Domenico Rizzo

• Head of Department at Servizio Fitosanitario, Regione Toscana

Molecular assays based on qPCR TaqMan Probes were developed to identify three species of the genus Xylosandrus, X. compactus, X. crassiusculus and X. germanus (Coleoptera Curculionidae Scolytinae). These ambrosia beetles are xylopha-gous species alien to Europe, causing damages to many ornamental and fruiting trees as well as shrubs. DNA extraction...

Anoplophora glabripennis (Motschulsky 1853) (Coleoptera: Cerambycidae), the Asian Longhorned Beetle, is native to temperate and subtropical areas of China and the Korean peninsula. Due to its wide range of host plants, it is considered among the most economically important invasive plant pests. The morphological identification of A. glabripennis la...

The Japanese beetle ( Popillia japonica Newman) (Coleoptera Scarabaeidae) is a polyphagous invasive pest included in quarantine lists in Europe and in many countries of Africa, Asia and America. No specific molecular methods were available for the identification of adults and preimaginal stages. To bridge this gap, a rapid method (<2 h) based on SY...

Walnut species (Juglans spp.) are multipurpose trees, widely employed in plantation forestry for high-quality timber and nut production, as well as in urban greening as ornamental plants. These species are currently threatened by the thousand cankers disease (TCD) complex, an insect–fungus association which involves the ascomycete Geosmithia morbid...

The use of molecular tools to identify insect pests is a critical issue, especially when rapid and reliable tests are required. We proposed a protocol based on qPCR with SYBR Green technology to identify Philaenus italosignus (Hemiptera, Aphrophoridae). The species is one of the three spittlebugs able to transmit Xylella fastidiosa subsp. pauca ST5...

The diseases caused by the xylem‐inhabiting bacterium Xylella fastidiosa (Xf) are considered a consequence of the drastic reduction in water flow that this endophyte causes in some of its hosts. To date, within the EU, Xf has been found infecting several taxa that are common members of the Mediterranean maquis, a resource‐poor environment with very...

Pantoea stewartii subsp. stewartii (Pss) is a Gram-negative bacterium causing Stewart wilt, a severe disease in maize. Native to North America, it has spread globally through the maize seed trade. Resistant maize varieties and insecticides are crucial to mitigate the disease’s economic impact. Pss is a quarantine pest, requiring phytosanitary certi...

Funding: This work was partly funded by the 'Accordo di collaborazione per attività scientifica Regione Toscana-IPSP-CNR prot. RT n. 0562065 del 12/12/2023'. ABSTRACT In the present study, a biomolecular diagnostic assay based on qPCR-LNA (Locked Nucleic Acid) probes was developed for the identification of Agrilus anxius (Coleoptera: Buprestidae) f...

Euwallacea similis (Ferrari, 1867), an ambrosia beetle of Asian origin, is recorded for the first time in Europe on an ‘ornamental’ Ficus macrophylla tree, based on a series of specimens collected in Montechiarugolo (Parma, Italy), in May 2024. Morphological as well as molecular identifications of this species are presented and debated and the impl...

Aleurocanthus spiniferus (Quaintance) (Hemiptera: Aleyrodidae) has rapidly spread, mainly in the central and eastern Mediterranean coastal area, infesting various new host plants alongside known ones. This invasive species poses a significant threat to agricultural ecosystems, necessitating urgent action to monitor and control outbreaks in previous...

This study presents a qPCR-based molecular diagnostic method utilizing a TaqMan probe to ac-curately identify Agrilus auroguttatus Schäffer (Coleoptera: Buprestidae) from adult insects, ex-creta samples, and frass residues. The method demonstrated excellent DNA amplification across all sample types, highlighting strong analytical specificity. The p...

Monitoring of emerging plant diseases in the Apennine mountains (central Italy) identified Cryptostroma corticale as the cause of the disease sooty bark of maple trees. The identified sites were located in rural or forested areas, next to buildings, in villages or suburbs, and one site was in a forest. Samples of symptomatic tissues were taken from...

Flavescence dorée (FD) is a major grapevine disease in Europe, despite the quarantine status of its causal agent [FD phytoplasma (FDp)] and the mandatory monitoring and vector control practices. As alarming FD epidemic outbreaks continue to appear in Tuscany (Central Italy), a 4-year survey was carried out in the main wine-growing areas of the regi...

Maples species (Acer sp.p.) are commonly selected as urban trees due to their adaptability to various conditions. However, maples in central European cities are facing challenges from the emerging pathogen Cryptostroma corticale Ellis & Everhart) Gregory & Waller, the sooty bark disease agent. This fungus, believed to be introduced from America, is...

