• Title/Summary/Keyword: Root explant

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Transformation of Gourd through Leaf Explant Regeneration (잎 절편의 재분화에 의한 참박 형질전환)

  • Cho, Song-Mi;Moon, Sun-Jin;Chung, Soo-Jin;Kim, Mi-Seong;Kim, Young-Cheol;Yang, Kwang-Yeol;Choi, Yong-Soo;Sapkota, Kumar;Cho, Baik-Ho;Kim, Kwang-Sang
    • Korean Journal of Plant Resources
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    • v.19 no.5
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    • pp.634-639
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    • 2006
  • In order to develop a disease-resistant root stock for the growth of watermelon, an efficient regeneration system of the gourd(Lagenaria leucantha Duch.) inbred line GO701-2 via organogenesis was established in this experiment. Using proximal parts of cotyledon explant excised from germinated seedling in vitro, maximum adventitious shoot formation (39%) was achieved on MS medium where cytokinin (BA) and auxin (IAA) were added at a concentration of 3mg/L and 0.1mg/L, respectively. Roots of the elongated shoots were successfully formed on MS medium without adding any plant growth regulators. The cucumber CsGolS1 gene known as a resistance gene against biotic and abiotic stresses, was constructed into the binary vector pBI121 under the control of CaMV 35S promoter. When the gene was introduced into the genome of gourd by Agrobacterium-mediated transformation, putative transgenic plants were obtained with the transformation efficiency of approximately 20 percent.

Optimal conditions for adventitious root organogenesis from peony root explant callus cultures (작약(Paeonia lactiflora Pall.) 뿌리절편 유래 캘러스 배양으로부터 부정근발생을 위한 최적 배양조건)

  • Lee, Young Jin;Choi, Myung Suk;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.207-212
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    • 2022
  • The optimal culture conditions for root organogenesis from the callus of peonies (Paeonia lactiflora Pall.) were investigated. Root explants with vascular bundles were cultured in Murashige and Skoog (MS) medium combined with 0.5-4.0 mg/L auxins (indole acetic acid [IAA], naphthalene acetic acid [NAA], indolebutyric acid [IBA], and 2,4-dichlorophenoxyacetic acid [2,4-D]) and 0.0-2.0 mg/L cytokinins (kinetin, zeatin, and benzylaminopurine [BAP]) to induce callus formation. The callus was then cultured in MS medium combined with three concentrations (0.1, 0.5, and 1.0 mg/L) of IAA, NAA, IBA, kinetin, zeatin, and BAP in the dark for 6 weeks. Based on the results, the effects of dark and light conditions on the callus cultured in MS medium with combinations of 0.1-1.0 mg/L IBA and zeatin for 6 weeks were studied. Callus formation was most effective (>+++) in the medium with a combination of 1.0 mg/L NAA and 1.0 mg/L zeatin. A high number of long adventitious roots were observed in the mediums with 0.1 mg/L IBA (6.66 and 4.82 cm) and 0.5 mg/L zeatin (2.32 and 0.72 cm) among auxins and cytokinins, respectively. The highest number (14.06) of adventitious roots were formed from the callus cultured in light in the MS medium combined with 0.1 mg/L IBA and 0.5 mg/L zeatin. This same medium induced the formation of the longest adventitious root (5.45 cm) in the dark. Thus, optimization of in vitro culture conditions may be possible for the mass propagation of adventitious roots in peonies.

Effect of Genotype and Explant on Somatic Embryogenesis and Acclimatization of Acanthopanax senticosus (가시오갈피의 수집종과 배양조직에 따른 체세포배발생 및 재분화 식물체의 순화)

  • Lee, Cheng-Hao;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.3
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    • pp.217-221
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    • 2002
  • Callus induction and embryogenesis were studied in three different genotypes of Acanthopanax senticosus, to develop a protocol for somatic embryogenesis and acclimatization. Young leaf, stem, node, petiole, peduncle, flower and root explants were collected from 3-year old trees of A. senticosus accessions (Korea, Russia and Japan). Callus was obtained from all cultured explants but showed the higher rate of callus formation in flower cultured. For the three A. senticosus accessions, callus was well formd on MS media containing 2mg/ l of 2,4-D and 2mg/ l of TDZ, 4mg/ l of 2,4-D and 1mg/ l of TDZ than other treatments. For three A. senticosus accessions, when callus transferred to MS medium with 2,4-D, embryogenic cell formed. For A. senticosus accessions Korea, embryogenic cells were obtained on callus induced from petiole, stem, node and root explants, and induction rate was lower than 3%. 200mg of embryogenic callus was transferred to MS free liquid medium and somatic embryos of heart stage were obtained after 45days of culture. When somatic embryo of germination stage were transferred to solid medium, most of the embryos were regenerated into plantlets on 1/4 MS medium. Normal plants with both shoots and roots were transferred to greenhouse soil and were successfully acclimatized.

