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More than 50 isolates of Potato virus Y (PVY) with characteristics of strains that cause tobacco veinal necrosis (PVY

Beet leafhoppers were collected weekly on yellow sticky traps placed at 36 locations in Morrow and Umatilla Counties in northeastern Oregon in April through November 2007, 2008, and 2009. Insects were counted, collected, and a subset of the insects was tested for the presence of the beet leafhopper-transmitted virescence agent phytoplasma, the causal agent of potato purple top disease in this region. Beet leafhoppers were present throughout the sampling period and the number of insects collected peaked in June of each year with smaller peaks in July and October. Of the 804 insects tested for phytoplasma in 2007, 2008, and 2009, 21, 18, and 22% tested positive for phytoplasma, respectively. Most of the phytoplasma-positive insects were collected from mid-June through July. Positive insects, however, were collected as late as 13 November in 2007 and 2008. These data indicate that a relatively high proportion of the beet leafhoppers in this area are harboring the phytoplasma. Therefore, the potential for development of purple top disease of potatoes from migrating beet leafhoppers in this important potato producing region is quite high and measures to control this pest throughout the growing season are probably necessary in order to reduce disease pressure.

Root galls possibly caused by Spongospora subterranea were observed on hairy nightshade (Solanum sarrachoide; HNS). HNS galls and galls from potato were used to artificially inoculate potato and HNS. Eighty-three and 52% potato and HNS plants inoculated with potato inoculum had root galls, respectively. Ten and 31% potato and HNS plants inoculated with HNS inoculum had root galls, respectively. A S. subterranea-specific PCR significantly correlated (P < 0.0001, Phi Coefficient = 0.69) with visual assessment of gall incidence in 34 of 40 samples. Five of 40 samples had no galls, but gave positive PCRs. One of 40 samples had root galls, but gave a negative PCR. More sporosori were produced on potato than on HNS. Therefore, the root galls on HNS were caused by S. subterranea that is able to complete its life cycle on HNS and produce new generations of sporosori that are infectious on potato.

Hairy nightshade is a common weed in potato rotations in the western United States. As a close relative of potato, hairy nightshade can host numerous potato nematodes, diseases, and insect pests. Hairy nightshade hosts three common parasitic nematodes of potato, Columbia and northern root-knot nematodes, and stubby root nematode. Tubers of a potato breeding line with roots that are resistant to Columbia root-knot nematode—race 1, were damaged when grown in the presence of hairy nightshade. The weed provided an alternate host for the nematode, which then allowed the nematode to infect susceptible tubers. Stubby root nematodes transmit tobacco rattle virus (TRV), the causal agent for corky ringspot disease (CRS) of potato. CRS disease was maintained in soil when hairy nightshade was present in rotation crops of alfalfa or Scotch spearmint that otherwise eliminated the disease. Hairy nightshade also is a host of potato leaf roll virus (PLRV), which is transmitted by green peach aphids (GPA). GPA pre...

The ability of several weed species to serve as hosts for tobacco rattle virus (TKV), the causal agent of corky ringspot disease of potato (CRS), and its nematode vector,Paratrichodorus allius, was investigated in greenhouse studies. ViruliferousP. allius multiplied on 24 out of 37 weed species tested, indicating they were suitable hosts of the vector. However, only 11 of these weeds were infected with TRV, as determined by ELISA. The nonhost status of a given weed species was not changed whether the viruliferous vector population originated from CRS problem fields in WA, OR, or ID. Several weeds served as hosts for the vector and virus including kochia, prickly lettuce, henbit, nightshade species (black, hairy, and cutleaf), common chickweed, and annual sowthistle. Virus-freeP. allius acquired TRV from the three nightshade species, volunteer potato grown from TRV-infected tubers, and prickly lettuce, and subsequently transmitted the virus to ‘Samsun NN’ tobacco indicator plants. Thus, some weeds may play a role in the epidemiology of CRS by perpetuating TRV and its vector in a problem field.

Tobacco rattle virus (TRV), the causal agent of corky ringspot disease of potato (CRS), was detected by reverse transcription-polymerase chain reaction (RT-PCR) in root tissues of field- and greenhouse-grown wheat (two) and corn (10) cultivars. TRV was also detected in some of the same samples by enzyme-linked immunosorbent assay (ELISA), but in a low frequency. Compared to ‘Samsun NN’ tobacco, wheat and corn appeared to be poorer hosts of TRV. However, these crops served as inoculum reservoirs of TRV. A nonviruliferous population of the nematodeParatrichodorus allius, the vector of TRV, acquired the virus from infected wheat and corn roots, and transmitted it to potato. ‘Russet Norkotah’ potatoes grown following TRV-infected corn and wheat, were blemished with arcs, concentric rings and diffuse browning, characteristic symptoms of CRS. The observations presented here suggest that wheat and corn as rotational crops will sustain TRV in a CRS problem field and the virus may cause severe damage on subsequent potato crops.

