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Direct somatic embryogenesis as a rapid and efficient system for in vitro regeneration of Arabidopsis thaliana

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Abstract

A simple, rapid and effective system to regenerate Arabidopsis plants via direct somatic embryogenesis has been established. Somatic embryogenesis was induced directly during culture of immature zygotic embryos. The frequency of somatic embryogenesis was strongly influenced by the stage of development of the explants. Explants in different developmental stages were cultured on B5 agar medium containing 5 μM 2,4-dichlorophenoxyacetic acid and the highest frequency (up to 90%) of somatic embryogenesis was observed in zygotic embryos with fully-developed cotyledons. The first somatic embryos developing directly from explant tissue were noticed after 8 days of culture. Somatic embryogenesis of a high frequency (87–96%) was observed in cultures of the all six genotypes tested (Columbia, C-24, RLD, Wassilewskaja, Landsberg erecta and Wilna). Subculture of somatic embryos onto auxin-free medium resulted in their conversion into plants with an average frequency of 79.5%. The regenerates showed normal morphological characteristics and were fertile. All 56 analysed plants displayed a diploid number of chromosomes and two out of 96 (2.1%) tested plants carried a chlorophyll or embryo-lethal mutation.

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References

  • Acedo GN (1986) Regeneration of Arabidopsis callus in vitro. Plant Cell Tiss. Org. Cult. 6: 109–114

    Google Scholar 

  • Ahloowalia BS (1991) Somatic embryos in monocots. Their genesis and genetic stability. Rev. Cytol. Biol. Veget–Bot. 14: 223–235

    Google Scholar 

  • Akama K, Shiraishi H, Ohta S, Nakamura K, Okada K & Shimura Y (1992) Efficient transformation of Arabidopsis thaliana: comparison of the efficiencies with various organs, plant ecotypes and Agrobacterium strains. Plant Cell Rep. 12: 7–11

    Google Scholar 

  • Akashi R, Uchiyama T, Sakamato A, Kawamura O & Hoffmann F (1998) High–frequency embryogenesis from cotyledons of bird's foot trefoil (Lotus corniculatus) and its effective utilization in Agrobacterium tumefaciens–mediated transformation. J. Plant Physiol. 152: 84–91

    Google Scholar 

  • Bhojwani SS & Razdan MK (1996) Somatic embryogenesis. In: Bhojwani SS & Razdan MK (eds) Plant Tissue Culture: Theory and Practice, a Revised Edition (pp 125–166). Elsevier, Amsterdam, New York, Oxford

    Google Scholar 

  • Choi YE, Yang DC, Park JC, Soh WY & Choi KT (1998) Regenerative ability of somatic single and multiple embryos from cotyledons of Korean ginseng on hormone–free medium. Plant Cell Rep. 17: 544–551

    Google Scholar 

  • Cuenca B, San–Jose MC, Martinez MT, Ballester A & Vieitez AM (1999) Somatic embryogenesis from stem and leaf explants of Ouercus robur L. Plant Cell Rep. 118: 538–543

    Google Scholar 

  • Distabanjong K & Geneve RL (1997) Multiple shoot formation from normal and malformed somatic embryo explants of Eastern redbud (Cercis canadensis L.). Plant Cell Rep. 16: 334–338

    Google Scholar 

  • Dodeman VL, Ducreux G & Kreis M(1997) Zygotic embryogenesis versus somatic embryogenesis. J. Exp. Bot. 48: 1493–1509

    Google Scholar 

  • Duncan RR (1997) Tissue culture induced variation in crop improvement. In: Sparks DL (ed), Advances in Agronomy, Vol 58 (pp 201–240). Academic Press

  • Eady CC, Butler RC & Suo Y (1998) Somatic embryogenesis and plant regeneration from immature embryo cultures of onion (Alium cepa L.). Plant Cell Rep. 18: 111–116

    Google Scholar 

  • Engelborghs I, Swennen R & van Campenhout S (1998) The potential of AFLP to detect genetic differences and somaclonal variants in Musa spp. InfoMusa 7: 3–6

    Google Scholar 

  • Farooqui MA, Rao A, Jayasree V & Sadanandam A (1997) Induction of atrazine resistance and somatic embryogenesis in Solanum melongena. Theor. Appl. Genet. 95: 702–705

    Google Scholar 

  • Feldmann KA & Marks MD (1986) Rapid and efficient regeneration of plants from explants of Arabidopsis thaliana. Plant Science 47: 63–69

    Google Scholar 

  • Fouree JL, Barger P, Niquet L & Andre P (1997) Somatic embryogenesis and somaclonal variation in Norway spruce: morphogenetic, cytogenetic and molecular approaches. Theor. Appl. Genet. 94: 159–169

    Google Scholar 

  • Gaj MD & Maluszynski M (1987) Genetic variation in callusderived plants of Arabidopsis thaliana (L.) Heynh. Arabidopsis Inf. Serv 23: 1–8

    Google Scholar 

  • Gaj MD, Kucharska M, Maluszynski M & Polok K (1991) Isozyme variation in callus culture of Arabidopsis thaliana (L.) Heynh. Genetica Polonica 32: 217–225

    Google Scholar 

  • Gamborg OL, Miller RA & Ojima K (1968) Nutrient requirement of suspension cultures of soybean root cells. Exp Cell Res. 50: 151–158

    Google Scholar 

  • Goebel–Tourand I, Mauro MC, Sossontzov, Miginiac E & Deloire A (1993) Arrest of somatic embryo development in grapevine: histological characterization and the effect of ABA, BAP and zeatin in stimulating plant development. Plant Cell Tiss. Org. Cult. 33: 91–103

