First record of<i>Aleurocanthus</i><i>camelliae</i>Kanmiya &amp; Kasai, 2011 (Hemiptera, Aleyrodidae) from Italy, on ornamental<i>Camellia</i>spp. plants

This paper provides a first report of Aleurocanthus camelliae, the Camellia spiny whitefly, from Italy. The pest was found on plants spp. grown in nursery. Brief morphological and biological information is provided this as well some considerations phytosanitary measures to be adopted reduce potential risk its spread ornamental Europe EPPO region. Cet article présente le premier signalement de l’aleurode camelliae en Italie. organisme nuisible été signalé sur des cultivés pépinière. brièvement informations la morphologie et biologie cet aleurode, ainsi que considérations les mesures phytosanitaires à adopter pour réduire son risque dissémination plantes ornementales dans région OEPP. ? ?????? ?????? ???????????? ?????? ????????? ? ?????????? ?????????? ?????????? ?? ??????. ??????? ???????? ??? ?????? ?? ????????? spp., ???????????? ? ?????????. ?? ???? ???????????? ??????? ??????????????? ? ????????????? ??????????, ? ????? ????????? ??????????? ?? ?????????????? ?????, ??????? ?????? ???? ??????? ??? ???????? ?????????????? ????? ?? ??????????????? ???????????? ???????? ?????? ??????? ?????. insect group Aleyrodidae (commonly known whiteflies due white powdery wax secretion covering entire body wings adults) includes more than 1500 species worldwide (Mound & Halsey, 1978; Martin Mound, 2007), mostly having an intertropical distribution but with various extending also temperate regions. They all exclusively feed plants, causing both direct (suction phloem sap production honeydew, which sooty mould develops) indirect (virus transmission) damage (Martin al., 2000). Crop losses caused by whitefly transmitted virus diseases are difficult estimate because vast number crops they attack their diverse distribution. In particular, quarantine importance region worth special mention current global climate change, present world several pests host increasing plant trade, increases introduction noxious pests. Concerning movement among groups highest accidentally introduced into Italy during last decades (Rapisarda 2017). Among these recent invasions, record spiniferus (Quaintance) citrus southern Italian areas (Porcelli, 2008) noteworthy country Longo, 2021). genus (Quanitance Baker) presently around 80 (Kanmiya 2011; Dubey Ko, 2012; Gillespie, Jansen Porcelli, 2018), A. only one prior for fauna. September 2020, presence noted nursery-grown L. (Ericales, Theaceae); no were recorded before genus. morphology adults nymphs allowed it ascribed Aleurocanthus; however, characters raised suspicion that different spiniferus, therefore possible new hypothesis has been confirmed molecular study material, led identification Kanmiya Kasai, whitefly. Adults observed lower surface leaves eight nurseries Pistoia province (Tuscany, Italy) where sasanqua Thunb. outdoors plastic containers. inspected generally had although infestation scarce. From infested collected each nursery, preserved ethanol subsequent laboratory investigations. material identified basis at Department Agriculture, Food Environment (Di3A) University Catania through analysis Laboratory Phytopathological Diagnostics Molecular Biology, Plant Protection Service Tuscany. According established practice taxonomy, based fourth-instar (also puparia). About 50 cleaned treatment 95% gentle heating, then treated cold 10% solution KOH days until black cuticle bleached specimens light brown colour. Bleached nymphal rinsed 70% heating 3 min, placed glacial acetic acid subsequently clove oil about 15 min each, finally slide mounted Canada balsam. Morphological observations measurements slide-mounted carried out using phase contrast microscope (Olympus BX51TF, equipped digital camera Olympus Camedia C30-30 zoom). DNA extracted 10 according methodology described other Rizzo al. (2020a,b). extraction two pooled samples (each consisting five adults), technical replicates stage development (fourth-instar adults). quality quantity analysed QiaExpert spectrophotometer (Qiagen, Hilden, Germany). eluted 100 µL nuclease-free water used immediately qPCR or stored ?20°C use. addition, quantitatively assess suitability tests, amplified 18S uni-F/18S uni-R primers (5?-GCAAGGCTGAAACTTAAAGGAA-3?/5?