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To understand the role of the MIKC-type  dormancy-associated MADS-box  ( DAM ) genes in the regulation of endodormancy in japanese pear ( Pyrus pyrifolia ), we isolated two  DAM  genes from ‘Kosui’ and characterized their expression throughout the seasonal endodormancy phases in ‘Kosui’, as well as in TP-85–119 taiwanese pear ( P .  pyrifolia ), which is a less dormant type. Several copies of the corresponding  DAM  genes are present in the  P .  pyrifolia  genome. Rapid amplification of cDNA ends enabled the isolation of two full-length cDNAs, designated as  PpMADS13–1  and  PpMADS13–2 , with complete open reading frames encoding 227 and 234 amino acids, respectively. Multialignment of the two ‘Kosui’ and the database DAM genes (based on the deduced amino acid sequences) showed that PpMADS13–1 and PpMADS13–2 were highly identical to the Rosaceae DAM genes and encoded the conserved domains characteristic of other MIKC-type MADS-box genes. The phylogenetic relationships showed that PpMADS13–1 and PpMADS13–2 were more closely related to the  Prunus  DAM, though they formed a unique subclade. The specific expression analysis of  PpMADS13–1  and  PpMADS13–2  by real-time polymerase chain reaction showed that both  DAM  genes are gradually down-regulated concomitant with endodormancy breaking.  PpMADS13–1  and  PpMADS13–2  showed similar fluctuations in expression patterns, although  PpMADS13–2  was more highly expressed relative to  PpMADS13–1 . The expression of  PpMADS13–1  and  PpMADS13–2  in the less dormant taiwanese pear, TP-85–119, was quite low (nearly zero level), which is consistent with a down-regulated pattern of expression of the  DAM  genes in japanese pear, peach ( Prunus persica ), and japanese apricot ( Prunus mume ). Differential genomic DNA methylation patterns detected in  PpMADS13–1  and  PpMADS13–2  were not concomitant with seasonal endodormancy transition phases, suggesting that DNA methylation in these loci under investigation may not be linked to endodormancy progression in ‘Kosui’.

journal of the american society for horticultural science

Molecular Cloning of Dormancy-associated MADS-box Gene Homologs and Their Characterization during Seasonal Endodormancy Transitional Phases of Japanese Pear