First Report of Natural Infection by 'Candidatus Liberibacter solanacearum' in Bittersweet Nightshade (Solanum dulcamara) in the Columbia Basin of Eastern Oregon.

Potatoes are a major crop in the Columbia Basin of Oregon and Washington, representing an annual farm gate value of almost $750 million. Zebra chip disease (ZC), a new and economically important disease of potato, was first reported in Oregon and Washington in 2011 (1). The disease is caused by the bacterium 'Candidatus Liberibacter solanacearum' (Lso, also referred to as 'Ca. L. psyllaurous'), which is vectored by the potato psyllid (Bactericera cockerelli Sulc) (1,2). Identifying alternative hosts for Lso may facilitate management of ZC disease, which has increased potato production costs in the region. The perennial weed, bittersweet nightshade (Solanum dulcamara L.), is a year-round host of the potato psyllid (3) and is also a suspected host of Lso. However, little is known about the role of this weed in ZC epidemiology. Naturally occurring bittersweet nightshade plants (n = 21) were sampled at six different locations near Hermiston, Oregon, between May and October in 2012. These plants exhibited several symptoms associated with Lso, ranging from asymptomatic to slight purpling, chlorosis, or scorching of the foliage. However, S. dulcamara exhibits similar symptoms under a variety of environmental conditions (drought stress, etc.); therefore, it was difficult to identify potentially infected plants based solely on symptomology. Leaf and stem tissue (n = 21) was analyzed with high-fidelity PCR using species-specific primers for the 16S rDNA gene, CLipoF, and OI2c (2,4). Approximately 27.3% of the plants tested positive for Lso using these primers, including plants from the following locations on 16 April, 16 May, and 24 May, respectively: Hat Rock, OR (45°55.033' N, 119°10.495' W), Irrigon, OR (45°54.560' N, 119°24.857' W), and Stanfield, OR (45°46.971' N, 119°13.203' W). Three plants were selected for further PCR analysis with primers for the outer membrane protein gene, 1482f and 2086r (1). Amplicons obtained with both sets of PCR primers were directly sequenced. A BLAST analysis showed that the 16S rDNA gene sequence (993 to 1,000 bp) shared 99 to 100% identity with several Lso accessions, including JN848751.1 (from Washington) and JN848753.1 (from Oregon). Likewise, the outer membrane protein gene sequence (600 to 601 bp) shared 99 to 100% identity with 'Ca. L. solanacearum' accession KC768330.1 (from Honduras). All six sequences were deposited in GenBank (Accession Nos. KJ854199 to KJ854204). According to these findings, bittersweet nightshade may be an important annual source of Lso in the region, particularly since it serves as a host for the potato psyllid. Potato psyllids were also detected at two of the locations with infected S. dulcamara: Irrigon, OR, and Stanfield, OR. A subsample of the psyllids collected in 2012 were analyzed with PCR and Lso was detected in a sample from Stanfield, OR (5). Identifying perennial hosts of Lso promotes a better understanding of both ZC disease epidemiology and management. To our knowledge, this is the first report of Lso causing natural infections in S. dulcamara in the United States. References: (1) J. M. Crosslin et al. Plant Dis. 96:452, 2012. (2) S. Jagoueix et al. Mol. Cell. Probes 10:43, 1996. (3) A. F. Murphy et al. Am. J. Pot. Res. 90:294, 2013. (4) G. A. Secor et al. Plant Dis. 93:574, 2009. (5) K. D. Swisher et al. Am. J. Pot. Res. 90:570, 2013.

First Report of Ustilago cynodontis Causing Smut of Cynodon dactylon in Washington State, United States.

