RESUMO
'Candidatus Liberibacter solanacearum' (CaLsol), the etiological agent of potato zebra chip (ZC), is transmitted to potato plants by the psyllid Bactericera cockerelli (Sulc, 1909) in North and Central America and New Zealand. The risk of the dispersion of ZC in Spain depends on the presence of an efficient vector. This work studies the presence and abundance of ZC symptoms and CaLsol in potato plants, as well as the presence and abundance of psyllid species associated with potato crops in the main producing areas in Spain. Eighty-eight plots were surveyed punctually to detect ZC symptoms and psyllid species in the main potato-producing areas. Furthermore, fourteen potato plots were surveyed by different sampling methods during the cropping season to detect psyllid species from 2016 to 2018. Very few symptomatic and CaLsol-positive plants were detected in Mainland Spain, and any positive plant was detected in the Canary Islands. Most of the adult psyllids captured were identified as Bactericera nigricornis (Foerster, 1848), and some of them as Bactericera trigonica, but no B. cockerelli was detected. B. nigricornis was found widely distributed in the northern half of the Iberian Peninsula; however, this psyllid does not seem sufficient to pose a threat to potato production, due to the scarce number of specimens and because the frequency of B. nigricornis specimens that were CaLsol+ was very low.
RESUMO
In September 2019, symptoms resembling those of bacterial leaf blight were observed on carrot plants (Daucus carota L. subsp. sativus Hoffm.) cv. Romance cultivated in commercial plots in Chañe (Segovia), Spain. Symptoms were observed in two plots surveyed representing three hectares, with an incidence greater than 90%, and also in some plots in other nearby municipalities sown with the same batch of seeds. The lesions observed at the ends of the leaves were initially yellow that develop dark brown to black with chlorotic halos on leaflets that turned necrotic. Yellow, Xanthomonas-like colonies were isolated onto YPGA medium (Ridé 1969) from leaf lesions. Two bacterial isolates were selected and confirmed by real-time PCR using a specific primer set for Xanthomonas hortorum pv. carotae (Temple et al. 2013). All isolates were gram-negative, aerobic rods positive for catalase, able of hydrolyzing casein and aesculin and growing at 2% NaCl, while were negative for oxidase and urease tests. Sequences of 16S rRNA gene showed 100% similarity with Xanthomonas campestris, X. arboricola, X. gardneri, X. cynarae strains (GenBank accession numbers: MW077507.1 and MW077508.1 for the isolates CRD19-206.3 and CRD19-206.4, respectively). However, the resulting phylogeny of multilocus sequence analysis (MLSA) of a concatenation of the housekeeping genes atpD, dnaK, and efp (Bui Thi Ngoc et al. 2010), by using neighbour-joining trees generated with 500 bootstrap replicates, grouped the two isolates with the X. hortorum pv. carotae M081 strain (Kimbrel et al. 2011) (GenBank accession numbers: MW161270 and MW161271 for atpD for the two isolates, respectively; MW161268 and MW161269 for dnaK; MW161272 and MW161273 for efp). A pairwise identity analysis revealed a 100% identity between all three isolates. Pathogenicity of the isolates was tested by spray inoculation (Christianson et al. 2015) with a bacterial suspension (108 CFU/ml) prepared in sterile distilled water at 3 to 4 true-leaf stage (six plants per isolate). Sterile distilled water was used as negative control. The inoculated plants were incubated in a growth chamber (25°C and 95% relative humidity [RH]) for 72 h, and then transferred to a greenhouse at 24 to 28°C and 65% RH. Characteristic leaf blight symptoms developed on inoculated carrot plants, while no symptoms were observed on the negative control plants 20 days after inoculation. The bacterium was re-isolated from symptomatic tissue and the identity confirmed through PCR analysis. Based on PCR, morphological and phenotypic tests, sequence analysis, and pathogenicity assays, the isolates were identified as X. hortorum pv. carotae. To our knowledge, this is the first report of bacterial leaf blight of carrot caused by X. hortorum pv. carotae in Spain, and the first molecular and pathological characterization. It is important to early detect this pathogen and take suitable measures to prevent its spread, since it could cause yield losses for a locally important crop such as carrot.