Pear plants (Pyrus pyrifolia var. Hengshen) showing symptoms of chlorotic spots on leaves were observed in orchards in central Taiwan in 2004. The sap of diseased leaves reacted positively to Apple chlorotic leaf spot virus (ACLSV) antiserum. A purified virus isolate (LTS1) from pear was characterized by host range, electron microscopy, phylogenetic analyses, serological property, and back-inoculation experiments to pear. Fifteen of 28 species of tested plants were susceptible to this virus after mechanical inoculation. Pathogenicity of ACLSV isolate LTS1 was verified by back-inoculating to pear seedlings. Filamentous virions of ≈12 × 750 nm were observed in the preparations of purified virus. Virus particles accumulated in the cytoplasm were observed in the ultrathin sections of LTS1-infected pear leaf tissue. Sequence analyses of the coat protein (CP) gene of LTS1 and the CP gene of LTS2, which originated from a distinct symptomatic pear sample, shared 81.4% to 92.6% nucleotide and 87.6% to 98.4% amino acid identities with those of the CP of 35 ACLSV isolates available in GenBank. ACLSV isolates were grouped into four clusters, i.e., Asia I, II, III, and Europe, and isolates LTS1 and LTS2 were classified as members of cluster Asia II and Asia I, respectively, based on phylogenetic data. Moreover, the variability of amino acid sequences of the CP gene of 37 ACLSV isolates showed geographically associated clustering in the phylogenetic tree. To our knowledge, this is the first study on the characterization of ACLSV causing the leaf chlorotic spot disease of pear in Taiwan. This study also provides the phylogenetic relationships among ACLSV populations based on amino acid sequences of CPs, which are correlated with their geographic origins.
Zhong-Bin Wu, Hsin-Mei Ku, Yuh-Kun Chen, Chung-Jan Chang and Fuh-Jyh Jan
Chung-Jan Chang, Ruth Donaldson, Phil Brannen, Gerard Krewer and Robert Boland
Since 2004, growers and scientists have observed a disorder described as “yellow twig” or “yellow stem” affecting a major selection of southern highbush blueberry, FL 86-19, in the south Georgia blueberry production region. The initial symptom observed was leaf marginal chlorosis and subsequent necrosis, which eventually progressed throughout the whole leaf resulting in early leaf fall. Thin, yellow twigs or yellow stems became evident on some cultivars. The described symptoms on blueberry were similar to those exhibited on grapes with Pierce's disease and on plum with leaf scald disease. This prompted the enzyme-linked immunosorbent assay (ELISA) tests and isolations of Xylella fastidiosa, which is the causal agent of the previously mentioned grape and plum diseases. Two leaf and two root tissue samples were collected from a diseased FL 86-19 plant for isolation and ELISA testing on 2 Mar. 2006. ELISA results indicated all four tissues tested positive for the bacterial pathogen, X. fastidiosa, whereas only the two root tissues provided positive isolations. One leaf and one root tissue sample were later collected from each of five additional diseased plants for isolation and ELISA testing. Both isolation and ELISA testing methods obtained positive results. Cultures were multiplied to inoculate seedlings of three cultivars: ‘Southern Belle’ (eight plants), ‘Premier’ (six), and ‘Powderblue’ (six) on 23 May 2006 and one selection, FL 86-19 (eight), on 31 May 2006. Two FL 86-19 plants started to show symptoms of marginal necrosis 54 days postinoculation, whereas one plant each of ‘Southern Belle’ and ‘Powderblue’ started to show symptoms of marginal necrosis 63 days postinoculation and ‘Premier’ stayed symptomless. All eight culture-inoculated FL 86-19 plants (100%) showed symptoms 72 days postinoculation, but no symptoms were observed on the control plants. One hundred twenty-six days postinoculation, two ‘Powderblue’ and four ‘Southern Belle’ plants showed mild symptoms, whereas all ‘Premier’ plants were asymptomatic. Positive reisolations of the bacteria from the inoculated symptomatic plants, not from asymptomatic plants, fulfilled Koch's postulates, which confirmed X. fastidiosa was the causal bacterium of the new blueberry disorder, the bacterial leaf scorch of blueberry.