First report of Flavescence dorée phytoplasma identification and characterization in three species of leafhoppers

Belgeri, Elena; Rizzoli, Attilio; Jermini, Mauro; Angelini, Elisa; Filippin, Luisa; Rigamonti, I.E.

doi:10.1007/s42161-021-01012-y

Cited by 7 publications

(6 citation statements)

References 21 publications

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“…This could be an indication of introduction by planting material, but we find it strange that the same genotype has not induced epidemic outbreak in Italy while in north Serbia it became the dominant one with potential for competitive exclusion with regard to M51, as was experimentally evidenced for FD-C to outcompete FD-D [70]. This is why endemic emergence from alders or clematis still remains a possibility, especially because the same genotype of M12 was recently identified in Orienthus ishidae on Alnus glutinosa bordering vineyards of southern Swiss Alps [58], and in another invasive leafhopper Hishimonus hamatus in southern Switzerland on the edges and woods surrounding cultivated vineyards in 2017 [71]. This is an indication of the adaptation of the same genotype on a wider geographic area and the possibility of its simultaneous emergence in vineyard ecosystems.…”

Section: Discussionmentioning

confidence: 91%

Genetic Diversity of Flavescence Dorée Phytoplasmas in Vineyards of Serbia: From the Widespread Occurrence of Autochthonous Map-M51 to the Emergence of Endemic Map-FD2 (Vectotype II) and New Map-FD3 (Vectotype III) Epidemic Genotypes

et al. 2022

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Flavescence dorée (FD) is a European quarantine disease of grapevine caused by FD phytoplasma (FDp) transmitted by the leafhopper of North American origin Scaphoideus titanus. The disease affects the most important viticultural regions of Europe and all wine-growing regions of Serbia. Unlike the insect vector, the pathogen is native to Europe and associated with several wild host plants among which alder trees as the main source of two out of three map genetic clusters of pathogen variants (Map-FD1 and FD2). Heretofore, the FDp epidemic in Serbian vineyards was thought to be monotypic, i.e., caused by the single genotype of the Map-FD3 cluster, M51, and correlated with clematis as the natural source plant. This study aimed to provide data on genetic diversity, through map and vmpA gene typing, and insights into ecological properties of epidemiological cycles driving the epidemic outbreaks of FD in Serbia today. Map genotyping of 270 grapevine isolates collected from 2017 to 2019 confirmed M51 as autochthonous genotype widespread in all wine producing regions of Serbia and the dominant FDp epidemic genotype in most of the districts (75%, 202/270 isolates), except in north Serbia where multiple outbreaks of M12 Map-FD3 were recorded (54 isolates). Tree of heaven is reported as a new FDp plant reservoir for the Serbian vineyards, hosting the M51 genotype, along with clematis. An outbreak of a new endemic Map-FD3 genotype M144 was documented in grapevine samples from east Serbia (5 isolates), correlating with previous findings of the same genotype in clematis. In addition, single grapevine infections with five new Map-FD3 genotypes (M150-M154) were recorded in central Serbia, thus indicating high endemic potential for new outbreaks. The vmpA typing placed all Map-FD3 isolates into the VmpA-III cluster, i.e., Vectotype III. Finally, we found direct evidence that at least two FDp endemic genotypes, M89 and M148, of the Map-FD2/VmpA-II have escaped from alders and propagated in the grapevine-S. titanus pathosystem in Serbia (Vectotype II). Our findings confirm the high complexity of the FDp ecological cycle and provide evidence of a unique, autochthonous Balkan epidemiology sourced endemically.

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Section: Discussionmentioning

confidence: 91%

Genetic Diversity of Flavescence Dorée Phytoplasmas in Vineyards of Serbia: From the Widespread Occurrence of Autochthonous Map-M51 to the Emergence of Endemic Map-FD2 (Vectotype II) and New Map-FD3 (Vectotype III) Epidemic Genotypes

et al. 2022

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“…Interestingly, Osbornellus auronitens is also a Nearctic species recently recorded in Europe [ 38 ] and this specimen was collected by the first author in a sample representing the first European record in a woody patch surrounded by vineyards. The vineyard close to the patch is heavily affected by Flavescence dorée phytoplasma 16SrV-D, and other exotic species were found harboring both strains 16SrV-C and -D [ 39 , 40 , 41 ]. However, it is not possible to speculate further on these results because specific strains of Flavescence dorée phytoplasmas cannot be distinguished based on the 16S gene alone.…”

Section: Resultsmentioning

confidence: 99%

Comparison of Traditional and Next-Generation Approaches for Uncovering Phytoplasma Diversity, with Discovery of New Groups, Subgroups and Potential Vectors

