• Title/Summary/Keyword: induction of callus

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In vitro micropropagation of radish (Raphanus sativus L.) using callus induction and plant regeneration (캘러스 유기와 식물체 재분화를 이용한 무의 기내 대량증식)

  • You Kyoung Kim;Sug Youn Mo;Su Bin Choi;Han Yong Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.155-162
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    • 2023
  • Radish (Raphanus sativus L.), a root vegetable grown worldwide, is consumed in several ways. In the cross between parental lines to produce F1 seeds of radish, the problem of low purity may arise because of pollen contamination. Therefore, we aimed to establish conditions for callus induction and regeneration so that in vitro cultured plants could be used for the propagation of stock seeds. The most effective hormone combination containing various concentrations of 2,4-D, TDZ, and kinetin was selected for callus induction using radish hypocotyl, and the induced calli were transferred to two types of hormone media to investigate the optimal conditions for shoot regeneration of the callus. The combination of 1 mg/L 2,4-D + 0.05 mg/L kin was the most effective for callus induction of RA2 and RA10, 1 mg/L 2,4-D + 0.1 mg/L kin + 0.025 mg/L TDZ of RA4, and 1 mg/L 2,4-D + 0.2 mg/L kin of RA30. Shoot regeneration of the RA4 callus occurred in both shoot regeneration media, but the frequency was much higher in the 5H+1B medium (1 mg/L NAA + 0.1 mg/L 2,4-D + 1 mg/L IPA + 0.02 mg/L GA3 + 2 mg/L zeatin + 1 mg/L BA). For the in vitro micropropagation of radish, the conditions selected in this study can assist in the propagation and maintenance of stock seeds to produce F1 seeds.

Embryogenic Callus Induction and Plant Regeneration in Kentucky bluegrass (Poa pratensis L.) Native to Korea (자생 왕포아풀(Poa pratensis L.)의 배발생 캘러스 유도 및 식물체 재분화)

  • 이재신;심상렬;안병준
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.277-281
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    • 2001
  • Embryogenic callus induction and plant regeneration methods were developed for native Kentucky bluegrass (Poa pratenes L.) ecotypes. Mature caryopses and immature inflorescences (20 mm in length) of 4 native ecotypes and 5 foreign cultivars were plated on MS medium (30 g/L sucrose, 3 g/L Phytagel) supplemented with 1 mg/L 2,4-D, and cultured in the dark at 24$^{\circ}C$. Most explants formed calli, but more embryogenic calli were induced from the explants of immature inflorescences than caryopses which produced mostly non-embryogenic rooty calli. In P77 ecotypes, immature inflorescence explants formed embryogenic calli with the rate of 62~95%, and those of field-grown plants were more efficient than greenhouse-grown ones in embryogenic callus induction. Plantlets were regenerated from the embryogenic calli when they were transferred to hormone-free MS medium, and grew to maturity without morphological variations in greenhouse.

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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.

Plantlet Regeneration via Somatic Embryogenesis from Hypocotyls of Common Buckwheat (Fagopyrum esculentum Moench.)

  • Kwon, Soo-Jeong;Han, Myong-Hae;Huh, Yoon-Sun;Roy, Swapan Kumar;Lee, Chul-Won;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.4
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    • pp.331-335
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    • 2013
  • Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micro-propagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species, Fagopyrum esculentum which differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) $2.0mg{\cdot}L^{-1}$, benzyladenine (BA) $1.0mg{\cdot}L^{-1}$ and 3% sucrose. Friable callus was transferred to solidified MS media containing BA ($1.0mg{\cdot}L^{-1}$) with various concentrations of 2,4-dichlorophenoxyacetic acid for the induction of embryogenesis. The optimum concentrations of growth regulators (for regeneration of plantlet) were indole-3-acetic acid ($2.0mg{\cdot}L^{-1}$), Kinetin ($1.0mg{\cdot}L^{-1}$), BA ($1.0mg{\cdot}L^{-1}$). Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5% to 20%. Whole plants were obtained at high frequencies when the embryogenic calli with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. The main objective of this research was to develop an efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.

