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Molecular cloning and characterization of anthocyanin biosynthesis genes in eggplant (Solanum melongena L.)

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Abstract

Eggplant is rich in anthocyanins, which are one of the important secondary metabolites and beneficial to human health. In this study, four anthocyanin biosynthesis genes, including chalcone synthase (SmCHS), chalcone isomerase (SmCHI), flavanone 3-hydroxylase (SmF3H) and dihydroflavonol 4-reductase (SmDFR), were isolated from eggplant. Their expression profiles were investigated along with other two structural genes (SmF3′5′H and SmANS) in different tissues, bagging, and low-temperature treatments. The highest expression levels were observed in peels except for SmF3H which was detected in stems, and SmF3H was also the exclusive gene that did not show correlation with anthocyanin content. Unlike purple peels of the control, the bagged fruits displayed white peels and had no anthocyanin accumulation, because of the expression of SmCHI, SmF3′5′H, SmDFR, and SmANS totally depended on light. As low temperature stimulates anthocyanin accumulation, all the six anthocyanin biosynthesis genes were up-regulated in cold stress experiment, and SmCHS increased most obviously. Moreover, early biosynthesis genes (SmCHS, SmCHI, and SmF3H) responded earlier than late biosynthesis genes (SmF3′5′H, SmDFR, and SmANS) under low temperature. Subcellular localization suggested that the enzymes encoded by the genes cloned in this study were all located in cytosol and nucleus. To further characterize the function, their ectopic expression in Arabidopsis was performed, and overexpression lines displayed higher anthocyanin accumulation in stems and siliques. The present study provides insight into anthocyanin biosynthesis in eggplant and may facilitate genetic engineering for improvement of the anthocyanin content in plants.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31471870), the National Science Foundation for Young Scholars of China (31301770), and the Shanghai Seed Industry of Agricultural Science Project (2013, No. 5).

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  1. School of Agriculture and Biology, Shanghai Jiao Tong University, NO. 800, Rd. Dong Chuan, Shanghai, 200240, China

    Mingmin Jiang, Yang Liu, Li Ren, Hongli Lian & Huoying Chen

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  1. Mingmin Jiang
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Correspondence to Huoying Chen.

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Communicated by A. Chandra.

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11738_2016_2172_MOESM1_ESM.tif

Supplementary Fig. 1 A schematic representation of the exon-intron architecture of SmCHS, SmCHI, SmF3H, and SmDFR. SmCHS consists of two exons (black box) and one intron (fold line); SmCHI consists of four exons (black box) and three introns (fold line); SmF3H consists of three exons (black box) and two introns (fold line); SmDFR consists of six exons (black box) and five introns (fold line). The scale bar indicates 100 bp. (TIFF 154 kb)

11738_2016_2172_MOESM2_ESM.tif

Supplementary Fig. 2 Phylogenetic trees based on the amino acid sequences of CHSs CHIs, F3Hs, and DFRs. These trees were constructed using the MEGA 6.0 and Neighbor-Joining method with 1000 bootstrap replicates. (TIFF 1815 kb)

11738_2016_2172_MOESM3_ESM.tif

Supplementary Fig. 3 Expression of SmCHS, SmCHI, SmF3H, and SmDFR in different transgenic Arabidopsis lines. (A) SmCHS; (B) SmCHI; (C) SmF3H; and (D) SmDFR. WT, wild-type Arabidopsis; L1-L10, transgenic Arabidopsis lines, respectively. Quantifications were normalized by the expression of Arabidopsis actin (NM_179953). Data represent mean ± SD of three biological replicates. (TIFF 664 kb)

Supplementary Table 1 List of primer sequences used in this study (DOC 46 kb)

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Jiang, M., Liu, Y., Ren, L. et al. Molecular cloning and characterization of anthocyanin biosynthesis genes in eggplant (Solanum melongena L.). Acta Physiol Plant 38, 163 (2016). https://doi.org/10.1007/s11738-016-2172-0

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