Departamento de Biotecnología Microbiana y de Plantas, Centro de Investigaciones Biológicas Margarita Salas (CIB-CSIC), Madrid, Spain.
Methods Mol Biol. 2020;2156:85-97. doi: 10.1007/978-1-0716-0660-5_8.
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.
低温是自然植物群落构成的重要决定因素，决定了重要作物的分布和生长范围。一些植物，包括拟南芥，已经进化出复杂的适应机制来耐受冰冻温度。适应的核心是低温驯化过程。通过这个过程，许多来自温带地区的植物能够在应对低温、非冰冻温度时发展或增加其抗冻能力。鉴定和描述与抗冻性相关的因素对于理解冷驯化反应的分子机制至关重要，并且对于提高作物对冰冻温度的耐受性具有潜在的兴趣。通过分子方法和反向遗传学分析，已经在许多植物物种中鉴定出许多与冷驯化相关的基因。然而，令人惊讶的是，直接的遗传分析并没有被充分利用来鉴定在适应反应中具有关键作用的基因。在本章中，我们描述了一种评估非驯化和低温驯化拟南芥植物抗冻能力的方案。该方案允许对突变体文库进行准确而简单的筛选，以鉴定与抗冻性和低温驯化相关的新因子。
