Marzachì, Cristina

Pear decline (PD), associated with ‘Candidatus Phytoplasma pyri’, is a major disease of pear in Europe and the United States. Several psyllid species are involved in the tritrophic system of PD as vectors of phytoplasmas belonging to the 16SrX group. Four years after the first detection of PD in Sicily, an integrated approach was applied to investigate the epidemic in a major pear-growing area. Visual surveys and molecular analyses were conducted over two years in eight orchards. A total of 115 plant samples and 101 Cacopsylla spp. specimens selected from a total of 1,435 collected individuals were analysed, confirming ‘Ca. P. pyri’ in 69% of symptomatic plants and in 4.6% of C. pyri individuals. Multilocus sequence typing (MLST) revealed high genetic similarity among 16SrX isolates. Remote sensing analyses since 2018, combined with vector monitoring, confirmed the epidemic nature of PD and the persistence of a risk of further pathogen spread within the region, proving, inter alia, to be a valid method for identifying the syndrome even on a large scale.

ABSTRACT Two bacterial symbionts of the European pest leafhopper, Macrosteles quadripunctulatus (Hemiptera: Cicadellidae), were fully sequenced. “ Candidatus Sulcia muelleri” and “ Ca . Nasuia deltocephalinicola” represent two of the smallest known bacterial genomes at 190 kb and 112 kb, respectively. Genome sequences are nearly identical to strains reported from the closely related host species, M. quadrilineatus .

To characterize potentially important surface-exposed proteins of the phytoplasma causing chrysanthemum yellows (CY), new primers were designed based on the conserved regions of 3 membrane protein genes of the completely sequenced onion yellows and aster yellows witches’ broom phytoplasmas and were used to amplify CY DNA. The CY genes secY, amp, and artI, encoding the protein translocase subunit SecY, the antigenic membrane protein Amp and the arginine transporter ArtI, respectively, were cloned and completely sequenced. Alignment of CY-specific secY sequences with the corresponding genes of other phytoplasmas confirmed the 16S rDNA-based classification, while amp sequences were highly variable within the ‘Candidatus Phytoplasma asteris’. Five CY partial sequences were cloned into the pRSetC expression vector, and 3 of the encoded protein fragments (Amp 64/651, Amp 64/224, ArtI 131/512) were expressed as fusion antigens for the production of CY-specific polyclonal antibodies (A416 against Amp 64/224; A407 against ArtI 131/512). A416 recognized, in Western blots, the full-length Amp from CY-infected plants (periwinkle, daisy) and insect vectors ( Euscelidius variegatus , Macrosteles quadripunctulatus ). A416 also reacted to European aster yellows, to primula yellows phytoplasmas, to northern Italian strains of ‘Ca. Phytoplasma asteris’ from lettuce and gladiolus, but it did not react to American aster yellows phytoplasma.