Ripersiella hibisci (Kawai & Takagi, 1971) is a root mealybug included in the EPPO A1 list of pests recommended for regulation and an EU A1 Quarantine pest list (Annex IIA). This pest causes damage to several species of monocotyledonous and dicotyledonous ornamental and foliage potted plants, especially bonsais. The underground location of R. hibis...

A molecular tool has been developed for the molecular identification of Ips sexdentatus (Börner 1776) (Coleoptera Curcu-lionidae Scolytidae), the well-known six thooted bark beetle, widely distributed in Eurasia, where it infests several species of the genus Pinus and occasionally a few conifer species of the genera Abies, Larix and Picea. The deve...

In Europe, the recently reported plant pathogen Xylella fastidiosa subsp. multiplex affects several wild, ornamental, and cultivated trees causing scorch diseases. In 2018, the sequence type 87 was reported in Tuscany on Mediterranean shrubs and trees. Although spittlebugs (Hemiptera: Aphrophoridae) were already identified as main vectors of this b...

Thousand cankers disease (TCD) of walnut is an emerg- ing new disease of walnut caused by the ascomycete fun- gus Geosmithia morbida (Hypocreales, Bionectriaceae) and its vector insect, the walnut twig beetle Pityophthorus juglandis (Coleoptera, Curculionidae, Scolytinae). molecular diagnostic tools have been developed to be used in phytosanitary s...

A molecular tool has been set up for the unambiguous molecular identification of the ambrosia beetle, Xyleborinus saxesenii (Ratzeburg, 1837) (Coleoptera Curculionidae Scolytinae), the cosmopolitan fruit‐tree pinhole borer, widely distributed in temperate regions of the five continents, where it infests a wide range of hardwoods and softwoods. The...

Simple Summary The accurate identification of the Oriental fruit fly, Bactrocera dorsalis, is complicated by its similarities to other species and taxonomic uncertainties. This represents a significant threat to fruit crops as it is already present in Europe, and this is a cause for great concern. To expedite identification, a reliable method using...

Leaf disk processing technique to enhance DNA extraction and sample storage for “flavescence dorée” phytoplasma detection by real-time LAMP assay

The overall accuracy was around 70%, with some misclassifications only between the most symptomatic classes. Furthermore, variations of spectral vegetation indices and leaf functional traits derived from spectra by partial least squares regression-models allowed to elucidate the effects of FD to leaf physiology and biochemistry. Overall, the presen...

Anoplophora chinensis (Forster, 1771), the citrus longhorned beetle, and Anoplophora glabripennis (Motschulsky, 1853), the Asian longhorned beetle, are wood borers of great economic importance, infesting a wide range of ornamental trees and shrubs. Native to Asia, their presence has been detected in several countries on different continents. Both a...

In 2022, a test performance study (TPS) assessing the influence of different master mixes on the performance of the tetraplex real-time PCR (TqPCR) assay was organized. TqPCR allows for the specific detection and identification of Xylella fastidiosa (Xf) subspecies in a single reaction. Eighteen official laboratories of the Italian National Plant P...

After its first discovery in Europe, in Apulia (Italy) in 2008, the citrus spiny whitefly (CSW), Aleurocanthus spiniferus (Quaintance, 1903) (Hemiptera Aleyrodidae) spread throughout the Italian peninsula, as well as in Albania, Bulgaria, Croatia, Greece and Montenegro. The species has been established mainly in urban areas, infesting ornamental...

Light and electron microscopy observations of Rhamnus alaternus L. infected by Xylella fastidiosa subsp. multiplex. Poster al 98 Convegno della Società di Botanica Italiana.

Thousand Cankers Disease (TCD) is a complex disease involving the ascomycete fungus Geosmithia morbida and the vector Pityophthorus juglandis (Coleoptera Curculionidae Scolytinae), a bark beetle known to attack plants of the genera Juglans (J. ailantifolia, J. californica, J. cinerea, J. hindsii, J. major, J. mandshurica, J. microcarpa, J. mollis,...

Kiwifruit is inhabited by a heterogeneous community of bacteria belonging to the Pseudomonas syringae species complex (Pssc). Only a few of its members, such as the specialist Pseudomonas syringae pv. actinidiae biovar 3 (Psa3), are known as pathogens, but for most of the species, such as P. viridiflava (Pv), a generalist with high intraspecific va...

Hydrangeas (Hydrangea L.) are popular ornamental plants used in urban landscapes and gardens worldwide for the beauty of their large flowers. In June 2022, dark brown/purple and irregular water-soaked spots coalescing into large areas of necrosis were observed on the leaves of potted Hydrangea quercifolia Bartr. plants growing in two ornamentals nu...

