• Title/Summary/Keyword: Plantlet Regeneration

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Somatic Embryogenesis from In Vitro Grown Leaf Explants of Rosa hybrida L.

  • Kim Chang-Kil;Chung Jae-Dong;Jee Sun-Ok
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.169-172
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    • 2003
  • Somatic embryogenesis was initiated from in vitro grown leaf explants of rose following an induction period of four weeks on MS basal medium supplemented with auxin and several subcultures on MS medium with cytokinin. '4th of July' showed the highest regeneration frequencies on 1 mg/L NAA followed by culture on medium with 4 mg/L zeatin. The embryogenic callus was propagated on MS medium with NAA, zeatin and $GA_3$. Germination of somatic embryos was achieved on MS medium with 1 mg/L BA. Somatic embryo derived plantlets were hardened and successfully transferred to the greenhouse.

The Factors on Somatic Embryogenesis of Soybean [Glycine max. (L.) Merrill]

  • Kim, Kyong-Ho;Kim, Hag-Sin;Oh, Young-Jin;Suh, Sug-Kee;Kim, Tae-Soo;Park, Ho-Kee;Park, Moon-Soo;Kim, Seok-Dong;Yeo, Up-Dong
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.123-128
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    • 2000
  • To enhance in vitro plantlet regeneration efficiency of soybean through embryogenesis, the culture conditions such as material part and size of immature seed, 2,4-D, pH and solidifying agents for somatic embryogenesis were investigated. Somatic embryogenesis was induced from the immature embryo, immature cotyledon and embryonic axis explants of the immature seed on MS medium supplemented with 2.0 mg/L 2,4-D. The highest rate (up to 22.9%) of somatic embryogenesis was obtained from the immature cotyledon, following embryonic axis and the immature embryo. The rate varied with the developmental stages of seed. The maximum rate (25.4%) of embryogenesis was obtained from 3-4 mm length of the seed (after 25 days of flowering). The optimum concentration of 2,4-D for embryogenesis was 10 mg/L. The optimum pH was at 5.8 and solidifying agent for medium was better with 0.4% gelrite than with agar. For rapid multiplication of shoot tips from the germinating somatic embryos, they were cultured on MS medium containing 2 mg/L indole-3-butyyic acid (IBA) and 1 mg/L 6-benzyladenine (BA). After then somatic embryos with one and three cotyledons were transferred to the growth regulator free medium. The medium exhibited the higher rate (ca. 50%) of development than the multiplication medium.

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Plant Regeneration by Anther Culture of Lilium asiatic hybrid 'Gran Paradiso' (아시아틱 백합 ( Lilium asiatic hybrid 'Gran Paradiso' )의 약배양에 의한 식물체 재생)

  • 고정애
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.1-6
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    • 1999
  • In order to obtain plantlet derived by anthers, the anthers of Lilium asiatic hybrid 'Gran Paradiso' were cultured on Murashige and Skoog's medium supplemented with various combinations of auxin and cytokinin. The most suitable pollen stage of anther culture for the callus induction was 3 days before anthesis at the early to late binucleate stage. Organogenic calli were induced on MS medium supplemented with 5.0 mg/L 2,4-D alone and the combination of 1.0 mg/L 2,4-D and 1.0 mg/L kinetin, however, the combination of NAA and BA was more effective than that of 2,4-D and kinetin on plant regeneration through organogenesis. Shoots were formed from the induced callus on the medium with 0.5 mg/L NAA and 1.0 mg/L BA after 180 days of culture. Multiple shoots with 3-4 leaves, roots, and bulblets were formed on the medium with the combination of 2.0 mg/L NAA and 2.0 mg/L BA after 250 days of culture. The chromosome from root tip of the regenerated plantlet showed the diploid (2n=2x=24). Diploid plants were transferred to the pots and all plants were flowered in two years.

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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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In vitro plantlet regeneration of "dwarf" Indian olive (Elaeocarpus robustus Roxb.): a fruit plant of Bangladesh

  • Rahman, Md. Mahabubur;Amin, Muhammad Nurul;Ishiguri, Futoshi;Yokota, Shinso;Sultana, Rubaiyat Sharmin;Takashima, Yuya;Iizuka, Kazuya;Yoshizawa, Nobuo
    • Plant Biotechnology Reports
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    • v.3 no.3
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    • pp.259-266
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    • 2009
  • A plantlet regeneration protocol was developed on pot-grown mature plants of Elaeocarpus robustus Roxb. cv. Dwarf from nodal and leaf explants. The best yield of adventitious shoots was achieved from the leaf-derived calli in a modified MS ($MMS_1$, half strength of major salts, full strength of minor salts, and vitamins) medium containing $4.0{\mu}M$ BA + $4.0{\mu}M$ Kn + $0.5{\mu}M$ NAA + 15% coconut water (CW). The shoot multiplication rate was amplified about twofold per culture after the addition of 15% CW to the medium. The rate of shoot multiplication reached maximum at the 5th subculture, and it maintained this rate throughout the 3 subsequent subcultures. The best rooting in vitro was investigated by subculturing the microcuttings in an $MMS_2$ (half strength of both major salts and minor salts and full strength of vitamins) medium containing $1.0{\mu}M$ IBA in the dark for one initial week at $30^{\circ}C$, followed by subculturing them in a plant-growth regulator (PGR)-free medium in the light. The plantlets raised in vitro were successfully established under ex vitro conditions.