Soil samples from corky ringspot (CRS) problem fields of potato in the states of Washington and Oregon were collected and planted withNicotiana tabacum ‘Samsun NN’ tobacco to bait tobacco rattle virus (TRV) and to increaseParatrichodorus allius populations, the vector of TRV. Pathogenicity of three isolates of TRV was assessed on Russet Burbank and Russet Norkotah plants usingP. allius as the vector. The most severe CRS tuber symptoms were with TRV from Pasco, WA, followed by Umatilla, OR, and Mattawa, WA, indicating a distinct variation in virulence among virus isolates. The lowest number ofP. allius that transmitted TRV resulting in CRS symptoms on Russet Norkotah was three nematodes per 250 cm3 soil. When potato plants were exposed toP. allius at different times in their growth, tubers on older plants were more resistant than younger tubers to CRS. Severity of CRS tuber symptoms was correlated with age of potato plants in pot culture. Reproduction ofP. allius on potato roots did not influence the incidence and severity of tubers symptoms. Under similar conditions, Bintje, a variety known to be resistant to CRS, served as a host forP. allius, yet remained asymptomatic. The protocol employed in these studies is appropriate for testing potato germplasm for resistance to CRS.

The stubby root nematode, Paratrichodorus allius, is important to the potato industry in the Pacific Northwest of USA, because it vectors Tobacco rattle virus (TRV), the causal agent of corky ringspot disease. The current method for determining if nematodes are viruliferous for TRV takes several weeks, requiring a glasshouse bioassay followed by a serological test. To overcome this drawback, a rapid and affordable molecular test was developed using reverse transcription polymerase chain reaction (RT-PCR) to identify viruliferous P. allius nematodes within 48 hours. Primers from the 16 kDa gene of TRV were used to detect TRV in both greenhouse-reared and field collected P. allius. TRV RNA can be detected consistently in nucleic acids equivalent to one quarter of a viruliferous adult nematode reared in the greenhouse. In order to reduce the time and expense of processing individual nematodes from field samples, viral RNA was consistently and affordably detected in extracts from 5 field-collected adult P. allius.

A method to establish two experimental corky ringspot disease (CRS) plots that had no prior CRS history is described. CRS is a serious disease of potato in the Pacific Northwest caused by tobacco rattle virus (TRV) and transmitted primarily by Paratrichodorus allius. ‘Samsun NN’ tobacco seedlings were inoculated with viruliferous P. allius in the greenhouse before they were transplanted into the field soil at the rate of 3,000 plus seedlings/ha. Care was taken to keep soil around plants in the greenhouse and transplants in the field moist to avoid vector mortality. The vector population in the soil of one of the fields was monitored by extraction, examination under microscope and bioassay on tobacco seedlings to ascertain that they were virus carriers. Presence of virus in tobacco bioassay plants was determined by visual symptoms on tobacco leaves and by testing leaves and roots using ELISA. Although TRV transmission was rapid, there was loss of infectivity in the first winter which necessitated a re-inoculation. After two years of planting infected tobacco seedlings, 100% of soil samples collected from this field contained viruliferous P. allius. In the second field, all five commercial potato cultivars, known to be susceptible, expressed symptoms of CRS disease indicating that the procedure was successful.

Experiments were conducted to determine whether the beet leafhopper, Circulifer tenellus (Baker) (Hemiptera: Cicadellidae), transmits the purple top phytoplasma to potato, Solanum tuberosum L.; beets, Beta vulgaris L.; and selected weed hosts. The beet leafhopper-transmitted virescence agent (BLTVA) phytoplasma was identified as the causal agent of the potato purple top disease outbreaks that recently occurred in the Columbia Basin of Washington and Oregon. The phytoplasma previously was found to be associated almost exclusively with the beet leafhopper, suggesting that this insect is the probable vector of BLTVA in this important potato-growing region. Eight potato cultivars, including 'Russet Burbank', 'Ranger Russet', 'Shepody', 'Umatilla Russet', 'Atlantic', 'FL-1879', 'FL-1867', and 'FL-1833', were exposed for a week to BLTVA-infected beet leafhoppers. After exposure, the plants were maintained outdoors in large cages and then tested for BLTVA by using polymerase chain reaction after 6 to 7 wk. The leafhoppers transmitted BLTVA to seven of the eight exposed potato cultivars. Sixty-four percent of the exposed plants tested positive for the phytoplasma. In addition, 81% of the BLTVA-infected potato plants developed distinct potato purple top disease symptoms. Beet leafhoppers also transmitted BLTVA to beets and several weeds, including groundsel, Senecio vulgaris L.; shepherd's purse, Capsella bursa-pastoris (L.) Medik); kochia, Kochia scoparia (L.) Schrad; and Russian thistle, Salsola kali L. This is the first report of transmission of BLTVA to potatoes, beets, and the above-mentioned four weed species. Results of the current study prove that the beet leafhopper is a vector of the potato purple top disease.

