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Identification of a viroid associated with fruit disorder of Japanese pear


Bulletin of the Fruit Tree Research Station 0(27): 65-77
Identification of a viroid associated with fruit disorder of Japanese pear
Recently, abnormal fruit disorders of Japanese pear cultivars, Niitaka and Yoshino, have been observed in Chiba, Ibaraki and Oita Prefectures. The characteristic symptoms consist of a rough fruit shape. In order to determine whether viroids are the causal agents of this disorder, RNA was isolated from Japanese pear with the disorder. Electrophoresis on polyacrylamide gels showed two bands (RNA1, RNA2) which were not detected in extracts from healthy trees. RNA1 was susceptible to RNaseA in 0.1 times SSC (0.15M NaCl, 0.015M sodium citrate) and 2.0 times SSC. RNA2 was susceptible to RNaseA in 0.1 times SSC but was resistant to RNaseA in 2.0 times SSC. These reactions to RNaseA indicated that RNA1 displayed a single-stranded structure and RNA2 was double-stranded (ds) RNA. The mobility of RNA1 and RNA2 on PAGE was similar to that of apple scar skin viroid (ASSVd) and apple scar skin associated RNA2 (ASSARNA2), respectively. RNA1 was detected by return, bidirectional and two-dimensional PAGE. RNA1 purified by electrophoresis was transmitted to pear and apple seedlings by the carborundum or razor-slash method and multiplied in them. The 32P-labeled ASSVd cDNA probe hybridized with RNA1. Healthy apple trees of cv. Starking Delicious, crab apple NY11894 were inoculated by grafting with Japanese pear's bark or bud with the fruit disorder. Dappling and scaring appeared in cv. Starking Delicious fruits. Dappling and size reduction appeared in NY11894 fruits. These results indicated that RNA1 is a viroid similar to ASSVd. Our results suggest that this viroid is strongly associated with the Japanese pear fruit disorder. However it remains to be determined whether this viroid is the causal agent of Japanese pear fruit disorder.

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Accession: 002863262



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