-CCACCACCCATAGAATCAAGA-3?) uni-P probe (HEX-ACGGAAGGGCACCACCAGGAGT-BHQ1) targeting highly conserved ribosomal (Ioos 2009). A specific SybrGreen test designed diagnosis camelliae. primer pairs OligoArchitectTM Primers Probe Design Online software (Sigma-Aldrich, St Louis, USA) ATP6 synthase mitochondrial gene (Fig. 1 Table 1). design considered melting temperatures, amplicon length absence secondary structures. specificity BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi), searching most closely related nucleotide sequences aligning them MAFFT package (Katoh Standley, 2013) implemented within Geneious® 10.2.6 (Biomatters, http://www.geneious.com). results shown Fig. 2. unrooted phylogenetic tree created Geneious 10.2.4, neighbour-joining method Tamura-Nei model, 1000 bootstrap 3). reactions 20 volume containing 0.4 µM primer, SsoAdvanced Universal SYBR Green Supermix (Biorad, Hercules, CA, 10–20 ng DNA. To determine optimal annealing temperature, thermal gradient 50°C 60°C performed. amplification reaction consisted initial denaturation 2 95°C followed 40 cycles s combined elongation step 55°C s. At end protocol, means curve 65°C (0.5°C increments per read hold time). peak 78 ± 0.5°C. reaction, Dialeurodes citri taken nontarget references. Considering relevant genetic proximity, particular organisms made not morphology, biomolecular analysis, amplifications (starting extracts D. citri) endpoints (LCO1490/HCO2198) cytochrome oxidase (COI) subunit 1. real-time performed CFX96 thermocycler (Biorad). Reactions duplicate including negative control (no DNA) set reactions. Three amplicons purified QIAquick PCR Purification kit (Qiagen) sequenced (CIBIACI, Florence, Italy). confirms study. gave good result point view amplificability, fact mean values Cq (18S rDNA) originated equal 18.26 1.6. No occurred 5). sequence analyses show (accession KU761949.1) (https://blast.ncbi.nlm.nih.gov/Blast.cgi) sequences. support results, TaqMan being developed. investigations, whole, samples. Based literature main below. strongly dimorphic, males 0.90–1.10 mm females 1.25–1.40 long. When mature, orange colour, covered greyish powder coating. Head, dorsal view, 240–275 µm wide, 260–320 long antennae compound eyes upper part connected three ommatidia. Forewings 0.84–0.90 (males) 1.1–1.2 (females) long, dark grey colour nine small whitish-grey spots. Vasiform orifice almost rounded, 45–60 nearly 1.2–1.3 times largely closed 20–35 operculum, covers 23–28 lingula. Nymphs settle leaf underside; flattened, oval shape, gradually darkening after egg hatching become black, marginal waxy fringe. nymph (puparium) 0.65–0.86 0.98–1.24 0.39–0.57 0.62–0.86 wide. Margin crenulated, 6–10 teeth/100 (total teeth: 158–196). Dorsum glandular spines arranged follows: 10–11 along submargin; 16 subdorsum, cephalothorax (arranged lines, external internal four pairs) seven abdomen single row); submedian abdominal segments third. Cephalic eyespots clearly visible, ovoid located close base third submarginal spine. Abdominal tergite eighth 42–56 elevated cone, 80–88 63–68 entirely conceals probably native China; here likely Japan, Saito 2012), Indonesia (Samsu Adi Susanti, 2020). Apart East Asian countries, till now Netherlands (Jansen especially interceptions camellia imported China Japan potted few localities scattered over country. report, Italy, allows us hypothesize wider diffusion currently known, suggesting care required controls material. scarce available date Yamashita 2016), lives genera (Theaceae): C. japonica L., sinensis (L.) Kuntze tea plant), Cleyera (Theaceae) (C. Thunb.), Eurya (E. Illicium (Illiciaceae) (I. anisatum L.) Zanthoxylum (Rutaceae) [Z. piperitum DC]. All Asia, highlights pathway via trade many significant interest, widening cultivation production. biology life cycle poorly known. It reported have generations year (Kasai 2012). Vibratory signals emitted attract studied (2011). Negative effects Asia (Uesugi 2016; may cause vigour reduction degradation. tea-producing areas, heavy infestations farmers unpleasant inhalation flying field work harvesting operations On contrary, cases when (in plants) 2018). finding reported, detected low population density, so consequent reduced levels toxic saliva injection, suction honeydew did any plants. important pest, cultivations Asia. European environments recently detected, represented exotic Theaceae should assessed define adopted. As whiteflies, cannot fly distances, thus international spread. Appropriate must transferring could develop prevent eventual further natural enemies useful Encarsia smithi (Silvestri) (Hymenoptera: Aphelinidae) active parasitoid parasitism rate reaching up 97% (Ozawa Uchiyama, 2013). (puparia) parasitized, showing typical emergence hole endophagous adult parasitoid. Unfortunately, parasitisation examined prevented adequate identification. still out. However, case species, Europe, eradication outbreaks, involving use chemical control, suggested future Open Access Funding Universita degli Studi di CRUI-CARE Agreement. [Correction added May 2022, online publication: CRUI funding statement added.]