ArticleJournal of experimental botany2020
Measuring Rubisco activity: challenges and opportunities of NADH-linked microtiter plate-based and 14C-based assays.
Article in Journal of experimental botany, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed, 32 citations in OpenAlex.
- Rubisco dark inhibition in angiosperms shows a complex distribution pattern.Journal of experimental botany · 2026Article
- Strigolactone-mediated regulation of chromium uptake, redox homeostasis, and photosynthetic carbon metabolism during Cr (VI) stress tolerance in Capsicum annuum L.Plant cell reports · 2026Article
- Antisense reduction in NADP-ME in the CThe New phytologist · 2026Article
- Leaf Ontogeny Shapes Divergent Physiological and Metabolic Responses to Contrasting Nitrogen Forms in Chinese Fir (International journal of molecular sciences · 2026Article
- Inhibitory binding to plastoquinone B site on D1 protein of photosystem II leading to decreased population growth rate via disruption of cellular energy metabolism.Frontiers in plant science · 2026Review
- Shining light on photosynthesis.Journal of experimental botany · 2025Article
- In vivo transformations of positively charged nanoparticles alter the formation and function of RuBisCO photosynthetic protein corona.Nature nanotechnology · 2025Article
- Robust Synthetic Biology Toolkit to Advance Carboxysome Study and Redesign.ACS synthetic biology · 2025Article
- Dynamic photosynthetic labeling and carbon-positional mass spectrometry monitor in vivo RUBISCO carbon assimilation rates.Plant physiology · 2025Article
- Increasing Rubisco as a simple means to enhance photosynthesis and productivity now without lowering nitrogen use efficiency.The New phytologist · 2025Review
- Critical stages in pea photosynthesis impaired by tetracycline as an environmental contaminant.Journal of plant research · 2024Article
- RuBisCO activity assays: a simplified biochemical redox approach for in vitro quantification and an RNA sensor approach for in vivo monitoring.Microbial cell factories · 2024Article
- Extraction of Soluble Proteins from Leaves.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Phenotypic variation in photosynthetic traits in wheat grown under field versus glasshouse conditions.Journal of experimental botany · 2022Article
- The Contrivance of Plant Growth Promoting Microbes to Mitigate Climate Change Impact in Agriculture.Microorganisms · 2021Review
- Magnesium Application Promotes Rubisco Activation and Contributes to High-Temperature Stress Alleviation in Wheat During the Grain Filling.Frontiers in plant science · 2021Article
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Authors and funding
3 authors at 2 institutions in 2 countries.
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Abstract
Rubisco is central to carbon assimilation, and efforts to improve the efficiency and sustainability of crop production have spurred interest in phenotyping Rubisco activity. We tested the hypothesis that microtiter plate-based methods provide comparable results to those obtained with the radiometric assay that measures the incorporation of 14CO2 into 3-phosphoglycerate (3-PGA). Three NADH-linked assays were tested that use alternative coupling enzymes: glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and glycerolphosphate dehydrogenase (GlyPDH); phosphoenolpyruvate carboxylase (PEPC) and malate dehydrogenase (MDH); and pyruvate kinase (PK) and lactate dehydrogenase (LDH). To date there has been no thorough evaluation of their reliability by comparison with the 14C-based method. The three NADH-linked assays were used in parallel to estimate (i) the 3-PGA concentration-response curve of NADH oxidation, (ii) the Michaelis-Menten constant for ribulose-1,5-bisphosphate, (iii) fully active and inhibited Rubisco activities, and (iv) Rubisco initial and total activities in fully illuminated and shaded leaves. All three methods correlated strongly with the 14C-based method, and the PK-LDH method showed a strong correlation and was the cheapest method. PEPC-MDH would be a suitable option for situations in which ADP/ATP might interfere with the assay. GAPDH-GlyPDH proved more laborious than the other methods. Thus, we recommend the PK-LDH method as a reliable, cheaper, and higher throughput method to phenotype Rubisco activity for crop improvement efforts.
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