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首页> 外文期刊>Journal of Experimental Botany >A high throughput gas exchange screen for determining rates of photorespiration or regulation of C-4 activity
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A high throughput gas exchange screen for determining rates of photorespiration or regulation of C-4 activity

机译：高通量气体交换屏，用于确定光呼吸速率或调节C-4活性

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Large-scale research programmes seeking to characterize the C-4 pathway have a requirement for a simple, high throughput screen that quantifies photorespiratory activity in C-3 and C-4 model systems. At present, approaches rely on model-fitting to assimilatory responses (A/C-i curves, PSII quantum yield) or real-time carbon isotope discrimination, which are complicated and time-consuming. Here we present a method, and the associated theory, to determine the effectiveness of the C-4 carboxylation, carbon concentration mechanism (CCM) by assessing the responsiveness of V-O/V-C, the ratio of RuBisCO oxygenase to carboxylase activity, upon transfer to low O-2. This determination compares concurrent gas exchange and pulse-modulated chlorophyll fluorescence under ambient and low O-2, using widely available equipment. Run time for the procedure can take as little as 6 minutes if plants are pre-adapted. The responsiveness of V-O/V-C is derived for typical C-3 (tobacco, rice, wheat) and C-4 (maize, Miscanthus, cleome) plants, and compared with full C-3 and C-4 model systems. We also undertake sensitivity analyses to determine the impact of R-LIGHT (respiration in the light) and the effectiveness of the light saturating pulse used by fluorescence systems. The results show that the method can readily resolve variations in photorespiratory activity between C-3 and C-4 plants and could be used to rapidly screen large numbers of mutants or transformants in high throughput studies.
机译：试图表征C-4途径的大规模研究计划要求有一个简单，高通量的筛查方法，以量化C-3和C-4模型系统中的光呼吸活性。目前，方法依赖模型拟合以适应同化响应（A / C-i曲线，PSII量子产率）或实时碳同位素判别，这既复杂又费时。在这里，我们介绍一种方法和相关的理论，通过评估VO / VC的响应度，RuBisCO加氧酶与羧化酶活性的比值（转移至低浓度时）来确定C-4羧化，碳浓缩机制（CCM）的有效性O-2。该测定使用广泛可用的设备比较了在环境和低O-2下同时进行的气体交换和脉冲调制的叶绿素荧光。如果植物预先适应，则该过程的运行时间可能仅需6分钟。 V-O / V-C的响应性是针对典型C-3（烟草，水稻，小麦）和C-4（玉米，Miscanthus，Cleome）植物得出的，并与完整的C-3和C-4模型系统进行了比较。我们还进行灵敏度分析，以确定R-LIGHT（光中的呼吸）的影响以及荧光系统使用的光饱和脉冲的有效性。结果表明，该方法可以轻松解决C-3和C-4植物之间的光呼吸活性变化，并可以用于高通量研究中快速筛选大量突变体或转化体。

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