Bactericera cockerelli (tomato/potato psyllid).

B. cockerelli is one of the most destructive potato pests in the western hemisphere. It was recognized in the early 1900s that B. cockerelli had the potential to be an invasive and harmful insect, particularly in western United States and Mexico (Šulc, 1909; Crawford, 1914; Compere, 1915; 1916; Essig, 1917). By the 1920s and 1930s, B. cockerelli had become a serious and destructive pest of potatoes in most of the southwestern United States, giving rise to the description of a new disease that became known as 'psyllid yellows' (Richards, 1928; 1931; 1933; Binkley, 1929; Richards and Blood, 1933; List and Daniels, 1934; Pletsch, 1947; Wallis, 1955). In recent years, other solanaceous crops, including tomato, pepper, eggplant, tobacco and tamarillo in a number of geographic areas have suffered extensive economic losses associated with B. cockerelli outbreaks (Trumble, 2008, 2009; Munyaneza et al., 2007a, b; 2008; 2009a, b, c, d; Liefting et al., 2008; 2009; Secor et al., 2009; Espinoza, 2010; Munyaneza, 2010; Crosslin et al., 2010; Rehman et al., 2010; Crosslin et al., 2012a, b; Munyaneza, 2012). Despite being a native of North America, B. cockerelli is also found in Central America and has recently invaded New Zealand, where it has caused extensive damage to indoor and outdoor solanaceous crops (Teulon et al., 2009; Thomas et al., 2011). B. cockerelli has recently been placed on the list of quarantine pest in EPPO region (EPPO, 2012).

“Candidatus Liberibacter solanacearum” infection of commercial tomatillo, Physalis ixocarpa Brot. (Solanales: Solanaceae) in Saltillo, Mexico

The potato psyllid, Bactericera cockerelli (Šulc), (Hemiptera: Triozidae) is a pest of Solanaceous crops (Solanales) including potato (Solanum tuberosum L.) and tomato (Solanum lycopersicum L.). Feeding by high populations of nymphs causes psyllid yellows while adults and nymphs are vectors of the plant pathogen, “Candidatus Liberibacter solanacearum” (Lso). Foliar symptoms that were consistent with either Lso-infection or psyllid yellows were observed in 2019 on tomatillo (Physalis ixocarpa Brot.; Solanaceae) grown within an experimental plot located near Saltillo, Mexico. This study had three primary objectives: 1) determine whether the foliar symptoms observed on tomatillo were associated with Lso infection, 2) identify the haplotypes of Lso and potato psyllids present in the symptomatic plot, and 3) use gut content analysis to infer the plant sources of Lso-infected psyllids. Results confirmed that 71% of symptomatic plants and 71% of psyllids collected from the plants were infected with Lso. The detection of Lso in plants and psyllids, and the lack of nymphal populations associated with psyllid yellows strongly suggests that the observed foliar symptoms were caused by Lso infection. All infected plants and insects harbored the more virulent Lso haplotype B, but one psyllid was also co-infected with haplotype A. The potato psyllids were predominantly of the central haplotype, but one psyllid was identified as the western haplotype. Molecular gut content analysis of psyllids confirmed the movement of psyllids between non-crop habitats and tomatillo and indicated that Lso infection of psyllids was associated with increased plant diversity in their diet.

First Report of “Candidatus Liberibacter psyllaurous” (synonym “Ca. L. solanacearum”) Associated with ‘Tomato Vein-Greening’ and ‘Tomato Psyllid Yellows’ Diseases in Commercial Greenhouses in Arizona

