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Oxidative stress induced in chloroplasts or mitochondria promotes proline accumulation in leaves of pea (Pisum sativum): another example of chloroplast-mitochondria interactions

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Abstract

Oxidative stress can occur in different parts of plant cells. We employed two oxidants that induce reactive oxygen species (ROS) in different intracellular compartments: methyl viologen (MV, in chloroplasts) and menadione (MD, in mitochondria). The responses of pea (Pisum sativum) leaf discs to MV or MD after 4-h incubation in dark or moderate (300 μE m−2 s−1) or high light (1200 μE m−2 s−1) were examined. Marked increase in ROS levels was observed, irrespective of compartment targeted. The levels of proline, a compatible solute, increased markedly much more than that of ascorbate or glutathione during oxidative/photo-oxidative stress, emphasizing the importance of proline. Further, the activities and transcripts of enzymes involved in biosynthesis or oxidation of proline were studied. An upregulation of biosynthesis and downregulation of oxidation was the basis of proline accumulation. Pyrroline-5-carboxylate synthetase (P5CS, involved in biosynthesis) and proline dehydrogenase (PDH, involved in oxidation) were the key enzymes regulated under oxidative stress. Since these two enzymes—P5CS and PDH—are located in chloroplasts and mitochondria, respectively, we suggest that proline metabolism can help to mediate inter-organelle interactions and achieve redox homeostasis under photo-oxidative stress.

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Abbreviations

ASC:

Ascorbate

DAB:

3,3′-Diaminobenzidine

GSH:

Glutathione

HL:

High light

MD:

Menadione

MDA:

Malondialdehyde

ML:

Moderate light

MV:

Methyl viologen

NBT:

Nitro blue tetrazolium chloride

P5CDH:

Pyrroline-5-carboxylate dehydrogenase

P5CR:

Pyrroline-5-carboxylate reductase

P5CS:

Pyrroline-5-carboxylate synthetase

PDH:

Proline dehydrogenase

PVDF:

Polyvinylidene difluoride

ROS:

Reactive oxygen species

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Acknowledgments

Authors thank Prof. Maria Elena Alvarez, Faculty of Chemistry, National University of Cordoba, Argentina for kindly providing us with proline dehydrogenase antibodies for our experiments.

Funding

This work was supported by grants to ASR from Council of Scientific and Industrial Research (No. 38(1404)/15/EMR-II), JC Bose National Fellowship (No. SR/S2/JCB-06/2006). VA, PR, and RBB were all supported by Research Fellowships from University Grants Commission, New Delhi, India. We also thank grants from DST-FIST, UGC-SAP-CAS, and DBT-CREBB, from New Delhi, India for support of infrastructure in Department/School.

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Authors and Affiliations

  1. Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, 500046, India

    Vetcha Aswani, Pidakala Rajsheel, Ramesh B. Bapatla, Bobba Sunil & Agepati S. Raghavendra

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  1. Vetcha Aswani
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  2. Pidakala Rajsheel
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  3. Ramesh B. Bapatla
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  4. Bobba Sunil
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  5. Agepati S. Raghavendra
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Contributions

ASR designed the study. VA performed most of the experiments. PR, RBB, and BS performed some experiments. VA and ASR analyzed results and wrote the manuscript. VA, PR, RBB, BS, and ASR revised and finalized the manuscript. All the authors read and approved the final manuscript.

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Correspondence to Agepati S. Raghavendra.

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Aswani, V., Rajsheel, P., Bapatla, R.B. et al. Oxidative stress induced in chloroplasts or mitochondria promotes proline accumulation in leaves of pea (Pisum sativum): another example of chloroplast-mitochondria interactions. Protoplasma 256, 449–457 (2019). https://doi.org/10.1007/s00709-018-1306-1

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