Abstract
Cell cultures of chili pepper (Capsicum annuum L.) were established from callus tissue inoculated in MS liquid medium supplemented with 6.25 μM 2,4-d and 0.44 μM BA. Cell clones were isolated by plating the cell suspension on filter paper discs supported by polyurethane foam that were bathed with culture medium containing 15% PEG. The cell clones T6 and T7 were chosen based on their characteristics of growth and friability. These cell clones were established as cell suspensions in the presence of 15% PEG and subsequently subcultured in increasing concentrations of osmoticum. By this approach the cell clones T7 and T6 were capable of growing in the presence of 20 and 25% PEG, respectively. The cell clone T7 was found to grow better in the presence of 5–10% PEG after a period of subculturing in the absence of osmoticum indicating that the tolerance trait was stable. The tolerant cell clones exhibited a 3 to 3.5-fold decrease in the osmotic potentials in comparison with the nonselected cells suggesting that osmotic adjustment occurred. K+ was the major contributing solute to the osmotic potential in all the cell cultures among those tested and was found to be higher in concentration in the PEG-tolerant clones (1.3–3 times higher than nonselected cells). Proline and glycine betaine levels showed a positive correlation with the degree of tolerance to water deficit in the PEG-tolerant cell clones. The levels of proline in the cell clone T7 subcultured in the absence of PEG in the culture medium decreased to values similar to those of nonselected cells, whereas the contents of glycine betaine in the same conditions were maintained at high levels.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- BA:
-
benzyladenine
- 2,4-d :
-
2,4-dichlorophenoxyacetic acid
- MS:
-
Murashige and Skoog medium
- PEG:
-
polyethylene glycol
References
Bates LS, Waldren RP & Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39: 205–207
Ben-Hayyim G (1987) Relationship between salt tolerance and resistance to polyethylene glycol-induced water stress in cultured citrus cells. Plant Physiol. 85: 430–433
Borkird C, Simoens C, Villarroel R & Van Montagu M (1991) Gene expression associated with water-stress adaptation of rice cells and identification of two genes as hsp 70 and ubiquitin. Physiol. Plant. 82: 449–457
Bressan RA, Hasegawa PM & Handa AK (1981) Resistance of cultured higher plant cells to polyethylene glycol-induced water stress. Plant Sci. Lett. 21: 23–30
Bressan RA, Handa AK, Handa S & Hasegawa PM (1982) Growth and water relations of cultured tomato cells after adjustment to external water potentials. Plant Physiol. 70: 1303–1309
Cali JP, Bowers Jr. GN & Young DS (1973) Referee method for the determination of total calcium in serum. Clinical Chem. 19: 1208–1213
Conner AJ & Meredith CP (1984) An improved polyurethane support system for monitoring growth in plant cell cultures. Plant Cell Tiss. Org. Cult. 3: 59–68
Corchete P & Guerra H (1986) Effect of NaCl and polyethylene glycol on solute content and glycosidase activities during germination of lentil seeds. Plant Cell Environm. 9: 589–593
Grieve CM & Gratan SR (1983) Rapid assay for determination of soluble quaternary ammonium compounds. Plant Soil 70: 303–307
Gulati A & Jaiwal PK (1993) Selection and characterization of mannitol-tolerant callus lines of Vigna radiata (L.) Wilczek. Plant Cell Tiss. Org. Cult. 34: 35–41
Handa AK, Bressan RA, Handa S & Hasegawa PM (1982) Characteristics of cultured tomato cells after prolonged exposure to medium containing polyethylene glycol. Plant Physiol. 69: 514–521
Handa AK, Bressan RA, Handa S & Hasegawa PM (1983) Clonal variation for tolerance to polyethylene glycol-induced water stress in cultured tomato cells. Plant Physiol. 72: 645–653
Hanson AD & Hitz WD (1982) Metabolic responses of mesophytes to plant water deficit. Ann. Rev. Plant Physiol. 33: 163–203
Harms CT & Oertli JJ (1985) The use of osmotically adapted cell cultures to study salt tolerance in vitro. J. Plant Physiol. 120: 29–38
Heyser JW & Nabors MW (1981) Growth, water content, and solute accumulation of two tobacco cell lines cultured on sodium chloride, dextran, and polyethylene glycol. Plant Physiol. 68: 1454–1459
