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A single amino acid change at position 96 (Arg to His) of the sweetpotato Orange protein leads to carotenoid overaccumulation

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Abstract

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IbOr-R96H resulted in carotenoid overaccumulation and enhanced abiotic stress tolerance in transgenic sweetpotato calli.

Abstract

The Orange (Or) protein is involved in the regulation of carotenoid accumulation and tolerance to various environmental stresses. Sweetpotato IbOr, with strong holdase chaperone activity, protects a key enzyme, phytoene synthase (PSY), in the carotenoid biosynthetic pathway and stabilizes a photosynthetic component, oxygen-evolving enhancer protein 2-1 (PsbP), under heat and oxidative stresses in plants. Previous studies of various plant species demonstrated that a single-nucleotide polymorphism (SNP) from Arg to His in Or protein promote a high level of carotenoid accumulation. Here, we showed that the substitution of a single amino acid at position 96 (Arg to His) of wild-type IbOr (referred to as IbOr-R96H) dramatically increases carotenoid accumulation. Sweetpotato calli overexpressing IbOr-WT or IbOr-Ins exhibited 1.8- or 4.3-fold higher carotenoid contents than those of the white-fleshed sweetpotato Yulmi (Ym) calli, and IbOr-R96H overexpression substantially increased carotenoid accumulation by up to 23-fold in sweetpotato calli. In particular, IbOr-R96H transgenic calli contained 88.4-fold higher levels of β-carotene than those in Ym calli. Expression levels of carotenogenesis-related genes were significantly increased in IbOr-R96H transgenic calli. Interestingly, transgenic calli overexpressing IbOr-R96H showed increased tolerance to salt and heat stresses, with similar levels of malondialdehyde to those in calli expressing IbOr-WT or IbOr-Ins. These results suggested that IbOr-R96H is a useful target for the generation of efficient industrial plants, including sweetpotato, to cope with growing food demand and climate change by enabling sustainable agriculture on marginal lands.

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Abbreviations

CCD:

Carotenoid cleavage dioxygenases

H2O2 :

Hydrogen peroxide

IbOr:

Ipomoea batatas orange

MDA:

Malondialdehyde

Or:

Orange

PSII:

Photosystem II

PSY:

Phytoene synthase

ROS:

Reactive oxygen species

Ym:

Yulmi

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Acknowledgements

This work was supported by grants from the Systems & Synthetic Agrobiotech Center (PJ01318401 and PJ01318402), the Biogreen 21 Project for the Next Generation, Rural Development Administration, Korea, the National Natural Science Foundation of China (31700335), and the KRIBB initiative program.

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  1. So-Eun Kim and Ho Soo Kim contributed equally to this work.

Authors and Affiliations

  1. Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Daejeon, 34141, Korea

    So-Eun Kim, Ho Soo Kim, Chan-Ju Lee, Sul-U Park, Ye-Hoon Lim & Sang-Soo Kwak

  2. Department of Environmental Biotechnology, KRIBB School of Biotechnology, University of Science and Technology (UST), 217 Gajeong-ro, Daejeon, 34113, Korea

    So-Eun Kim, Chan-Ju Lee, Sul-U Park, Ye-Hoon Lim & Sang-Soo Kwak

  3. Institute of Soil and Water Conservation, Northwest A & F University, Yangling, 712100, Shaanxi, China

    Zhi Wang & Qingbo Ke

  4. College of Pharmacy and Research Institute of Life Sciences, Gyeongsang National University, 501 Jinjudae-ro, Jinju, 52828, Korea

    Woo Sung Park & Mi-Jeong Ahn

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  1. So-Eun Kim
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Contributions

SEK, HSK, and SSK contributed to the research design. SEK, HSK, ZW, QK, CJL, SUP, and YHL performed site-directed mutagenesis, transformation, qRT-PCR, and stress-tolerant assays. WSP and MJA performed HPLC for the measurement of carotenoid contents. SEK, HSK and SSK primarily wrote the manuscript.

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Correspondence to Sang-Soo Kwak.

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Communicated by Salim Al-Babili.

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Kim, SE., Kim, H.S., Wang, Z. et al. A single amino acid change at position 96 (Arg to His) of the sweetpotato Orange protein leads to carotenoid overaccumulation. Plant Cell Rep 38, 1393–1402 (2019). https://doi.org/10.1007/s00299-019-02448-4

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