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Expression level of Rubisco activase negatively correlates with Rubisco content in transgenic rice

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Abstract

The relationship between ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and Rubisco activase (Rca) levels was studied using transgenic rice overexpressing maize Rca (OX-mRca) and knockdown transgenic rice expressing antisense Rca (KD-Rca). The ratio of Rubisco to total soluble protein was lower in OX-mRca, whereas it was higher in KD-Rca than in WT, indicating that Rca expression was negatively correlated with Rubisco content. The expressions of other Calvin–Benson–Bassham cycle enzymes such as sedoheptulose-1,7-bisphosphatase and phosphoribulokinase analyzed by immunoblotting did not show such a negative correlation with Rca, suggesting that the effect of Rca on protein expression may be specific for Rubisco. Although Rubisco content was decreased in OX-mRca, the transcript levels of the Rubisco large subunit (OsRbcL) and the Rubisco small subunit mostly increased in OX-mRca as well as in KD-Rca. Additionally, polysome loading of OsRbcL was slightly higher in OX-mRca than it was in WT, suggesting that the OsRbcL translation activity was likely stimulated by overexpression of Rca. 35S-methionine labeling experiments demonstrated that there was no significant difference in the stability of newly synthesized Rubisco among genotypes. However, 35S-methionine-labeled Rubisco was marginally decreased in OX-mRca and increased in KD-Rca compared to the WT. These results suggest that Rca negatively affects the Rubisco content, possibly in the synthesis step.

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Acknowledgements

This work was supported by a Grant-in-Aid for Japan Society for the Promotion of Science KAKENHI (Grant Nos. 15H04443 and 24580021 to H.F.).

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Authors and Affiliations

  1. Laboratory of Tropical Crop Science, Graduate School of Agricultural Science, Kobe University, Kobe, 657-8501, Japan

    Hiroshi Fukayama, Akina Mizumoto, Ryutaro Morita & Tetsushi Azuma

  2. Laboratory of Crop Science, Graduate School of Agricultural Science, Kobe University, Kobe, 657-8501, Japan

    Chiaki Ueguchi, Daisuke Sasayama & Tomoko Hatanaka

  3. Laboratory of Crop Science, Faculty of Agriculture, Kobe University, Kobe, 657-8501, Japan

    Jun Katsunuma

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  1. Hiroshi Fukayama
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Correspondence to Hiroshi Fukayama.

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Fukayama, H., Mizumoto, A., Ueguchi, C. et al. Expression level of Rubisco activase negatively correlates with Rubisco content in transgenic rice. Photosynth Res 137, 465–474 (2018). https://doi.org/10.1007/s11120-018-0525-9

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