    Google Scholar 

  • Graaff E & Hooykaas PJJ (1996) Improvements in the transformation of Arabidospsis thaliana C24 leaf–discs by Agrobacterium tumefaciens. Plant Cell Rep. 15: 572–577

    Google Scholar 

  • Henry RJ (1998) Molecular and biochemical characterization of somaclonal variation. In: Jain SM, Brar DS & Ahloowalia BS (eds) Somaclonal Variation and Induced Mutations in Crop Improvement (pp 485–499). Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Henry RJ, Nato A & De Buyser J (1998) Genetic fidelity of plants regenerated from somatic embryos of cereals. In: Jain SM, Brar DS & Ahloowalia BS (eds) Somaclonal Variation and Induced Mutations in Crop Improvement (pp 65–80). Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Huang BC & Yeoman MM (1983) Formation of somatic embryos in tissue cultures of Arabidopsis thaliana. Arab Inf. Service 20: 73–78

    Google Scholar 

  • Kaldenhoff R, Henningsen U & Richter G (1994) Gene activation in suspension–cultured cells of Arabidopsis thaliana during bluelight–light dependent plantlet regeneration. Planta 195: 182–187

    Google Scholar 

  • Kintzios S, Manos C & Makri O (1999) Somatic embryogenesis from mature leaves of rose (Rosa sp.). Plant Cell Rep. 18: 467–472

    Google Scholar 

  • Luo Y & Koop HU (1997) Somatic embryogenesis in cultured immature zygotic embryos and leaf protoplasts of Arabidopsis thaliana ecotypes. Planta 202: 387–396

    Google Scholar 

  • Meijer EA, de Vries SC & Mordhorst AP (1999) Co–culture with Daucus carota somatic embryos reveals high 2,4–D uptake and release rates of Arabidopsis thaliana cultured cells. Plant Cell Rep. 18: 656–663

    Google Scholar 

  • Meinke DW (1991) Perspectives on genetic analysis of plant embryogenesis. The Plant Cell 3: 857–866

    Google Scholar 

  • Meinke DW (1995) Molecular genetics of plant embryogenesis. Annu. Rev. Physiol. Plant Mol. Biol. 46: 369–394

    Google Scholar 

  • Mordhorst AP, Toonen MAJ & de Vries SC (1997) Plant Embryogenesis. Crit. Rev. Plant Sci. 16: 535–576

    Google Scholar 

  • Mordhorst AP, Voerman KJ, Hartog MV, Meijer EA, VanWent J, Koornneef M & deVries SC (1998) Somatic embryogenesis in Arabidopsis thaliana is facilitated by mutation in genes repressing meristematic cell divisions. Genetics 149: 549–563

    Google Scholar 

  • Müller AJ (1963) Embryonentest zum Nachweis rezessiver Letafaktoren bei Arabidopsis thaliana. Biologischen Zentralblatt 82: 133–163

    Google Scholar 

  • Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 437–497

    Google Scholar 

  • O'Neill CM & Mathias RJ (1993) Regeneration of plants from protoplasts of Arabidopsis thaliana L. cv.Columbia (C24) via direct embryogenesis. J. Exp. Bot. 44: 1579–1585

    Google Scholar 

  • Pillon E, Terzi M, Baldan B, Mariani P & Schiavo FL (1996) A protocol for obtaining embryogenic cell lines from Arabidopsis. The Plant J. 9: 573–577

    Google Scholar 

  • Plader W, Malepszy S, Burza W & Rusinowski Z (1998) The relationship between the regeneration system and genetic variability in the cucumber (Cucumis sativus L.) Euphytica 103: 9–15

    Google Scholar 

  • Rival A, Bertrand L, Beule T, Combes MC, Trouslot P & Lashermes P (1998) Suitability of RAPD analysis for the detection of somaclonal variants in oil palm (Elaeis guineesis Jacq). Plant Breeding 117: 73–76

    Google Scholar 

  • Sangwan RS, Bourgeois Y, Dubois F & Sangwan–Norreel BS (1992) In vitro regeneration of Arabidopsis thaliana from cultured zygotic embryos and analysis of regenerates. J. Plant. Physiol. 140: 588–595

    Google Scholar 

  • Thakur RC, Goto S, Ishii K & Jain SM (1999) Monitoring genetic stability in Quercus serrata Thunb. somatic embryogenesis using RAPD markers. J. For. Res, 4: 157–160

    Google Scholar 

  • Valvekens D, van Montagu M & van Lijsebettens M (1988) Agrobacterium tumefaciens–mediated transformation of Arabidopsis thaliana root explants by using kanamacin selection. Proc. Natl. Acad. Sci. USA 85: 5536–5540

    Google Scholar 

  • Vendrame WA, Kochert G & Wetzstein HY (1999) AFLP analysis of variation in pecan somatic embryos. Plant Cell. Rep. 18: 853–857

    Google Scholar 

  • Wu Y, Haberland G, Zhou C & Koop HK (1992) Somatic embryogenesis, formation of morphogenic callus and normal development in zygotic embryos of Arabidopsis thaliana in vitro. Protoplasma 169: 89–96

    Google Scholar 

  • Yasutani I, Shoichi O, Nishida T, Sugiyama M & Kamamine A (1994) Isolation of tempereture–sensitive mutants of Arabidopsis thaliana that are defective in the redifferentation of shoots. Plant Physiol. 105: 815–822

    Google Scholar 

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Gaj, M.D. Direct somatic embryogenesis as a rapid and efficient system for in vitro regeneration of Arabidopsis thaliana. Plant Cell, Tissue and Organ Culture 64, 39–46 (2001). https://doi.org/10.1023/A:1010679614721

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