Flavescence dorée (FD) is the most threatening grapevine yellows (GY) disease in Europe. Despite strict control measures, alarming signs of the spread of the disease in viticultural areas continue to be detected. FD is attributed to infection by phytoplasma strains of an incidentally cited species, 'Candidatus Phytoplasma vitis'. In 2017, a GY fiel...

The sap-sucking insect Corythucha arcuata (Say) (Hemiptera Tingidae), also known as the oak lace bug (OLB), is an invasive pest mainly associated with oaks. Native to North America, it was introduced in Europe in 2000 (Northern Italy). The OLB is now reported in several other European countries, mostly from the Balkan region. At the time of its fir...

Spartium junceum L. is a typical species of Mediterranean shrubland areas, also grown in gardens and parks as an ornamental. In recent years in Europe, S. junceum has been recurrently found to be infected by different subspecies and genotypes of the quarantine regulated bacterium Xylella fastidiosa (Xf). This work presents for the first time the an...

DNA extraction from plant samples is very important for a good performance of diagnostic molecular assays in phytopathology. The variety of matrices (such as leaves, roots, and twigs) requires a differentiated approach to DNA extraction. Here we describe three categories of matrices: (a) symptomatic bark/wood tissue; (b) residues of frass resulting...

2-years-old plants of Pyrus communis showing symptoms of fire blight disease were sampled in an orchard in Tuscany (Italy) during Autumn 2020. Plants were obtained the previous spring from a commercial nursery located in a region where the disease is present since 1994. The collected material was processed in the lab in order to verify the presence...

en This paper provides a first report of Aleurocanthus camelliae, the Camellia spiny whitefly, from Italy. The pest was found on plants of Camellia spp. grown in the nursery. Brief morphological and biological information is provided on this whitefly, as well as some considerations on the phytosanitary measures to be adopted to reduce the potential...

A molecular diagnostic method using TaqMan probe qPCR is presented for the identification of Anoplophora chinensis (Förster) (Coleoptera: Cerambycidae) from whole body insects (adults and larvae) and frass samples stored under different conditions. The results showed a perfect amplification of DNA from all samples; the re-peatability and reproducib...

Nuts of the sweet chestnut ( Castanea sativa ) are a widely appreciated traditional food in Europe. In recent years producers and consumers reported a drop of nut quality due to the presence of rot diseases caused by Gnomoniopsis smithogilvyi. Early detection of this pathogen is fundamental to the economic viability of the chestnut industry. In the...

Three molecular protocols using qPCR TaqMan probe, SYBR Green, and loop-mediated isothermal amplification (LAMP) methods were set up for the identification of larvae and adults of an African invasive moth, Thaumatotibia leucotreta (Meyrick, 1913) (Lepidoptera: Tortricidae). The DNA extracts from larval and adult samples of T. leucotreta were perfec...

The walnut twig beetle Pityophthorus juglandis is a phloem-boring bark beetle responsible, in association with the ascomycete Geosmithia morbida, for the Thousand Cankers Disease (TCD) of walnut trees. The recent finding of TCD in Europe prompted the development of effective diagnostic protocols for the early detection of members of this insect/fun...

The vector‐borne bacterium Xylella fastidiosa(Wells and Raju) causes several serious diseases to plants. Recently, different subspecies of X. fastidiosa were reported in some European countries. The risk of the bacterium’s spread on the entire European territory is very high; therefore, it has been added into the priority pest list (2019/1702/EU Re...

Tomato brown rugose fruit virus (ToBRFV) represents an emerging viral threat to the productivity of tomato and pepper protected cultivation worldwide. This virus has got the status of quarantine organism in the European Union (EU) countries. In particular, tomato and pepper seeds will need to be free of ToBRFV before entering the EU and before comi...

The red-necked longhorn beetle Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae) is native to east Asia, where it is a major pest of cultivated and ornamental species of the genus Prunus . Morphological or molecular discrimination of adults or larval specimens is required to identify this invasive wood borer. However, recovering larval st...

The first report of Stephanitis lauri Rietschel, 2014 (Heteroptera, Tingidae) in Tuscany and Liguria (Italy). The species has been observed in Pisa (Pisa, Tuscany) and Piano di Vezzano (La Spezia, Liguria) on bay laurel plants. S. lauri has been firstly described in Greece and it was recorded in Cote d’Azur (France) in 2017.

The cultivation of walnuts (Juglans sp.) in Europe retains high economic, social, and environmental value. The recent reporting of the Thousand Cankers Disease (TCD) fungus, Geosmithia morbida, and of its vector, Pityophthorus juglandis, in walnut trees in Italy is alarming the whole of Europe. Although Italy is at present the only foothold of the...

