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Plant and Cell Physiology, 2001, Vol. 42, No. 5 492-498
© 2001 Oxford University Press

Molecular Cloning of ß-Galactosidase from Japanese Pear (Pyrus pyrifolia) and its Gene Expression with Fruit Ripening

Akira Tateishi1,3, Hiroaki Inoue1, Hajime Shiba1 and Shohei Yamaki2

1 College of Bioresource Sciences, Nihon University, Fujisawa, Kanagawa, 252-8510 Japan 2 Graduate School of Bioagricultural Science, Nagoya University, Chikusa, Nagoya, 464-8601 Japan

We have cloned a cDNA fragment encoding a ß-galactosidase from Japanese pear (Pyrus pyrifolia) fruit (JP-GAL). It contained an untranslated sequence of 182 nucleotides at the 5' end, a presumptive coding sequence of 2,193 nucleotides and an untranslated sequence of 268 nucleotides including a polyadenylation signal and a poly (A) tail at the 3' end. It encoded a protein with a calculated molecular weight of 80.9 kDa which consists of 731 amino acids. Both the nucleotide and the deduced amino acid sequences showed a 98% sequence identity with that obtained from the apple ß-galactosidase cDNA. The peptide sequence obtained from the purified Japanese pear ß-galactosidase III matched the deduced amino acid sequence of SVSYDHKAIIINGQKRILISG (amino acid 25–45). Northern blot analysis showed that the probe derived from JP-GAL hybridized to a single 2.6 kb RNA. The mRNA was detected solely in the fruit; none was detected in the buds, leaves, roots or shoots of the Japanese pear. The steady-state level of the ß-galactosidase mRNA was measured during fruit ripening in three cultivars, Housui, Kousui (early ripening) and Niitaka (late ripening). The results showed that regardless of the cultivar, no JP-GAL mRNA was detected in the immature fruit. Increment of the mRNA level with fruit ripening coincided with the increase in the ß-galactosidase III activity. Our results showed that the expression of JP-GAL correlated with fruit softening and JP-GAL may be ß-galactosidase III.

3 Corresponding author: E-mail, tateishi@brs.nihon-u.ac.jp; Fax, +81-466-84-3622.


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