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Evolution of endogenous hormone concentration in embryogenic cultures of carrot during early expression of somatic embryogenesis

  • Physiology and Biochemistry
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Abstract

Embryogenic callus and suspension cultures of carrot (Daucus carota L., cv. Nantaise), growing on/in medium including 1 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D), were transferred to medium with or without this plant growth regulator, to impair or induce, respectively, further development of somatic embryos. The endogenous hormone levels of the cultures were determined over 7 days by means of radio-immunoassay, to characterize their evolution in the initial stages of embryo development. In general, levels of indoleacetic acid (IAA) and abscisic acid (ABA) showed only short-lived differences among treatments during this time in both types of tissue analyzed (i.e., a peak of IAA in callus cultures in the absence of 2,4-D, 48 h after medium change, and higher ABA contents 144 h after subculture of suspension cultures in the presence of 2,4-D). Gibberellins (1, 3 and 20) were detected only in suspension cultures devoid of 2,4-D, starting 24 h after subculture. Concerning the evaluated cytokinins—zeatin/zeatin riboside and N62-isopentenyl) adenine/N62-isopentenyl) adenosine—the most remarkable observation is that high levels of the former generally coincided with low concentrations of the latter, indicating a shift from precursor to the active form, and vice versa.

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References

  • Bertling I, Bangerth F (1995) Changes in hormonal pattern of the new growth of Sclerocarya birrea after rejuvenation treatment with GA3 and ‘heading back’. Gartenbauwissenschaft 60:119–124

    CAS  Google Scholar 

  • Centeno ML, Rodríguez R, Berros B, Rodríguez A (1997) Endogenous hormonal content and somatic embryogenic capacity of Corylus avellana L. cotyledons. Plant Cell Rep 17:139–144. DOI 10.1007/s002990050366

    Article  CAS  Google Scholar 

  • Chen C-M (1997) Cytokinin biosynthesis and interconversion. Physiol Plant 101:665–673. DOI 10.1034/j.1399-3054.1997.1010402.x

    Article  CAS  Google Scholar 

  • Dodeman VL, Ducreux G (1996) Total protein pattern expression during induction and development of carrot somatic embryos. Plant Sci 120:57–69. DOI 10.1016/S0168-9452(96)04487-1

    Article  CAS  Google Scholar 

  • Einset JW (1986) Zeatin biosynthesis from N6-(Δ2-isopentenyl) adenine in Actinidia and other woody plants. Proc Natl Acad Sci USA 83:972–975

    CAS  Google Scholar 

  • Fehér A, Pasternak TP, Dudits D (2003) Transition of somatic plant cells to an embryogenic state. Plant Cell Tissue Organ Cult 74:201–228. DOI 10.1023/A:1024033216561

    Article  Google Scholar 

  • Fujii N (1997) Pattern of DNA binding of nuclear proteins to the proximal Agrobacterium rhizogenes rolC promoter is altered during somatic embryogenesis of carrot. Gene 201:55–62. DOI 10.1016/S0378-1119(97)00427-7

    Article  CAS  PubMed  Google Scholar 

  • Fujimura T, Komamine A (1979) Synchronization of somatic embryogenesis in a carrot suspension culture. Plant Physiol 64:162–164

    Google Scholar 

  • Fujimura T, Komamine A (1980) Mode of action of 2,4-D and zeatin on somatic embryogenesis in a carrot cell suspension culture. Z Pflanzenphysiol 99:1–8

    CAS  Google Scholar 

  • Giuliano G, Rosellini D, Terzi M (1983) A new method for purification of the different stages of carrot embryoids. Plant Cell Rep 2:216–218

    Google Scholar 

  • Grieb B, Schäfer F, Imani J, Mashayekhi KN, Arnholdt-Schmitt B, Neumann KH (1997) Changes in soluble proteins and phytohormone concentrations of cultured carrot petiole explants during induction of somatic embryogenesis (Daucus carota L.). J Appl Bot 71:94–103

    CAS  Google Scholar 

  • Guzzo F, Baldan B, Levi M, Sparvoli E, Lo Schiavo F, Terzi M, Mariani P (1995) Early cellular events during induction of carrot explants with 2,4-D. Protoplasma 185:28–36

