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Loss of Function of the Carotenoid Isomerase Gene BrCRTISO Confers Orange Color to the Inner Leaves of Chinese Cabbage (Brassica rapa L. ssp. pekinensis)

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An Erratum to this article was published on 21 September 2014

Abstract

Chinese cabbage varieties with orange inner leaves are of interest due to their attractive appearance and high carotenoid content. Previously, a recessive locus, Br-or, that confers this trait was mapped to linkage group A09 in Brassica rapa, and Bra031539 was identified as the candidate gene. We used a backcross population consisting of 1,392 individuals for fine mapping; three linked markers delimited the Br-or locus to a region of 9.47 kb, which contained only one predicted functional gene, Bra031539 (the carotenoid isomerase gene BrCRTISO). We identified two bacterial artificial chromosome (BAC) clones carrying Br-or and confirmed that this allele of BrCRTISO confers the mutant phenotype by sequencing. Many DNA sequence variations are present in Br-or, including a 90-bp DNA deletion in the promoter and a 501-bp insertion at the 3′ end of the gene. GUS staining showed that the spatial expression of BrCRTISO was perfectly coincident with the site of carotenoid synthesis and that the 90-bp deletion in the promoter slightly downregulated expression of the gene. Using an in vitro enzyme assay, we showed that the mutant Br-or protein cannot convert prolycopene to all-trans-lycopene. Light has a positive effect on the expression of carotenoid biosynthesis genes, except for beta-ring carotene hydroxylase (CHXB), and this coincided with the increase in total lycopene and β-carotene in the Br-or mutant. We also demonstrated that loss of BrCRTISO function, upregulation of the upstream genes, and downregulation of downstream genes lead to the accumulation of prolycopene and confer an orange color to the inner head leaves in Chinese cabbage.

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Abbreviations

CHXB:

Beta-ring carotene hydroxylase

DH:

Doubled haploid

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

IPI:

Isopentenyl pyrophosphate isomerase

LCYB:

Lycopene beta-cyclase

LCYE:

Lycopene epsilon-cyclase

PDS:

Phytoene desaturase

PSY:

Phytoene synthase

VDE:

Violaxanthin de-epoxidase

ZDS:

Zeta-carotene desaturase

ZEP:

Zeaxanthin epoxidase

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Acknowledgments

We would like to thank Dr. Qiuju Yu and Professor Peter Beyer for providing us with helpful suggestions and the tangerine tomato fruits for the prolycopene purification experiment. We thank Dongtao Ren and Luo Zhao (China Agricultural University, China) for their help with the HPLC experiments. This work was supported by grants from the National High Technology Research and Development Program of China (863 Program, Nos. 2012AA020103 and 2012AA100101), the National “Twelfth Five-Year” Plan for Science & Technology Support (2012BAD50G01), and the Program of National Natural Science Foundation (C150203).

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Authors and Affiliations

  1. Beijing Vegetable Research Center (BVRC), Beijing Academy of Agriculture and Forestry Science (BAAFS), 100097, Beijing, China

    Tongbing Su, Shuancang Yu, Yangjun Yu, Deshuang Zhang, Xiuyun Zhao & Weihong Wang

  2. Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (North China), Ministry of Agriculture, 100097, Beijing, China

    Jiao Wang Fenglan Zhang

Authors
  1. Tongbing Su
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  2. Shuancang Yu
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  3. Jiao Wang Fenglan Zhang
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  4. Yangjun Yu
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  5. Deshuang Zhang
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  6. Xiuyun Zhao
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  7. Weihong Wang
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Correspondence to Jiao Wang Fenglan Zhang.

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Tongbing Su and Shuancang Yu contributed equally to this work.

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Su, T., Yu, S., Zhang, J.W.F. et al. Loss of Function of the Carotenoid Isomerase Gene BrCRTISO Confers Orange Color to the Inner Leaves of Chinese Cabbage (Brassica rapa L. ssp. pekinensis). Plant Mol Biol Rep 33, 648–659 (2015). https://doi.org/10.1007/s11105-014-0779-0

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