Isolation and Analysis of Anthocyanin Pathway Genes from <i>Ribes</i> Genus Reveals MYB Gene with Potent Anthocyanin-Inducing Capabilities
Pavel Starkevič,
Aušra Ražanskienė,
Urtė Starkevič,
Vaiva Kazanavičiūtė,
Erna Denkovskienė,
Vidmantas Bendokas,
Tadeušas Šikšnianas,
Rytis Rugienius,
Vidmantas Stanys,
Raimundas Ražanskas
Affiliations
Pavel Starkevič
Department of Eukaryotic Gene Engineering, Institute of Biotechnology, Vilnius University, 10257 Vilnius, Lithuania
Aušra Ražanskienė
Department of Eukaryotic Gene Engineering, Institute of Biotechnology, Vilnius University, 10257 Vilnius, Lithuania
Urtė Starkevič
Department of Eukaryotic Gene Engineering, Institute of Biotechnology, Vilnius University, 10257 Vilnius, Lithuania
Vaiva Kazanavičiūtė
Department of Eukaryotic Gene Engineering, Institute of Biotechnology, Vilnius University, 10257 Vilnius, Lithuania
Erna Denkovskienė
Department of Eukaryotic Gene Engineering, Institute of Biotechnology, Vilnius University, 10257 Vilnius, Lithuania
Vidmantas Bendokas
Department of Orchard Plant Genetics and Biotechnology, Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Babtai, Lithuania
Tadeušas Šikšnianas
Department of Orchard Plant Genetics and Biotechnology, Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Babtai, Lithuania
Rytis Rugienius
Department of Orchard Plant Genetics and Biotechnology, Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Babtai, Lithuania
Vidmantas Stanys
Department of Orchard Plant Genetics and Biotechnology, Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, 54333 Babtai, Lithuania
Raimundas Ražanskas
Department of Eukaryotic Gene Engineering, Institute of Biotechnology, Vilnius University, 10257 Vilnius, Lithuania
Horticultural crops of the Ribes genus are valued for their anthocyanin-rich fruits, but until now, there were no data about the genes and regulation of their flavonoid pathway. In this study, the coding sequences of flavonoid pathway enzymes and their putative regulators MYB10, bHLH3 and WD40 were isolated, and their expression analyzed in fruits with varying anthocyanin levels from different cultivars of four species belonging to the Ribes genus. Transcription levels of anthocyanin synthesis enzymes and the regulatory gene RrMYB10 correlated with fruit coloration and anthocyanin quantities of different Ribes cultivars. Regulatory genes were tested for the ability to modulate anthocyanin biosynthesis during transient expression in the leaves of two Nicotiana species and to activate Prunus avium promoters of late anthocyanin biosynthesis genes in N. tabacum. Functional tests showed a strong capability of RrMyb10 to induce anthocyanin synthesis in a heterologous system, even without the concurrent expression of any heterologous bHLH, whereas RrbHLH3 enhanced MYB-induced anthocyanin synthesis. Data obtained in this work facilitate further analysis of the anthocyanin synthesis pathway in key Ribes species, and potent anthocyanin inducer RrMyb10 can be used to manipulate anthocyanin expression in heterologous systems.