    CAS  Google Scholar 

  • Guzzo F, Cantamessa K, Portaluppi P, Levi M (2002) Flow cytometry and sorting of protoplasts from carrot cell cultures reveal two cell subpopulations with different morphogenetic potential. Plant Cell Rep 21:214–219. DOI 10.1007/s00299-002-0519-z

    Article  CAS  Google Scholar 

  • Hatzopoulos P, Fong F, Sung ZR (1990) Abscisic acid regulation of DC8, a carrot embryogenic gene. Plant Physiol 94:690–695

    CAS  Google Scholar 

  • Jiménez VM, Bangerth F (2001) Endogenous hormone levels in explants and in embryogenic and non-embryogenic cultures of carrot. Physiol Plant 111:389–395. DOI 10.1034/j.1399-3054.2001.1110317.x

    Article  PubMed  Google Scholar 

  • Jiménez VM, Guevara E, Herrera J, Bangerth F (2001) Endogenous hormone levels in habituated nucellar Citrus callus during the initial stages of regeneration. Plant Cell Rep 20:92–100. DOI 10.1007/s002990000280

    Article  Google Scholar 

  • Kamada H, Harada H (1981) Changes in the endogenous level and effects of abscisic acid during somatic embryogenesis of Daucus carota L. Plant Cell Physiol 22:1423–1429

    CAS  Google Scholar 

  • Kitamiya E, Suzuki S, Sano T, Nagata T (2000) Isolation of two genes that were induced upon the initiation of somatic embryogenesis on carrot hypocotyls by high concentrations of 2,4-D. Plant Cell Rep 19:551–557. DOI 10.1007/s002990050772

    Article  CAS  Google Scholar 

  • Kiyosue T, Nakajima M, Yamaguchi I, Satoh S, Kamada H, Harada H (1992) Endogenous levels of abscisic acid in embryogenic cells, non-embryogenic cells and somatic embryos of carrot (Daucus carota L.). Biochem Physiol Pflanzen 188:343–347

    CAS  Google Scholar 

  • Kiyosue T, Satoh S, Kamada H, Harada H (1993) Somatic embryogenesis in higher plants. J Plant Res 3(Special Issue):75–82

    Google Scholar 

  • Komamine A, Kawahara R, Matsumoto M, Sunabori S, Toya T, Fujiwara A, Tsukuhara M, Smith J, Ito M, Fukuda H, Nomura K, Fujimura T (1992) Mechanisms of somatic embryogenesis in cell cultures: physiology, biochemistry, and molecular biology. In Vitro Cell Dev Biol Plant 28P:11–14

    CAS  Google Scholar 

  • Michalczuk L, Cooke TJ, Cohen JD (1992a) Auxin levels at different stages of carrot embryogenesis. Phytochemistry 31:1097–1103. DOI 10.1016/0031-9422(92)80241-6

    Article  CAS  Google Scholar 

  • Michalczuk L, Ribnicky DM, Cooke TJ, Cohen JD (1992b) Regulation of indole-3-acetic acid biosynthetic pathways in carrot cell cultures. Plant Physiol 100:1346–1353

    CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    CAS  Google Scholar 

  • Nagamori E, Omote M, Honda H, Kobayashi T (2001) Enhanced and prolonged production of plantlets regenerated from carrot callus in a viscous additive-supplemented medium. J Biosci Bioeng 91:283–287. DOI 10.1016/S1389-1723(01)80135-3

    Article  CAS  Google Scholar 

  • Noma M, Huber J, Ernst D, Pharis RP (1982) Quantitation of gibberellins and the metabolism of [3H]gibberellin A1, during somatic embryogenesis in carrot and anise cell cultures. Planta 155:369–376

    CAS  Google Scholar 

  • Nomura K, Komamine A (1985) Identification and isolation of single cells that produce somatic embryos at a high frequency in a carrot suspension culture. Plant Physiol 79:988–991

    CAS  Google Scholar 

  • Nuti-Ronchi V, Giorgetti L (1995) The cell’s commitment to somatic embryogenesis. In: Bajaj YPS (ed) Somatic embryogenesis and synthetic seed. I. Biotechnology in agriculture and forestry, vol 30. Springer, Berlin Heidelberg New York, pp 3–19