During the winter of 2006–2007, plants in commercial tomato greenhouses (GH-1 and GH-2; total 320 acres [129.5 ha]) in Arizona were infested with the potato psyllid Bactericera cockerelli (Sulc) and more than 60% and ~20% of the plants, respectively, exhibited leaf curling, chlorosis, and shortened internodes. In addition, some plants in GH-1 developed an unusual ‘vein-greening’ phenotype. Nucleic acids were isolated from 10 symptomatic and three asymptomatic plants from each greenhouse. PCR primers designed to amplify a phytoplasma-like 16S rDNA (850 bp) yielded the expected size product from GH-1 samples, whereas samples from GH-2 and the asymptomatic samples from both greenhouses did not. Several 16S rDNA PCR products (3 of 60) when cloned and sequenced, surprisingly shared 97% homology with ‘Candidatus Liberibacter asiaticus’ (GenBank No. GQ926917). PCR primers PSY680F 5′-GTTCGGAATAACTGGGCGTA-3′ and PSY1R 5′-CCCATAAGGGCCATGAGGACT-3′, based on the resultant 16S rDNA sequences, were used to amplify a 680-bp fragment from plant DNA extracts and psyllid lysates (1). A robust PCR product (~680 bp) was obtained from 10 of 10 GH-1 plant extracts (GQ926918) and from a GH-1-derived psyllid colony (28 of 35 adults) (GQ926919) and the tomato plants on which they were reared. In contrast, no 680-bp product was obtained from GH-1 asymptomatic plants (0 of 3), GH-2 plants (0 of 10 symptomatic; 0 of 3 asymptomatic), GH-2-derived psyllid colonies (0 of 35 adults), or psyllid colony tomato plants (data not shown). At least three 680-bp amplicons for each sample type were cloned and 8 to 10 inserts were sequenced for each. BLAST analysis revealed that all 680-bp sequences shared 99 to 100% nt identity with the analogous 16SrDNA from “Ca. Liberibacter psyllaurous” (2) and synonym “Ca. L. solanacearum” (3). A second molecular marker was obtained with the 1611F and 480R primers (2) to amplify the 16SrDNA-23S-ITS (980 bp) from >3 plant extracts and psyllid lysates that tested positive for liberibacter. Clustal W alignment of the 16S-23S-ITS sequences from GH-1 original tomato plants and psyllid colony plants (GQ926920) and psyllids (GQ926921) indicated they were 100% identical to each other and BLAST analysis indicated 99 to 100% shared identity with “Ca. L. psyllaurous” (EU812558) (synonym “Ca. L. solanacearum”). Transmission electron microscopy examination of GH-1 and GH-2 psyllids revealed rod and pleomorphic-shaped bacteria (0.5 to 2.0+ μm) at the brain-salivary gland interface in psyllids from the GH-1 liberibacter-positive colony. No such bacteria were observed in GH-2 liberibacter-negative psyllids. These results support an etiological role of a new liberibacter spp. in the development of the ‘vein-greening’ symptom phenotype. In contrast, the GH-2 ‘yellows’ phenotype is reminiscent of ‘psyllid toxicity’ in tomato colonized by B. cockerelli (4). To our knowledge, this is the first report of distinct psyllid-associated diseases in greenhouse tomato in Arizona, one associated with a new ‘Ca. Liberibacter’ spp., manifest as ‘vein-greening’ disease, and the other associated with psyllid feeding, in which liberibacter is undetectable in plants and psyllids, and is manifest as the ‘tomato psyllid yellows’ disease. References: (1) D. R. Frohlich et al. Mol. Ecol. 8:1683, 1999. (2) A. K. Hansen et al. Appl. Environ. Microbiol. 74:5862, 2008. (3) L. W. Liefting et al. Plant Dis. 93:208, 2009. (4) H. J. Pack. Utah Agric. Exp. Stn. Bull. 209, 1929.

First Report of “Candidatus Liberibacter psyllaurous” Associated with Psyllid Yellows of Tomato in Colorado