Bermudagrass (Cynodon dactylon) is an important warm-season perennial turf and forage grass that is typically grown in warm, tropical and subtropical climates. Smutted inflorescences of bermudagrass were observed and collected in Benton County, Washington, United States, in October of 2012 in an unmanaged, naturalized area located near the banks of the Columbia River and adjacent to large expanses of managed turf containing bermudagrass. The climate in this area is favorable to bermudagrass due to the relatively mild winters and hot, dry summers that usually occur in this region. The infected plants occurred in patches alongside healthy plants and several disease foci were observed along a 100-m transect of non-contiguous bermudagrass. The disease was severe wherever it occurred. Diseased inflorescences were covered with black-brown teliospores, distorted, and frequently failed to fully emerge and develop. Teliospores (n = 80) were irregularly globose to subglobose, 5.3 to 7.0 × 4.5 to 6.2 µm (mean 6.4 × 5.9 µm) and 6.2 to 8.8 × 5.3 to 7.0 µm (mean 7.0 × 6.5 µm), with a smooth wall approximately 1 µm thick, and were consistent with previous descriptions of Ustilago cynodontis teliospores (1,3). Teliospores germinated within 24 h when plated on 0.2% malt agar at 16°C and produced 4-celled basidia in a 3+1 arrangement, also consistent with U. cynodontis (3). Basidia gave rise to lateral and terminal, ovoid to long ellipsoidal basidiospores. Basidiospores budded or germinated by hyphae from which lateral or terminal aerial sporidia developed as previously described (3,4). DNA was extracted from sporidia of three single-spored isolates grown in malt extract broth. Complete nucleotide sequences of the 5.8S ribosomal RNA coding region and partial sequences of the internal transcribed spacer (ITS) regions 1 and 2 were obtained from the three isolates using ITS1 and ITS4 primers. The corresponding regions of the three aligned sequences (GenBank Accession Nos. KC920742 to KC920744) were identical and exhibited 99 to 100% identity with U. cynodontis strains previously deposited in GenBank (HM143013, AY740168, AF038825, and AY345000). Representative specimens were deposited in the WSU Mycological Herbarium as WSP 72345 to WSP 72348. This is the first report of U. cynodontis causing smut on bermudagrass in Washington State and represents the northernmost record of this fungus in North America (2). The occurrence of U. cynodontis in Washington State suggests that the pathogen may exist in other hot and dry areas of northwestern North America where bermudagrass is found associated with turf in recreational, landscape, or natural settings. References: (1) S. D. Brook. Trans. R. Soc. N. Z. 84:643, 1957. (2) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory, ARS, USDA. Online. Retrieved from http://nt.ars-grin.gov/fungaldatabases , April 18, 2013. (3) C. T. Ingold. Trans. Br. Mycol. Soc. 83:251, 1984. (4) C. T. Ingold. Trans. Br. Mycol. Soc. 89:471, 1987.

First Report of Zebra Chip Disease and "Candidatus Liberibacter solanacearum" on Potatoes in Oregon and Washington State.

In August of 2011, potato (Solanum tuberosum) tubers grown in the lower Columbia Basin of southern Washington State and northern Oregon were observed with internal discolorations suggestive of zebra chip disease (ZC). Symptoms included brown spots, streaks, and stripes in and near the vascular tissue, typical of ZC (1). Symptoms were observed in cvs. Alturas, Russet Norkotah, Pike, Ranger Russet, Umatilla Russet, and Russet Burbank. Foliar symptoms on plants that produced symptomatic tubers included purple discoloration in upper leaves, leaf rolling, axial bud elongation, chlorosis, leaf scorch, and wilt. Tissue was taken from two symptomatic tubers each of cvs. Alturas and Russet Norkotah, three tubers of cv. Umatilla Russet, and one tuber of cv. Pike. These tubers were tested by PCR for "Candidatus Liberibacter solanacearum", an unculturable alphaproteobacterium associated with ZC (1,4). Primers specific for the 16S rDNA were CLipoF (4) and OI2c (3), and primers OMB 1482f and 2086r were specific for the outer membrane protein (2). All of these samples, except one Umatilla tuber, were positive for the bacterium. The 16S rDNA and OMB amplicons from one symptomatic tuber each of Alturas (from Washington) and Pike (from Oregon) were cloned and three clones of each were sequenced. BLAST analysis of the consensus sequences confirmed "Ca. L. solanacearum". The 16S sequences (1,071 bp) from the two tubers were identical and showed 99 to 100% identity to a number of 16S rDNA sequences of "Ca. L. solanaceaum" in GenBank (e.g., Accession Nos. HM246509 and FJ957897). The 16S rDNA sequences were deposited in GenBank as Accession Nos. JN848751 and JN848753. Consensus sequences of the two OMB clones (605 bp; deposited in GenBank as Accession Nos. JN848752 and JN848754) were identical and showed 97% identity to the two "Ca. L. solanacearum" OMB sequences in GenBank (Accession Nos. CP002371 and FJ914617). Potato psyllids (Bactericera cockerelli Sulc), the vector of "Ca. L. solanacearum", were present in ZC-affected fields in Oregon and Washington and the bacterium was confirmed by PCR in 5 to 10% of 128 adult psyllids collected from two fields. On the basis of foliar and tuber symptoms, specific PCR amplification with two primer pairs, sequence analyses, and the presence of Liberibacter-infected potato psyllids, ZC and "Ca. L. solanacearum" are present in potatoes in Oregon and Washington State. Washington and Oregon together grow ~80,000 ha of potatoes. ZC has caused significant economic damage to potatoes in Texas, Mexico, Central America, and New Zealand (1). Therefore, ZC may pose a risk to agriculture in Oregon, Washington, and neighboring states. However, the potential for development of widespread and serious disease will depend upon the arrival time and number of infective potato psyllids entering the region. References: (1) J. M. Crosslin et al. Online publication. doi:10.1094/PHP-2010-0317-01-RV, Plant Health Progress, 2010. (2) J. M. Crosslin et al. Southwest. Entomol. 36:125, 2011. (3) S. Jagoueix et al. Mol. Cell. Probes 10:43, 1996. (4) G. A. Secor. Plant Dis. 93:574, 2009.

