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Plant Physiol, April 2003, Vol. 131, pp. 1800-1807
Freezing Sensitivity in the sfr4 Mutant of
Arabidopsis Is Due to Low Sugar Content and Is Manifested by Loss of
Osmotic Responsiveness1
Matsuo
Uemura,*
Gareth
Warren, and
Peter L.
Steponkus2
Department of Crop and Soil Sciences, Cornell University, Ithaca,
New York 14853 (M.U., P.L.S.); Cryobiosystem Research Center, Faculty
of Agriculture, Iwate University, Morioka 020-8550, Japan (M.U.); and
School of Biological Sciences, Royal Holloway, University of London,
Egham TW20 0EX, United Kingdom (G.W.)
Protoplasts were tested to determine whether the freezing
sensitivity of the sfr4 (sensitive to freezing) mutant
of Arabidopsis was due to the mutant's deficiency in soluble sugars
after cold acclimation. When grown under nonacclimated conditions,
sfr4 protoplasts possessed freezing tolerance similar to
that of wild type, with the temperature at which 50% of protoplasts
are injured (LT50) of  4.5°C. In both wild-type and
sfr4 protoplasts, expansion-induced lysis was the
predominant lesion between 2°C and 4°C, but its incidence was
low (approximately 10%); below 5°C, loss of osmotic responsiveness
(LOR) was the predominant lesion. After cold acclimation, the
LT50 was decreased to only 5.6°C for
sfr4 protoplasts, compared with 9.1°C for wild-type
protoplasts. Although expansion-induced lysis was precluded in both
types of protoplasts, the sfr4 protoplasts remained
susceptible to LOR. After incubation of seedlings in Suc solution in
the dark at 2°C, freezing tolerance and the incidence of
freeze-induced lesions in sfr4 protoplasts were
examined. The freezing tolerance of isolated protoplasts
(LT50 of 9°C) and the incidence of LOR were now similar
for wild type and sfr4. These results indicate that the
freezing sensitivity of cold-acclimated sfr4 is due to
its continued susceptibility to LOR (associated with lyotropic
formation of the hexagonal II phase) and associated with the low sugar
content of its cells.
1
This work was supported by the U.S. Department
of Agriculture/National Research Initiative Competitive Grants Program
(grant no. 96-35100-3163 to P.L.S.), by the U.S. Department of Energy (grant no. DE-FG01-84ER13214 to P.L.S.), by the UK Biotechnology and
Biological Sciences Research Council (grant to G.W.), and by the
Bio-oriented Technology Research Advancement Institution, Japan (grant
to M.U.).
2
Professor Steponkus died before the final
drafting of this manuscript.
*
Corresponding author; e-mail uemura{at}iwate-u.ac.jp; fax
81-19-621-6253.
© 2003 American Society of Plant Biologists
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