Greenhouse tomato growers from Fort Lupton, CO contacted the USDA-ARS-USHRL in 2002 regarding plant symptoms resembling “psyllid yellows” associated with Bactericera cockerelli (Sulc) infestations that initially begin as retarded growth, erectness of new growth, chlorosis, and purpling of leaves followed by widespread chlorosis and production of many small, poor-quality fruit (1). Symptoms appeared ≈6 weeks after psyllids were observed and were generally restricted to the top half of the plant. Leaf cuttings from beefsteak tomatoes cv. Quest were immediately placed in RNAlater (Applied Biosystems, Austin, TX). Samples from symptomatic and asymptomatic plants were collected in September and December of 2002. At each date, leaves were sampled from multiple plants and placed in separate RNAlater bottles. September samples exhibited initial “psyllid yellows” symptoms and December samples exhibited severe symptoms. Samples remained at 4°C in RNAlater for 6 years until recent findings suggested that a new species of bacteria, named either “Candidatus Liberibacter psyllaurous” (2) or “Ca. L. solanacearum” (3), may be the causal agent of “psyllid yellows”. The Qiagen (Valencia, CA) DNeasy Plant Kit and recommended protocols were used for four separate DNA isolations from each of the four tomato samples that had previously remained unopened. Five PCR primer pairs designed to amplify three distinct genetic regions within the “Ca L. psyllaurous” rrn operon (16S rRNA, 16S-23S rRNA intergenic region, and 23S rRNA) were used and one primer pair specific to the tomato DNA (18S rRNA gene) that successfully amplified from all samples was used as a positive control. Bacterial primers included one pair designed specifically for 16S rRNA sequences of ‘Ca. L. asiaticus’, ‘americanus’, and ‘africanus’ species (USHRL-CL1) and four sets, Lp-1 through Lp-4, previously described (2) that amplify nonoverlapping regions of the 16S-23S rRNA operon. The USHRL-CL1 primers (USHRL-CL1f: 5′-CTTACCAGCCCTTGACATGTATAGGA-3′, and USHRL-CL1r: 5′-TCCCTATAAAGTACCCAACATCTAGGTAAA-3′) amplify a 195-bp fragment from bp 895 to 1,089 of the ‘Ca. Liberibacter’ sp. 16S rRNA Genbank Accession No. L22532. Only samples from severe symptomatic plants collected in December 2002 yielded amplicons that were purified and sequenced (Genbank: USHRL-CL1, FJ871062; Lp-1, FJ871058; Lp-2, FJ871059; Lp-3, FJ871060; Lp-4, FJ871061). For each bacterial primer pair, the fragment amplified was highly homologous (98 to 100% identity) to “Ca. L. psyllaurous” rRNA gene/intergenic space sequences. The 16S rRNA coding region was identical to two GenBank ‘Ca. Liberibacter’ sp. entries: EU921627 and EU921626 from B. cockerelli samples collected in Dalhart, TX and zebra chip potato samples from Garden City, KS, respectively; however, the whole 2,500 bp amplified and sequenced from our sample contained 11 to 14 polymorphisms when compared to nine “Ca. L. psyllaurous” sequences. Our results clearly indicate that “Ca. L. psyllaurous” isolates were associated with tomato “psyllid yellows” symptoms in Colorado as early as 2002 and significant sequence variation exists within the 16S/23S rRNA intergenic region and 23S rRNA coding region to allow analysis of genetic diversity among “Ca. L. psyllaurous” isolates. References: (1) L. B. Daniels. Ph.D. diss. University of Minnesota, St. Paul, 1954. (2) A. K. Hansen et al. Appl. Environ. Microbiol. 74:5862, 2008. (3) L. W. Liefting et al. Plant Dis. 93:208, 2009.

First Report of ‘Candidatus Liberibacter psyllaurous’ in Zebra Chip Symptomatic Potatoes from California

