MdMYBL2 helps regulate cytokinin-induced anthocyanin biosynthesis in red-fleshed apple (Malus sieversii f. niedzwetzkyana) callus
Yicheng Wang
A , Jingjing Sun B , Nan Wang A , Haifeng Xu A , Changzhi Qu A , Shenghui Jiang A , Hongcheng Fang A , Mengyu Su A , Zongying Zhang A and Xuesen Chen A C
+ Author Affiliations
- Author Affiliations
A State Key Laboratory of Crop Biology, College of Horticulture Science and Engineering, Shandong Agricultural University, Tai-An, Shandong, China.
B College of Forestry, Shandong Agricultural University, Tai-An, Shandong, China.
C Corresponding author. Email: chanyetixi@163.com
Functional Plant Biology 46(2) 187-196 https://doi.org/10.1071/FP17216
Submitted: 2 August 2017 Accepted: 20 September 2018 Published: 18 October 2018
Abstract
Anthocyanin biosynthesis is induced by cytokinins, and is regulated by MYB transcription factors. However, the underlying molecular mechanisms have not been fully characterised. In the present study, red-fleshed apple callus were induced from the leaves of an R6/R6 homozygous line, which was the hybrid offspring of Malus sieversii f. niedzwetzkyana and ‘Fuji’. We analysed the callus anthocyanin contents in response to different cytokinin concentrations. We observed that cytokinin treatments upregulated the expression of anthocyanin structural genes MdDFR and MdUFGT and transcription factor genes MdMYB10 and MdbHLH3. Additionally, the expression of MdMYBL2, which encodes the bHLH and EAR motifs, was inhibited by cytokinin treatments. The MdMYBL2-overexpressing callus had lower anthocyanin contents than the wild-type controls. We noted that the expression levels of anthocyanin biosynthesis structural genes MdDFR and MdUFGT and transcription factor genes MdMYB10 and MdbHLH3 were strongly suppressed in the transgenic callus. Subsequent yeast two-hybrid, bimolecular fluorescence complementation, and pull-down assays indicated that MdMYBL2 interacts with MdbHLH3, which may influence the expression of anthocyanin biosynthesis-related genes. Our findings may provide new insights into how MYB transcription factors influence the cytokinin-regulated anthocyanin biosynthesis in red-fleshed apples.
Additional keywords: anthocyanin, cytokinins, MdbHLH3, MdMYBL2.
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