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Updated: Nov 4, 2025

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Rubisco Engineering by Plastid Transformation and Protocols for Assessing Expression.

Spencer M Whitney1, Robert E Sharwood2

  • 1Plant Sciences, Research School of Biology, College of Science, The Australian National University, Acton, ACT, Australia. spencer.whitney@anu.edu.au.

Methods in Molecular Biology (Clifton, N.J.)
|May 24, 2021
PubMed
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Transcriptome and metabolome insights into closely related upland cotton (Gossypium hirsutum) genotypes during differing responses to progressive soil drying.

BMC plant biology·2026
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Reprogramming encapsulins into modular carbon-fixing nanocompartments.

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<i>In Vivo</i> Directed Evolution of an Ultra-Fast Rubisco from a Semi-Anaerobic Environment Imparts Oxygen Resistance.

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Proceedings of the National Academy of Sciences of the United States of America·2025
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Journal of experimental botany·2025

Researchers engineered the Rubisco enzyme in tobacco plastids to improve its efficiency. This work presents methods for Rubisco engineering and content analysis in plants.

Area of Science:

  • Plant molecular biology
  • Biochemistry
  • Photosynthesis research

Background:

  • Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes CO2 assimilation in chloroplasts.
  • Rubisco's inefficiency, due to slow turnover and oxygenase activity, leads to carbon loss via photorespiration.
  • Engineering Rubisco's carboxylation efficiency and CO2/O2 selectivity is a key goal in plant science.

Purpose of the Study:

  • To present methodologies for efficiently engineering Rubisco within the plastids of a tobacco master line.
  • To describe methods for analyzing leaf Rubisco content post-engineering.

Main Methods:

  • Utilizing plastid transformation techniques for Rubisco engineering in tobacco.
  • Developing and applying experimental protocols for Rubisco content analysis in plant leaves.
Keywords:
CABP bindingImmunodetectionRubiscoTobacco master linerbcL gene

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Main Results:

  • Demonstrated efficient engineering of Rubisco in tobacco plastids.
  • Established reliable methods for quantifying leaf Rubisco content.

Conclusions:

  • Plastid transformation is an effective strategy for improving Rubisco function in plants.
  • The presented methodologies facilitate advancements in Rubisco engineering and analysis for enhanced crop productivity.