A disease that severely affects processing potatoes (Solanum tuberosum L.), termed zebra chip (ZC), has been identified in several locations in the United States (Texas, Nebraska, Colorado, Kansas, New Mexico, Arizona, and Nevada), Mexico, and Central America (4). The disease name comes from the rapid oxidative darkening of freshly cut tubers and the dark stripes and blotches that occur in chips processed from infected tubers. Recently, the disorder has been associated with a new ‘Candidatus Liberibacter’ species in New Zealand (3). Also, a bacterium designated ‘Candidatus Liberibacter psyllaurous’ has been identified recently in potato plants with “psyllid yellows” symptoms that resemble foliar symptoms of ZC (2). In the fall of 2008, 10 tubers were received at the Prosser laboratory from a commercial potato grower and five had symptoms characteristic of ZC. The tubers, cv. Dakota Pearl, were grown near Lancaster in southern California. The tubers showed rapid oxidation upon slicing and the sunken stolon attachment characteristic of ZC (4). Nucleic acid was extracted from symptomatic tubers (1) and tested by PCR for ‘Ca. Liberibacter’ species with primer pairs OA2/OI2c (5′-GCGCTTATTTTTAATAGGAGCGGCA-3′ and 5′-GCCTCGCGACTTCGCAACCCAT-3′) and CL514F/R (5′-CTCTAAGATTTCGGTTGGTT-3′ and 5′-TATATCTATCGTTGCACCAG-3′), which amplify from the 16S rDNA and rplJ and rplL ribosomal protein genes, respectively (3). Four of the five tubers with distinct ZC symptoms yielded the expected amplicons with both primer pairs. Two tubers with mild internal discoloration yielded correctly sized amplicons but in lesser amounts than from the severely affected tubers. Nucleic acid from healthy potato tubers yielded no product with these primers. One clone of the 1,168-bp OA2/OI2c amplicon and two clones of the 669-bp CL514F/R amplicon from a strongly positive sample were sequenced in both directions (ACGT, Inc., Wheeling, IL). BLAST alignments of the consensus sequences of the OA2/OI2c and CL514F/R amplicons (GenBank Accessions Nos. FJ498802 and FJ498803, respectively) revealed 100% identity with analogous ‘Ca. Liberibacter’ sequences reported from ZC-symptomatic potatoes in New Zealand (GenBank Accession Nos. EU849020 and EU919514). The OA2/OI2c amplicon was also identical to a sequence of ‘Ca. Liberibacter psyllaurous’ (GenBank Accession No. EU812559) from psyllid yellows-affected potatoes in the United States (2) and also showed a 99% identity with sequences from a ‘Ca. Liberibacter’ species reported in ZC tubers from Kansas (GenBank Accession No. EU921626). Potato crops with symptoms of ZC have been observed previously in California (4), but this is the first identification of ‘Ca. Liberibacter psyllaurous’ from diseased potatoes grown in California. Since ZC was first reported in the mid- to late-1990s, information from potato growers and processors suggests that ZC is becoming more important. The disease has caused millions of dollars in losses, particularly in south Texas (4). The identification of ‘Ca. Liberibacter psyllaurous’ in California provides additional evidence that the disease is increasing in importance in other potato-growing regions. References: (1) J. M. Crosslin et al. Plant Dis. 90:663, 2006. (2) A. K. Hansen et al. Appl. Environ. Microbiol. 74:5862, 2008. (3) L. W. Liefting et al. Plant Dis. 92:1474, 2008. (4) J. E. Munyaneza et al. Subtrop. Plant Sci. 59:30, 2007.

Ovipositional preferences, damage thresholds, and detection of the tomato–potato psyllid Bactericera cockerelli (Homoptera: Psyllidae) on selected tomato accessions

The tomato–potato psyllid Bactericera [Paratrioza] cockerelli (Sulc) has recently caused losses exceeding 50% on fresh market tomatoes in California and Baja, Mexico by injecting a toxin that results in a condition known as ‘psyllid yellows’. The objectives of this study were to: (i) document oviposition preferences on a range of tomato cultivars; (ii) determine threshold levels for psyllid densities that would cause psyllid yellows on tomatoes within the first three weeks following transplanting; and (iii) identify the most important ‘psyllid yellows’ symptoms that might be used in surveying and monitoring for this pest. Plant lines tested included the commonly-planted commercial cultivars ‘Shady Lady’ and ‘QualiT 21’, an older, previously commercial cultivar ‘7718 VFN’, a common cultivar planted by consumers ‘Yellow Pear’, and a wild type plant accession, PI 134417. When given a choice, psyllids significantly preferred ‘Yellow Pear’ and avoided PI 134417 for oviposition. Under no-choice conditions psyllids laid significantly fewer eggs on PI 134417, but all the other plant lines were equally good substrates for laying eggs. Thus, oviposition preference is not likely to provide a functional management strategy in large plantings. On ‘Shady Lady’, psyllids preferred to oviposit on plants already infested with adults. On both ‘Shady Lady’ and ‘7718 VFN’ oviposition was significantly greater on plants previously infested by nymphs as compared to uninfested control plants. This suggests that, at least for some cultivars, there is a physiological change in plant attractiveness following psyllid feeding. ‘Yellow Pear’ and ‘QualiT 21’ were relatively tolerant of psyllids, requiring 18 nymphs per plant to produce the disease symptoms. Only eight nymphs per plant were needed on ‘Shady Lady’ and ‘7718 VFN’. For all cultivars, the pest density showed strong correlations with measurements such as the number of yellowing leaves and leaflets and distorted leaves, which were as good as or better than the first factor extracted from principal component analysis. Therefore, such measurements have the potential to simplify field surveys.