A high throughput gas exchange screen for determining rates of photorespiration or regulation of C4 activity.
A high throughput gas exchange screen for determining rates of photorespiration or regulation of C4 activity.
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DOI：
10.1093/jxb/eru238
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发表时间：
2014-07
影响因子：
6.9
通讯作者：
Hibberd JM
中科院分区：
文献类型：
--
作者：
Bellasio C;Burgess SJ;Griffiths H;Hibberd JM
一种快速测定植物光呼吸活性的方法，适用于分析光合作用突变体、C3-C4中间体以及具有弱甘氨酸穿梭的品系。寻求表征C4途径的大规模研究计划需要一种简单、高通量的筛查来量化C3和C4模型系统中的光呼吸活动。目前，对同化响应(A/CI曲线、PSII量子产率)或实时碳同位素判别的方法依赖于模型拟合，这些方法既复杂又耗时。在这里，我们提出了一种方法和相关的理论，以确定C4羧化，碳浓缩机制(CCM)的有效性，通过评估VO/VC的响应，即Rubisco加氧酶与羧基酶活性的比率，在转移到低O2时。这项测定使用广泛可用的设备，比较了环境和低氧气条件下的并行气体交换和脉冲调制的叶绿素荧光。如果植物是预先适应的，该程序的运行时间可能只需6分钟。推导了典型C3(烟草、水稻、小麦)和C4(玉米、芒属、玉米)植物的VO/VC响应度，并与完整的C3和C4模型系统进行了比较。我们还进行了灵敏度分析，以确定R光(光中的呼吸作用)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，该方法可以很好地解决C3和C4植物之间光呼吸活性的差异，可用于高通量研究中快速筛选大量突变体或转化子。
A rapid approach to determine photorespiratory activity in plants suitable for analysis of photosynthetic mutants, C3–C4 intermediates, as well as lines with a weak glycine shuttle. Large-scale research programmes seeking to characterize the C4 pathway have a requirement for a simple, high throughput screen that quantifies photorespiratory activity in C3 and C4 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 C4 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 O2. This determination compares concurrent gas exchange and pulse-modulated chlorophyll fluorescence under ambient and low O2, 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 C3 (tobacco, rice, wheat) and C4 (maize, Miscanthus, cleome) plants, and compared with full C3 and C4 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 C3 and C4 plants and could be used to rapidly screen large numbers of mutants or transformants in high throughput studies.
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影响因子：
7.3
作者：
Kromdijk, Johannes;Griffiths, Howard;Schepers, Hans E.
通讯作者：
Schepers, Hans E.
影响因子：
7.4
作者：
Chang, Yao-Ming;Liu, Wen-Yu;Ku, Maurice S. B.
通讯作者：
Ku, Maurice S. B.
影响因子：
3.7
作者：
Earl, HJ;Ennahli, S
通讯作者：
Ennahli, S
影响因子：
3.7
作者：
Bellasio C;Fini A;Ferrini F
通讯作者：
Ferrini F
影响因子：
6.4
作者：
Gupta, Kapuganti J.;Zabalza, Ana;van Dongen, Joost T.
通讯作者：
van Dongen, Joost T.