Reducing tuber damage by potato tuberworm (Lepidoptera: Gelechiidae) with cultural practices and insecticides.

Cultural practices and insecticide treatments and combinations were evaluated for effect on tuber damage by potato tuberworm, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae) in the Columbia basin of eastern Oregon and Washington. A range of intervals between initial application of several insecticides and vine-kill were tested to determine how early to implement a program to control potato tuberworm tuber damage. Esfenvalerate, methamidophos, and methomyl were applied at recommended intervals, with programs beginning from 28 to 5 d before vine-kill. All insecticide treatments significantly reduced tuber damage compared with the untreated control, but there was no apparent advantage to beginning control efforts earlier than later in the season. Esfenvalerate and indoxacarb at two rates and a combination of the two insecticides were applied weekly beginning 4 wk before and at vine-kill, and indoxacarb was applied at and 1 wk postvine-kill as chemigation treatments. Application of insecticides at and after vine-kill also reduced tuberworm infestation. 'Russet Norkotah' and 'Russet Burbank' plants were allowed to naturally senesce or were chemically defoliated. They received either no irrigation or were irrigated by center-pivot with 0.25 cm water daily from vine-kill until harvest 2 wk later. Daily irrigation after vine-kill reduced tuber damage, and chemical vine-kill tended to reduce tuber damage compared with natural senescence. Covering hills with soil provides good protection but must be done by vine-kill. Data from these trials indicate that the most critical time for initiation of control methods is immediately before and at vine-kill.

Spatial and temporal dynamics of potato tuberworm (Lepidoptera: Gelechiidae) in the Columbia Basin of the Pacific Northwest.

A landscape-scale study from 2004 to 2006 investigated the spatial and temporal dynamics of a new pest to the Columbia Basin of the Pacific Northwest, the potato tuberworm, Phthorimaea operculella (Zeller). Male P. operculella were monitored in spring, summer, and fall each year with a pheromone-baited trapping network in Oregon and Washington. The objectives of the study were to (1) describe the temporal and spatial dynamics of the recent outbreak of P. operculella in the region and (2) examine the relationship of the spatial and temporal distribution of the outbreak and weather (air temperature, precipitation, and dew point) and geographic variables (elevation and latitude). Weather data during the P. operculella outbreak were compared with a reference period (1993-1999) that occurred before the outbreak. The outbreak in 2004, which caused the first widespread tuber damage in the region, was positively associated with warmer temperatures in the preceding fall and in the spring, summer, and fall of the growing season. October and November 2003 and March 2004 were also drier than the reference period. However, the winter of 2003/2004 was colder than the reference period and thus mild winter conditions did not explain the outbreak. The importance of environmental variables on the seasonal spatial distribution of the pest each year was examined using nonparametric multiplicative regression. Locations with higher spring, summer, or fall temperatures were associated with increased trapping rates in most seasons. Elevation and latitude seemed to play a constraining role, because low trapping rates of P. operculella were associated with higher elevations and latitudes.

First Report of Garlic virus B and Garlic virus D in Garlic in the Pacific Northwest.

With the recent report of several viruses infecting garlic (Allium sativum L.) grown in the Pacific Northwest (1-3), studies were initiated on cloves planted in the fall of 2006 to determine the presence of additional viruses infecting plants exhibiting mosaic and/or chlorotic leaves. Cloves from symptomatic plants of the cultivar 'Early' from two seed production fields in Benton County, WA and two seed production fields in Morrow County, OR were tested by two-step reverse transcription (RT)-PCR using primers specific to the coat protein (CP) of the allexiviruses (4), since garlic infected with this group had similar symptoms in Asia and South America (4). Of the 87 cloves tested, 84 were positive, and four representative samples of the RT-PCR amplicons from each location were cloned and sequenced. Sequence comparisons indicated that the cloves from both locations were infected with Garlic virus D (GarV-D), also known as Japanese garlic virus (JGV), since they shared 98% identity with known isolates (GenBank Accession Nos. L388922.1, AF519572.1, and AB010303.1). In addition, sequences of isolates from the Oregon cloves shared a 96% identity with a known isolate of Garlic virus B (GarV-B; GenBank Accession No. AF543829.1). Because no antiserum specific to these viruses was available, primers specific to the CP genes of GarV-D (JGV-F2/JGV-R2 5'-GCTCACTCRGATGTGTTAGC-3' and 5'-CGCGTGGACATAAGTTGTTG-3') and GarV-B (GVB-F1/GVB-R2 5'-GAGGAGAACTAACGCCACAC-3' and 5'-ACGACCTAGCTTCCTACTTG-3') were designed and the cloves were retested by RT-PCR using these virus-specific primers. With the GarV-D specific primers, 98 and 63% of the cloves were positive from Washington and Oregon, respectively, and 52% of the cloves from Oregon were positive using the GarV-B specific primers. None of the cloves tested from Washington were positive for GarV-B. The identity of the amplicons was verified by cloning and sequencing (GarV-D, GenBank Accession No. FJ643476; GarV-B, GenBank Accession No. FJ643475). Incidence of the two viruses differed between Oregon and Washington was likely due to the expansion of the seed lots in two different locations (California and Nevada) prior to planting in 2006. With such high infection rates, studies should be conducted to determine the impact of these viruses on yield when plants are singly infected as well as in combination with the other viruses known to infect garlic in this region. These and the other viruses (1) are likely to impact yield. To our knowledge, this is the first report of GarV-D (JGV) and GarV-B in garlic in the Pacific Northwest. References: (1) S. L. Gieck et al. Plant Dis. 91:461, 2007. (2) H. R. Pappu et al. Plant Dis. 89:205, 2005 (3) H. R. Pappu et al. Online publication. doi:10.1094/PHP-2008-0919-01-RS. Plant Health Progress, 2008. (4) T. Tsuneyoshi et al. Phytopathology 86:253, 1996.