    Google Scholar 

  • Osuga K, Masuda H, Komamine A (1999) Synchronization of somatic embryogenesis at high frequency using carrot suspension cultures: model systems and application in plant development. Methods Cell Sci 21:129–140. DOI 10.1023/A:1009884806166

    Article  CAS  PubMed  Google Scholar 

  • Ribnicky DM, Ilic N, Cohen JD, Cooke TJ (1996) The effects of exogenous auxins on endogenous indole-3-acetic acid metabolism. Plant Physiol 112:549–558

    CAS  PubMed  Google Scholar 

  • Ribnicky DM, Cohen JD, Hu W-S, Cooke TJ (2002) An auxin surge following fertilization in carrots: a mechanism for regulating plant totipotency. Planta 214:505–509. DOI 10.1007/s004250100639

    Article  CAS  PubMed  Google Scholar 

  • Ross JJ (1998) Effects of auxin transport inhibitors on gibberellins in pea. J Plant Growth Regul 17:141–146

    CAS  Google Scholar 

  • Sasaki K, Shimomura K, Kamada H, Harada H (1994) IAA metabolism in embryogenic and non-embryogenic carrot cells. Plant Cell Physiol 35:1159–1164

    CAS  Google Scholar 

  • Sato-Nara K, Fukuda H (2000) The rates of deceleration of nuclear and organellar DNA syntheses differ in the progenitor cells of the apical meristems during carrot somatic embryogenesis. Planta 211:457–466. DOI 10.1007/s004250000317

    Article  CAS  PubMed  Google Scholar 

  • Schiavone FM, Cooke TJ (1985) A geometric analysis of somatic embryo formation in carrot cell cultures. Can J Bot 63:1573–1578

    Google Scholar 

  • Schrader S, Kaldenhoff R, Richter G (1997) Expression of novel genes during somatic embryogenesis of suspension-cultured carrot cells (Daucus carota). J Plant Physiol 150:63–68

    CAS  Google Scholar 

  • Seitz HU, Seitz U, Alfermann W (1985) Pflanzliche Gewebekultur: ein Praktikum. Gustav Fischer, Stuttgart, p 114

    Google Scholar 

  • Sharma AK (1999) Synchronization in plant cells—an introduction. Methods Cell Sci 21:73–78. DOI 10.1023/A:1009828419370

    Article  CAS  PubMed  Google Scholar 

  • Stasolla C, Loukanina N, Ashihara H, Yeung EC, Thorpe TA (2003) Changes in deoxyribonucleotide biosynthesis during carrot somatic embryogenesis. Plant Physiol Biochem 41:779–785. DOI 10.1016/S0981-9428(03)00122-0

    Article  CAS  Google Scholar 

  • Tokuji Y, Kuriyama K (2003) Involvement of gibberellin and cytokinin in the formation of embryogenic cell clumps in carrot (Daucus carota). J Plant Physiol 160:133–141. DOI 10.1078/0176-1617-00892

    CAS  PubMed  Google Scholar 

  • Vu JCV, Niedz RP, Yelenosky G (1993) Glycerol stimulation of chlorophyll synthesis, embryogenesis and carboxylation and sucrose metabolism enzymes in nucellar callus of ‘Hamlin’ sweet orange. Plant Cell Tissue Organ Cult 33:75–80

    CAS  Google Scholar 

Download references

Acknowledgements

The authors thank the German Academic Exchange Service (DAAD) for financial support in the form of short-term scholarships. The skilled work of A. Azofeifa, E. Tavares and S. Masís, conducting part of the in vitro culture, is gratefully acknowledged.

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Correspondence to V. M. Jiménez.

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Communicated by D. Dudits

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Jiménez, V.M., Guevara, E., Herrera, J. et al. Evolution of endogenous hormone concentration in embryogenic cultures of carrot during early expression of somatic embryogenesis. Plant Cell Rep 23, 567–572 (2005). https://doi.org/10.1007/s00299-004-0869-9

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  • DOI: https://doi.org/10.1007/s00299-004-0869-9

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