Search

A field isolate of Potato virus Y (PVY) was collected in New Brunswick, Canada in 2007 due to unusual symptoms observed on different potato cultivars. To unveil the PVY strain identity, tobacco and potato bioassays, PVYO and PVYN-specific antibody-based enzyme-linked immunosorbent assays, and reverse-transcription polymerase chain reaction (PCR)-based genotyping were carried out. All the assays demonstrated that the isolate, designated as PVYO-FL in this study, belonged to the PVYO strain group. Greenhouse tests with the potato cvs. FL 1533 and Jemseg confirmed the severe nature of infection by PVYO-FL. The complete genome sequences of PVYO-FL and PVYO-RB, the latter a mild PVYO isolate, were determined. BLAST analysis revealed that the two isolates shared 97 and 98% sequence identities at the nucleotide and polyprotein levels, respectively. Further BLAST analysis unveiled that PVYO-FL shared 99.7% nucleotide sequence identity with PVYO-Oz, an isolate reported in New York, United States, whereas the PVYO-RB isolate shared 99.2% sequence identity with PVYO-139, a PVYO isolate reported in New Brunswick, Canada. A phylogenetic tree of available, full-length sequences of PVY isolates demonstrated two subgroups within the PVYO branch, one clustered with PVYO-RB and the other with PVYO-FL. Group-specific sense primers for differentiation of the two subgroups were developed and evaluated. A limited survey of potato tubers collected from a field plot at the Potato Research Centre, Agriculture and Agri-Food Canada, using the newly developed PCR primers, indicated that 65.3 and 2.4% of the PVYO-positive tubers were infected with PVYO isolates belonging to the PVYO-FL and PVYO-RB subgroups, respectively. Assessment of the pathogenicity of three representative isolates from each subgroup on the potato cv. Jemseg demonstrated that severe and mild symptoms were induced by the PVYO-FL-like and PVYO-RB-like isolates, respectively.

Potato (Solanum tuberosum L.) cv. Shepody is highly resistant to Potato virus A (PVA), yielding no visible symptoms after rub inoculation. In 'Shepody' rootstocks graft-inoculated by PVA-infected scions from a susceptible host, we found a resistance consisting of traces of necrosis (necrotic streaks) in stems and chlorosis in newly emerged leaves. The response was temperature dependent, appearing at 15 to 18 degrees C but not at 28 to 31 degrees C. Necrosis was also observed in tubers, appearing first in the bud end and spreading randomly throughout the majority of the tuber. Trace amounts of PVA in plant tissues were detected by a combination of reverse transcription polymerase chain reaction and Southern blot procedures. The virus concentration was significantly higher in visibly necrotic areas than in nonnecrotic areas, suggesting that presence of the necrosis may be concentration dependent. Pathogenesis-related gene expression showed that chitinase A and B, glucanase B, and PR-10a were associated with chlorotic or necrotic symptoms in leaves, stems, and tubers. Gene expression was markedly more evident in tuber tissues than in leaves and stems. Furthermore, generation of oxidants was also observed within the necrotic areas. Although PVA was detected in necrotic areas of tubers, newly emerged plants from the same tubers were PVA-free.

Columnea latent viroid, originating from ornamental plants, is known to be harmful to crop plants (2). Despite the potential threat to crop plants, the importance of ornamental plants in viroid evolution is not fully appreciated. Availability of a Pospiviroid genus-specific primer pair (1) to detect the most prevalent viroids in ornamental plants and a simplified nucleic acid preparation protocol (3) for use in reverse-transcription polymerase chain reaction (RT-PCR) have facilitated surveys of ornamental plants for pospiviroids. Using the above protocol in India, leaf and shoot samples were collected randomly from roadside beds consisting of ground covers or creepers/trailing plants at the IARI campus, New Delhi. These were extracted in 50 mM NaOH + 2.5 mM EDTA solution, centrifuged to sediment the coarse debris from sap, and 10 μl of the supernatant was spotted on a nitrocellulose membrane. Individual spots were eluted with distilled sterile water (30 μl) and the eluates were used for RT-PCR detection of viroids (3). Amplified fragments or subsequently cloned plasmids were also purified using NaOH-EDTA membrane protocol. Cloning and sequencing of amplicons (195 to 224 bp) revealed a very high sequence identity with specific viroids from the viroid sequence database (NCBI). Among the 19 plant samples assayed, only three plant species were infected by viroids, although none of them exhibited any symptoms. The three plant species included: (i) moss verbena, Glandularia puchella (Verbenaceae, native to Argentina and Chile, now established in several regions of the world), infected with a viroid (Accession No. DQ846884) having 99% sequence identity to Citrus exocortis viroid (CEVd) (Accession No. S67446); (ii) trailing verbena, Verbena × hybrida (Verbenaceae, ornamental plant), doubly infected with a viroid (Accession No. DQ846885) having 95% sequence identity to CEVd (Accession No. DQ094297) and infected with another viroid (Accession No. DQ846883) having 98% sequence identity to Tomato chlorotic dwarf viroid (TCDVd) (Accession No. AF162131); and (iii) red joyweed, Alternanthera sessilis (Amaranthaceae, a perennial weed herb) infected with a viroid (Accession No. DQ846886) having 96% sequence identity to Iresine viroid (IrVd) (Accession No. DQ094293). CEVd and TCDVd were mechanically transferred to tomato seedlings causing reduced growth of plants, smaller leaves, and bunchy-top appearance of plants, symptoms similar to those typically observed with other isolates of these viroids. As expected from previous studies, IrVd was not transmitted to tomato plants. Natural infection of verbena with CEVd has been detected in North America (2) but this was a novel observation in India. Additional novel observations include: A. sessilis as a new host for IrVd; and TCDVd is the first crop viroid to be isolated from a naturally infected ornamental plant. The significance of these viroid findings in ground cover and widely grown ornamental plants may lie in their potential role in spreading the viroids to citrus plants in citrus-growing countries such as India. References: (1) H. Bostan et al. J. Virol. Methods 116:189, 2004, (2) R. P. Singh and J. A. Teixeira da Silva. Floriculture, Ornamental Plant Biotechnol. 3:531, 2006. (3) R. P. Singh et al. J. Virol. Methods 132:204, 2006.

Potato spindle tuber viroid (PSTVd) was detected in pollen grains from PSTVd-infected potato cultivar Katahdin by return polyacrylamide gel electrophoresis. Pollination of the flowers of healthy Katahdin plants with pollen from PSTVd-infected plants resulted in infection of leaves at the base of inflorescences, apical leaves, and tubers. Electrophoretic analysis of fruits indicated sporadic PSTVd infection of sepals, fruit skin, and fruit pulp. True potato seed from each fruit was 35-66% infected with PSTVd (.)

(...) Second-generation potato plants infected with a single strain and grown from greenhouse or field-infected tubers were completely protected against reinoculation with another strain irrespective of the cultivar resistance, and no challenge strain was detected. However, protection was readily broken by changing the method of challenge inoculation. Studies of cross-protection with eight experimental host plants indicated that protection was more complete in those hosts in which the protecting strain multiplied rapidly and spread to the entire plant rather than in those where PSTV infection was difficult or the spread of the viroid was erratic. These studies point out that the extent of cross-protection depends highly on the host plant