Abstract
Low temperature is an important determinant in the configuration of natural plant communities and defines the range of distribution and growth of important crops. Some plants, including Arabidopsis thaliana, have evolved sophisticated adaptive mechanisms to tolerate freezing temperatures. Central to this adaptation is the process of cold acclimation. By means of this process, many plants from temperate regions are able to develop or increase their freezing tolerance in response to low, nonfreezing temperatures. The identification and characterization of factors involved in freezing tolerance is crucial to understand the molecular mechanisms underlying the cold acclimation response and has a potential interest to improve crop tolerance to freezing temperatures. Many genes implicated in cold acclimation have been identified in numerous plant species by using molecular approaches followed by reverse genetic analysis. Remarkably, however, direct genetic analyses have not been conveniently exploited in their capacity for identifying genes with pivotal roles in that adaptive response. In this chapter, we describe a protocol for evaluating the freezing tolerance of both nonacclimated and cold acclimated Arabidopsis plants. This protocol allows for the accurate and simple screening of mutant collections for the identification of novel factors involved in freezing tolerance and cold acclimation.
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References
Levitt J (1980) Responses of plants to environmental stresses: chilling, freezing and high temperatures stresses, 2nd edn. Academic, New York
Steponkus PL, Uemura M, Joseph RA et al (1998) Mode of action of the COR15a gene on the freezing tolerance of Arabidopsis thaliana. Proc Natl Acad Sci U S A 95:14570–14575
Guy CL (1990) Cold acclimation and freezing stress tolerance: role of protein metabolism. Annu Rev Plant Physiol Plant Mol Biol 41:187–223
Lightner J, Caspar T (1998) Seed mutagenesis of Arabidopsis. In: Martínez-Zapater JM, Salinas J (eds) Methods in molecular biology, vol 82. Humana, Totowa, NJ, pp 91–103
Kim YS, Schumaker KS, Zhu JK (2006) EMS mutagenesis of Arabidopsis. In: Salinas J, Sanchez-Serrano JJ (eds) Methods in molecular biology, vol 323. Humana, Totowa, NJ, pp 101–103
Weigel D, Glazebrook J (2006) EMS mutagenesis of Arabidopsis seed. CSH Protoc 28. https://doi.org/10.1101/pdb.prot4621
Warren G, McKown R, Marin AL et al (1996) Isolation of mutations affecting the development of freezing tolerance in Arabidopsis thaliana (L.) Heynh. Plant Physiol 111:1011–1019
McKown R, Kuroki G, Warren G (1996) Cold responses of Arabidopsis mutants impaired in freezing tolerance. J Exp Bot 47:1919–1925
Xin Z, Browse J (1998) eskimo1 mutants of Arabidopsis are constitutively freezing-tolerant. Proc Natl Acad Sci U S A 95:7799–7804
Llorente F, Oliveros JC, Martínez-Zapater JM et al (2000) A freezing-sensitive mutant of Arabidopsis, frs1, is a new aba3 allele. Planta 211:648–655
Koornneef M, Dellaert LWM, van der Veen JH (1982) EMS- and radiation-induced mutation frequencies at individual loci in Arabidopsis thaliana (L.) Heynh. Mutat Res 93:109–123
Alonso JM, Stepanova AN (2003) T-DNA mutagenesis in Arabidopsis. In: Grotewold E (ed) Methods in molecular biology, vol 236. Humana, Totowa, NJ, pp 177–188
Jander G (2006) Gene identification and cloning by molecular marker mapping. In: Salinas J, Sanchez-Serrano JJ (eds) Methods in molecular biology, vol 323. Humana, Totowa, NJ, pp 115–126
Austin RS, Vidaurre D, Stamatiou G et al (2011) Next-generation mapping of Arabidopsis genes. Plant J 67:715–725
Uchida N, Sakamoto T, Kurata T et al (2011) Identification of EMS-induced causal mutations in a non-reference Arabidopsis thaliana accession by whole genome sequencing. Plant Cell Physiol 52:716–722
Wachsman G, Modliszewski JL, Valdes M, Benfey PN (2017) A SIMPLE pipeline for mapping point mutations. Plant Physiol 174:1307–1313
O’Malley RC, Alonso JM, Kim CJ et al (2007) An adapter ligation-mediated PCR method for high-throughput mapping of T-DNA inserts in the Arabidopsis genome. Nat Protoc 2:2910–2917
McCourt P, Keith K (1998) Sterile techniques in Arabidopsis, Methods in molecular biology, vol 82. Humana, Totowa, NJ, pp 13–17
Mikkelsen MD, Thomashow MF (2009) A role for circadian evening elements in cold-regulated gene expression in Arabidopsis. Plant J 60:328–339
Dong MA, Farré EM, Thomashow MF (2011) Circadian clock-associated 1 and late elongated hypocotyl regulate expression of the C-repeat binding factor (CBF) pathway in Arabidopsis. Proc Natl Acad Sci U S A 108:7241–7246
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Perea-Resa, C., Catalá, R., Salinas, J. (2020). Identification of Arabidopsis Mutants with Altered Freezing Tolerance. In: Hincha, D., Zuther, E. (eds) Plant Cold Acclimation. Methods in Molecular Biology, vol 2156. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0660-5_8
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DOI: https://doi.org/10.1007/978-1-0716-0660-5_8
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