Anoplophora glabripennis (Motschulsky, 1853), native to eastern Asia, is a destructive woodborer of many ornamental species, leading to the decline and the death of the attacked trees. In outbreak areas as Europe or North America, this pest is usually identified using morphological or molecular analyses of adult or larval specimens. However, the pr...

Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae), the red-necked longhorn beetle is native to eastern Asia, where it is an important wood-borer of fruit and ornamental species of the genus Prunus. A. bungii is a quarantine pest in the European Union, following its accidental introduction and establishment in Germany and Italy, and is cur...

Grapevine yellows (GY) are an important threat to grapevine industry in Europe. In Italy, GY are mostly represented by Flavescence dorée (FD) and Bois noir (BN), respectively associated with infection by 16SrV and 16SrXII phytoplasma ribosomal groups. In the frame of a regional strategy aimed to monitor GY presence, an extensive survey was carried...

In data 15 gennaio 2019 si è proceduto all'abbattimento di piante di noce da legno attaccate da Geosmithia morbida e Pytophtorus juglandis come prescritto dalla Misura ufficiale 159/2018 emessa dal Servizio Fitosanitario Regionale. Questo ritrovamento rappresenta il primo focolaio del patogeno e del suo insetto vettore in Toscana ed è stato comunic...

An outbreak of Xylella fastidiosa was discovered in late 2018 in northern Italy affecting several plant species. Multilocus sequence typing analyses detected the presence of strains clustering in X. fastidiosa subsp. multiplex and harboring a hitherto uncharacterized sequence type, ST87. Three cultured strains (TOS4, TOS5, and TOS14) were subjected...

The vector-borne bacterial pathogen Xylella fastidiosa is widely distributed in the Americas; in the last decade it has emerged as a serious threat for agricultural crops, natural environment and landscape in Europe. Following the first EU outbreak in 2013 in southern Italy, associated with a severe disease in olive trees, annual mandatory surveys...

Bois Noir (BN) is a Grapevine Yellows (GY) disease largely spread in the main viticultural areas worldwide. 'Candidatus Phytoplasma solani' is the BN etiological agent. Multi-Locus Sequence Typing (MLST), based on tufB, secY, vmp1 and stamp genes, contributes to increase knowledge about its genetic variability. In this study, MLST analysis on the b...

Due to its complex epidemiological cycle, including several polyphagous insect vectors and host plants, and the absence of efficient control strategies, Bois Noir (BN) disease of grapevine is encroaching wider territories in the main viticultural areas worldwide. Molecular approaches allowed to increase the knowledge about its etiological agent (Bo...

Quantitative PCR protocols for phytoplasma detection were used to monitor grapevine yellows (GY) in 373 vineyards located in nine Tuscan districts. Among more than 70,000 plants visually monitored, 1.867 plants were sampled and "flavescence dorée" phytoplasmas (FD) were detected in 122 plants and mainly identified as trains belonging to 16SrV-C sub...

Bois Noir (BN) is the most widespread disease of the grapevine yellows complex in the Euro-Mediterranean area. BN is caused by 'Candidatus Phytoplasma solani' (BNp), transmitted from herbaceous plants to grapevine by polyphagous insect vectors. In this study, genetic diversity among BNp strains, their prevalence and possible association with grapev...

Quantitative PCR protocols for phytoplasma detection were used to monitor grapevine yellows (GY) in 373 vineyards located in nine Tuscan districts. Among more than 70,000 plants visually monitored, 1.867 plants were sampled and “flavescence dorée” phytoplasmas (FD) were detected in 122 plants and mainly identified as trains belonging to 16SrV-C sub...

Early trans-plasma membrane behavior after in vivo mechanical inoculation of tobacco plants with Tobacco mosaic virus (TMV) was investigated and compared to virus quantification in leaf tissues. In order to identify early events related to virus/host interaction, the systemic virus TMV was used to infect lower leaves and tests were carried out on u...

Citrus tristeza closterovirus (CTV) has been found several times in the last decades in Italy, and plant protection services are involved in monitoring and surveillance. Although orchards linked to the citrus industry are well monitored, there is an underestimated risk of viruses or virus-like diseases in ornamental nurseries. Our aim was to modify...

In order to evaluate the health status of Calla and Peruvian lily in Tuscan nurseries, 18 viruses belonging to six families and one unassigned virus were assayed. Tests were carried out on 90 Zantedeschia aethiopica plants and 48 Alstroemeria spp. plants collected from 12 Tuscan nurseries in two years, via RT-PCR tests. Z. aethiopica was mainly aff...

