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 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.

Potato Psyllid (Hemiptera: Triozidae) Response to Insecticides Under Controlled Greenhouse Conditions.

Bactericera cockerelli (Sulc) is a common pest of solanaceous crops largely known for vectoring "psyllid yellows" in potatoes. In recent years, however, this pest has attracted considerable attention for vectoring Zebra Chip, a devastating bacterial disease that was first reported in the United States in 2004 and has spread across the southern and northwestern states, causing significant economic losses to potato growers and the industry. Management of the disease is mainly achieved by reducing the psyllid population using insecticides, including pyrethroids and neonicotinoids; however, new insecticides with different modes of action are needed to avoid the acquisition of resistance. In the current study, the immediate and residual effects of conventional and new materials were tested under greenhouse conditions against adults and nymphs. Cyantraniliprole at a high rate and spinetoram showed the highest immediate and residual activity against adults, whereas cyantraniliprole and oxamyl (low and high rates) were effective against nymphs, with no differences between rates. Soil applications of cyantraniliprole, oxamyl, and imidacloprid were still toxic to adults and nymphs 7 and 14 d after treatment. Also, cyantraniliprole and tolfenpyrad were detrimental to psyllid reproduction. The incorporation of new chemistry in rotation programs may provide a useful management tool against this pest.

Population structure of Phthorimaea operculella (Lepidoptera: Gelechiidae) in the United States.

The potato tuberworm, Phthorimaea operculella Zeller, is one of the most damaging pests of potatoes worldwide. Although tuberworm was recorded in California as early as 1856, it was first reported in damaging numbers in Oregon and Washington in the early 2000s. The objective of this study was to provide baseline information on the population structure of potato tuberworm in the United States. Potato tuberworm adults were collected from potato fields in the major production regions in the United States. Amplified fragment length polymorphisms (AFLP) were used to determine the genetic population structure of potato tuberworm. We used 219 individuals and 335 polymorphic AFLP bands to infer the geographic population structure of potato tuberworm. Posterior probabilities calculated using the model-based clustering method implemented in STRUCTURE, and F(st) values calculated using AFLP-SURV confirmed the existence of two distinct populations in the United States (one mostly associated to the western United States and the other one mostly associated to the eastern United States). This study provides baseline data for the molecular characterization of potato tuberworm populations, which will aid in tracking the origin of future invasions within the United States.

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.