Differential expression of members of the β-galactosidase gene family during japanese pear (Pyrus pyrifolia L.) fruit growth and on-tree ripening

Galactosidases are thought to play a key role in cell wall metabolism during fruit growth and ripening. In this study we cloned seven β-galactosidase (β-Gal) cDNAs from Japanese pear fruit and designated them PpGAL2, PpGAL3, Pp-GAL4, PpGAL5, PpGAL6, PpGAL7, and PpGAL8, in addition to the previously described JP-GAL hereinafter termed PpGAL1. mRNA expression patterns of these clones were characterized throughout fruit growth and on-tree ripening, and in leaves and shoots in three Japanese pear cultivars, 'Housui', 'Kousui', and 'Niitaka'. The shared amino acid sequence identity among the eight Japanese pear β-Gal (PpGAL) clones ranged from 50% to 60%. They all contained the putative active site containing consensus sequence pattern G-G-P-[LIVM](2)-x(2)-Q-X-E-N-E-[FY] belonging to glycoside hydrolase family 35. Expression of all the clones was both development- and tissue-specific. PpGAL1 and PpGAL4 were only expressed in the ripe fruit while PpGAL2 and PpGAL3 were expressed in both expanding and ripening fruit with their abundance being highest in the ripe fruit. The abundance of PpGAL5, PpGAL6, and PpGAL7 mRNAs was highest in expanding fruit but decreased drastically upon the onset of ripening. PpGAL8 was only detected in very young fruit (15 days after full bloom) and not in expanding and ripening fruit. These results indicate that in japanese pear fruit β-Gal is encoded by a multigene family whose members show distinct and overlapping expression during the various phases of fruit development. Some of the members are not only fruit-specific but also ripening-specific and, therefore, may play a crucial role in cell wall disassembly during japanese pear fruit softening.