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Pest categorisation of the non‐EU phytoplasmas of Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L.

Abstract Following a request from the European Commission, the EFSA Panel on Plant Health performed a pest categorisation of nine phytoplasmas of Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L. (hereafter “host plants”) known to occur only outside the EU or having a limited presence in the EU. This opinion covers the (i) reference strains of ‘Candidatus Phytoplasma australiense’, ‘Ca. P. fraxini’, ‘Ca. P. hispanicum’, ‘Ca. P. trifolii’, ‘Ca. P. ziziphi’, (ii) related strains infecting the host plants of ‘Ca. P. aurantifolia’, ‘Ca. P. pruni’, and ‘Ca. P. pyri’, and (iii) an unclassified phytoplasma causing Buckland valley grapevine yellows. Phytoplasmas can be detected by available methods and are efficiently transmitted by vegetative propagation, with plants for planting acting as a major entry pathway and a long‐distance spread mechanism. Phytoplasmas are also transmitted in a persistent and propagative manner by some insect families of the Fulgoromorpha, Cicadomorpha and Sternorrhyncha (order Hemiptera). No transovarial, pollen or seed transmission has been reported. The natural host range of the categorised phytoplasmas varies from one to more than 90 plant species, thus increasing the possible entry pathways. The host plants are widely cultivated in the EU. All the categorised phytoplasmas can enter and spread through the trade of host plants for planting, and by vectors. Establishment of these phytoplasmas is not expected to be limited by EU environmental conditions. The introduction of these phytoplasmas in the EU would have an economic impact. There are measures to reduce the risk of entry, establishment, spread and impact. Uncertainties result from limited information on distribution, biology and epidemiology. All the phytoplasmas categorised here meet the criteria evaluated by EFSA to qualify as potential Union quarantine pests, and they do not qualify as potential regulated non‐quarantine pests, because they are non‐EU phytoplasmas.

Background and Terms of Reference as provided by the requestor 1.1.1. Background Council Directive 2000/29/EC 1 on protective measures against the introduction into the Community of organisms harmful to plants or plant products and against their spread within the Community establishes the present European Union plant health regime. The Directive lays down the phytosanitary provisions and the control checks to be carried out at the place of origin on plants and plant products destined for the Union or to be moved within the Union. In the Directive's 2000/29/EC annexes, the list of harmful organisms (pests) whose introduction into or spread within the Union is prohibited, is detailed together with specific requirements for import or internal movement.
Following the evaluation of the plant health regime, the new basic plant health law, Regulation (EU) 2016/2031 2 on protective measures against pests of plants, was adopted on 26 October 2016 and will apply from 14 December 2019 onwards, repealing Directive 2000/29/EC. In line with the principles of the above mentioned legislation and the follow-up work of the secondary legislation for the listing of EU regulated pests, EFSA is requested to provide pest categorisations of the harmful organisms included in the annexes of Directive 2000/29/EC, in the cases where recent pest risk assessment/ pest categorisation is not available.

Terms of Reference
EFSA is requested, pursuant to Article 22(5.b) and Article 29(1) of Regulation (EC) No 178/2002 3 , to provide scientific opinion in the field of plant health.
EFSA is requested to prepare and deliver a pest categorisation (step 1 analysis) for each of the regulated pests included in the appendices of the annex to this mandate. The methodology and template of pest categorisation have already been developed in past mandates for the organisms listed in Annex II Part A Section II of Directive 2000/29/EC. The same methodology and outcome is expected for this work as well.
The list of the harmful organisms included in the annex to this mandate comprises 133 harmful organisms or groups. A pest categorisation is expected for these 133 pests or groups and the delivery of the work would be stepwise at regular intervals through the year as detailed below. First priority covers the harmful organisms included in Appendix 1, comprising pests from Annex II Part A Section I and Annex II Part B of Directive 2000/29/EC. The delivery of all pest categorisations for the pests included in Appendix 1 is June 2018. The second priority is the pests included in Appendix 2, comprising the group of Cicadellidae (non-EU) known to be vector of Pierce's disease (caused by Xylella fastidiosa), the group of Tephritidae (non-EU), the group of potato viruses and virus-like organisms, the group of viruses and virus-like organisms of Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L. and the group of Margarodes (non-EU species). The delivery of all pest categorisations for the pests included in Appendix 2 is end 2019. The pests included in Appendix 3 cover pests of Annex I part A section I and all pests categorisations should be delivered by end 2020.