Detection, Quantification, and Vegetative Compatibility of Verticillium dahliae in Potato and Mint Production Soils in the Columbia Basin of Oregon and Washington.

Soil samples from 87 fields intended for potato production in the Columbia Basin of Washington and Oregon and 51 fields intended for mint production in Washington were assayed on a semiselective medium to quantify populations of Verticillium dahliae. The pathogen was isolated from 77 (89%) of the fields intended for potato production and 41 (80%) of the fields intended for mint production. Population densities ranged from 0 to 169 propagules/g of air-dried soil in fields intended for potato production and 0 to 75 propagules/g of air-dried soil in fields intended for mint production. Isolates of V. dahliae were recovered from soil assay plates and pure cultures were prepared to provide a collection of isolates for vegetative compatibility analysis. Among fields intended for potato production from which isolates of the fungus were assigned to a vegetative compatibility group (VCG), 93% of the fields were found to contain one or more isolates assigned to VCG 4A, nine (23%) contained one or more isolates assigned to VCG4B, and only one (3%) contained any isolates assigned to VCG 2B. In the case of fields planned for mint production in 1999 from which isolates of the fungus were assigned to a VCG, 13 fields (81%) were found to contain one or more isolates assigned to VCG 4A, 7 (44%) contained one or more isolates assigned to VCG 4B, and 5 (31%) contained one or more isolates assigned to VCG 2B. VCG 4A isolates of V. dahliae are widespread and numerous, particularly following potato production, but cause only mild to moderate symptoms in mint; therefore, this pathotype is unlikely to seriously endanger subsequent plantings of mint. However, planting potato in a field recently used to produce mint may pose a significant risk to the potato crop if high populations of the VCG4A pathotype (highly aggressive to potato) predominate. Preplant assessment of soil populations of V. dahliae without regard for the relative populations of various pathotypes present in a particular sample may lead to information not fully useful in integrated pest management systems.

First Report of Corky Ringspot Caused by Tobacco rattle virus on Potatoes (Solanum tuberosum) in Michigan.

Potato is the fourth most important agricultural commodity in Michigan and is grown for table stock, chip processing, and seed. Tubers are either processed or fresh packed immediately following harvest or sent to storage. Tubers of potato cv. FL1879 representing two separate fields in Huron County were removed from separate storage facilities in February and March of 2007, and substantial internal necrosis was observed in 1 to 2% of the tubers. Symptoms included arcs similar to those caused by Tobacco rattle virus (TRV). This virus is a member of the genus Tobravirus and is transmitted by a number of species of stubby-root nematodes (Paratrichodorus or Trichodorus spp.). Stubby-root nematodes have been reported previously from Michigan (1). To confirm the presence of TRV, nucleic acid extractions were made from these symptomatic tubers (3). Samples were initially tested for TRV by reverse transcription (RT)-PCR using primers specific to the 16 kDa open reading frame on genomic RNA-1 (2). Samples from both storage facilities were positive. The 463-bp RT-PCR product, amplified with these primers, was sequenced (GenBank Accession No. EU315227) and was 99.6% identical to the corresponding region of two TRV isolates from Florida and Washington (GenBank Accession Nos. AF055912 and EU315226, respectively). In addition, sap from cv. FL1879 tuber samples was used to transmit the virus mechanically to tobacco cv. Samsun NN, which produced typical TRV symptoms following inoculation, and sap extracts from the tobacco leaves also tested positive with antiserum specific to TRV upon subsequent ELISA testing. Corky ringspot can result in substantial losses, with entire potato fields being rejected because of internal tuber damage. Once found, fields must be considered permanently at risk to this disease due to the large host range of both the virus and the nematode vector. This disease has been previously found in the United States in California, Colorado, Florida, Idaho, Washington, Oregon, and it is likely present in Indiana. To our knowledge, this is the first report of corky ringspot and TRV on potato in Michigan. References: (1) G. W. Bird and N. Knobloch. Plant Dis. Rep. 60:76, 1976. (2) J. M. Crosslin and P. E. Thomas. Am. Potato J. 72:605, 1995. (3) G. G. Presting et al. Phytopathology 85:436, 1995.