Plantlet Regeneration by Tissue Cultures of Cyclamen persicum Mill. (시클라멘 (Cyclamen persicum Mill.)의 조직배양에 의한 식물체 재분화)

  • 은종선;김영선;한상권
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.479-484
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    • 2000
  • This study was conducted to examine the effects of explant sources and plant growth regulators on mass propagation of Cyclamen persicum. Tuber, cotyledon, and petiole tissues were cultured on MS medium supplemented with different concentrations and combinations of auxins and cytokinins. Shoots were not induced from calli on cotyledon and petiole explants cultured on MS medium containing various concentrations of 2,4-D or NAA. However, multiple shoots were formed directly from tuber explants cultured on the medium containing 0.5 and 1.0 mg/L 2,4-D or NAA. In MS medium with cytokinin alone, TDZ was more effective in shoot formation than BA or kinetin in all explants. The combinations of NAA and BA was found to be most effective in shoot formation from tuber, cotyledon and petiole explants. Especially, shoots were formed from all the tuber explants on the medium containing 0.5 mg/L of NAA and BA. Hormonal effects on root formation were examined by subculturing single shoots on MS medium containing NAA or IBA. The medium with 0.5 mg/L IBA was most effective in root induction and subsequent plantlet regeneration.

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Effect of Growth Regulators and Osmoticums on Somatic Embryogenesis and Plants Regeneration in Aralia elata Cultivar 'Zaoh' (두릅나무 '자오'의 체세포배 유도와 식물체 형성에 미치는 생장조절제 및 삼투압제 효과)

  • Kim Ji-Ah;Moon Heung-Kyu;Kim Yong-Wook
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.129-134
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    • 2005
  • Effective micropropagation system via somatic embryognesis was established for a Phytophthora resistant Aralia elata cultivar. Different kinds of growth regulators were needed to induce embryogenic callus with different explant sources. When leaf explants were used, a combination of 2,4-D, TDZ and L-glutamine was needed, whereas when petiole and root explants needed only 1.0 mg/L 2,4-D. Embryogenic callus induction rate under the optimum culture condition was 75.0%, 67.0% and 83.0% from leaf, petiole and root segment, respectively. Somatic embryo germination and plantlet conversion rate appeared to be influenced greatly by various osmoticums. More than 90% of embryos germinated when treated with sucrose, glucose and maltose. However, the highest conversion rate (72%) was recorded on medium with 2% sucrose only. The converted plantlets grew normally on 1/2MS basal medium, were acclimatized on artificial soil mixture and survived more than 95% in the greenhouse condition. The results suggest that the species can be clonally propagated through in vitro culture system via somatic embryogenesis.

High-frequency regeneration of plants in vitro from seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim

  • Kim, Tae-Dong;Kim, Nam-Ho;Park, Eung-Jun;Lee, Na-Nyum
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.54-61
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    • 2021
  • This work describe an efficient method for the shoot induction and plant regeneration of seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim. The highest rate of shoot induction (82.2%) was obtained when apical bud explants from juvenile seedlings (5 months old) were cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BAP). However, apical bud explants obtained from mature trees (12 years old) did not produce any shoots, even with BAP supplementation. Among the three cytokinins tested for shoot multiplication (BAP, zeatin, and kinetin), BAP was the most effective; the highest number of shoots per explant (2.1) was observed on MS medium supplemented with 1.0 mg/L BAP. In contrast, the longest average shoot length (3.0 cm) was observed after growth on MS medium with 2.0 mg/L zeatin. No multiplication occurred when apical bud explants were cultured with kinetin-supplemented media. During rooting of in vitro-elongated shoots, the highest rooting rate (100%) was observed in half-strength MS medium supplemented with 0.5 ~ 1.0 mg/L indole-3-butyric acid (IBA) or 3.0 mg/L 1-naphthaleneacetic acid (NAA). During the acclimatization process, plantlets that were rooted on the IBA (0.5 mg/L)-supplemented medium had the highest survival rate (100%) and maximum root length (18.5 cm). These findings suggest that a low concentration (0.5 mg/L) of IBA is appropriate for the rooting and acclimatization of T. mandshurica. Plants were successfully transferred to the greenhouse with a 100% survival rate. This protocol will be useful for the large-scale propagation of Tilia species.

Effects of Explant Parts and Plant Growth Regulators on the in vitro Propagation of Lycoris squamigera (상사화의 기내증식에 미치는 배양부위와 생장조절물질의 영향)

