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Kumar S.,CSIR - Central Electrochemical Research Institute | Singh L.,CSIR - Central Electrochemical Research Institute | Ferretti L.,Cra Centro Of Ricerca Per La Patologia Vegetale | Barba M.,Cra Centro Of Ricerca Per La Patologia Vegetale | And 2 more authors.
Indian Journal of Virology | Year: 2013

During a survey conducted in the grapevine orchards of Himachal Pradesh, variety of symptoms ranging from leaf yellowing, vein greening, reduced leaf size, downward rolling/cup shaped leaves to reduced fruit bearing were observed. Symptomatic leaf samples were collected and analyzed by serological (DAS-ELISA) and molecular methods (RT-PCR, PCR) for viruses and phytoplasma known worldwide on grapevine. DAS-ELISA was used for detection of Grapevine leafroll associated virus 1, 2 and 3 (GLRaV-1, 2 & 3), Grapevine virus A (GVA), Grapevine fan leaf virus (GFLV), Grapevine fleck virus (GFkV) and successfully detected GLRaV-1 & 3 and GFkV. All these samples were complemented with RT- PCR along with GVb and phytoplasma (additional to ELISA) using specific primers. Specific amplification in RT-PCR for GLRaV-1 (∼232 bp), GLRaV-3 (∼300 bp), GFkV (∼179 bp) and GVB (∼440 bp) confirmed the presence of these pathogens. Overall, ELISA and RT-PCR results confirmed the presence GLRaV-3 (66.7 %), GLRaV-1& GFkV (50 %), and Grapevine virus B (GVB) (12.5 %) in symptomatic plants. None of the samples were found positive for GFLV, GLRaV-2 and phytoplasma. Mixed infection was common and none of the plants were found virus free. To the best of our knowledge this is the first report of detection of GFkV and GVB in India. © 2013 Indian Virological Society. Source

Gallelli A.,Cra Centro Of Ricerca Per La Patologia Vegetale | Talocci S.,Cra Centro Of Ricerca Per La Patologia Vegetale | L'Aurora A.,Cra Centro Of Ricerca Per La Patologia Vegetale | Loreti S.,Cra Centro Of Ricerca Per La Patologia Vegetale
Phytopathologia Mediterranea | Year: 2011

Pseudomonas syringae pv. actinidiae (Psa), the causal agent of bacterial canker of kiwifruit, was monitored in symptomless fruits, twigs and pollen of the host using bacterial isolation and DNA-extraction followed by two PCR-assays (direct-PCRs). A procedure for Psa detection from symptomless twigs was established. Out of 16 symptomless twigs samples, Psa was detected in 12 samples by isolation and 13 samples by direct-PCR. Thirteen pollen samples were treated using two different procedures; Psa was detected in eight samples by isolation and ten samples by direct-PCR. By washing 108 samples of fruits, Psa was detected by isolation in only two samples, collected from severely affected orchards. However, one of these samples contained wilted fruits, whereas for the other, only one colony was isolate. From 60 bulk-samples of fruits, endophytic Psa was detected in six samples by isolation and ten samples by direct-PCRs. A Psa-positive bulk-sample of fruits was analyzed separately as individual fruits: there was a faint signal in five or seven fruits out of 50 depending on the PCR assay used. Isolation was negative for these samples. Presence of the pathogen on bulk-fruit samples could be due to low amounts of inoculum distributed over many fruits: as a consequence, there is a negligible risk of introducing the pathogen into countries free of bacterial canker by symptomless fruits. This integrated approach (isolation plus PCR) is proposed as a tool for the analysis of symptomless kiwifruit material for the presence of Psa. Source

de Simone D.,Cra Centro Of Ricerca Per La Patologia Vegetale | D'Amico L.,Cra Centro Of Ricerca Per La Patologia Vegetale | Bressanin D.,Cra Centro Of Ricerca Per La Patologia Vegetale | Motta E.,Cra Centro Of Ricerca Per La Patologia Vegetale | Annesi T.,Cra Centro Of Ricerca Per La Patologia Vegetale
Mycological Progress | Year: 2011

Thirteen isolates of Inonotus rickii/Ptychogaster cubensis, from different geographic provenances, were analyzed by sequencing ITS1, ITS 2 and 5,8S ribosomal RNA region. A phylogenetic tree, also including sequences available in Genbank database, showed that the strains enclosed in this study fall into two well-separated groups, one formed by isolates from Florida (USA) and the other one by isolates from Europe, Argentina and China.Differences were also highlighted on the growth rate of mycelial cultures at different temperatures. In fact, although the tested isolates generally attained the best growth at 30°C, isolates from Europe seem well adapted to higher temperatures and went on growing at 40°C whilst the growth of isolates from Florida significantly decreased at 35°C. Since the teleomorph I. rickii was never detected in Florida, and in this study noticeable differences were detected by analysis of ITS region, the existence of two possible distinct species, not discriminated solely on the basis of morphological characters, could be suggested. © 2010 German Mycological Society and Springer. Source

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