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Embryo induction and regeneration from root explants of Medicago truncatula after osmotic pre-treatment

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Embryo induction and regeneration from suspension culture of two Medicago truncatula cvs. (cv. R 108 1 and cv. Jemalong) have been studied. The influence of osmotic pre-treatment (1 M solution of sucrose for 48 h and 72 h) of roots as an initial explant, on embryogenic efficiency of the suspension culture was assessed. In comparison to the control, the level of abscisic acid (ABA) increased significantly after osmotic stress. The increased ABA level did not correlate with the induction of embryogenesis neither with the improved embryogenic potential of cv. R 108 1. The shortest regeneration period and the highest percent of conversion to plants were found in cv. R 108 1 after 72-h pre-treatment of roots. The efficiency of somatic embryo conversion was less after 48-h pre-treatment and much less for the untreated control. Osmotic stress did not positively affect the process of embryogenesis from root explants of cv. Jemalong, confirming its cultivar dependence. A single cell suspension fraction was produced in both Medicago trunacatula cvs. during the somatic embryo maturation stage. A higher embryogenic potential than the initial suspension culture was established only for the cell suspension originating from 72-h pre-treated roots of cv. R 108 1. The data confirms that the process of somatic embryo induction and embryo conversion from root explants of cv. R 108 1 could be promoted by osmotic stress pre-treatment.

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abscisic acid


dichlorophenoxyacetic acid




  1. A Avjioglu RB Knox (1989) ArticleTitleStorage lipid accumulation by zygotic and somatic embryos in culture Ann. Bot. 63 409–420

  2. DCW Brown A Atanassov (1985) ArticleTitleRole of genetic background in somatic embryogenesis in Medicago Plant Cell Tiss. Org. Cult. 4 111–122

  3. M Chabaud MC Larsonneau T Huguet (1996) ArticleTitleTransformation of barrel medicsMedicago truncatula Gaertn. by Agrobacterium tumefaciens and regeneration via somatic embryogenesis of transgenic plants with the MtENOD 12 nodulin promoter fused to the gus reporter gene Plant Cell Rep. 15 305–310

  4. HH Chen H Li ML Brenner (1983) ArticleTitleInvolvement of abscisic acid in potato cold acclimation Plant Physiol. 71 362–365

  5. K Chengalrayan VB Mhaske S Hazra (1998) ArticleTitleGenotypic control of peanut somatic embryogenesis Plant Cell Rep. 17 522–525

  6. GC Churchill B Ewan MJT Reaney SR Abrams LV Gusta (1992) ArticleTitleStructure-activity relationships of abscisic acid analogs based on the induction of freezing tolerance in bromegrass (Bromus enermis Leyss) cell cultures Plant Physiol. 100 2024–2029

  7. DR Cook (1999) ArticleTitleMedicago truncatula – a model in the making! Current Opinion in Plant Biol. 2 301–304

  8. ML Dahmer DF Hildebrand GB Collins (1992) ArticleTitleComparative protein accumulation pattern in soybean somatic and zygotic embryos In Vitro Cell Dev. Biol. 28 106–111

  9. E Epstein JD Cohen (1981) ArticleTitleMicroscale preparation of pentafluorobenzyl esters: electron-capture gas chromatographic detection of Indole-3-acetic Acid from plants J. Chromatogr. 209 413–420

  10. H Etienne P Montorro N Michaux-Ferriere MP Carron (1993) ArticleTitleEffect of desiccation, medium osmolarity and abscisic acid on the maturation of Hevea brasilensis somatic embryos J. Exp. Bot. 44 1613–1619

  11. OL Gamborg RA Miller K Ojima (1968) ArticleTitleNutrient requirements of suspension cultures of soybean root cells Exp. Cell Res. 50 151–158

  12. G Gangopadhyay S Basu S Gupta (1997) ArticleTitleIn vitro selection and physiological characterization of NaCl – and manitol-adapted callus lines in Brassica juncea Plant Cell Tiss. Org. Cult. 50 161–169

  13. MA Huhhes MA Dunn RS Pearce AJ White L Zhang (1992) ArticleTitleAn abscisic- acid –response, low temperature barley gene has homology with a maize phospholipid transfer protein Plant Cell Environ. 15 861–865

  14. Iantcheva A, Thrin H, Brown S & Atanassov A (2001b) Early events in direct somatic embryo formation from single cells in Medicago truncatula observed using green fluorescent protein. Second meeting of the COST 843 WG3. Quality Enhancement of Plant Production through Tissue Culture. April 19–21, Carcavelos, Portugal (pp. 1–2)

  15. A Iantcheva M Vlahova E Bakalova E Kondorosi MC Elliott A Atanassov (1999) ArticleTitleRegeneration of diploid annual medics via direct somatic embryogenesis promoted by thidiazuron and benzylaminopurine Plant Cell Rep. 18 904–910

  16. A Iantcheva M Vlahova H Trinh S Brown A Slater M Elliott A Atanassov (2001a) ArticleTitleAssessment of polysomaty, embryogenic potential, embryo formation and regeneration for various species of diploid annual Medicago Plant Sci 160 4621–4627

  17. TL Kahn SE Fender EA Bray MA ÓConnell (1993) ArticleTitleCharacterization of expression of drought and abscisic acid – regulated tomato genes in the drought – resistant species Lycopersicon pennellii Plant Physiol. 103 597–605