In vitro Induction of Tetraploid Roots by Various Pretreatments from Anther of Panax ginseng C. A. Meyer

  • Lee, Jung-Hye;Kim, Yu-Jin;Jung, Dae-Young;Shim, Ju-Sun;Kim, Ik-Hwan;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.33 no.1
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    • pp.65-71
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    • 2009
  • This experiment was done to determine the optimum conditions for the induction of tetraploidy in Panax ginseng C. A. Meyer using bud length, temperature and plant growth regulator pretreatments. Highest callus formation was obtained when the medium was inoculated with flower bud in the size of 2-3 mm in length. The optimum temperature for the callus formation was high when treated at $4^{\circ}C$ for 4-5 days. Among the treatments of growth regulators and different concentration, highest callus formation was observed in combination of 5 mg/L 2,4-D and 1 mg/L kinetin for P. ginseng. As a result of flow cytometer analysis, all 7 adventitious roots were confirmed as tetraploidys. Cytological analysis revealed that the chromosome number of tetraploid roots was 96, while that of diploid roots was 48. Tetraploid ginseng roots were inoculated to flower bud size of 2-3 mm in length. The callus formation was optimum when treated with 1 mg/L 2,4-D at $4^{\circ}C$ for 5 days. Compared with control roots, tetraploid roots were thicker and longer and had few lateral branches. Fresh weight of tetraploid roots was relatively higher than the control roots.

Plant Regeneration Through Adventitious Bud Formation and Callus Induction from Scales of Lilium lancifolium Thunb. (참나리 (Lilium lancifolium Thunb.) 인편으로부터 부정아 발생과 캘러스 유도를 통한 식물체 재생)

  • Nam, Sang-Wook;Kim, Hei-Young
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.53-58
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    • 2003
  • This study was performed to investigate the effects of various media compositions in regeneration of Lilium lancifolium. The adventitious bud initiation from microscale was the best on MS medium supplemented with BAP 1.0 mg/L and NAA 0.1 mg/L after 4 weeks of culture. However, from bulbscales, adventitious bud initiation was the best in dark condition on MS medium supplemented with BAP 0.5 mg/L and NAA 0.1 mg/L. On the other hand, callus induction was found to be the best from the microscales incubated in complete dark condition for 8 weeks on MS medium supplemented with 2,4-D 1.0 mg/L and BAP 0.1 mg/L. The highest plantlet regeneration from callus was obtained after incubation in the light condition for 8 weeks on MS medium supplemented with NAA 0.5 mg/L and BAP 0.1 mg/L. Rooting of shoots was obtained easily on MS medium and the plantlets were transferred to soil pots after 8 weeks. The chromosome analysis of the root tip cells was revealed that the callus-derived plantlets had normal chromosome number, 2n=24. No variation was observed in the morphology of the plantlets.

Effect of Plant Growth Regulators on Callus Induction and Plant Regeneration from Mature Seed Culture of Miscanthus sinensis (억새 성숙종자로부터 캘러스 유도 및 식물체 재분화에 있어서 식물생장호르몬의 영향)

  • Park, Choong-Hoon;Kim, Yong-Goo;Kim, Kyung-Hee;Alam, Iftekhar;Lee, Hyo-Jin;Sharmin, Shamima Akhtar;Lee, Ki-Won;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.4
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    • pp.291-298
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    • 2009
  • In order to optimize tissue culture conditions for genetic transformation of Miscanthus sinensis, we investigated the effects of different plant growth regulators on callus induction and plant regeneration using mature seeds as explant. Dehusked mature seeds were cultured on MS medium supplemented with 3 to 10 mg/L 2,4-D, dicamba or NAA, 30 g/L sucrose and 750 mg/L $MgCl_2{\cdot}6H_2O$. A number of combinations of auxin and cytokinin (BA, kinetin) were also used. MS medium containing 3 mg/L 2,4-D was found optimal for embryogenic callus induction (75.7%) from mature seed. The highest number of plants were regenerated (44.6%) upon transferring the embryogenic callus to MS medium supplemented with 1 mg/L 2,4-D plus 2 mg/L BA. This high efficient plant regeneration system could be useful to use for molecular breeding of new cultivars by genetic transformation.