At least 16 taxa of cicadellids and delphacids were tested by polymerase chain reaction (PCR) for the presence of a phytoplasma in the clover proliferation group, designated 16SrVI. Nucleic acid extracts from individual insects or groups of 5-10 were tested using PCR primers designed from the DNA sequence of 16S-23S rRNA or ribosomal protein genes of the pathogen. The beet leafhopper, Circulifer tenellus (Baker), was most often associated with the phytoplasma, with approximately 16% of the insects testing positive. The phytoplasma was occasionally found associated with Ceratagallia spp. Leafhopper species that were not associated with the phytoplasma included Macrosteles spp., Dikraneura spp., Colladonus montanus (Van Duzee), Circulifer geminatus (Van Duzee), Ballana spp., Amplysellus spp., Paraphlepsius spp., Texananus spp., Balclutha spp., Latalus spp., Erythroneura spp., Exitianus exitiosus (Uhler), and unidentified delphacids. The detected phytoplasma was similar to, or synonymous with, the beet leafhopper-transmitted virescence agent that is associated with the potato purple top disease in the Columbia Basin region of Washington and Oregon. This is in contrast to the phytoplasma associated with potato purple top disease in Mexico that is related to aster yellows (group 16SrI). The association of the group 16SrVI phytoplasma almost exclusively with the beet leafhopper suggests that this insect is the major vector of the phytoplasma in this region.

More than 50 isolates of Potato virus Y (PVY) with characteristics of strains that cause tobacco veinal necrosis (PVYN) were obtained from potatoes (Solanum tuberosum L.) grown in the northwestern United States. These isolates are being characterized at the biological and molecular levels. Isolate RR1 was obtained from leaves of potato cv. Ranger Russet showing distinct mottling and leaf deformity, which is in contrast to the leaf-drop and necrosis usually observed with ordinary strains of PVY (PVYO) in this variety. Isolate AL1 was obtained from tubers of potato cv. Alturas showing distinct internal light brown rings and blotches. When RR1 and AL1 were transmitted to tobacco (Nicotiana tabacum L. cvs. Samsun NN and 423), they caused systemic veinal necrosis, including stem and petiole lesions typical of PVYN strains (2). Symptoms induced by RR1 and AL1 on tobacco appeared 9 to 11 days after inoculation, whereas some other isolates caused delayed veinal necrosis. All isolates that produced veinal necrosis on tobacco were detectable with PVY polyclonal antisera. Potato virus X was not detected by enzyme-linked immunosorbent assay in tobacco plants showing veinal necrosis. Some isolates, including AL1, failed to react in serological tests using PVYN-specific monoclonal antibodies obtained from three commercial sources. Other isolates, including RR1, were detectable with these monoclonal antibodies. Reverse transcription-polymerase chain reaction (RT-PCR) products obtained with primers specific for the coat protein (CP) open reading frame (ORF) were cloned and sequenced. AL1 possesses a CP more closely related to PVYO type isolates, which would account for its failure to react with PVYN monoclonal antibodies. In this regard, AL1 is similar to the PVYN-Wilga isolate (1). Other isolates that are detectable with the PVYN monoclonal antibodies possess a CP more consistent with N strains of the virus. Results of RT-PCR tests using primers derived from the P1 ORF sequence (3), and the restriction enzyme analysis and sequencing of the RT-PCR products, all suggest that AL1 and RR1 are related to European-type members of PVY tuber necrotic (NTN) or N strains. However, other isolates under investigation appear to be more closely related to previously reported North American NTN types (3). The symptomatology of these viruses on tobacco and potato, and the serological and molecular data clearly show that at least two distinct variants of PVYN have been found for the first time in a major potato production area of the United States, and pose a potential threat to the potato industry. References: (1) B. Blanco-Urgoiti et al. Eur. J. Plant Pathol. 104:811, 1998. (2) J. A. de Bokx and H. Huttinga. Potato virus Y. Descriptions of Plant Viruses. No. 242, CMI/AAB, Surrey, England, 1981. (3) R. P. Singh et al. Can J. Plant Pathol. 20:227, 1998.