• Title/Summary/Keyword: Callus Induction

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Effect of nitrogen sources and 2, 4-D treatment on indirect regeneration of ginger (Zingiber officinale Rosc.) using leaf base explants

  • Mehaboob, Valiyaparambath Musfir;Faizal, Kunnampalli;Raja, Palusamy;Thiagu, Ganesan;Aslam, Abubakker;Shajahan, Appakan
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.17-21
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    • 2019
  • Ginger is an important monocotyledonous plant belonging to the family Zingiberaceae. The objective of this study was to investigate the regeneration potential of ginger using leaf base explants. Auxins such as 2, 4-D and NAA in combination with BA were used for initiation of callus. Different combinations of both ammonium ($NH^{4+}$) and nitrate ($NO^{3-}$) were also studied for efficient callus production. High frequency of white friable calli was observed on modified Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 2, 4-D, 0.5 mg/L NAA and 0.5 mg/L BA. The highest shoot induction (92.33%), shootlets number ($7.33{\pm}0.33$) and length ($88.33{\pm}4.40$) mm were achieved on MS media containing 0.5 mg/L BA. Regenerated shoots were transferred to in vitro rooting media containing 1.0 mg/L IBA. Afterwards, plantlets with well-developed root and shoot system were subjected to a twostep hardening process. 71% of plantlets survived after secondary hardening without any abnormal morphology.

Chromosome Doubling of Allium wakegi Araki by In Vitro Cultures (기내배양에 의한 쪽파의 체세포 염색체 배가)

  • Yim, Sun-Hee;Ahn, Chang-Soon;Jeong, Chang-Nam;Han, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.259-264
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    • 2002
  • Induction of embryogenic callus from Allium wakegi Araki explants was promoted on medium containing 2,4-D, and production of abnormal embryos from the embryogenic callus increased as 2,4-D concentration was raised. Shoot tip was found to be the best explant source for embryogenic callus formation followed in the order by bulb scale and leaf section. Medium containing 0.09 M sucrose was effective for embryogenic callus production. The regenerants from embryogenic callus on medium containing 2,4-D and BA at different concentrations was consisted of diploids, tetraploids and a few mixaploids of 2n+4n, and their chromosomal aberration rate ranged from 8.0 to 33.3%. Frequency of chromosomal aberrants was the highest (18.7%) in the regenerants obtained from bulb scale-derived embryogenic callus among others. Plant regeneration rate was high (33.5%) from the shoot tip-derived embryogenic callus and the frequency of chromosomal aberrants was very low (7.0%). The plantlets regenerated on medium containing 0.26 M sucrose resulted in high chromosomal aberrants. The regenerants on medium containing sucrose at 0.09∼0.20 M produced chromosomal aberrants at around 15.2∼16.6%.

In Vitro Production of Indian Citrs Ringspot Virus-Free Plants of Kinnow Mandarin (Citrus nobilis Lour X C. deliciosa Tenora) by Ovule Culture

  • Singh B.;Sharma S.;Rani G.;Zaidi A.A.;Hallan V.;Nagpal A.;Virk G.S.
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.259-265
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    • 2005
  • Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.

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.

Somatic Embryogenesis and Plant Regeneration from Poinsettia (Euphorbia pulcherrima L.) Stem Explants (포인세티아 줄기조직배양에 의한 재분화체 제조.)

  • Hee-Sung Park
    • Journal of Life Science
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    • v.8 no.6
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    • pp.623-626
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    • 1998
  • Conditions for somatic embryogenesis and plant regeneration from stem tissues of Euphorbia pulcherrima were esta-blished. Explants from leaf, petiole, stem were examined for their embryogenesis on MS solid medium supplemented with plant growth hormones in combination at various concentrations. From leaf or petiole explants, callus was indu-ced well but never proceeded to the embryonic stage in our expermental conditions. From stem explants, however, multiple shoots following callus induction emerged in about 6 to 8 weeks on MS agar medium supplemented with 1.5 mg/L of benzyladenine. Upon transfer, roots were developed on hormone-free MS solid medium.

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Determination Times for Induction of Adventitious Shoots, Roots, Trichomes, and Calli from Segments of Arabidopsis thaliana by NAA and BA (애기장대(Arabidopsis thaliana)잎 절편에서 NAA와 BA에 의한 신초, 부정근, 모용 및 캘러스 형성 결정 시기)

  • Kim, Song-Lim;Han, Tae-Jin
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.161-165
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    • 2003
  • The effect of NAA and Benzyladenine(BA) for determination times on the formation of adventitious shoots, roots trichmoes and calli in MS basal medium was investigated in leaf segments from ecotype 'Nosses' of Arabidopsis thalliana. Adventitious shoots, roots, trichomes and calli were formed fromed from leaf segments in a wede range of NAA and BA. The optimal combination of hormones for adventitious shoots formation, 20mg/L NAA for trichome formation, 100mg/L for callus formation. Inductive times for formation of adventitious shoots, roots, trichomes and calli were determined at 14, 4, 6 and 18 days respectively by periodical transfer of leaf segments from hormines containing media to hormone free medium.