Control of Meloidogyne chitwoodi in Potato with Shank-injected Metam Sodium and other Nematicides.

Metam sodium (MS) is often applied to potato fields via sprinkler irrigation systems (water-run, WR) to reduce propagules of soil-borne pathogenic fungi, particularly Verticillium dahliae, to prevent yield loss from potato early dying disease. However, this procedure has not been effective for controlling quality defects in tubers caused by Columbia root-knot nematode (Meloidogyne chitwoodi). In five trials from 1996 to 2001, application of MS by soil shank injection (SH) provided better control and tuber quality than that generally obtained by WR MS, in three of five trials. Results were similar when SH MS was injected at one (41-45 cm), two (15 and 30 cm) or three (15, 30 and 45 cm) depths. In the two trials where SH metam potassium was tested, culls were reduced to 3% and 0% and were equivalent to those resulting from a similar rate in kg a.i./ha of SH MS. A shank-injected tank mix of MS plus ethoprop EC and SH MS plus in-season chemigation applications of oxamyl provided acceptable control in trials where SH MS alone was inadequate. In-furrow application of aldicarb at planting following SH MS did not appear to increase performance. Most consistent control (0-2% culled tubers in five trials) occurred when SH MS at 280 liters/ha was used together with 1,3- dichloropropene (140 liters/ha), applied simultaneously or sequentially. This was similar to combinations of 1,3-D and WR MS, but SH MS may be preferred under certain conditions.

Control of Paratrichodorus allius and Corky Ringspot Disease in Potato with Shank-injected Metam Sodium.

Corky ringspot disease (CRS) of potato produces necrotic areas in tubers that are considered quality defects that can lead to crop rejection. CRS is caused by tobacco rattle virus that is vectored by stubby-root nematodes (Paratrichodorus spp., Trichodorus spp.) at very low population densities, making disease management difficult and expensive. Fumigation with metam sodium (MS) is a common practice to control soil-borne fungi and increase potato yield. MS is generally applied in water via chemigation (water-run, WR) but is ineffective at controlling CRS when WR-applied, even at high rates. Therefore, WR MS is often used in combination with 1,3-dichloropropene (1,3-D), aldicarb or oxamyl to attain adequate CRS control. Between 1996 and 2000, fields with a history of CRS were treated with WR MS, shank-injected MS, and/or 1,3-D, and tubers were evaluated for symptoms of CRS. Shank injection of MS (SH MS) at depths of 41 cm, 15 and 30 cm, or 15, 30 and 45 cm controlled CRS over 3 years of testing. All rates of 280 liters/ha or greater were effective. Shank injection of metam potassium (MP) at rates of 448 liters/ha was also effective. 1,3-D controlled CRS alone or in combination with WR or SH MS. Proper shank application of MS or MP may adequately control CRS without the additional cost of other nematicides at low (<10 P. allius/250 g soil) to moderate (10 to 30 P. allius/250 g soil) populations of the nematode vector. Although SH MS was superior to WR MS, additional research is necessary to determine if this practice would be sufficient at higher CRS disease pressure or if addition of other nematicides would be necessary.

First Report of Soilborne wheat mosaic virus on Wheat (Triticum aestivum) in the Columbia Basin of Oregon.

