nontranslated region of AlMV through an improved particle bombardment system. The bar gene was used as a selection marker. More than 300 independent transgenic FEC lines were obtained. Among these, 155 lines resistant to phosphinothricin (PPT) were selected under low stringent conditions. After increasing the stringency of PPT selection, 44 transgenic lines remained, and 710 somatic embryos from these lines germinated and developed into shoots. These transgenic shoots were then transferred to the greenhouse and challenged with AlMV. In total, 25 of the 44 lines showed some degree of resistance. PCR analysis confirmed the presence of the viral sequence. Virus resistance was observed at various levels. Establishment of an efficient transformation system for Alstroemeria will allow inserting transgenes into this plant to confer resistance to viral and fungal pathogens. Accordingly, this is the first report on the production of a transgenic virus-resistant Alstroemeria and lays the foundation for alternative management of viral diseases in this plant.

Micorpropagation of Corylopsis coreana by Thidiazuron Treatment (Thidiazuron 처리에 의한 히어리나무의 기내번식)

  • Kang, Hyo-Jin;Moon, Heung-Kyu;Yi, Jae-Seon
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.263-267
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    • 2003
  • This study was conducted to evaluate the effect of thidazuron(TDZ) on shoot proliferation and growth from axillary buds of 20-years-old Corylopsis coreana. Shoots proliferation was effectively achieved on WPM(Woody Plant Medium) supplemented with 0.03∼0.1mg/L TDZ. The highest shoot number(6.5$\pm$0.7) was obtained on 0.1mg/L TDZ treatment. On the TDZ medium shoots formed as clusters less than 1cm in height and therefore needed to subculture on GA$_{3}$ containing medium to induce elongation. In consecutive cultures, phenolic compounds were excreted at the proximal part of the explants and inhibited growth of the explants. Growth inhibition by the compounds was overcome using liquid and paper bridge culture system. About 60% of the elongated shoots rooted on half- strength MS medium containing IBA. Generally, IBA was mire effective on in vitro rooting than NAA with optimal range of 0.5mg/L to 1.0mg/L. Rooted plantlets were transferred in an artificial soil(vermculite) and acclimatized in high humidity greenhouse condition. Survival rate differed greatly depending on rooting types of the explants. Two types of rooting were observed. The first type was direct rooting from the explants. The second type was callus formation followed by rooting from the callus. The explants showing the 1st type rooting survived can be multiplicated in vitro by TDZ treatment followed by elongation with GA$_{3}$ and rooting with IBA.

A Selection of Korean Native Sedum spp. for Evergreen Ground Covers of the Central Districts of Korea (중부지방 상록지피용 기린초 속의 조경용 소재 선발)

  • Lee, Sun-A;Ha, Yoo-Mi;Han, In-Song
    • Journal of the Korean Institute of Landscape Architecture
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    • v.37 no.5
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    • pp.90-97
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    • 2009
  • Sedum takesimenses among the Korean Native Genus Sedum was selected as evergreen ground covers during winter in the central districts of Korea. In order to obtain basic information for production as a pot flower with good quality, the effects of pinching frequencies on growth and flowering of S. takesimense were investigated. S. takesimense plant height became much shorter and numbers of lateral shoots were increased with increased pinching. Shoots of S. takesimense treated with 30% pinching during 2004 and 2005 showed greater length and less number of lateral shoots than those with 60% pinching. Leaves of S. takesimense treated with 70% shading showed a dark green color and had a longer shoot length and a lower number of shoots than cultivation under full sunlight. Therefore, Korean native S. takesimense will be utilized as ground cover treated with 70% shading or under a tree. Therefore, S. takesimense will be promising as ground cover materials in the central districts of Korea.

Regeneration from Storage Root Disk Culture of Purple Sweet Potato

  • Park, Hyejeong;Park, Hyeonyong
    • Korean Journal of Plant Resources
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    • v.28 no.3
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    • pp.363-369
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    • 2015
  • Sweet potato has low regeneration capacity, which is a serious obstacle for the fruitful production of transgenic plants. Simple and rapid regeneration method from storage root explants of purple sweet potato (Ipomoea batatas L.) was investigated. The embryogenic callus was observed from 4 cultivars and its highest rate was induced at 1 μM 2,4-D after 5 weeks of culture. Result revealed that a low concentration of 2,4-D and low light intensity was important factors for embryogenic callus formation. After subculture on medium with 5 μM ABA for 4 days, subsequently, occurred the regeneration of shoots within 4 weeks when these embryogenic callus was transferred onto the MS hormone free medium. Regenerated shoots were developed into platelets, and grown normal plants in the greenhouse. We developed a simple and quickly protocol to regenerate plantlets in storage root explants of purple sweet potato. This regeneration system will facilitate tissue culture and gene transfer research of purple sweet potato.