For the above-mentioned groups, each covering a large number of pests, the pest categorisation will be performed for the group and not the individual harmful organisms listed under "such as" notation in the Annexes of the Directive 2000/29/EC. The criteria to be taken particularly under consideration for these cases, is the analysis of host pest combination, investigation of pathways, the damages occurring and the relevant impact.
Finally, as indicated in the text above, all references to 'non-European' should be avoided and replaced by 'non-EU' and refer to all territories with exception of the Union territories as defined in Article 1 point 3 of Regulation (EU) 2016/2031.

(b) Viruses and virus-like organisms
Beet necrotic yellow vein virus

Interpretation of the Terms of Reference
Non-EU phytoplasmas of Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L. (from now on: "the host plants") are pests listed in the Appendices to the Terms of Reference (ToR) to be subject to pest categorisation to determine whether they fulfil the criteria of quarantine pests or those of regulated non-quarantine pests (RNQPs) for the area of the EU excluding Ceuta, Melilla and the outermost regions of Member States (MS) referred to in Article 355(1) of the Treaty on the Functioning of the European Union (TFEU), other than Madeira and the Azores.
The EFSA PLH Panel decided to address the pest categorisation of this group of infectious agents in two steps: first, a list of the non-EU phytoplasmas of the host plants (EFSA PLH Panel, 2020) and second, the present pest categorisation.
The process is described in EFSA PLH Panel (2020), in which a systematic approach identified 27 phytoplasmas reported to naturally infect one or more of host plants. "Among these phytoplasmas, based on information on distribution and prevalence both inside and outside the EU, the Panel identified 10 non-EU phytoplasmas, known to occur only outside the EU or having only a limited presence in the EU. The remaining 17 phytoplasmas, non-EU phytoplasmas known to occur only outside the EU or having only a limited presence in the EU, whose ability to infect host species is not fully confirmed by available literature (3 phytoplasmas), or phytoplasmas which have a substantial presence in the EU or are so far reported from the EU only (14 phytoplasmas), will not be categorised within the current mandate." One non-EU phytoplasma ('Candidatus Phytoplasma phoenicium', PHYPPH) was excluded from further categorisation, as a recent pest risk assessment is available (EPPO, 2017). The same statements and definitions reported above also apply to the current opinion.
This opinion provides the pest categorisation of the 9 non-EU phytoplasmas with confirmed presence in at least one of the host plants, that have been listed in EFSA PLH Panel (2020). Although phytoplasmas have not yet been cultivated in vitro, phylogenetic analyses based on various conserved genes have shown that they represent a distinct, monophyletic clade within the class Mollicutes. Phytoplasmas are therefore accommodated within the 'Candidatus Phytoplasma' genus. Within this genus, several sub-taxa have been described to accommodate organisms sharing less than 97.5% similarity among their 16S rRNA gene sequences. Additional species are described to accommodate organisms that, despite their 16S rRNA gene sequence being > 97Á5 % similar to those of other 'Ca. Phytoplasma' species, are characterized by distinctive biological, phytopathological and genetic properties. Conversely, some organisms, despite their 16S rRNA gene sequence being < 97Á5 % similar to that of any other 'Ca. Phytoplasma' species, are not presently described as Candidatus species, due to their poor overall characterization (IRPCM, 2004). The current opinion covers only phytoplasma strains infecting at least one of the host plants within their officially described 'Ca. P. species'. To this purpose, pathogens were identified according to the list of strains/-related strains within the original 'Ca. P. species' description, when available. Otherwise, affiliation to a 'Ca. P. species'-related strain was based on the identity of the 16S rRNA subgroup. In one case, in the absence of 'Ca. P. species' description, the pathogen strain is addressed as 'unclassified'.
