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In vitro selection and physiological characterization of NaCl- and mannitol-adapted callus lines in Brassica juncea

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Abstract

Na Cl (salt)- and mannitol (drought)- tolerant callue lines of Brassica juncea (L) Czern. Var. RW-85-59 were isolated by a plated cell suspension culture technique against 43 mM NaCl and 165 mM mannitol, respectively. Callus lines, adapted to a high concentration of Na Cl (171 mM) and mannitol (329 mM) were then established bv direct adaptation procedures. In the initial passages, the calluses showed severe reduction in tissue growth when grown on NaCl/mannitol-containing media but growth of adapted calluses recovered and was sustainable in the subsequent passages. Adapted calluses showed considerable accumulation of free proline in NaCl-/mannitol- containing media compared to the control callus grown on stress-free medium. A significant increase of intensity of one particular acid phosphatase isozymic band in the adapted calluses, irrespective of NaCl or mannitol stress, indicated that it may be used as an osmotic stress-marker in this system. Short-term salt/osmotic-shock-treatment with high concentrations of osmotica revealed that only the adapted lines retained the maximum amount of free proline within the cells for osmoregulation. This response probably helped the cells to restore their normal growth when the stress was withdrawn.

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References

  • Bates LS, Waldren RP & Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39: 205–207

    Article  CAS  Google Scholar 

  • Chandler SF & Thorpe TA (1987) Characterization of growth, water relations and proline accumulation in sodium sulfate tolerant callus of Brassica napus L. cv. Westar (Canola). Plant Physiol. 84: 106–111

    PubMed  CAS  Google Scholar 

  • Collin HA & Dix PJ (1990) Culture system and selection procedure. In: Dix PJ (ed.) Plant Cell Line Selection (pp 3–18). VCH, Weinheim, VCH, NY

    Google Scholar 

  • Croughan TP, Stavarek SJ & Rains DW (1981) In vitro development of salt resistant plants. Env. Exptl. Bot. 21: 311–324

    Google Scholar 

  • Cushman JC, DeRocher EJ & Bohnert HJ (1990) Gene expression during adaptation to salt stress. In: Katterman F (ed) Environmental Injury to Plants. (pp 173–203). Academic Press, Inc., San Diego

    Google Scholar 

  • Eberhardt HJ & Wegmann K (1989) Effects of abscisic acid and proline on adaptation of tobacco callus cultures to salinity and osmotic shock. Physiol. Plant. 76: 283–288

    CAS  Google Scholar 

  • Fallon KM & Phillips R (1989) Responses to water stress in adapted and unadapted carrot cell suspension cultures. J. Exptl. Bot. 40(215): 681–687

    CAS  Google Scholar 

  • Gangopadhyay G, Basu S, Mukherjee SP, Poddar R, Gupta S & Mukherjee BB (1995) Water, salt and freezing stresses: Effect on relative water content, viability and banding patterns of some isozymes in Brassica juncea (L) Czern. callus. Ind. J. Plant Physiol. XXXVIII (1): 41–44

    Google Scholar 

  • Gomes-Filho E & Sodek L (1988) Effect of salinity on ribonuclease activitv of Vigne unguiculata cotyledons during germination. J. Plant Physiol. 132: 307–311

    CAS  Google Scholar 

  • Greensway H & Munns R (1980) Mechanism of salt tolerance in non-halophytes. Ann. Rev. Plant Physiol. 31: 149–190

    Article  Google Scholar 

  • Harrington HM& Alm DM(1988) Interaction of heat and salt shock in cultured tobacco cells. Plant Physiol. 88: 618–625

    PubMed  CAS  Google Scholar 

  • Jain RK, Jain S, Nainawatee HS & Chowdhury JH (1990) Salt tolerance in Brassica juncea L. 1. In vitro selection, agronomic evaluation and genetic stability. Euphytica 48: 141–152

    Article  Google Scholar 

  • Jain RK, Jain S & Chowdhury JB (1991a) In vitro selection for salt tolerance in Brassica juncea L. using cotyledon explants, callus and cell suspension cultures. Annals Bot. 67: 517–519

    Google Scholar 

  • Jain S, Nainawatee HS, Jain RK & Chowdhury JB (1991b) Proline status of genetically stable salt-tolerant Brassica juncea L. somaclone and their parent cv. Prakash. Plant Cell Rep. 9: 684–687

    Article  CAS  Google Scholar 

  • Kavi Kishor PB, Hong Z, Miao G-H, Hu C-AA & Verma DPS (1995) Overexpression of -Δ1Pyrrilone-5-carboxylate synthetase increases proline productlon and confers osmotolerance in transgenic plants. Plant Physiol. 108: 1387–1394