Wild herbaceous dicotyledonous plants (dicots) showing symptoms ascribable to phytoplasma disorders were found to be widely distributed in organic vineyards in central Tuscany (Italy) affected by bois noir, a grapevine yellows disease caused by "Candidatus Phytoplasma solani". In 2010 symptomatic dicots were tentatively identified to species level...

RT-real time PCR was used for the detection of 13 viruses in 768 samples of Vitis vinifera cv. Sangiovese collected from nine Tuscanian districts (Italy). Analysis of virus distribution was carried out on 482 samples infected with viruses responsible for infectious degeneration or associated with leafroll, fleck and rugose wood diseases that are in...

Leafroll is one of the most harmful viral diseases affecting grapevine worldwide. Historically, a dozen of viruses, named grapevine leafroll-associated viruses (GLRaVs), belonging to genera Closterovirus and Ampelovirus (Family Closteroviridae), have been found associated with the disease. Recent studies showed that GLRaV-4, -5, -6, -Pr, De- and -C...

In order to enhance the Italian lily bulb production, a breeding program was carried out at CRA-VIV in Pescia (PT - Italy) during the last years. Asiatic hybrids (Lilium x elegans Thunb.), lily cultivars and other native species were involved in the program. The obtained lily lines, which presented interesting traits, were preserved in a collection...

Calla lily (Zantedeschia aethiopica, Araceae Family) has become one of the most popular cut flowers worldwide for its ornamental characteristics, such as spathe beauty and postharvest shelf-life. Calla lily is mainly used as cut flower, but also as potted plant. Tospovirus (Bunyaviridae Family) causes severe agronomic and economic damages to variou...

Phytophthora ramorum L. è il fungo patogeno responsabile di una malattia conosciuta come "Sudden oak death" (SOD) che è stata segnalata a partire dal 1995 in California e Oregon (USA) su alcune specie di querce tipiche di quell'areale. L'elevata polifagia di questo patogeno unita alla capacità di attaccare gli organi epigei (come foglie e rami), co...

In April 2012 the presence of hyperplastic outgrowths on trunks, branches, and twigs of sweet olive plants, Osmanthus fragrans Lour (Fam. Oleaceae), was recorded in two ornamental hedges made up of five and four plants, respectively, in the city center of Montecatini (Pistoia-Italy). All sweet olive plants were seriously affected by the disease wit...

Con la continua espansione del commercio in-ternazionale, anche il numero di fitofagi che riescono a stabilizzarsi al di fuori del loro are-ale originario è in continuo aumento. I fitofagi parassiti del legno sono, tra le migliaia di specie di insetti esotici, quelli con maggior diffusione in tutte le aree del mondo. Due dei più distruttivi coleott...

Lily mottle virus (LMoV), a member of the genus Potyvirus, is one of the main viruses infecting lily. Symptoms on lily differ according to the susceptibility and sensitivity of different cultivars and hybrids. They range from leaf mottle or mosaic, vein clearing, chlorotic and yellow streaking, leaf curling, and necrotic spots, to milder forms of l...

In 2011, the methods to perform phytosanitary tests to check for viruses in grapevine nurseries were reassessed and new regulations were defined (DM 13 December 2011). The mandatory tests require serological assays for the diagnosis of five viruses in grapevine mother plants transplanted in Tuscany in 2001 or before. The aim of the present paper is...

The spread of 'bois noir' disease in grapevine was monitored in three organic vineyards in Tuscany (Italy). Annual incidence, infec-tion and recovery rates were calculated to describe changes in the epidemic with time. In general symptomatic plants showed the re-covery in a relatively short time, meanwhile the infection rate decreased rapidly after...

Virus diseases represent some of the most important diseases of Lilium, because of the devastating effects caused to the crops and the absence of effective treatments. The viruses causing the most significant damage in Lilium are: LSV (Lily symptomless virus), CMV (Cucumber mosaic virus) and TBV (Tulip breaking virus). Other viruses identified in L...

Calla lily (Zantedeschia aethiopica) Araceae Family, has become one of the most popular cut flowers worldwide because of its ornamental characteristics, such as spathe beauty and post-harvest shelf-life. Calla lily is mainly used for cut flowers, but also for potted plants production. Tospoviruses, which belong to the family Bunyaviridae, are agric...

Il legno nero della vite (LN) è una fitoplasmosi presente in maniera endemica in tutte le coltivazioni viticole del mondo. Recentemente in alcune aree di coltivazione italiane ha mostrato una incidenza epidemica che sembra possa essere correlata alla prevalente presenza di uno dei due ceppi del fitoplasma 16SrXII-A che si possono distinguere median...