Viruses, virus-like diseases of unknown aetiology or diseases caused by other graft-transmissible bacteria of the host plants are not addressed in this opinion.
The new Plant Health Regulation (EU) 2016/2031 4 , on the protective measures against pests of plants, will be applying from December 2019. The regulatory status sections (3.3.) of the present opinion are still based on Council Directive 2000/29/EC, as the document was adopted in November 2019.

Literature search
A literature search on non-EU phytoplasmas infecting the host plants was conducted at the beginning of the categorisation in the Web of Science (WoS) database, using the scientific name of the pests as search term. Relevant papers were reviewed and further references and data were obtained from experts, as well as from citations within the references and grey literature.

Database search
Pest information, on host(s) and distribution, was retrieved from the EPPO Global Database (EPPO GD) (EPPO, 2019) and relevant publications. Data kindly provided by National Plant Protection Organisations of the EU MS were also considered.
Information on pest vectors was retrieved from the Hemiptera-Phytoplasma-Plant biological interaction database (Trivellone, 2019). Data on the EU distribution of pest vectors were retrieved from the Fauna Europaea database (de Jong et al., 2014), and the Catalogue of Life 2019 checklist (Roskov et al., 2019). To ensure appropriate screening of the literature on the EU distribution of the species/ genera, a WoS search was performed using the species name as a search string. All results were individually checked. When more than 300 items were retrieved, the search was refined by including 'Europe' as search string.
Data about the area of hosts grown in the EU were obtained from EUROSTAT (Statistical Office of the European Communities).
The Europhyt database was consulted for pest-specific notifications on interceptions and outbreaks. Europhyt is a web-based network run by the Directorate General for Health and Food Safety (DG SANT E) of the European Commission, and is a subproject of PHYSAN (Phyto-Sanitary Controls) specifically concerned with plant health information. The Europhyt database manages notifications of interceptions of plants or plant products that do not comply with EU legislation, as well as notifications of plant pests detected in the territory of the EU MS and the phytosanitary measures taken to eradicate or avoid their spread.

Methodologies
The Panel performed the pest categorisation for the non-EU phytoplasmas of the host plants following guiding principles and steps presented in the EFSA guidance on quantitative pest risk assessment (EFSA PLH Panel, 2018) and in the International Standard for Phytosanitary Measures (ISPM) No 11 (FAO, 2013) and No 21 (FAO, 2004).
This work was started following an evaluation of the EU plant health regime. Therefore, to facilitate the decision-making process, in the conclusions of the pest categorisation, the Panel addresses explicitly each criterion for a Union quarantine pest and for a Union RNQP in accordance with Regulation (EU) 2016/2031 on protective measures against pests of plants, and includes additional information required in accordance with the specific terms of reference received by the European Commission. In addition, for each conclusion, the Panel provides a short description of its associated uncertainty. Table 1 presents the Regulation (EU) 2016/2031 pest categorisation criteria on which the Panel bases its conclusions. All relevant criteria have to be met for the pest to potentially qualify either as a quarantine pest or as a RNQP. If one of the criteria is not met, the pest will not qualify. A pest that does not qualify as a quarantine pest may still qualify as a RNQP that needs to be addressed in the opinion. For the pests regulated in the protected zones only, the scope of the categorisation is the territory of the protected zone; thus, the criteria refer to the protected zone instead of the EU territory.