Iraki NM, Singh NK, Bressan RA & Carpita NC (1989) Cell walls of tobacco cells and changes in composition associated with reduced growth upon adaptation to water and saline stress. Plant Physiol. 91: 48–53
Irigoyen JJ, Emerich DW & Sánchez-Díaz M (1992) Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa) plants. Physiol. Plant. 84: 55–60
Ladyman JAR, Hitz WD & Hanson AD (1980) Translocation and metabolism of glycine betaine by barley plants in relation to water stress. Planta 150: 191–196
Larkin PJ & Scowcroft WR (1981) Somaclonal variation-a novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet. 60: 197–214
Lerner HR (1985) Adaptation to salinity at the plant cell level. Plant Soil 89: 3–14
Meredith CP (1983) On being selective: mutants from cultured cells. Plant Mol. Biol. Rep. 1: 105–110
Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497
Pahlich E, Kerres R & Jäger H-J (1983) Influence of water stress on the vacuole/extravacuole distribution of proline in protoplasts of Nicotiana rustica. Plant Physiol. 72: 590–591
Paleg LG, Stewart GR & Bradbeer JW (1984) Proline and glycine betaine influence protein solvation. Plant Physiol. 75: 974–978
Paleg LG, Stewart GR & Starr R (1985) The effect of compatible solutes on proteins. Plant Soil 89: 83–94
Parsons LR & Howe TK (1984) Effects of water stress on the water relations of Phaseolus vulgaris and the drought resistant Phaseolus acutifolius. Physiol. Plant. 60: 197–202
Pollard A & Wyn Jones RG (1979) Enzyme activities in concentrated solutions of glycine betaine and other solutes. Planta 144: 291–298
Premachandra GS, Saneoka H, Kanaya M & Ogata S (1989) Responses of relative growth rate, water relations and solute accumulation to increasing water deficits in maize. J. Plant Physiol. 135: 257–260
Sabbah S & Tal M (1990) Development of callus and suspension cultures of potato resistant to NaCl and mannitol and their response to stress. Plant Cell Tiss. Org. Cult. 21: 119–128
Salgado-Garciglia R, López-Gutierrez F & Ochoa-Alejo N (1985) NaCl-Resistant variant cells isolated from sweet potato cell suspensions. Plant Cell Tiss. Org. Cult. 5: 3–12
Salgado-Garciglia R & Ochoa-Alejo N (1990) Increased capsaicin content in PFP-resistant cells of chili pepper (Capsicum annuum L.). Plant Cell Rep. 8: 617–620
Schobert B & Tschesche H (1978) Unusual solution properties of proline and its interaction with proteins. Biochim. Biophys. Acta 541: 270–277
Schwab KB & Gaff DF (1986) Sugar and ion content in leaf tissues of several drought tolerant plants under water stress. J. Plant Physiol. 125: 257–265
Schwab KB & Gaff DF (1990) Influence of compatible solutes on soluble enzymes from dessication-tolerant Sporobulus stapfianus and dessication-sensitive Sporobolus pyramidalis. J. Plant Physiol. 137: 208–215
Singh NK, Handa AK, Hasegawa PM & Bressan RA (1985) Proteins associated with adaptation of cultured tobacco cells to NaCl. Plant Physiol. 79: 126–137
Singh NK, LaRosa C, Handa AK & Hasegawa PM (1987) Hormonal regulation of protein synthesis associated with salt tolerance in plant cells. Proc. Natl. Acad. Sci. USA 84: 739–743
Singh NK, Nelson DE, Kuhn D, Hasegawa PM & Bressan RA (1989) Molecular cloning of osmotin and regulation of its expression by ABA and adaptation to low water potential. Plant Physiol. 90: 1096–1101
Sumaryati S, Negrutiu I & Jacobs M (1992) Characterization and regeneration of salt-and water-stress mutants from protoplast culture of Nicotiana plumbaginifolia (Viviani). Theor. Appl. Genet. 83: 613–619
Trivedi S, Galiba G, Sankhla N & Erdei L (1991) Responses to osmotic and NaCl stress of wheat varieties differing in drought and salt tolerance in callus cultures. Plant Sci. 73: 227–232
Yancey PH, Clark ME, Hand SC, Bowlus PD & Somero GN (1982) Living with water stress: evolution of osmolyte systems. Science 217: 1214–1222
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Santos-Díaz, M.d.S., Ochoa-Alejo, N. PEG-tolerant cell clones of chili pepper: Growth, osmotic potentials and solute accumulation. Plant Cell Tiss Organ Cult 37, 1–8 (1994). https://doi.org/10.1007/BF00048110
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00048110