Efficient Procedures for Direct Shoots Regeneration from Leaf Explants of Rehmania glutinosa Lib. (지황 잎조직 절편으로부터 신초 형성)

  • Hwang, Sung-Jin
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.6
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    • pp.273-277
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    • 2005
  • Adventitious shoots were directly induced from leaf explants of R. glutinosa, an important medicinal plant. Proliferating shoot cultures were obtained by culturing leaf discs on Murashige and Skoog(MS) medium alone or combination with auxins and cytokinins. MS medium supplemented with 1 $mg/{\ell}\;BA\;and\;2\;mg/{\ell}$ IAA was the most effective, providing high shoot bud formation frequency without formation of intervening callus. The effect of leaf age on adventitious shoot formation was also investigated. The maximum shoot bud production (93.4%) was achived using 3rd leaf from apex of 6 weeks old plantlets after seed germination. Plantlet were rooted on an half-strength MS (1/2MS) medium containing 0.1 $mg/{\ell}\;IBA$. This prtocol is useful for clonal propagation and Agrobacterium-mediated transforamtion in R. glutinosa.

Plantlet Regeneration Cotyledon and Petiole Cultures of Cyclamen Persicum Mill (시클라멘(Cyclamen Persicum Mill.)의 자엽과 엽병배양에 의한 식물체 재분화)

  • 은종선;고정애;김영선
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.212-216
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    • 1995
  • These experiments were carried out to examine the effect of explant sources and plant growth regulators on callus induction and plantlet differentiation. Cotyledon and petiole explants of Cyclamen persium were cultured on MS medium supplemented with different concentrations of auxins and cytokinins. Cotyledon cultured on medium containing 2, 4-D and kinetin did not form callus or shoots. But when calli induced from petiole explants on medium with 1.0 mg/L 2, 4D and 1.0 mg/L kinetin were subcultured on the same medium the formation of shoots from calli occurred after 150 days of culture. The combination of NAA and BA were more effective than that of 2, 4 D and kinetin in the formation of shoots from calli, cotyledon culture was most effective on medium with 0.2 mg/L NAA and 0.5 mg/L BA. Shoots excised from calli were rooted on medium with 1.0 mg/L NAA. The plantlets were subsequently transplanted to potting soil.

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Influence of plant growth regulators on adventitious shoot formation of Glehnia littoralis Fr. Schmid (갯방풍의 기내부정아 형성에 미치는 식물생정조절물질의 영향)

  • Choo, Byung-Kil;Ji, Yun-Ui;Moon, Byeong-Cheol;Choi, Go-Ya;Lee, Hye-Won;Lee, A-Yeong;Kim, Ho-Kyoung
    • Korean Journal of Oriental Medicine
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    • v.14 no.1
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    • pp.113-116
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    • 2008
  • Petiole explant of Glehnia littoralis Fr. Schmidt was in vitro cultured MS plant medium(DUCHEFA co.) supplemented with various plant growth regulators and examined to find out their optimum combination and concentration for plantlet regeneration. We investigated optimal condition for efficient plant regeneration through adventitious shoot formation on MS plant medium with various kinds of plant growth regulators. Embryogenic calli and adventitious shoot formation were greatly influenced by plant growth regulators. Embryogenic calli induction showed a good response on MS medium supplemented with NAA and BA than others. Especially, combination of 1.0 mg/L NAA and 0.5 mg/L BA on MS medium led to the greatest frequency in adventitious shoot. The results suggest that plant regulator selection be important factor to achieve an efficient regeneration.

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Thidiazuron Induced High Frequency Adventitious Shoot Formation and Plant Regeneration in Capsicum annuum L.

  • VENKATAIAH PEDDABOINA;CHRISTOPHER THAMIDALA;SUBHASH KARAMPURI
    • Journal of Plant Biotechnology
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    • v.5 no.4
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    • pp.245-250
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    • 2003
  • An efficient procedure was developed for adventitious shoot bud induction and plantlet regeneration from various explants of the ten genotypes of Pepper (Capsicum annuum L.) using Thidiazuron (TDZ). Among various treatments at 1.0-3.0 mg/L TDZ Induced maximum number of adventitious shoots depending upon the explant type and genotype compared to other treatments. Among the explants tested, leaf induced maximum number of adventitious shoots than the cotyledons. TDZ-mediated organo-genesis was possible in 10 pepper cultivars, the extent of the response being genotype-dependent. Of the ten genotypes tested, C. annuum cvs CA960, $G_4$ and X-235 were produced maximum number of adventitious shoots and Sell was the least, and all other genotypes gave moderate response. Elongation of multiple shoots was observed on medium supplemented with SA (0.05 mg/L) in combination of IAA (0.05 mg/L). Differences in ability for in vitro shoot regeneration and elongation depend upon the variety and explant type. The elongated shoots were success. Fully rooted on MS medium containing at 1.0 mG/L IAA. Plantlets regenerated from different explants of ten genotypes were found to be diploid (2n=24) and were devoid of any chromosomal aberrations. Regenerated plants were successfully established in soil where 85-90% of them developed into morphologically normal and fertile plants.

Efficient plant regeneration from immature embryo cultures of Jatropha curcas, a biodiesel plant

  • Varshney, Alok;Johnson, T. Sudhakar
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.139-148
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    • 2010
  • Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.