It should be noted that the Panel's conclusions are formulated respecting its remit and particularly with regard to the principle of separation between risk assessment and risk management (EFSA founding regulation (EU) No 178/2002); therefore, instead of determining whether the pest is likely to have an unacceptable impact, the Panel will present a summary of the observed pest impacts. Economic impacts are expressed in terms of yield and quality losses and not in monetary terms, whereas addressing social impacts is outside the remit of the Panel. The Panel will not indicate in its conclusions of the pest categorisation whether to continue the risk assessment process, but following the agreed two-step approach, will continue only if requested by the risk managers. However, during the categorisation process, experts may identify key elements and knowledge gaps that could contribute significant uncertainty to a future assessment of risk. It would be useful to identify and highlight such gaps so that potential future requests can specifically target the major elements of uncertainty, perhaps suggesting specific scenarios to examine.

3.
Pest categorisation 3. Are there measures available to prevent the entry into, establishment within or spread of the pest within the protected zone areas such that the risk becomes mitigated?
Is it possible to eradicate the pest in a restricted area within 24 months (or a period longer than 24 months where the biology of the organism so justifies) after the presence of the pest was confirmed in the protected zone?
Are there measures available to prevent pest presence on plants for planting such that the risk becomes mitigated?

Conclusion of pest categorisation (Section 4)
A statement as to whether (1) all criteria assessed by EFSA above for consideration as a potential quarantine pest were met and (2) if not, which one(s) were not met A statement as to whether (1) all criteria assessed by EFSA above for consideration as potential protected zone quarantine pest were met, and (2) if not, which one(s) were not met A statement as to whether (1) all criteria assessed by EFSA above for consideration as a potential RNQP were met, and (2) if not, which one(s) were not met Phytoplasmas are accommodated within the 'Candidatus Phytoplasma' genus. Within this genus, several species have been described based on their 16S rRNA gene sequences. Within the species, strains officially included in the species description share a common signature at this locus. For each species, a reference strain is described, and its 16S rRNA sequence determined. Strains with minimal differences in the 16S rRNA gene (≥ 97.5% identity) are considered as related strains. In the presence of minimal differences of the 16S rRNA gene, if the two phytoplasmas are transmitted by different vectors, have a different natural plant host (or, at least, their behaviour is significantly different in the same plant host), and there is evidence of significant molecular diversity, the description of a new species is recommended (IRPCM, 2004). The current opinion covers pathogens at the strain level, infecting at least one of the host plants within their officially described 'Ca. P. species'. In one case (Buckland valley grapevine yellows) the phytoplasma has not yet been assigned to a 'Ca. P.' species.
Key information on the identity of the phytoplasmas categorised in the present opinion is reported in Table 2.  (Zreik et al., 1995). The phytoplasma belongs to the 16SrII ribosomal group (IRPCM, 2004). Many other diseases have been associated with phytoplasmas within the 16SrII group, and those infecting the host plants are categorised here, including 'Ca. P. australasia' [PHYPAA], as this species is not officially accepted (IRPCM, 2004). Phytoplasmas of the 16Sr-II group are the causal agents of sweet potato little leaf (SPLL) [PHYP39], crotalaria witches'-broom phytoplasma (CrWB), pear decline Taiwan II (PDTWII), papaya mosaic, peanut witches' broom phytoplasma, sunn hemp witches' broom phytoplasma, Australian big bud of tomato, dieback of papaya, mosaic of papaya, yellow crinkle of papaya, tomato big bud (TBB) [PHYP01], dwarf disease of sweet potato, little leaf of sweet potato. Inclusion of 16Sr-II phytoplasma strains papaya yellow crinkle (PpYC), papaya mosaic (PpM), and tomato big bud (TBB) into the 'Ca. P. australasia' taxon (White et al., 1998) has not been further