Fonseca Jose P, Griffiths Marcus, York Larry M, Mysore Kirankumar S
Noble Research Institute, LLC., Ardmore, OK, USA.
Institute for Agricultural Biosciences, Oklahoma State University, Ardmore, OK, 73401, USA.
Bio Protoc. 2021 Oct 5;11(19):e4181. doi: 10.21769/BioProtoc.4181.
Dark respiration refers to experimental measures of leaf respiration in the absence of light, done to distinguish it from the photorespiration that occurs during photosynthesis. Dark aerobic respiration reactions occur solely in the mitochondria and convert glucose molecules from cytoplasmatic glycolysis and oxygen into carbon dioxide and water, with the generation of ATP molecules. Previous methods typically use oxygen sensors to measure oxygen depletion or complicated and expensive photosynthesis instruments to measure CO accumulation. Here, we provide a detailed, step-by-step approach to measure dark respiration in plants by recording CO fluxes of shoot and root tissues. Briefly, plants are dark acclimated for 1 hour, leaves and roots are excised and placed separately in airtight chambers, and CO accumulation is measured over time with standard infrared gas analyzers. The time-series data is processed with R scripts to produce dark respiration rates, which can be standardized by fresh or dry tissue mass. The current method requires inexpensive infrared gas analyzers, off-the-shelf parts for chambers, and publicly available data analysis scripts.
暗呼吸是指在无光条件下对叶片呼吸作用进行的实验测量，这样做是为了将其与光合作用过程中发生的光呼吸区分开来。黑暗中的有氧呼吸反应仅在线粒体中发生，它将细胞质糖酵解产生的葡萄糖分子和氧气转化为二氧化碳和水，并产生ATP分子。以前的方法通常使用氧传感器来测量氧气消耗，或者使用复杂且昂贵的光合作用仪器来测量二氧化碳积累。在这里，我们提供了一种详细的、逐步的方法，通过记录茎和根组织的二氧化碳通量来测量植物的暗呼吸。简而言之，将植物在黑暗中适应1小时，切除叶片和根并分别置于气密小室中，然后使用标准红外气体分析仪随时间测量二氧化碳积累。利用R脚本处理时间序列数据以得出暗呼吸速率，该速率可以通过新鲜或干组织质量进行标准化。当前方法需要廉价的红外气体分析仪、小室的现成部件以及公开可用的数据分析脚本。
