Almeida Diego M, Almadanim M Cecília, Lourenço Tiago, Abreu Isabel A, Saibo Nelson J M, Oliveira M Margarida
Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Genomics of Plant Stress Unit, Av. da República, 2780-157, Oeiras, Portugal.
IBET, Apartado 12, 2781-901, Oeiras, Portugal.
Methods Mol Biol. 2016;1398:155-82. doi: 10.1007/978-1-4939-3356-3_14.
Rice (Oryza sativa) is the primary source of food for more than half of the world population. Most rice varieties are severely injured by abiotic stresses, with strong social and economic impact. Understanding rice responses to stress may help breeding for more tolerant varieties. However, papers dealing with stress experiments often describe very different experimental designs, thus making comparisons difficult. The use of identical setups is the only way to generate comparable data. This chapter is organized into three sections, describing the experimental conditions established at the Genomics of Plant Stress (GPlantS) unit of ITQB to assess the response of rice plants to three different abiotic stresses--high salinity, cold stress, and drought. All sections include a detailed description of the materials and methodology, as well as useful notes gathered from the GPlantS team's experience. We use rice seedlings as plants at this stage show high sensitivity to abiotic stresses. For the salt and cold stress assays we use hydroponic cultures, while for the drought assay plants are grown in soil and subjected to water withholding. All setups enable visual score determination and are suitable for sample collection along the imposition of stress. The proposed methodologies are simple and affordable to implement in most labs, allowing the discrimination of several rice genotypes at the molecular and phenotypic level.
水稻(Oryza sativa)是世界上一半以上人口的主要食物来源。大多数水稻品种会受到非生物胁迫的严重伤害,产生强烈的社会和经济影响。了解水稻对胁迫的反应可能有助于培育更具耐受性的品种。然而,涉及胁迫实验的论文往往描述非常不同的实验设计,因此难以进行比较。使用相同的设置是生成可比较数据的唯一方法。本章分为三个部分,描述了在ITQB的植物胁迫基因组学(GPlantS)单元建立的实验条件,以评估水稻植株对三种不同非生物胁迫——高盐度、冷胁迫和干旱的反应。所有部分都包括对材料和方法以及从GPlantS团队经验中收集的有用注释的详细描述。我们使用水稻幼苗,因为这个阶段的植株对非生物胁迫表现出高度敏感性。对于盐胁迫和冷胁迫试验,我们使用水培法,而对于干旱试验,植株种植在土壤中并进行停水处理。所有设置都能进行视觉评分测定,并且适合在施加胁迫的过程中进行样本采集。所提出的方法简单且在大多数实验室中实施成本较低,能够在分子和表型水平上区分几种水稻基因型。