The Columbia Basin of Oregon consists of six counties (Gilliam, Hood River, Morrow, Sherman, Wasco, and Umatilla) and is the major wheat-producing region in the state. In 2005, these counties produced 300,277 ha of mostly fall-planted wheat. While primarily a dryland production area, wheat (approximately 8,094 ha) is also grown as a rotational crop under irrigation. Stunted and chlorotic winter wheat plants with leaves exhibiting a mosaic pattern consistent with that caused by Soilborne wheat mosaic virus (SBWMV) were observed in March 2005. These plants originated from four center-pivot irrigated fields in Umatilla County with each field approximately 50.6 ha. One-half of one field was planted with cv. Western Breeders 470 (WB470) and the other half with cv. Tubbs, while the three other fields were planted to Tubbs. In the split-planted field, symptoms were widespread in the WB470 half but only observed in low-lying areas planted with Tubbs. ELISA with a monoclonal antibody (Agdia Inc., Elkhart, IN) confirmed the presence of SBWMV, which is transmitted by the soilborne organism Polymyxa graminis. Electron microscopy confirmed rigid, rod-shaped particles that were 19 nm wide and of two size classes, 138 to 222 and 416 to 471 nm long. Presence of SBWMV was further verified by reverse transcription (RT)-PCR using SBWMV RNA-2 specific primers (sense 5'-AAAGAGTCTIGCGTRTARCAYTC-3' and antisense 5'-AACGGTGTTAGTAARYTRGGKGA-3'), which amplified the predicted 338-bp product from the coat protein gene (1). Additional positive samples were found in 2006 from fall-planted wheat cvs. WB 528 and MJ9 from two additional 50.6-ha fields in Umatilla County. In 2005, yield of WB470 in the split-planted field was reduced by approximately 15% compared with yields obtained in similar fields planted with WB470 not exhibiting symptoms. SBWMV has been reported previously in Oregon (2) but nearly 322 km to the west in an area that is not the major wheat-producing region in Oregon. Because of the apparent reduced susceptibility of Tubbs, which is an older cultivar, as compared with WB470, WB528, and MJ9, which are three new high-yielding cultivars, additional research is needed to identify the reaction to SBWMV of cultivars adapted for production in the Pacific Northwest, particularly if this disease becomes widely distributed in both irrigated and dryland production areas. References: (1) G. R. G. Clover et al. Plant Pathol. 50:761, 2001. (2) M. L. Putman et al. Plant Dis. 78:102, 1994.

First Report of Onion yellow dwarf virus, Leek yellow stripe virus, and Garlic common latent virus in Garlic in Oregon.

A general mosaic and yellowing of leaves of three cultivars of garlic (Allium sativum L., Late, Early, and Germinador) were observed in two seed-production fields in Morrow County, OR in June 2005. Approximately 50% of plants within the 50-ha fields were symptomatic. With recent findings of Onion yellow dwarf virus (OYDV), Leek yellow stripe virus (LYSV), and Garlic common latent virus (GCLV) in Washington (2), 45 composite samples of 10 leaves each from symptomatic (mosaic and yellowing) and nonsymptomatic plants were analyzed with a GCLV-specific antiserum (Agdia Inc., Elkhart, IN). All samples of 'Germinador' were infected regardless of symptoms, whereas 6.7% of all 'Late' and 'Early' samples were positive. GCLV infection was verified by reverse transcription (RT)-PCR using primers specific to the coat protein gene of GCLV followed by cloning and sequencing of the cloned amplicon. To determine the presence of a potyvirus, all composite samples were also tested with a general potyvirus antiserum (Agdia) and all samples from symptomatic plants were found to be positive. Representative positive samples from each cultivar were then tested by RT-PCR using degenerate, potyvirus group specific primers (3), and an amplicon of the expected size was obtained. To confirm which potyvirus was present, amplicons were cloned and sequenced, and sequence comparisons indicated that the representative samples were infected with OYDV. All symptomatic samples from the three cultivars were positive for OYDV when tested by RT-PCR using primers specific to its coat protein gene (1). Additionally, 53.3 and 6.7% of 'Early' and 'Late' samples, respectively, were also positive when tested with LYSV-specific primers (4). LYSV infection was further verified through cloning and sequencing of the cloned amplicon. Because this garlic is grown for seed, studies are being initiated to determine if current season spread occurs and yields are reduced. To our knowledge, this is the first report of OYDV, LYSV, and GCLV in garlic in Oregon. References: (1) P. Lunello et al. J. Virol. Methods 118:15, 2004. (2) H. R. Pappu et al. Plant Dis. 89:205, 2005. (3) S. S. Pappu et al. J. Virol. Methods 41:9, 1993. (4) T. Tsuneyoshi et al. Phytopathology. 86:253, 1996.

Whole genome characterization of Potato virus Y isolates collected in the western USA and their comparison to isolates from Europe and Canada.

Potato virus Y (PVY) is a serious potato pathogen that affects potato seed and commercial production crops. In recent decades, novel PVY strains have been described that cause necrotic symptoms on tobacco foliage and/or potato tubers. The major PVY strains that affect potato include PVY(O) and PVY(N), which have distinct serotypes that can be differentiated by immunoassay. Other economically important strain variants are derived from recombination events, including variants that cause tuber necrotic symptoms (PVY(NTN)) and PVY(O) serotypes that cause tobacco veinal necrosis (PVY(N)-W, PVY(N:O)). Although the PVY(NTN) and PVY(N)-W variants were first reported in Europe, apparently similar strains have been appearing in North America. Confirmation of the existence of these recombinant strains in North America is important, as is whether they spread from a common source or were derived by independent recombination. Whole genome sequencing can be used to positively identify strain variants and begin to address the issue of origins. Symptomology, serology, RT-PCR, and partial sequencing of the coat protein region were used to identify isolates of the PVY(NTN), PVY(N), PVY(NA-N), and PVY(N:O) for whole-genome sequencing. Sequencing confirmed the presence of PVY(NTN) and PVY(N) isolates that were >99% identical to European sequences deposited in GenBank in the 1990's. Sequences of the PVY(NA-N) and PVY(N:O) types were 99.0% and 99.5% identical to known sequences, respectively. There was no indication that recombinant strains PVY(NTN) or PVY(N:O) had different parental origins than recombinant strains previously sequenced. This is the first confirmation by whole-genome sequencing that "European"-type strain variants of PVY(N) and PVY(NTN) are present in North America, and the first reported full-length sequence of a tuber necrotic isolate of PVY(N:O).