supported (IRPCM, 2004) 'Ca. P. australiense' (reference strain) The 'Ca. P. australiense' species includes different phytoplasma genetic lineages. They are the causal agents of: Australian grapevine yellows (AUSGY; AGY); Strawberry lethal yellows; papaya dieback (PpDB); phormium yellow leaf (PYL), Australian lucerne yellows. Within the species (Davis et al., 1997a), three distinct subgroups are proposed based on sequence analyses of the tuf gene: tuf 1, tuf 2, and tuf 3 . A related strain is reported in Australia (Getachew et al., 2007). The phytoplasma is listed as PHYPAU in the EPPO GD 'Ca. P. fraxini' (reference strain) The 'Ca. P. fraxini' species (Griffiths et al., 1999) includes members of the AshY phytoplasma group (16SrVII-A). They are the causal agents of Ash yellows (AshY) and lilac witches' broom (LWB) diseases in North America . Phytoplasmas of the same 16Sr group (subgroup -C) have been reported to infect Fragaria  in Argentina, although their status within the 'Ca. P. fraxini' is uncertain. The phytoplasma is listed as PHYPFR in the EPPO GD 'Ca. P. hispanicum' (reference strain) The 'Ca. P. hispanicum' species includes different phytoplasma genetic lineages. They are the causal agents of: Mexican periwinkle virescence (MPV) which was indicated as reference strain, strawberry multiplier (STRAWB1), Mexican potato purple top (MPPT-SINPV), papaya apical curl necrosis (PACN-Br04) and strawberry red leaf (StrawRL-Tc1) . Other diseases recently described and associated with this phytoplasma species are: broccoli stunt in Brazil , strawberry green petal disease in Mexico , and strawberry fruit phyllody (SFP-Br02)) in Brazil (Melo et al., 2018). The Strawberry multiplier disease phytoplasma is listed as PHYP75 in the EPPO GD. The phytoplasma listed as Strawberry witches' broom mycoplasm (SYWB00) in Annex IAI was detected before the development of molecular identification tools, therefore its designation as related strain of 'Ca. P. hispanicum' is uncertain. Phytoplasmas identified in Melia azedarach  are now included in 'Ca. P. meliae' . The 'Ca. P. hispanicum' is listed as PHYP07 in the EPPO GD Phytoplasma name, reference strain/ related strain name Justification 'Ca. P. pruni'-related strain (North American grapevine yellows, NAGYIII) 16SrIII NAGY phytoplasma strains are closely related to, but distinct from, strains of 'Ca. Phytoplasma pruni', cause of Prunus X-disease, based on deduced 3-dimensional structure of SecY proteins, and SNPs (single nucleotide polymorphisms) in 16S rRNA, secY, and ribosomal protein (rp) genes. They differ from 'Ca. Phytoplasma pruni' in regions of the 16S rRNA gene corresponding to three segments described as speciesunique for 'Ca. Phytoplasma pruni'  'Ca. P. pyri'-related strain (peach yellow leaf roll, PYLR) This disease peach yellow leaf roll (PYLR) was first observed in 1948 in the Sacramento Valley, California, USA. The incidence of PYLR remained relatively low until an epidemic outbreak in the late 1970s and early 1980s (Marcone et al., 2014). A disease named Almond brown line and decline (ABLD) may also be caused by PYLR phytoplasma (Uyemoto et al., 1992). The PYLR phytoplasma is phylogenetically closely related to, but not identical to, the 'Ca. P. pyri' reference strain agent of the pear decline disease, PD. In most molecular analyses based on ribosomal and non-ribosomal DNA sequences, PYLR is indistinguishable from the PD phytoplasma (Seemuller and Schneider, 2004). Comparison of genes coding the immunodominant membrane protein (IMP) clearly indicates that PYLR and PD are different (Morton et al., 2003). For this reason, the PYLR is considered as a related strain of the 'Ca. P. pyri' (Seemuller and Schneider, 2004) 'Ca. P. trifolii' (reference strain) The 'Ca. P. trifolii' phytoplasma includes Clover proliferation (CPR) as the reference strain, and alfalfa witches'-broom (AWB), brinjal little leaf (BLL), beet leafhopper-transmitted virescence (BLTV), Illinois elm yellows (ILEY), potato witches'-broom (PWB), potato yellows (PY), tomato big bud in California (TBBc) and phytoplasmas from Fragaria multicipita (FM) (Hiruki and Wang, 2004). The latter is an invalid taxon, as F. multicipita, thought to be a rare plant with unusual vegetative morphology, is actually a phytoplasmadiseased aberrant growth form of F. virginiana (Jomantiene et al., 1998a). The vegetative morphology of F. multicipita is typical