Factors Affecting Callus Culture and Plant Regeneration in Kentucky Bluegrass (켄터키 블루그래스에 있어서 캘러스 배양 및 식물체 재분화에 미치는 요인의 영향)

  • Lee, K.W.;Lee, S.H.;Lee, D.G.;Woo, H.S.;Kim, D.H.;Choi, M.S.;Won, S.H.;Seo, S.;Lee, B.H.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.1067-1074
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    • 2005
  • In order to optimize tissue culture conditions of Kentucky bluegrass(Poa pratensis L.), effects of culture medium supplements, media and cultivars on embryogenic callus induction and regeneration of plants were investigated. MS medium containing 3mg/L 2,4-D and 0.1mg/L BA was optimal for embryogenic callus induction from mature seeds. The highest plant regeneration frequency(57.7%) was observed when the embryogenic calli were cultured on N6 medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Among several basic media, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. Genotype was an important factor in plant regenerability. ‘Newport’ showed to have higher regeneration frequency of 53.4%. Regenerated plants were grown normally when shoots transplanted to the soil. A short tissue culture period and high-frequency regeneration system would be beneficial for molecular breeding of Kentucky bluegrass through genetic transformation.

Regeneration and Acclimatization of Plants Derived from Anther Cultures in Carrot (Daucus carota L.) (당근 약배양에 의한 식물체 재분화 및 순화)

  • Cho, Moon-Soo;Juang, Ue-Dong;Park, Sang-Gyu;Park, Yong
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.47-52
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    • 2003
  • Anthers from several lines of carrot (Daucus carota L.) were plated on the semi-solid B$_{5}$, basal medium supplemented with 2,4-D and NAA at two concentrations, 1.0 and 2.0 mg/L plus 0.2 mg/L BAP (benzylaminop-urine). Anthers of the most lines on the B$_{5}$ basal medium with 2,4-D showed higher percentages of callus formation than those with NAA. Particularly, in line 45477, highest percentages of callus formation (50%) were observed on B$_{5}$ medium with 1.0 mg/L 2,4-D plus 0.2 mg/L BAP. With 1.0 mg/L 2,4-D, two months was sufficient for initiation of callus development. Calli were regenerated into plantlets through embryogenesis onto regeneration medium without any growth regulators. When callus showing yellowish and soft structure was cultured, it yielded green plants at high regeneration rates, The response of anthers in callus induction and plant regeneration was different among lines investigated. Optimal callus induction and plant regeneration could be obtained through manipulating the concentration of growth regulators. Plantlets after transfer to perlite were grown successfully in greenhouse conditions. Anther culture of carrot will be used as a useful breeding tool in future.

Efficient Plant Regeneration from Alfalfa Callus by Osmotic Stress Treatment (알팔파 캘러스로부터 삼투압 스트레스 처리에 의한 효율적인 식물체 재분화)

  • Kim, J.S.;Lee, D.G.;Lee, S.H.;Woo, H.S.;Lee, B.H.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.879-886
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    • 2004
  • Effects of culture mediwn supplements and osmotic stress treatment on embryogenic callus induction and somatic embryogenesis were investigated in order to optimize tissue culture conditions of alfalfa(Medicago sativa L.). SH mediwn containing 5mgIL 2,4-D and 0.2mgIL kinetin was optimal for embryogenic callus induction from cotyledon tissue of alfalfa. Somatic embryos were formed when the embryogenic callus was cultured on SH mediwn supplemented with ImgIL 2,4-D and 2mgIL BA. Supplementation of 5mM L-proline and IgIL casein hydrolysate into the regeneration mediwn further increased plant regeneration frequency. Osmotic stress treatment of callus appeared to improve the frequency of somatic embryo formation, but the frequency of somatic embryo formation differed by the osmotic stress treatment using different osmotic stressors. The highest plant regeneration frequency of 30.7% was observed when embryogenic callus was treated with 0.7M sucrose for 18h. Efficient regeneration system established in this study will be useful for molecular breeding of alfalfa through genetic transformation.