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Exogenous ATP enhance signal response of suspension cells of transgenic rice (Oryza sativa L.) expressing maize C 4 -pepc encoded phosphoenolpyruvate carboxylase under PEG treatment

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Abstract

The aim of this study was to analyze the biochemical and molecular responses of suspension cells of transgenic rice (Oryza sativa L.) over-expressing the maize C 4 -pepc gene encodes phosphoenolpyruvate carboxylase (PEPC) (PC line) to drought stress by exogenous ATP. Suspension cells of PC and wild-type (WT) rice cultured in N6 basic medium were subjected to the following treatments: a 24-h drought treatment imposed by 20% (m/v) polyethylene glycol 6000 (PEG6000 treatment); an exogenous adenosine 5′-triphosphate treatment (ATP; 1 mmol L−1); and a combined exogenous ATP + PEG6000 treatment. We measured DNA fragmentation, cell viability, lipid peroxidation, antioxidant enzyme activities, PEPC activity, ATP content, and the levels of Ca2+, H2O2, and NO using spectrophotometric methods and fluorescent dyes, and quantified the transcript levels of stress-related genes such as NAC1 (one of the transcription factors from NAC (NAM/ATAF/CUC) family), cytochrome P450 (P450) and programmed cell death (PCD) under these treatments. Compared with WT cells, PC cells showed more stable cellular activity, lower lipid peroxidation, enhanced antioxidant enzyme activity, higher levels of endogenous ATP level, PEPC activity, and transcript levels of C 4 -pepc and the transcription factor NAC1 under drought stress. While the content of H2O2 and the transcript levels of P450 and PCD were decreased and kept at the low levels during the same treatments. Adding ATP, PC reinforced these early signaling effects response to PEG6000 treatment. Based on the results, the exogenous ATP triggers a series of signaling cascade reactions for enhancing antioxidant protection in PC cells during the drought treatment, conferring to the drought-tolerance.

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Acknowledgements

This work was supported by Grants from the National Natural Science Foundation of China (No. 31371554; 31571585), the Agricultural Science and Technology Innovation Fund of Jiangsu in China [cx(14)5004] and the Basic Scientific Support Project of Jiangsu Academy of Agricultural Sciences (ZX[16]2002). The authors thank the anonymous reviewers and editorial staff for their time and attention.

Author contributions

Prof. Li supervised the project, designed and guided experiments, and drafted the manuscript; Mr. Huo conducted experiments; Mr. He conducted experiments and analyzed some of the data; Dr. Wei helped the tissue culture. Mr. Lu supervised some of the experiments; Dr. Zhao supervised the qRT-PCR experiments; Prof. Wang provided suggestions on the manuscript. All authors read and approved the final.

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Authors and Affiliations

  1. Provincial Key Laboratory of Agrobiology, Nanjing Branch, Jiangsu High Quality Rice Research and Development Center, China National Center for Rice Improvement, Institute of Food Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, People’s Republic of China

    K. Huo, X. Li, Y. F. He, X. D. Wei, C. F. Zhao & C. L. Wang

  2. College of Life Science, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    K. Huo & W. Lu

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  1. K. Huo
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Huo, K., Li, X., He, Y.F. et al. Exogenous ATP enhance signal response of suspension cells of transgenic rice (Oryza sativa L.) expressing maize C 4 -pepc encoded phosphoenolpyruvate carboxylase under PEG treatment. Plant Growth Regul 82, 55–67 (2017). https://doi.org/10.1007/s10725-016-0238-z

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