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Ito T.,National Institute of Fruit Tree Science NIFTS | Furuta T.,Hiroshima Institute of Technology | Namba N.,Agriculture and Forestry Technical Development Center
Journal of General Plant Pathology | Year: 2011

Citrus psorosis virus (CPsV) was detected from citrus trees for the first time in Japan. The diagnosis was confirmed by molecular, serological, and biological indexing. RT-PCR detected CPsV from two citrus trees among ca. 200 tested. Both trees were variety Shiranui of [Citrus unshiu Marc. × C. sinensis (L.) Osb.] × C. reticulata Blanco, and neither had the bark scaling symptom typical of CPsV. The CPsV isolate could be genetically related to those from Spain, Italy, Florida, and California. © 2011 The Phytopathological Society of Japan and Springer. Source


Ito T.,National Institute of Fruit Tree Science NIFTS | Ohta S.,Okitsu Citrus Research Station
Journal of General Plant Pathology | Year: 2010

We tested citrus samples from Japan using reverse transcription-polymerase chain reaction (RT-PCR), and for the first time, Citrus viroid V (CVd-V) was detected in Japan. Using nucleotide sequence analysis, we identified two new CVd-V variants, which shared 91-96% nucleotide sequence identities with those reported previously and 89% with each other. The RT-PCR detected CVd-V from 44 of 275 citrus trees tested, showing that CVd-V can be spread to some extent within commercial orchards in Japan. © 2010 The Phytopathological Society of Japan and Springer. Source


Fujikawa T.,National Institute of Fruit Tree Science NIFTS | Iwanami T.,National Institute of Fruit Tree Science NIFTS
Molecular and Cellular Probes | Year: 2012

Citrus greening disease is caused by " . Candidatus Liberibacter spp.," including " . Candidatus Liberibacter asiaticus (Las)." For detecting this disease, we designed new primers from the Las 16S rDNA and used a very small DNA template for PCR. More Las-infected tissues were detected with our primers than with the common primers. © 2012 Elsevier Ltd. Source


Terakami S.,National Institute of Fruit Tree Science NIFTS | Nishitani C.,National Institute of Fruit Tree Science NIFTS | Yamamoto T.,National Institute of Fruit Tree Science NIFTS
Acta Horticulturae | Year: 2013

Single-nucleotide polymorphisms (SNPs) are desirable DNA markers for marker-assisted selection because of their high abundance, codominant inheritance, locus specificity and potential for automated high-throughput analysis. The average number of SNPs in introns has been reported to be several times higher than that in exons in many organisms. Potential intron polymorphism (PIP) markers, designed from expressed sequence tag (EST) sequences (exon regions), could amplify potential flanking intron regions without requiring genome sequence information. In this study, we used PIP markers designed from apple (Malus × domestica) ESTs to identify intron regions and to detect SNPs in pear (Pyrus spp.). We evaluated 170 PIP markers for Japanese pear P. pyrifolia cultivar 'Housui' and the European pear P. communis cultivar 'Bartlett'. Of the 170 PIP markers, 142 markers amplified fragments in both cultivars. The fragment size was larger than the size predicted from the apple EST sequences, suggesting the existence of introns in the amplified fragments. We sequenced the amplified fragments and detected candidate bi-allelic SNPs in intron regions of both cultivars. The genome sequences obtained from pear were aligned to the apple EST sequences, revealing that the sequences included donor and acceptor sites, and contained deduced exons and flanking introns in pear. SNP analysis showed that 55 and 54 markers were mapped on the genetic linkage maps of 'Bartlett' and 'Housui', respectively. The PIP markers designed from apple ESTs were applicable in pear and could be used to detect SNPs in intron regions and to develop SNP markers in pear. The PIP markers will facilitate marker-assisted selection for disease resistance and other important traits in pear breeding programs. Source

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