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Enhanced tolerance of transgenic sweetpotato plants that express both CuZnSOD and APX in chloroplasts to methyl viologen-mediated oxidative stress and chilling

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Abstract

Oxidative stress is one of the major factors causing injury to plants exposed to environmental stress. Transgenic sweetpotato [Ipomoea batatas (L.) Lam. cv. Yulmi] plants with an enhanced tolerance to multiple environmental stresses were developed by expressing the genes of both CuZn superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) under the control of an oxidative stress-inducible SWPA2 promoter in the chloroplasts of sweetpotato plants (referred to as SSA plants). SSA plants were successfully generated by the particle bombardment method and confirmed by PCR analysis. When leaf discs of SSA plants were subjected to 5 μM methyl viologen (MV), they showed approximately 45% less damage than non-transformed (NT) plants. When 200 μM MV was sprayed onto the whole plants, SSA plants showed a significant reduction in visible damage compared to leaves of NT plants, which were almost destroyed. The expression of the introduced CuZnSOD and APX genes in leaves of SSA plants following MV treatment was significantly induced, thereby reflecting increased levels of SOD and APX in the chloroplasts. APX activity in chloroplast fractions isolated from SSA plants was approximately 15-fold higher than that in their counterparts from NT plants. SSA plants treated with a chilling stress consisting of 4°C for 24 h exhibited an attenuated decrease in photosynthetic activity (Fv/Fm) relative to NT plants; furthermore, after 12 h of recovery following chilling, the Fv/Fm of SSA plants almost fully recovered to the initial levels, whereas NT plants remained at a lower level of Fv/Fm activity. These results suggest that SSA plants would be a useful plant crop for commercial cultivation under unfavorable growth conditions. In addition, the manipulation of the antioxidative mechanism in chloroplasts can be applied to the development of various other transgenic crops with an increased tolerance to multiple environmental stresses.

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Abbreviations

APX:

Ascorbate peroxidase

CuZnSOD:

CuZn superoxide dismutase

MV:

Methyl viologen

NT:

Non-transformed

POD:

Peroxidase

SWPA2 :

Sweetpotato peroxidase anionic 2

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Acknowledgements

This research was supported by grants from the BioGreen21 Program, Rural Development Administration, Korea, from the Environmental Biotechnology National Core Research Center, KOSEF/MOST, Korea, and from the International Collaboration Project, Ministry of Science and Technology (MOST), Korea. We are grateful to Prof. Ray A. Bressan, Purdue University for his valuable comments to the manuscript.

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Authors and Affiliations

  1. Environmental Biotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Oun-dong 52, Yuseong-gu, Daejeon, 305-806, Korea

    Soon Lim, Yun-Hee Kim, Sun-Hyung Kim, Suk-Yoon Kwon, Haeng-Soon Lee & Sang-Soo Kwak

  2. Biofunction Research Team, Bioorganic Science Division, Korea Research Institute of Chemical Technology (KRICT), P.O. Box 107, Yuseong-gu, Daejeon, 305-600, Korea

    Jin-Seog Kim & Kwang-Yun Cho

  3. Department of Horticulture, Chungbuk National University, Cheongju, 361-763, Korea

    Soon Lim & Kee-Yoeup Paek

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  1. Soon Lim
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  2. Yun-Hee Kim
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  3. Sun-Hyung Kim
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  4. Suk-Yoon Kwon
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  9. Sang-Soo Kwak
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Correspondence to Sang-Soo Kwak.

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Lim, S., Kim, YH., Kim, SH. et al. Enhanced tolerance of transgenic sweetpotato plants that express both CuZnSOD and APX in chloroplasts to methyl viologen-mediated oxidative stress and chilling. Mol Breeding 19, 227–239 (2007). https://doi.org/10.1007/s11032-006-9051-0

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