• Title/Summary/Keyword: leaf explant

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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.

Induction and Proliferation of Adventitious Roots in Codonopsis spp. (더덕 속 식물의 부정근 유도와 증식)

  • Kim, Ji Ah;Park, Eung Jun;Choi, Yong Eui
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.493-499
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    • 2012
  • We have established adventitious root culture systems of Codonopsis lanceolata, Codonopsis pilosula and Codonopsis ussuriensis. Root segments of C. lanceolata were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with $0.5mg/{\ell}$ NAA and produced $18.8{\pm}1.9$ roots per explant. Root segments of C. pilosula were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with $1.0mg/{\ell}$ NAA and produced $8.5{\pm}1.8$ roots per explant. Leaf segments of C. ussuriensis were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with $0.5mg/{\ell}$ NAA and produced $7.8{\pm}0.4$ roots per explant. In liquid culture, the best production of adventitious root (fresh weight) was obtained in 1/2 MS medium with $1.0mg/{\ell}$ NAA. This study demonstrated for the first time to produce adventitious roots in C. pilosula and C. ussuriensis.

Effect of Cell Source and pH of Culture Medium on the Production of Canthin-6-one Alkaloids from the Cell Cultures of Tongkat Ali (Eurycoma longifolia Jack)

  • Mahmud, Luthfi-Aziz;Chan;Boey
    • Journal of Plant Biotechnology
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    • v.6 no.2
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    • pp.125-130
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    • 2004
  • Callus and cell suspension cultures of Eurycoma longifolia Jack could be an alternative supply of 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one. The callus tissues were initiated from leaves of different trees. The friable calli were used for the preparation of the cell suspension cultures of E. longifolia. The leaf explant of tree Eu-9 produced the most callus and also induced high cell biomass in the cell suspension culture, but it produced low quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. The leaf explant from tree Eu-8 produced low quantity of callus and cell biomass, but produced the highest quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. Optimum production of cell biomass was obtained on cell culture medium with pH 5.75 prior to autoclaving, but high alkaloids content could be induced in culture medium in acidic condition with pH 4.75 and 5.25 prior to autoclaving.

Factors for high frequency plant regeneration in tissue cultures of Indian mustard (Brassica juncea L.)

  • Bhuiyan, Mohammed Shafi Ullah;Min, Sung-Ran;Choi, Kwan-Sam;Lim, Yong-Pyo;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.137-143
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    • 2009
  • An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

Effect of Morphological Factors, Antibiotics and Agrobacterium Co-cultivation in the Efficiency of Somatic Embryogenesis of Eggplant (Solanum melongena L.)

  • Claudia Magioli;Erika Tarre;Krul, William-Robert;Elisabeth Mansur
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.19-25
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    • 2001
  • Induction of somatic embryogenesis from Brazilian eggplant variety F-100 was studied in response to four auxin types. NAA, at the optimal concentration of 54 $\mu\textrm{m}$, was the only one that resulted in the induction of somatic embryos in either leaf and cotyledon explant and, at murk lower intensity and frequency, in hypocotyl and epicotyl explants. The optimal temperatures for embryo induction were 28 and 35$^{\circ}C$ for cotyledon and leaf explants. Incubation at 22$^{\circ}C$ caused a significant reduction both in the frequency and intensity of induction. This system was used to study the effects of position and orientation of the tissue on the culture medium as well as of antibiotics and explant co-cultivation with Agrobacterium on the efficiency of somatic embryo induction. The intensity of embryo induction was greater in the midsections of cotyledons relative to apical and basal regions, when the abaxial surface was in contact with the culture medium. The presence of antibiotics resulted in approximately 40-60% reduction of embryo induction relative to control explants, which originated 335$\pm$26.6 embryos. Co-cultivation with Agrobacterium before treatment with antibiotics caused a more drastic reduction (80-99%). Ampicilin treatment after cocultivalion with Agrobacterium caused the least inhibitory effect, allowing the production of 60 embryos/explant.

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Effect of Medium Component on Plant Regeneration via Adventitious Bud Formation from Leaf Explant Cultures of Strawberry (Fragaria ananassa Duch.) (딸기 (Fragaria ananassa Duch.) 잎 절편체 배양으로부터 부정아 형성을 통한 식물체 재생)

  • 조덕이;소웅영;정원일
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.171-178
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    • 2001
  • This study was investigated to establish a regeneration system of plant via adventitious bud formation from leaf explant cultures of strawberry (Fragaria ananassa Duch). Effects of plant growth regulators (2,4-D, BAP), agar sucrose and myo-inositol on adventitious bud formation were investigated. When the leaf explants were cultured on MS medium supplemented with 0.1 mg/L 2,4-D and 3 mg/L BAP, the adventitious bud formation was most promoted. The adventitious bud formation was not induced from leaf explants cultured on MS medium containing 2,4-D alone. Adventitious bud formation was enhanced to almost 3 times on medium with low level of agar concentration (0.4%) in comparison with those on the medium with high level of agar (1%), but almost of shoot was vitrificated on the medium. Therefore, the normal adventitious bud formation from leaf explants was most effective on the medium containing 0.05 mg/L 2,4-D,1 mg/L BAP, 0.8% agar, 30 g/L sucrose and 100 mg/L myo-inositol. Thus, the mass propagation of healthy strawberry could be established using leaf explants.

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High-frequency shoot regeneration from leaf explants through organogenesis in bitter melon (Momordica charantia L.)

  • Thiruvengadam, Muthu;Rekha, K.T.;Yang, Chang-Hsien;Jayabalan, Narayanasamypillai;Chung, Ill-Min
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.321-328
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    • 2010
  • An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro (15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g $1^{-1}$ sucrose, 2.2 g $1^{-1}$ Gelrite, and 7.7 lM naphthalene acetic acid (NAA) with 2.2 ${\mu}M$ thidiazuron (TDZ). Regeneration of adventitious shoots from callus (30-40 shoots per explant) was achieved on MS medium containing 5.5 ${\mu}M$ TDZ, 2.2 ${\mu}M$ NAA, and 3.3 ${\mu}M$ silver nitrate ($AgNO_3$). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 ${\mu}M$ gibberellic acid ($GA_3$). The elongated shoots were rooted in MS medium supplemented with 4.0 ${\mu}M$ indole 3-butyric acid (IBA). Rooted plants were acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an average of 40 plants per leaf explant with a culture period of 98 days.

Studies on the Culture of Haploid Tobacco Leaf (담배 반수성의 유엽배양에 관한 연구)

  • 한창열
    • Journal of Plant Biology
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    • v.15 no.1
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    • pp.28-32
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    • 1972
  • Young haploid leaf derived from the anthers of tobacco plant was cultuerd and plantlets of various ploidies were obtained. When the leaf was put on the medium supplemented with kinetin as growth regulator, plantlets developed directly from the leaf, and the plants coming out in early stage of culture were all haploid. Plants developing in later stage were mostly haploids with some exception of diploid and aneuploid. Leaves were also cultured on the callus-inducing media supplemented with 2,4-D and kinetiion, and the calluses were sub-cultured for six months. Plants developed from these calluses were mostly aneuploids of various chromosome numbers. In view of the fact that the plants directly developed from the leaf were all haploid, the tissue of the original leaf explant was assumed to be uniform as far as chromosome number was concerned. On the other hand, it seemed that the occurrence of various ploidies in the plants derived from the calluses of same origin was the result of the influence of in vitro culture. Apical meristem tissues and various multicellular bodies were formed in the epidermal and inner mesophyll tissues as well as in the sub-epidermal cells.

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