Establishment of a regeneration system for the production of Calla plants (Zantedeschia spp.) via embryogenic callus culture (배발생캘러스 배양에 의한 칼라 식물체 재분화 체계 확립)

  • Han, In-Song;Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.32-36
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    • 2019
  • Calla lilies (Zantedeschia spp.) are monocotyledonous ornamental plants which belongs to the Araceae family. After the release of elite calla cultivar, an efficient propagation system is needed for commercial use. Despite the use of conventional propagation methods such as splitting of tubers and rhizomes of calla, rapid and efficient propagation system should be developed. In order to achieve this goal, stem segments contained apical meristems derived from calla lily cultivar (cv. Gag-si) were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of cytokinin and auxin. This was aimed at inducing embryogenic calluses, shoots and multiple shoots. As a result, about 25% of induction rates of yellow embryogenic calluses were observed with MS medium containing both $0.5mg{\cdot}L^{-1}\;NAA$ and $1.5mg{\cdot}L^{-1}\;BA$ as growth regulators. In the experiments involving the regeneration from embryogenic calluses through shoot formation, MS medium supplemented with $0.5mg{\cdot}L^{-1}\;IAA$ and $2.0mg{\cdot}L^{-1}\;BA$ showed the highest rates at approximately 85 ~ 90% with regard to the formation of shoots in calla. Moreover, multiple shoots needed for rapid propagation were generated when explants were cultured on MS medium supplemented with $0.5mg{\cdot}L^{-1}\;IAA$ and $2.0mg{\cdot}L^{-1}\;BA$ with 40% of formation rate. In this study, the combination of auxin and cytokinin showed positive effects on both the induction of embryogenic calluses, the formation of shoots as well as multiple shoots in calla. The regeneration system described here can contribute to the development of breeding programs of calla in the future.

Adventitious root induction in Ophiorrhiza prostrata: a tool for the production of camptothecin (an anticancer drug) and rapid propagation

  • Martin, Kottackal Poulose;Zhang, Chun-Lai;Hembrom, Manoj Emanuel;Slater, Adrian;Madassery, Joseph
    • Plant Biotechnology Reports
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    • v.2 no.2
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    • pp.163-169
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    • 2008
  • Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

Introduction of two-step culture method for multiple seed bulb development from shoot tip culture of garlic (Allium sativium L.) (마늘의 경정배양에서 기내인경구 대량생산을 위한 2단계 배양의 도입)

  • Hwang, Hye-Yeon;Lee, Young-Bok
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.75-80
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    • 2008
  • In vitro culture of shoot tip of garlic (Allium sativium L. cv. Seosan) was carried out to find medium condition of the induction of multiple shoots and bulbing for muliproduction of virus-free seed bulbs. For this work, tank culture was introduced. In shoot tip culture on MS solid medium the induction of multiple shoots and bulbing were better by adding 3% sucrose than 8%. Supplementation with 2mg/L 2ip and 0.2 mg/L IAA in this medium was effective. Three point three shoots including 2.7 bulbs were formed from a shoot tip after cultivation for 30 days on this medium. Bulbing of garlic in liquid culture with plastic water tank of 20L supplied air at the side of the lower part was better by adding 3% sucrose than 8% by subculture for 45 days with shoots obtained from shoot tip culture for 30 days on soid MS medium. Shoot growth was vigorous at 3% sucrose however bulb growth was more effective on the medium of 8% sucrose. Because of the effectiveness on solid medium added 3% sucrose, 2 mg/L 2ip and 0.2 mg/L IAA for initial production of multi-shoot in stem tip culture and the effectiveness in liquid culture with water tank for growth of bulbs, the method of two-step culture could be introduced for the multiple production of seed bulb of high quality. It was more desirable by supply of 0.2 mg/L BA and 0.02 mg/L NAA at tank culture time. But growth of the bulbs became poor by increasing concentration of NAA of the medium.

Effects of Auxins end Cytokinins on Callus Induction from Leaf Blade, Petiole, and Stem Segments of in Vitro-grown 'Sheridan' Grape Shoots

  • Seung-Heui kim;Kim, Seon-Kyu
    • Journal of Plant Biotechnology
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    • v.4 no.1
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    • pp.17-21
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    • 2002
  • To establish an the mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of various combinations of auxins and cytokinins on friable callus production were studied. for friable callus production, 2,4-D was superior to other regulators. IAA at 2 mg/L induced callus from stem and petiole while NAA resulted in rooting. Callus induction rate increased with the 2,4-D level, and stem segments were superior to leaf blade or petiole, showing nearly 100% with 1 and 2 mg/L 2,4-D from petiole and stem. Combined treatments of 2,4-D + kinetin and NAA + BA also yielded friable callus from stem segments. In treatments with 1 mg/L 2,4-D + 1 mg/L kinetin and 1 mg/L NAA + 1 mg/L BA, callus induction rate was nearly 100%. The combination effect of 2,4-D and BA on anthocyanin production was not significant.