Studies on the Induction of Transformation and Multiplication in Orchid Plants I. Formation of Somatic Embryos and Regeneration from Immature Seeds of Bletilla striata (난과식물의 형질전환 유도 및 다량증식에 관한 연구 I. 자란 (Bletilla striata)의 미성숙 종자로부터 체세포배 형성 및 식물체 재분화)

  • 이정석
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.271-276
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    • 1990
  • Our study was carried out for plant regeneration via somatic embryogenesis from immature seeds of Bletilla striata. The highest frequency of embryogenic callus formation was obtained from the immature seeds (at 150 days after pollination) cultured on Hyponex and VW medium supplemented with 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg/l kinetin under the dark condition. Multiple somatic embryos were induced when embryogenic callus was transferred to VW medium without growth regulators under continued illumination. Somatic embryos were observed histologically with scanning electron microscopy. Regeneration of Bletilla striata was obtained from somatic embryos with a well-defined scutellum and coleoptile as well as with one or more shoot primordia and root primordia. We think that these methods for orchid multiplication must be useful to access clonal propagation of orchids.

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Characteristics Evaluation of a Zoysiagrass Line '232'in the Tissue Culture and Field (종자 다수확계통'232'의 조직배양 및 포장에서의 특성 평가)

  • 이재필;김종보;임성희;주영규;김두환
    • Asian Journal of Turfgrass Science
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    • v.11 no.4
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    • pp.321-326
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    • 1997
  • A high seed yield '232' was evaluated for morphological characteristics and tissue culture re-sponse. The effect of mowing and urea application on coverage in '232' was also studied and the results were as follows ;1.'232' produced high seed yield than any other lines, but its other characteristics such as coverage, establishment rate, stolon length, stolon number and colour change were not good. 2.Callus induction of '232'(52.9%) was relatively high, and higher than that of Z. sinica(49.0%). However, embryogenic callus formation of '232' was not as good as that of Z. japonica and Z. sinica. 3.Coverage of '232' was very much enhanced by mowing on July 18, but not at all by mowing on May 29 and September 4. 4.Urea application did not increase the coverage in '232',Therefore, it can be used as a breeding material rather than direct use.

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Growth Regulators and Colchicine Treatments for Embryo Culture Efficiency in Barley (보리 배배양 효율증진을 위한 생장조절제와 콜히친처리 효과)

  • Bong Yeon, Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.6
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    • pp.757-767
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    • 1995
  • This experiment was done to determine the optimum concentration of IAA for root development in plants regenerated from the callus culture of barley embryos. Two concentrations of 2,4-D, 3ppm and 5ppm selected as an optimum among five different concentrations in the previous experiment were used for callus induction and proliferation in this experiment. For callus induction, 3ppm of 2,4-D produced 35.6% in immature embryos and 4.4% in mature embryos, while 5ppm gave 33.8% in immature and 5.6% in mature embryos. Out of 320 immature embryos cultured, 111 embryos were induced to calli and 684 plants were produced from them, while only 16 embryos were induced to calli from 320 mature embryos and 92 plants were restored. The rates of callusing and plant regeneration were 34.7%, 214% in immature embryos and 5.0%, 28.7% in mature embryos, respectively. The average root lengths and root numbers of plants restored from callus at five different IAA concentrations of 0ppm, Ippm, 5ppm, l0ppm and 30ppm were 7.9mm, 3.6; 18.4mm, 5.2; 16.1mm, 3.9; 8.5mm, 3.5 and 6.4mm, 3.4, while plants directly obtained from mature embryos were 14.8mm, 4.9; 4.9mm, 3.6; 4.3mm, 3.1; 3.6mm, 2.6 and 3.2mm, 2.1, respectively. Therefore, 1ppm gave the best result for the root. promotion in callus, whle 0ppm, a control, gave the largest root developmemt in embryos. High concentration of lAA(30ppm) in callus and any exogeneous supplement of lAA in embryos negatively affected to the root lengths and root numbers. Genotypic effect was also observed in given four varieties, Bruce, Klages, Olbori and Albori. For chromosome doubling, when 0.1% colchicine was applied on 428 plants under three different conditions such as air circulation, temperatures and growth stages, 319 plants of doubled haploids were obtained so that the rate was 74.5%

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Varietal Response of Tobacco Plants Through Tissue Culture to Butachlor and Bialaphos Herbicides (조직배양(組織培養)에 의한 제초제(除草劑) Butachlor와 Bialaphos에 대(對)한 담배의 품종간반응(品種間反應))

  • Bae, Y.Z.;Kim, K.U.;Jeong, H.J.
    • Korean Journal of Weed Science
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    • v.8 no.1
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    • pp.53-58
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    • 1988
  • This study was carried out to determine effect of butachlor [N.-(buthoxymethyl)-2-chloro-N-(2,6-diethylphenyl) acetamide] and bialaphos [2-amino-4(hydroxy)(methyl) phosphionyl] butyryl-alanylalanine sodium salt on the germination of tobacco seed, induction and growth of callus from tobacco. Further, fatty acids and ammonia content of tobacco calli were determined. Bialaphos had no effect on tobacco seed germination, but the growth of seedling was markedly affected by an application of 10 ppm bialaphos. However, regardless of varieties tested, tobacco seed germination was completely inhibited by $5{\times}10^{-5}M$ of butachlor. At an application of $5{\times}10^{-5}M$ butachlor, tobacco seeds were to some extent germinated and showed further growth. Hyangcho among varieties tested, showed the most tolerant response to butachlor. In induction of callus from various tobacco varieties and their growth, aromatic type of tobacco varieties exhibited the most tolerance against bialaphos. However, no distinct varietal differences were determined in the treatment of butachlor. The major fatty acids identified in tobacco calli were palmitic, oleic and linoleic acid. No marked difference in terms of fatty acids was observed among tobacco varieties used, but it was observed that there was the higher ratio of quantity in unsaturated fatty acids over saturated one, bialaphos treatment accumulated about 9 times higher ammonia content than that of the untreated control, giving an evidence that bialaphos might inhibit glutamine synthetase activity.

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