Trivellone

Cao

Dietrich

2022

Biology

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Despite several decades’ effort to detect and identify phytoplasmas (Mollicutes) using PCR and Sanger sequencing focusing on diseased plants, knowledge of phytoplasma biodiversity and vector associations remains highly incomplete. To improve protocols for documenting phytoplasma diversity and ecology, we used DNA extracted from phloem-feeding insects and compared traditional Sanger sequencing with a next-generation sequencing method, Anchored Hybrid Enrichment (AHE) for detecting and characterizing phytoplasmas. Among 22 of 180 leafhopper samples that initially tested positive for phytoplasmas using qPCR, AHE yielded phytoplasma 16Sr sequences for 20 (19 complete and 1 partial sequence) while Sanger sequencing yielded sequences for 16 (11 complete and 5 partial). AHE yielded phytoplasma sequences for an additional 7 samples (3 complete and 4 partial) that did not meet the qPCR threshold for phytoplasma positivity or yielded non-phytoplasma sequences using Sanger sequencing. This suggests that AHE is more efficient for obtaining phytoplasma sequences. Twenty-three samples with sufficient data were classified into eight 16Sr subgroups (16SrI-B, I-F, I-AO, III-U, V-C, IX-J, XI-C, XXXVII-A), three new subgroups (designated as 16SrVI-L, XV-D, XI-G) and three possible new groups. Our results suggest that screening phloem-feeding insects using qPCR and AHE sequencing may be the most efficient method for discovering new phytoplasmas.

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“…Moreover, the possibility to carry out LAMP assays directly in the field, avoiding the transfer of insect and plant material from vineyards to laboratories, strongly reduces the risk of DNA degradation and accidental dispersion of S. titanus and of the hosted phytoplasma in FDp-free areas. Since FDp may also be transmitted by other leafhoppers such as Orientus ishidae (Matsumura), Oncopisis alni (Schrank), Dictyophara europaea (L.), Allygus modestus (F.), and Allygus mixtus (F.) [2,[50][51][52][53], it would be useful to optimize the LAMP assay in these insects. Nonetheless, in the light of the continuous updating of the taxonomic classification of FDp, it would be interesting to test whether the selected LAMP assay can detect the various FD genotypes, especially those responsible for epidemic outbreaks.…”

Section: Discussionmentioning

confidence: 99%

In-Field LAMP Detection of Flavescence Dorée Phytoplasma in Crude Extracts of the Scaphoideus titanus Vector

et al. 2022

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One of the most destructive diseases affecting grapevine in Europe is caused by Flavescence Dorée phytoplasma (FDp), which belongs to the 16Sr-V group and is a European Union quarantine pathogen. Although many molecular techniques such as loop-mediated isothermal amplification (LAMP) are widely used for the rapid detection of FDp in infected grapevine plants, there is no developed isothermal amplification assay for FDp detection in the insect vectors that are fundamental for the spread of the disease. For this reason, a simple in-field real-time LAMP protocol was optimized and developed for the specific detection of FDp in the insect vector Scaphoideus titanus. The LAMP assay was optimized to work with crude insect extracts obtained by manually shaking a single insect in a buffer for 5 min. Such a simple, sensitive, specific, economic, and user-friendly LAMP assay allowed the detection of FDp in S. titanus in less than half an hour, directly in the field. The developed insect tissue preparation procedure, combined with the LAMP protocol, promptly revealed the presence of FDp in infected S. titanus directly in the vineyards, allowing for monitoring of the spread of the pathogen in the field and to apply timely strategies required for the mandatory control of this pathogen.

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First report of Flavescence dorée phytoplasma identification and characterization in three species of leafhoppers

Cited by 7 publications

References 21 publications

Genetic Diversity of Flavescence Dorée Phytoplasmas in Vineyards of Serbia: From the Widespread Occurrence of Autochthonous Map-M51 to the Emergence of Endemic Map-FD2 (Vectotype II) and New Map-FD3 (Vectotype III) Epidemic Genotypes

Genetic Diversity of Flavescence Dorée Phytoplasmas in Vineyards of Serbia: From the Widespread Occurrence of Autochthonous Map-M51 to the Emergence of Endemic Map-FD2 (Vectotype II) and New Map-FD3 (Vectotype III) Epidemic Genotypes

Comparison of Traditional and Next-Generation Approaches for Uncovering Phytoplasma Diversity, with Discovery of New Groups, Subgroups and Potential Vectors

In-Field LAMP Detection of Flavescence Dorée Phytoplasma in Crude Extracts of the Scaphoideus titanus Vector

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