  18. H Kamada K Kobayashi T Kiyosue H Harada (1989) ArticleTitleStress induced somatic embryogenesis in carrot and its application to synthetic seed production In Vitro Cell. Dev. Biol Plant 25 1163–1166

  19. K Kamaté ID Rodrigues-Llorente M Scholte P Durand P Ratet E Kondorosi A Kondorosi TH Trinh (2000) ArticleTitleTransformation of floral organs with GFP in Medicago truncatula Plant Cell Rep. 19 647–653

  20. VD Kumar PB Kirti JKS Sachan VL Chopra (1994) ArticleTitlePlant regeneration via somatic embryogenesis in chickpea (Cicer arietinum L.) Plant Cell Rep. 13 468–472

  21. CG Lecouteux F Lai BD Mckersie (1993) ArticleTitleMaturation of alfalfa (Medicago sativa L.) somatic embryos by abscisic acid, sucrose and chilling stress Plant Sci. 94 207–213

  22. H Lou S Kako (1995) ArticleTitleRole of high sugar concentrations in inducing somatic embryogenesis from cucumber cotyledons Sci. Hort. 64 11–20

  23. NL McLean J Nowak (1998) ArticleTitleInheritance of somatic embryogenesis in red clover (Trifolium pratense L.) Theor. Appl. Genet. 97 557–562

  24. VB Mhaske S Hazra (1994) ArticleTitleAppearance of storage lipids (triglycerides) in somatic embryos of peanut (Arachis hypogaea L.) In Vitro Cell Dev. Biol. 30 113–116

  25. VB Mhaske K Chengalrayan S Hazra (1998) ArticleTitleInfluence of osmotica and abscisic acid on triglyceride accumulation in peanut somatic embryos Plant Cell Rep. 17 742–746

  26. T Murashige F Skoog (1962) ArticleTitleA revised medium for rapid growth and bioassays with tobacco tissue cultures Physiol. Plant 15 473–497

  27. JJJ Ooms KM Leon-Kloosterziel D Bartels M Koornneef CM Karssen (1986) ArticleTitleAcquisition of desiccation tolerance and longevity in seeds of Arabidopsis thaliana. A comparative study using abscisic acid-insensitive abi 3 mutants Plant. Physiol. 126 23–32

  28. VC Pence PM Hasegawa J Janik (1981) ArticleTitleSucrose mediated regulation of fatty acid composition in asexual embryos of Theobroma cacao Physiol. Plant. 53 378–384

  29. E Prinsen S Van Laer S Oden H Van Onckelen (2000) Auxin analysis G.A Tucker Roberts (Eds) Methods in Molecular Biology. Plant Hormone Protocols NumberInSeries141 Humana Press Inc. Totowa NJ

  30. MJT Reaney LV Gusta (1987) ArticleTitleFactors influencing the induction of freezing tolerance by abscisic acid in cell suspension culture of Bromus enermis Leyss and Medicago sativa L Plant Physiol. 83 423–427

  31. KGB Santos E Mudstock MH Bodanese-Zanettini (1997) ArticleTitleGenotype-specific normalization of soybean somatic embryogenesis through the use of ethylene inhibitor Plant Cell Rep. 16 859–864

  32. T Senaratna PK Saxena MV Rao J Afela (1995) ArticleTitleSignificance of the zygotic seed coat on quiescence and desiccation tolerance of Medicago sativa L. somatic embryos Plant Cell Rep. 14 375–379

  33. RE Sharp Y Wu GS Voetberg IN Saab ME LeNoble (1994) ArticleTitleConfirmation that abscisic acid accumulation is required for maize primary root elongation at low water potential J. Exp. Bot. 45 1743–1751

  34. K Shetty BD McKersie (1993) ArticleTitleProline, thioproline and potassium mediated stimulation of somatic embryogenesis in alfalfa (Medicago sativa L.) Plant Sci. 88 185–193

  35. RC Shoemaker EG Hammond (1988) ArticleTitleFatty acid composition of soybean (Glycine max (L.) Merr) somatic embryos In Vitro Cell Dev. Biol. 24 829–832

  36. AT Trieu MJ Harrisson (1996) ArticleTitleRapid transformation of Medicago truncatula via shoot organogenesis Plant Cell Rep. 16 6–11

  37. TH Trinh P Ratet E Kondorosi P Durand K Kamate P Bauer A Kondorosi (1998) ArticleTitleRapid and efficient transformation of diploid Medicago truncatula and Medicago sativa ssp. falcata lines improved in somatic embryogenesis Plant Cell Rep. 17 345–355

  38. DC Walton Y Li (1995) Abscisic acid biosynthesis and metabolism PJ Davies (Eds) Plant Hormones Kluwer Academic Publishers Dordrecht 140–157

  39. Z Xin PH Li (1993) ArticleTitleRelationship between proline and abscisic acid in the induction of chilling tolerance in maize suspension cultured cells Plant. Physiol. 103 607–613

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Correspondence to Anelia Iantcheva.

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Iantcheva, A., Slavov, S., Prinsen, E. et al. Embryo induction and regeneration from root explants of Medicago truncatula after osmotic pre-treatment. Plant Cell Tiss Organ Cult 81, 37–43 (2005). https://doi.org/10.1007/s11240-004-2774-x

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  • Medicago truncatula
  • osmotic stress
  • regeneration
  • root