    Google Scholar 

  • Kumar D (1984) The value of certain parameters as an index for salt tolerance in Indian mustard (Brassice juncea L.) Plant Soil 79: 261–271

    Article  CAS  Google Scholar 

  • Kumar V & Sharma DR (1989) Isolation and characterisation of sodium chloride-resistant callus cultures of Vigna radiata (L.) Wilezek-var. radiata. J. Exptl. Bot. 40(210): 143–147

    Google Scholar 

  • Leone A, Costa A, Tucci M & Grillo S (1994) Adaptation versus shock response to polyethylene glycol-induced low water potential in cultured potato cells. Physiol. Plant. 92: 21–30

    Article  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL & Randall RJ (1951) Protein measurements with the folin phenol reagent. J. Biol. Chem. 193: 265–275

    PubMed  CAS  Google Scholar 

  • McCoy TJ (1981) Characterization of alfalfa (Medicago sativa L.) plants regenerated from selected NaCl tolerant cell lines. Plant Cell Rep. 6: 411–422

    Google Scholar 

  • McHughen A (1987) Salt tolerance through increased vigour in a flax line (STS-II) selected for salt tolerance in vitro. Theor. Appl. Genet. 74: 727–732

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Panse VG & Sukhatme PT (1961) Statistical methods for agricultural workers. ICAR, New Delhi

    Google Scholar 

  • Pasteur N, Pasteur G, Bonhomne F, Catalan J & Britton-Davidian J (1988) Practical Isozyme Genetics (pp. 99). John Wilev & Sons, NY

    Google Scholar 

  • Rains DW (1989) Plant tissue and protoplast culture: Applications to stress physiology and biochemistry. In: Jones HG, Flowers TJ& Jones MB (eds.) Plants Under Stress. Society for Experimental Biology, Seminar series 39 (pp 181–196). Cambridge University Press

  • Rudulier DL, Strom AR, Dandekar AM, Smith LT & Valentine RC (198l) Molecular biology of osmoregulation. Science 224: 1064–1068

  • Skriver K & Mundy J (1990) Gene expresion in response to abscisic acid and osmotic stress. Plant Cell 2: 503–512

    Article  PubMed  CAS  Google Scholar 

  • Sumaryati S, Negretiu I & Jacobs M (1992) Characterization and regeneration of salt-and water-stress mutants from protoplast cultures of Nicotiana plumbaginifolia (viviani). Theor. Appl. Genet. 83: 613–619

    Article  CAS  Google Scholar 

  • Thomas JC, Armond RL & Bohnert HJ (1992) Influence of NaCl on growth, proline and phosphoenol pyruvate carboxylase levels in Mesembryanthemum crystallinum suspension cultures. Plant Physiol. 98: 627–631

    Article  Google Scholar 

  • Van Swaaij AC, Jacobsen E, Kiel JAKW & Feenstra WJ (1986) Selection, characterization and regeneration of hvdroxyprolineresistant cell lines of Solanum tuberosum: Tolerance to NaCl and freezing stress. Physiol. Plant. 68: 359–366

    Article  CAS  Google Scholar 

  • Verbruggen N, Villarroel R & Montagu MV (1993) Osmoregulation of a pyrrilone-5-carboxylate reductase gene in Arabidopsis thaliana. Plant Physiol. 103: 171–181

    Article  Google Scholar 

  • Watad A-EA, Reinhold L & Lerner HR (1983) Comparison between a stable NaCl-selected Nicotiana cell line and the wild type. Plant Physiol. 73: 624–629

    PubMed  CAS  Google Scholar 

  • Wetter L & Dyck J (1983) Isozyme analysis of cultured cells and somatic hybrids. In: Evans DA, Sharp WR, Ammirato PV & Yamada Y (eds.) Handbook of Plant Cell Culture Vol. 1 (pp 607–627). Macmillam Publishing Co., NY

    Google Scholar 

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Authors and Affiliations

  1. Deparpartment of Botany, Bose Institute, 93/1 A.P.C. Road, Calcutta-, 700 009, India

    Gaurab Gangopadhyay, Sangita Basu & Sukumar Gupta

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  1. Gaurab Gangopadhyay
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  2. Sangita Basu
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  3. Sukumar Gupta
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Gangopadhyay, G., Basu, S. & Gupta, S. In vitro selection and physiological characterization of NaCl- and mannitol-adapted callus lines in Brassica juncea. Plant Cell, Tissue and Organ Culture 50, 161–169 (1997). https://doi.org/10.1023/A:1005929316777

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