Plant Cell Reports

, Volume 38, Issue 11, pp 1393–1402 | Cite as

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

  • So-Eun Kim
  • Ho Soo Kim
  • Zhi Wang
  • Qingbo Ke
  • Chan-Ju Lee
  • Sul-U Park
  • Ye-Hoon Lim
  • Woo Sung Park
  • Mi-Jeong Ahn
  • Sang-Soo KwakEmail author
Original Article


Key message

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


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.


Carotenoid IbOr IbOr-R96H Metabolic engineering Sweetpotato 



Carotenoid cleavage dioxygenases


Hydrogen peroxide


Ipomoea batatas orange






Photosystem II


Phytoene synthase


Reactive oxygen species





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.

Author contribution statement

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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

299_2019_2448_MOESM1_ESM.docx (21 kb)
Supplementary material 1 (DOCX 20 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • So-Eun Kim
    • 1
    • 2
  • Ho Soo Kim
    • 1
  • Zhi Wang
    • 3
  • Qingbo Ke
    • 3
  • Chan-Ju Lee
    • 1
    • 2
  • Sul-U Park
    • 1
    • 2
  • Ye-Hoon Lim
    • 1
    • 2
  • Woo Sung Park
    • 4
  • Mi-Jeong Ahn
    • 4
  • Sang-Soo Kwak
    • 1
    • 2
    Email author
  1. 1.Plant Systems Engineering Research CenterKorea Research Institute of Bioscience and Biotechnology (KRIBB)DaejeonKorea
  2. 2.Department of Environmental Biotechnology, KRIBB School of BiotechnologyUniversity of Science and Technology (UST)DaejeonKorea
  3. 3.Institute of Soil and Water ConservationNorthwest A & F UniversityYanglingChina
  4. 4.College of Pharmacy and Research Institute of Life SciencesGyeongsang National UniversityJinjuKorea

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