Fernandes Telma, Melo Fredilson, Vieira Maria Beatriz, Lourenço Tiago F, Pucciariello Chiara, Saibo Nelson J M, Abreu Isabel A, Oliveira M Margarida
Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Genomics of Plant Stress Unit, Oeiras, Portugal.
PlantLab, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.
Methods Mol Biol. 2022;2494:161-194. doi: 10.1007/978-1-0716-2297-1_12.
Rice (Oryza sativa L.) is the staple food for over half of the world population. However, most rice varieties are severely injured by abiotic stresses, with strong social and economic impacts. Understanding rice responses to stress may guide breeding for more tolerant varieties. However, the lack of consistency in the design of the stress experiments described in the literature limits comparative studies and output assessments. The use of identical setups is the only way to generate comparable data. This chapter comprises three sections, describing the experimental conditions established at the Genomics of Plant Stress (GPlantS) unit of ITQB NOVA to assess the response of rice plants to different abiotic stresses-high salinity, cold, drought, simulated drought, and submergence-and their recovery capacity when intended. All sections include a detailed description of the materials and methodology and useful notes gathered from our team experience. We use seedlings since rice plants at this stage show high sensitivity to abiotic stresses. For the salt, cold, and simulated drought (PEG, polyethylene glycol) stress assays, we grow rice seedlings in a hydroponic system, while for the drought assay, plants are grown in soil and subjected to water withholding. For submergence, we use water-filled Magenta boxes. All setups enable visual score determination and are suitable for sample collection during stress imposition and also recovery. The proposed methodologies are affordable and straightforward to implement in most labs, allowing the discrimination of several rice genotypes at the molecular and phenotypic levels.
水稻(Oryza sativa L.)是世界上超过一半人口的主食。然而,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会指导培育更具耐受性的品种。然而,文献中描述的胁迫实验设计缺乏一致性,限制了比较研究和产量评估。使用相同的实验设置是生成可比数据的唯一方法。本章包括三个部分,描述了在ITQB NOVA的植物胁迫基因组学(GPlantS)单元建立的实验条件,以评估水稻植株对不同非生物胁迫(高盐度、寒冷、干旱、模拟干旱和淹水)的反应以及它们在必要时的恢复能力。所有部分都包括材料和方法的详细描述以及从我们团队经验中收集的有用注意事项。我们使用幼苗,因为这个阶段的水稻植株对非生物胁迫表现出高度敏感性。对于盐胁迫、冷胁迫和模拟干旱(PEG,聚乙二醇)胁迫试验,我们在水培系统中种植水稻幼苗,而对于干旱试验,植株在土壤中生长并进行控水。对于淹水试验,我们使用装满水的洋红色盒子。所有设置都能进行视觉评分测定,并且适合在胁迫施加期间以及恢复期间进行样本采集。所提出的方法经济实惠且在大多数实验室中易于实施,能够在分子和表型水平上区分几种水稻基因型。