  • Eun, Jong-Seon;Kim, Young-Seon;Park, Jong-Suk;JIN, Song Nan;CAO, Hounan
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.179-183
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    • 2002
  • This study was carried out to investigate the influence of medium composition for in vitro mass propagation of Lycoris squamigera Max. After the disks of short stems, segments of leaf within bulb and scale were cultured on MS basal medium supplemented with various plant growth regulators, they were examined for the extent of callus formation, shoot and root regeneration. In the culture of stem disks, adventitious shoots were regenerated from the basal tissue of bulb scales, and combined medium of 1.0 mg/L 2,4-D or NAA+2.0 mg/L BA or kinetin showed the the best response and 4∼6 shoots per explant formed. In the culture of leaf segments within bulbs, both MS medium supplemented with 1.0 mg/L NAA+2.0 mg/L TDZ and with 1.0 mg/L 2,4-D+1.0∼2.0 mg/L BA were produced callus profusely on the base of leaf tissue and 3∼6 shoots were regenerated per explant. In the scale segments culture, calli were produced on the basal tissue on medium with 1.0 mg/L 2,4-D+1.0∼2.0 mg/L BA. The best result were shown on MS medium with 1.0 mg/L NAA+2.0 mg/L TDZ, and 1.0 mg/L 2,4-D+1.0∼2.0 mg/L BA. Maximum number of regenerated shoots was up to 10∼12. Adventitious root formation from explants were formed profusely on MS medium with 1.0 mg/L NAA+2.0 mg/L kinetin. The most desirable method for mass propagation of plantlets was the shoot regeneration from scale segments then subsequently subcultured on medium for rooting.

Towards Conservation of Threatened Ceropegia Species Endemic to a Biodiversity Hotspot: In Vitro Microtuber Production and Proliferation, a Novel Strategy

  • Pandit, Sagar Subhash;Nair, Aneeshkumar;Naik, Dhiraj Dilip
    • Journal of Forest and Environmental Science
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    • v.24 no.2
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    • pp.79-88
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    • 2008
  • Twenty-eight of 44 Indian Ceropegia species are endemic and their survival is threatened. As a step towards conservation, we implied in vitro methods for the sustainable propagule production in C. hirsuta, C. lawii, C. maccannii, C. oculata and C. sahyadrica. Effects of explant, growth regulators, sucrose and photoperiod were studied. High frequency microtuber production was achieved with the seedling-apical buds, grown on MS medium containing 4-6 mg $1^{-1}$ BAP, 3-8% (w/v) sucrose, under continuous illumination. Each microtuber, when subcultured proliferated to form a cluster of secondary microtubers. Every primary and secondary microtuber bore at least one shoot-bud and a root primordium. Each tuber (formed with any of the significantly effective treatments) weighed more than 500 mg, enough to plant directly in non-sterilized soils. Microtubers could be produced and proliferated round the year. Proliferation could be solely attributed to in vitro procedures as these plants bear solitary tubers in vivo. Microtubers could be sprouted in vitro to prepare ready to pot plantlets. As, this novel method succeeded for all five species, though they belong to different eco-physiological backgrounds, we recommend its implementation in the conservation programs for a broader range of Ceropegia species, supported by other integrated strategies.

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In vitro Conservation of Coleus forskohlii- an Endangered Medicinal Plant

  • Rajasekharan P.E.;Ambika S.R.;Ganeshan S.
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.135-141
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    • 2005
  • Protocols for in vitro conservation was developed for Coleus forskohlii. Plants maintained both in field served as explant source. Shoot tips and single node cuttings were used to optimize protocols for in vitro multiplication. MS basal medium supplemented with $0.54\;{\mu}M$ naphthalene acetic acid (NAA) and $8.87\;{\mu}M$ benzy-ladenine (BA) induced multiple shoots in shoot tips and nodes. Shoot multiplication was amplified with a gradual decrease of BA concentration, leading to its final omission after 4 months. Concomitant rooting on multiplication media enabled successful establishment extra vitrum. For in vitro conservation studies, experiments were carried out with 2-3 week maintained in vitro plants under standard and reduced culture conditions (SCC, RCC). In vitro plants could be successfully conserved in full strength MS medium (FMS) under SCC for 6 months without subculture with full potential to regenerate, producing viable shoots and nodes. The root production remained unaffected due to conservation, showing high rooting activity in mannitol and low temperature treatments. Preset low temperature (15 and $10^{\circ}C$) and reduction in media constituents does not appear to favour conservation, although the former accomplished conservation levels equal to (FMS) under SCC.

Micropropagation of Aristolochia elegans(Mast.)

  • Lidia, Osuna T.;Alejandro, Mora I.;Elsa, Ventura Z.;Enrique, Jimenez F.;Crescencio, Bazaldua M.;Antonio, Jimenez A.
    • Journal of Crop Science and Biotechnology
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    • v.10 no.3
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    • pp.141-146
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    • 2007
  • The roots of Aristolochia elegans Mast.(Aristolochiaceae) are widely used in Mexican traditional medicine as a remedy for scorpion venom. Current experimental evidence supports its purported antidote properties. However, collection from the wilderness has lead to local extinction of natural populations. In order to contribute to species preservation, cultivation, and standardization of morphological and pharmacological properties, a micropropagation method was developed. This includes in-vitro germination of seeds to produce aseptic plantlets, induction of multiple budding, and acclimatization. The treatment with benzylamino purine(10 ${\mu}M$) induced the highest number of buds(3.1 on average) in both types of explants. On the other hand, indolebutyric acid(1.5 ${\mu}M$) caused the highest root index(11.8) per explant. One hundred percent of the micropropagated plantlets developed vigorously after the acclimatization process.

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