Summary
Protoplast-derived colonies of haploid N. plumbaginifolia leaves were used to select for resistance to NaCl, KCl and polyethylene glycol 6000 (PEG). Salt-and PEG-tolerant cell lines were isolated on the basis of growth in a culture medium containing inhibitory concentrations of either NaCl or KCl (200 mM) or PEG (25%). The frequency of resistant lines ranged from 10-5 to 10-6. One resistant line from each treatment was regenerated into plants. All resistant lines produced 10–25 times more proline than the wild type when grown on a non-selective medium. Similar values were also observed in the leaves of resistant progeny plants. In each mutant line, salt or PEG resistance was transmitted as a single dominant nuclear gene as shown by segregation ratios in progenies of crosses between resistant and wild-type plants. The latter observation demonstrates clearly the existence of a genetic basis for increased salt tolerance.
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References
Ben-Hayyim G (1985) Relationship between salt tolerance and resistance to polyethylene glycol-induced water stress in cultured citrus cells. Plant Physiol 85:430–433
Bhaskaran S, Smith RH, Schertz KF (1986) Progeny screening of sorghum plants regenerated from sodium chloride-selected callus for salt tolerance. J Plant Physiol 122:205–210
Bieleski RL, Turner NA (1966) Separation and estimation of amino acids in crude plant extracts by thin-layer electrophoresis and chromatography. Anal Biochem 17:278–293
Bressan RA, Handa AK, Handa S, Hasegawa PM (1982) Growth and water relation of cultured tomato cells after adjustment to low external water potentials. Plant Physiol 701:303–1309
Bright SWJ, Shewry PR, Miflin BJ (1978) Aspartate kinase and the synthesis of aspartate-derived amino acids in wheat. Planta 139:119–125
Caplan AB, Claes B, Dekeyser R, Van Montagu M (1990) Salinity and drought stress in rice. In: Sangwan RS, Sangwan-Norreel BS (eds) The impact of biotechnology in agriculture, vol 8. Kluwer Academic Publ, the Netherlands, pp 391–402
Dix PJ, Pearce RS (1981) Proline accumulation in NaCl-resistant and sensitive cell lines of N. sylvestris. Z flanzenphysiol 102:243–248
Dix PJ, Street HE (1975) Sodium chloride-resistant cultured cell lines from N. sylvestris and Capsicum annum. Plant Sci Lett 5:231–237
Epstein E, Norlyn JD, Rush DW, Kingsbury RW, Kelley DB, Cuningham GA, Wrona AS (1980) Saline culture of crops, a genetic approach. Science 210:399–403
Handa AK, Bressan RA, Handa S, Hasegawa PM (1983) Clonal variation for tolerance to polyetylene glycol-induced water stress in cultured tomato cells. Plant Physiol 72:645–653
Installé P, Negrutiu I, Jacobs M (1985) Protoplast-derived plants in N. plumbaginifolia Viviani: improving the regeneration response of wild type and mutant cultures. J Plant Physiol 119:443–454
Jacobs M, Negrutiu I, Dirks R, Cammaerts D (1987) Selection programmes for isolation and analysis of mutants in plant cell cultures. Plant Tissue Cell Cult 243–264
Le Rudulier D, Strom AR, Dandekar AM, Smith LT, Valentine RC (1984) Molecular biology of osmoregulation. Science 224:1064–1068
McHughen A, Swartz M (1984) A tissue-culture derived salt-tolerant line of Flax (Linum usitatissimum). J Plant Physiol 117:109–117
Nabors MW, Gibbs SE, Bernstein CS, Meis ME (1980) NaCl-tolerant tobacco plants from cultured cell. Z Pflanzenphysiol 97:13–17
Nabors MW (1990) Environmental stress resistance. In: Dix PJ (ed) Plant cell line selection procedures and applications. Weinheim, New York Basel Cambridge, pp 168–185
Negrutiu I (1990) Selection and choice of genetic markers. In: Dix PJ (ed) Plant cell line selection. Weinheim, New York Basel Cambridge, pp 217–229
Negrutiu I (1981) Improved conditions for large-scale culture, mutagenesis and selection of haploid protoplasts of N. plumbaginifolia. Z Pflanzenphysiol 104:431–442
Negrutiu I, Jacobs M, Cattoir-Reynaerts A (1984) Progress in cellular engineering of plants: Biochemical and genetic assessment of selectable markers from cultured cells. Plant Mol Biol 3:289–302
Rains DW, Croughn TP, Stavarek SJ (1980) Genetic engineering and tissue culture. In: Rains DW, Valentine RC, Hollaender A (ed) Genetic engineering of osmoregulation. Plenum Press, New York London, pp 290–292
Smith MK, McComb JA (1983) Selection for NaCl tolerance in cell cultures of Medicago sativa and recovery of plants from a NaCl-tolerant cell line. Plant Cell Rep 2:126–128
Stewart R, Larher F (1980) Accumulation of amino acids and related compounds in relation to environmental stress. In: Miflin BJ (ed) The biochemistry of plants, vol 5. Academic Press, New York, pp 609–635
Tal M (1984) Physiological genetics of salt resistance in higher plants, studies on the level of the whole plant and isolated organs, tissues and cells. In: Staples RC, Toenniessen (eds) Salinity tolerance in plants, strategies for crop improvement. John Wiley, New York, pp 443
Vajrabhaya M, Thanapaisal T, Vajrabhaya T (1989) Development of salt tolerant lines of KDML and LPT rice cultivars through tissue culture. Plant Cell Rep 8:411–414
Warne TR, Hickok LG (1987) Single gene mutants tolerant to NaCl in the Fern Ceratopteris, characterization and genetic analysis. Plant Sci 52:49–55
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Communicated by Yu. Gleba
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Sumaryati, S., Negrutiu, I. & Jacobs, M. Characterization and regeneration of salt- and water-stress mutants from protoplast culture of Nicotiana plumbaginifolia (Viviani). Theoret. Appl. Genetics 83, 613–619 (1992). https://doi.org/10.1007/BF00226906
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DOI: https://doi.org/10.1007/BF00226906