THE ANTHOCYANIN BIOSYNTHETIC REGULATOR MDMYB1 POSITIVELY REGULATES ASCORBIC ACID BIOSYNTHESIS IN APPLE

doi:10.15302/J-FASE-2020367

Front. Agr. Sci. Eng.

2021, Vol. 8

Issue (2) : 231-235 https://doi.org/10.15302/J-FASE-2020367

LETTER

THE ANTHOCYANIN BIOSYNTHETIC REGULATOR MDMYB1 POSITIVELY REGULATES ASCORBIC ACID BIOSYNTHESIS IN APPLE

Jianping AN, Xiaofei WANG, Chunxiang YOU, Yujin HAO(

)

State Key Laboratory of Crop Biology, Shandong Collaborative Innovation Center for Fruit and Vegetable Production with High Quality and Efficiency, College of Horticultural Science and Engineering, Shandong Agricultural University, Taian 271018, China.

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Abstract

• The contents of anthocyanin and AsA in red-flesh apples are higher than that in non-red-flesh apples.

• The anthocyanin biosynthetic regulator MdMYB1 directly activates the expression of dehydroascorbate reductase gene MdDHAR, thus promoting the activity of the DHAR enzyme and the accumulation of AsA.

• MdMYB1-MdDHAR module may play a key role in AsA-DHA homeostasis.

Ascorbic acid (AsA, vitamin C) is involved in the regulation of many aspects of plant growth and development. It is an essential micronutrient for humans and can prevent scurvy, maintain the health of gums and blood vessels, reduce the level of plasma cholesterol and enhance the immune systen. Apple cultivars Orin and Guanghui were crossed to obtain a group of hybrid offspring with and without red flesh in the course of assessing apple germplasm resources. Unexpectedly, the red-flesh apples had higher AsA contents than other apples. Further studies showed that the anthocyanin biosynthetic regulator MdMYB1 directly activates the expression of dehydroascorbate reductase gene MdDHAR, thus promoting the activity of the DHAR enzyme and the accumulation of AsA. This finding reveals the mechanism leading to high AsA levels in red-flesh apples and suggests a new idea to cultivate red-flesh apples with high AsA contents and produce AsA efficiently and without pollution.

Keywords anthocyanin apples ascorbic acid MdMYB1 vitamin C

Corresponding Author(s): Yujin HAO

Just Accepted Date: 17 November 2020 Online First Date: 05 January 2021 Issue Date: 13 July 2021

Cite this article:

Jianping AN,Xiaofei WANG,Chunxiang YOU, et al. THE ANTHOCYANIN BIOSYNTHETIC REGULATOR MDMYB1 POSITIVELY REGULATES ASCORBIC ACID BIOSYNTHESIS IN APPLE[J]. Front. Agr. Sci. Eng. , 2021, 8(2): 231-235.

URL:

https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2020367
https://academic.hep.com.cn/fase/EN/Y2021/V8/I2/231

Fig.1 MdMYB1 positively regulates AsA biosynthesis by activating MdDHAR. (a) Representative non-red-flesh and red-flesh apples. Apple crossbreeding groups (non-red-flesh and red-flesh apples) were harvested 140 days after full bloom. Measurement of (b) ascorbic acid (AsA) and (c) dehydroascorbic acid (DHA) contents. Error bars denote standard deviation. Different letters above the bars indicate significant differences (P<0.05) obtained by one-way analysis of variance. (d) Measurement of dehydroascorbic acid reductase activity. The experiments were repeated three times and each experiment contained 3–5 apple fruits per variety. A representative picture is shown here. (e) Detection of the expression levels of MdMYB1 and MdDHAR in non-red-flesh and red-flesh apples. The value for non-red-flesh apple was set to 1. qRT-PCR was conducted in three biological replicates and three technical replicates, and each sample contained 3–5 apple fruits. (f) Representative wild-type (GFP) and MdMYB1-overexpressing (MdMYB1-GFP) apple callus. Measurement of (g) AsA and (h) DHA contents. (i) Measurement of dehydroascorbic acid reductase activity. A representative picture is shown here. (j) Detection of the expression levels of MdMYB1, MdDHAR, MdMDHAR, MdAO and MdAPX1 in transgenic apple callus. The value for GFP was set to 1. (k) Chromatin immunoprecipitation (ChIP)-PCR assays of MdMYB1 binding to the promoter of the MdDHAR gene. The predicted 5′-CTGTTG-3′ sequences are indicated by the black line. Chromatin from the empty vector control (GFP) and 35S:MdMYB1-GFP apple callus (MdMYB1-GFP) were immunoprecipitated with and without anti-GFP antibodies. Two regions (P1 and P2) were examined by qRT-PCR. The enrichment of GFP was set to 1. (l) Electrophoretic mobility shift assay results showing that the MdMYB1-HIS fusion protein bound directly to the MdDHAR promoter. Unlabeled probes were used as competitors. In the mutated probe (Mut), the 5′-CTGTTG-3′ motif was replaced by 5′-CGGTGG-3′. (m) Schematic representation of the GUS reporter vector containing the MdDHAR promoter and the effector vector containing MdMYB1. (n) GUS activity detection of MdDHAR-promoter and MdDHAR-promoter/MdMYB1 transgenic apple callus. MdDHAR-promoter, MdDHAR-promoter transgenic apple callus; MdDHAR-promoter/MdMYB1, MdDHAR-promoter and 35S:MdMYB1-GFP co-transformed apple callus. The value for MdDHAR-promoter was set to 1. (o) Schematic representations of the effector vectors containing MdMYB1, and the fireﬂy luciferase reporter vectors containing the MdDHAR promoter. MdDHAR-promoter, MdDHAR-promoter-pGreen0800-LUC+ pGreen62-SK; MdDHAR-promoter/MdMYB1, MdDHAR-promoter-pGreen0800-LUC+ MdMYB1-pGreen62-SK. (p) Relative LUC/REN activity of effector plasmids and reporter plasmids. The value for MdDHAR-promoter was set to 1. (q) A working model of MdMYB1 functioning in AsA biosynthesis.

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