of strawberry plants affected by multiplier disease found in winter production strawberry fields in Florida, as also confirmed by molecular analyses . The phytoplasma is listed as PHYPTR in the EPPO GD, together with the following other names: Lucerne witches' broom phytoplasma, Potato witches' broom phytoplasma and proliferation of clover. Other microorganisms are listed in the EPPO GD as PHYP61 (Willow witches' broom phytoplasma) and PHYP62 (Brinjal little leaf phytoplasma and Eggplant little leaf phytoplasma). These can be assigned to 'Ca. P. trifolii' in agreement with the species description (Hiruki and Wang, 2004) 'Ca. P. ziziphi' (reference strain) The 'Ca. P. ziziphi' species includes the causal agents of Jujube witches' broom phytoplasma (or witches' broom of jujube), the sweet cherry virescence (SCV), a stem fasciation disease of persimmon trees, the cherry lethal yellows (CLY5) in China, the peach yellows in India (PY-In), and the Euonimus witches' broom in China, among others. These phytoplasmas form a homogenous ecological lineage (within the 16SrV-B) based on analysis of genetic loci that encode important phytoplasma cellular components, including an array of ribosomal proteins and preprotein translocase subunit SecY , although minor RFLP patterns are predicted by in silico restriction digestion of their 16s rRNA sequences . Sequence analyses of the 16S rRNA gene of several Chinese isolates of JWB confirm that the pathogen causes consistent symptoms from different regions and cultivars (Bu et al., 2016). The genome of 'Ca. P. ziziphi' has been recently sequenced . PHYPZI is the EPPO code for 'Ca. P. ziziphi'

Buckland valley grapevine yellows phytoplasma
This phytoplasma shows about 97,0% sequence identity to 'Candidatus Phytoplasma asteris' (16SrI), its closest relative, therefore it fulfills the requisite for being classified as a different species, although this has not been done so far (Fiona Constable, Microbiology, Agriculture Victoria Research, Department of Jobs, Precincts and Regions, AgriBio, 30/01/2019, personal communication). The Buckland valley grapevine yellows phytoplasma has been assigned as reference isolate of the 16SrXXIII group . No variation was detected between isolates of the phytoplasma when Heteroduplex Mobility Assay of the tuf gene was done and it is possible that this phytoplasma lacks molecular diversity  3.1.2. Biology of the pest All the phytoplasmas considered in the present pest categorisation are efficiently transmitted by grafting of infected scions on healthy plants, as well as by phloem feeder insect vectors. Phytoplasmas are transmitted by insects in the order Hemiptera. However, vector species are restricted to only a few families of the Fulgoromorpha and Cicadomorpha (most of the vector species belong to Cicadellidae and Cixiidae), and of Sternorrhyncha (Psyllidae) (Weintraub and Beanland, 2006)). Within a family, some species are known to be phytoplasma vectors, while others are not. Transmission is persistent and propagative, and insects are infective for life. No transovarial transmission has been reported for the phytoplasmas categorised here.
The phytoplasma transmission process consists of: i) acquisition of the pathogen during feeding on an infected plant, ii) a latent period in the insect, during which the phytoplasma crosses the midgut barrier, multiplies within the insect body and colonizes its salivary glands, and iii) inoculation of the bacterium during feeding on a healthy plant.
Details on the symptoms on the host plants, incubation period and epidemiology are listed in Table 3. Symptoms on other plants are listed in Appendix A. The known vector species are listed in Table 4. Phytoplasma infection is often due to a single phytoplasma strain/species, and insect vectors can acquire this phytoplasma and transmit it to other plants of the same species or other susceptible species. Therefore, the epidemiological cycle is simple, since a single phytoplasma is transmitted among susceptible plants of one or more botanical species. It appears that vectors can act in closed or open epidemiological cycles. A closed cycle is represented by a phytoplasma that circulates between the main, if not exclusive, host plant and the main, if not exclusive, vector species (Bosco and D'Amelio, 2010). No pollen and seed transmissions have been reported for the phytoplasmas considered in this opinion.
