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Cloning and Characterization of the RubisCO Activase Gene from Pinus massoniana

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Abstract

Pinus massoniana (P. massoniana) is an important fast-growing tree species in China. To explore photosynthesis-related gene resources and find possible targets for high photosynthetic-efficiency breeding of P. massoniana, we cloned two full-length cDNAs with conserved sequences of RubisCO activase (RCA) genes from P. massoniana. The RCA enzyme is important in maintaining the activity of RubisCO in vivo. Two genes, named PmRCA1 and PmRCA2, showed high similarity to RCA genes from other species and were identical to each other except for a 137 bp intervening sequence in the open reading frame (ORF) of PmRCA1, which led to different protein products, herein referred to as PmRCA1 and PmRCA2, respectively. PmRCA1 and PmRCA2 showed high homology to RCA of Pinus halepensis, and PmRCA1 had two cysteine residues in its carboxyl-terminal extension that were unique to the large RCA isoform. To characterize the function of PmRCA1 and PmRCA2, plant expression vectors driven by the constitutive 35S promoter were transformed in tobacco plants. Transgenic plants of PmRCA2 were grown better than wild tobacco plants and had significantly more leaves. Overexpression of PmRCA2 increased the maximum net photosynthetic rate and dark respiration rate of tobacco by 16.49% and 81.85%, respectively. Overexpression of PmRCA1 increased the maximum net photosynthetic rate and dark respiration rate of tobacco more than overexpression of PmRCA2, but it simultaneously enhanced photoinhibition of photosynthesis.

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Availability of Data and Material

The sequences of PmRCA1 and PmRCA2 were uploaded to GenBank and defined as Pinus massoniana ribulose-1,5-bisphosphate carboxylase/oxygenase activase large and small isoform (rca) mRNA respectively. Their accession numbers are KF420118 and KF420119, respectively.

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Acknowledgements

Thanks Professor Qiang Zhuge, Professor Jinhui Chen, and Professor Yinfeng Xie for providing us with plant material, test equipments and sincere advices.

Funding

The research was supported by the National Key Research and Development Program of China (2017YFD0600304) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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Authors and Affiliations

  1. Key Laboratory of Forestry Genetics & Biotechnology of Ministry of Education, Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China

    Ting Pan, Fengkai Zhang, Lu Sheng, Xiaofeng Wang, Weicheng Ruan, Peihuang Zhu, Tianwei Guo & Kongshu Ji

  2. Key Laboratory of State Forestry Administration on Prevention and Control of Pine Wood Nematode Disease, Anhui Forestry Academy, Hefei, 230000, China

    Ting Pan

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  1. Ting Pan
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  2. Fengkai Zhang
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  3. Lu Sheng
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Correspondence to Kongshu Ji.

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Key Message

This study showed a light on the gene sequences and function of two RubisCO activase from Pinus massoniana for the first time and pointed out that the small isoform RubisCO activase gene of Pinus massoniana could be a promising target for high light efficiency breeding.

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Pan, T., Zhang, F., Sheng, L. et al. Cloning and Characterization of the RubisCO Activase Gene from Pinus massoniana. Plant Mol Biol Rep 41, 81–91 (2023). https://doi.org/10.1007/s11105-022-01345-7

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  • DOI: https://doi.org/10.1007/s11105-022-01345-7

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