The members of class II are quite heterogeneous and can be further subdivided into the CIN and CYC/TB1 subclades ( Martin-Trillo and Cubas, 2010). The most striking difference between these two classes is a four-amino-acid deletion in the basic region of the TCP domain of class I relative to class II proteins ( Martin-Trillo and Cubas, 2010). According to the homology of the TCP domains, the members of the TCP family can be divided into two classes: class I (also named PCF or TCP-P) and class II (also named TCP-C Navaud et al., 2007). This class of transcription factors is characterized by a highly conserved 59-residue-long non-canonical basic helix-loop-helix (bHLH) structure at the N-terminus called the TCP domain, which is involved in DNA binding, protein nuclear localization, and protein–protein interactions ( Cubas et al., 1999 Kosugi and Ohashi, 2002). TCP transcription factors were discovered in 1999 and named after the first three characterized family members: TEOSINTE BRANCHED 1 (TB1) in maize ( Zea mays), CYCLOIDEA (CYC) in snapdragon ( Antirrhinum majus), and PROLIFERATING CELL FACTORS 1 and 2 (PCF1 and PCF2) in rice ( Oryza sativa Cubas et al., 1999). TEOSINTE BRANCHED 1, CYCLOIDEA, and PROLIFERATING CELL FACTORS (TCP) transcription factors constitute a small family of plant-specific transcription factors that play versatile functions in regulating diverse plant growth and development processes by controlling cell proliferation ( Martin-Trillo and Cubas, 2010). Taken together, the present study may provide the basis for functional studies to reveal the role of this gene famil y in strawberry growth and development. Notably, transient over-expression of FvTCP9 in strawberry fruits dramatically affected the expression of a series of genes implicated in fruit development and ripening. Subcellular localization analysis showed that six FvTCP-GFP fusion proteins showed distinct localizations in Arabidopsis mesophyll protoplasts. Transcripts of FvTCP genes also varied with different subcultural propagation periods and were induced by hormone treatments and biotic and abiotic stresses. Eleven FvTCP genes were down-regulated in different fruit developmental stages, while five FvTCP genes were up-regulated. Among them, 12 FvTCP genes exhibited distinct tissue-specific transcript accumulation patterns. We analyzed FvTCP gene transcript accumulation patterns in different tissues and fruit developmental stages. Promoter analysis revealed various cis-acting elements related to growth and development, hormone and/or stress responses. Phylogenetic analysis suggested that the FvTCP genes were classified into two main classes, with the second class further divided into two subclasses, which was supported by the exon-intron organizations and the conserved motif structures. In this study, 19 FvTCP genes were identified in the diploid woodland strawberry ( Fragaria vesca) accession Heilongjiang-3. However, no systematic study has been performed in strawberry. Plant-specific TEOSINTE BRANCHED 1, CYCLOIDEA, and PROLIFERATING CELL FACTORS (TCP) transcription factors play versatile functions in multiple processes of plant growth and development. The experiments were repeated three times and provided consistent results. Genes were hierarchically clustered based on average Pearson’s distance metric and ‘average linkage’ method. The color scale represents relative transcript levels with increased (red) or decreased (green) transcript abundance. The transcript accumulation profiles were generated by semi-quantitative PCR and were visualized as heat maps. (B) Hierarchical clustering of the transcript accumulation profiles of 19 FvTCP genes during different strawberry subcultural propagation periods (original results shown in Supplementary Figure S4). (A) Photos of strawberry subcultural propagation during the five different periods assessed (P1: original plantlet P2: plantlet with 1–2 branch crowns, approximately 2 weeks after subculture P3: plantlets with 3–4 branch crowns, approximately 3 weeks after subculture P4: plantlets with 5–7 branch crowns, approximately 4 weeks after subculture P5: plantlets with over 10 branch crowns, approximately 6 weeks after subculture). vesca) during different periods of subcultural propagation. FIGURE S3: Transcript accumulation pattern of 19 TCP genes in the diploid woodland strawberry ( F.
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