The Occurrence of PVYO, PVYN, and PVYN:O Strains of Potato virus Y in Certified Potato Seed Lot Trials in Washington and Oregon.

Totals of 960 and 286 certified potato seed lots from locations across North America were planted in trials in Washington and Oregon, respectively, in 2001 to 2003 and tested for strains of Potato virus Y (PVY). The incidence of PVYO-infected lots averaged 16.4 and 25.9% in the Washington and Oregon trials, respectively. There was a general trend of increasing incidence of the PVYO, PVYN:O, and PVYN strains during this period, as evidenced by more infected cultivars, sites of seed origin, and number of seed growers providing infected seed lots. In particular, there was a dramatic increase in seed lots with the PVYN:O strain from 2002 to 2003. PVYN:O, in contrast to PVYO, which only causes yield reduction, also causes internal and external damage to tubers, making them unmarketable. In 2003, PVYN:O occurred in seed lots originating in eight states and three Canadian provinces. The increased incidence of PVYN:O was likely due to the difficulty in differentiating this strain from PVYO. The prevalence of PVY in potato seed lots documented herein poses a threat to potato production in the United States and suggests that current measures to reduce the incidence of this virus are inadequate.

Association of beet leafhopper (Hemiptera: Cicadellidae) with a clover proliferation group phytoplasma in Columbia basin of Washington and Oregon.

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.

First Report of Stunting and Root Rot of Potato Associated with Pratylenchus penetrans in the Columbia Basin of Washington.

The root-lesion nematode, Pratylenchus penetrans (de Man, 1880) Filipjev, 1936, is a common pathogen of potato in the United States east of the Rocky Mountains and frequently interacts with Verticillium dahliae to aggravate early dying disease (4). West of the Rocky Mountains, P. penetrans is associated with numerous crops including mint, tree, and small fruits but is rarely recovered from potato fields. Pratylenchus neglectus is a common plant-parasitic nematode on potato in the west, but causes little loss in yield and does not usually interact with V. dahliae (1). Management of P. neglectus is generally unnecessary; although a population of P. neglectus from Ontario, Canada appears to be more pathogenic and does interact with V. dahliae (1). During May 2003 (6 weeks after planting), large areas of stunted plants were observed in field inspections and in aerial photographs of two fields (50.6 ha) of cv. Ranger Russet in Benton County, Washington. Lower roots and stolons had numerous, dark lesions that are typical of P. penetrans damage and were severely stunted, while long, white roots had formed abnormally near the soil surface. In early May 2003, lesion nematodes (65 nematodes per 250 g of dry soil and 810 nematodes per g of fresh root weight) recovered from these potato fields were identified as P. penetrans on the basis of morphological characters (2,3). The crop responded to oxamyl (four applications at 1.1 kg of a.i. per ha between early May and mid-July), but the grower estimated that yields were 1.62 tons/ha (4 tons/acre) less than in comparable unaffected fields. To our knoweldge, this is the first report of severe damage to potato from P. penetrans in the Colombia Basin potato-production area. Soil fumigation with Telone II (1,3-dichloropropene) is commonly used in the Columbia Basin to control root-knot (Meloidogyne chitwoodi and M. hapla) and stubby-root (Paratrichodorus allius) nematodes, and metam sodium is used to control V. dahliae. However, since the only nematode recovered from preplant samples was assumed to be P. neglectus, and because cv. Ranger Russet is relatively tolerant to V. dahliae, no fumigant was used in these fields. An increase in mint production in this area may be responsible for introducing P. penetrans into previously uninfested fields since mint is propagated vegetatively and lesion nematodes are commonly associated with mint and could be easily spread in planting material and adhering soil. Identification of P. penetrans in stunted corn from two nearby fields during 2004 suggests that this nematode may be a new and emerging problem in this area. The Columbia Basin is one of the largest potato-producing regions in the United States, and widespread introduction of P. penetrans could add substantial cost to potato production in this area. References: (1) K. Mukerji. No. 458 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1975. (2) H. Scheck and S. Koike. Plant Dis. 83:877, 1999.

First Report of the Necrotic Strain of Potato virus Y (PVYN) on Potatoes in the Northwestern United States.

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.

First Report of Rhizomania of Sugar Beet in the Columbia River Basin of Washington and Oregon.

Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV) and vectored by the soilborne fungus, Polymyxa betae Keskin, is one of the most economically damaging diseases affecting sugar beet (Beta vulgaris L.) worldwide and has been found in most sugar beet-growing areas of the United States (2). During harvest in October 2000, sugar beet plants exhibiting typical symptoms of rhizomania (1) were found in a field near Paterson, WA. Sugar beet had been planted in the field in 1999 and 2000, but prior to this, the field had not been planted with sugar beet for approximately 20 years. Symptomatic roots from the field exhibited stunting, vascular discoloration, and proliferation of lateral rootlets. Leaves of affected plants were chlorotic. Four soil samples were taken from symptomatic areas of the field and diluted with an equal amount of sterile sand. Seeds of rhizomania-susceptible sugar beet cv. Beta 8422 were planted in the soil and sand mix and maintained in a controlled environment at 24°C and 12 h of daylight at one location and in the greenhouse at another. After 8 weeks, enzyme-linked immunosorbent assay (ELISA) was performed on roots of plants grown at each location. Triple-antibody sandwich (TAS) ELISA (Agdia, Inc., Elkhart, IN) was conducted at the University of Idaho, Twin Falls, ID and double-antibody sandwich (DAS) ELISA was performed at USDA-ARS, Salinas, CA, with antiserum specific for BNYVV (2). Two of four samples were positive for BNYVV in the ELISA tests at both locations based on absorbance values at least three times those of healthy controls. TAS-ELISA tests were conducted on roots collected in July 2001 from a field in Washington, 12.9 km from the first field, as well as from a field across the Columbia River near Boardman, OR. Samples from both fields tested positive for BNYVV. All three fields are within 24 km of one another. Four additional fields have subsequently been confirmed to be infected with BNYVV in this region, based on symptomology and ELISA. There are approximately 3,240 ha of sugar beet grown in the region, and growers have been advised as a result of this confirmation to plant resistant cultivars and increase the sugar beet rotation interval with nonhost crops to a minimum of 4 years. References: (1) J. E. Duffus. Rhizomania. Pages 29-30 in: Compendium of Beet Diseases and Insects. E. D. Whitney and J. E. Duffus, eds. The American Phytopathological Society, St. Paul, MN, 1986. (2) G. C. Wisler et al. Plant Dis. 83:864, 1999.

Thyroid function changes related to use of iodinated water in the U.S. Space Program.

BACKGROUND: The National Aeronautics and Space Administration (NASA) has used iodination as a method of microbial disinfection of potable water systems in U.S. spacecraft and long-duration habitability modules. A review of thyroid function tests of NASA astronauts who had consumed iodinated water during spaceflight was conducted. METHODS: Thyroid function tests of all past and present astronauts were reviewed. Medical records of astronauts with a diagnosis of thyroid disease were reviewed. Iodine consumption by space crews from water and food was determined. Serum thyroid-stimulating hormone (TSH) and urinary iodine excretion from space crews were measured following modification of the Space Shuttle potable water system to remove most of the iodine. RESULTS: Mean TSH significantly increased in 134 astronauts who had consumed iodinated water during spaceflight. Serum TSH, and urine iodine levels of Space Shuttle crewmembers who flew following modification of the potable water supply system to remove iodine did not show a statistically significant change. There was no evidence supporting association between clinical thyroid disease and the number of spaceflights, amount of iodine consumed, or duration of iodine exposure. CONCLUSIONS: It is suggested that pharmacological doses of iodine consumed by astronauts transiently decrease thyroid function, as reflected by elevated serum TSH values. Although adverse effects of excess iodine consumption in susceptible individuals are well documented, exposure to high doses of iodine during spaceflight did not result in a statistically significant increase in long-term thyroid disease in the astronaut population.

Design and current status of the longitudinal study of astronaut health.

BACKGROUND: Information has been collected regarding the immediate physiological effects of spaceflight on humans. However, little is yet known regarding long-term effects. The purpose of this paper is to describe the Longitudinal Study of Astronaut Health (LSAH) and report current mortality data. METHODS: All astronauts selected for the United States Space Program are followed from selection throughout their lifetime or until the end of the study. Comparisons are ground-based Johnson Space Center (JSC) employees matched to the astronauts at a 3:1 ratio by sex-specific age and body mass index. They are followed in the same manner as astronauts. Morbidity and mortality data are obtained from medical records supplemented with study questionnaires. Checks for death certificates are made to ascertain death of participants who miss routine examinations. RESULTS: Current cause-specific mortality rates for astronauts selected from 1959 through 1991 are not statistically different from rates for comparison participants for cardiovascular (p = 0.8112), cancer (p = 0.2382), or other disease (p = 0.5040) mortality. Astronauts have a significantly higher mortality rate due to accidents and injuries (p < 0.0001). CONCLUSIONS: Astronauts have a similar risk of death due to chronic diseases as ground-based participants, but are at greater risk for occupational-related accidental death.