For this pest categorisation, two vector categories were identified (Trivellone, 2019): 1) An insect species is considered a competent vector if the phytoplasma capability to overcome the barriers of gut and salivary glands has been proven using classical acquisition/ inoculation experiments in the laboratory, or inoculation trials with caged infected specimens collected from the field. 2) An insect species is considered a potential vector if the phytoplasma has been detected in the insect body using standard molecular methods, or inoculated to artificial medium under laboratory conditions. The status as a potential vector does not prove the ability to transmit the phytoplasma from plant to plant. Fragaria: strawberry leaves from fruit (Streten et al., 2005c).
In Prunus (apricot), the minimum time between inoculation and symptom expression is of 21 months (Rasoulpour et al., 2019).
In Pyrus, diseased seedlings begin to exhibit the characteristic upward leaf curling symptoms of pear decline (PDTWII) three to six months after transmission .
In Carica papaya: the mean time from symptom appearance to plant death is about 4 months  Pyrus: leaf redness and curling followed by progressive weakening and wilt ; general dieback, poor terminal growth, and poorly developed root systems .
Vitis: decline, leaf yellowing and shortening of internodes (Ghayeb Zamharir et al., 2017), late season leaf curl, LSLC  papayae . The infectious period (incubation period plus the period from postincubation to time-to-death period) of infected papaya ranges from 6 to 9 months (Esker et al., 2006) 'Ca. P. australiense' (reference strain) Fragaria: plants growing flatter to the ground, purpling of older leaves, reduced leaf size, yellowing of younger leaves, and sometimes plant death .
Prunus: yellowing of leaf margins and rolling, drying and necrosis (leaves), proliferation of shoots along branches  In infected papaya, plant apical death occurs within 2 weeks of first visible external symptom expression, and 3 weeks of first detection of phytoplasma in host tissue (Guthrie et al., 2001); plants survive less than 3 months  No information was found 'Ca. P. fraxini' (reference strain) Fragaria: phyllody .
Prunus spp: decline, leaf reddening, yellowing, shortening of internodes, witches' broom and reduced fruit size (Zunnoon- . Vitis: severe yellows, decline, reduced internode size and leaf with lack of lignification (Ghayeb Zamharir et al., 2017;Zambon et al., 2018) Mortality of infected trees occurs within 2 years in young ash trees and up to 10 years after infection of old trees (reviewed in (Olivier et al., 2009)). Mortality (30%) over an 8-year study period is reported for Fraxinus velutina adult trees in Arizona, with a lag time between ash infection and symptom development of about 2 years (Bricker and Stutz, 2004). Symptom incubation up to 4 years has been reported in 20% of the ash trees of a US population (Sinclair and Griffiths, 1995) No information was found 'Ca. P. hispanicum' (reference strain) Fragaria: fruit phyllody, achenes' hypertrophy and leaf reddening (Cui et al., 2018), slow growth (Melo et al., 2018), deformation of fruits, large, elongated purple leaves and green petals (Avendano-Benequen et al., 2017), stunting, young leaves with yellowing at the edges, mature leaves with curling and a reddish colouration at the abaxial face, death  Fragaria Vitis: Symptoms of NAGYIII are similar to those of other grapevine yellows diseases that occur globally, and include leaf reddening in redfruited cultivars (cvs.), leaf chlorosis in white-fruited cvs., downward rolling of leaf margins, dieback of shoot tips, abortion of fruit clusters, and nonuniform maturation of shoot stem periderm (Wolf, 2015) NAGYIII affected vines often die within 2 or 3 years of symptom onset (Wolf, 2015) Infected Prunus: PYLR causes chlorosis, downward curling of leaf tips and rolling of leaf margins in mid-summer due to a cork layer deposition, while leaves remain normal in size One year (Uyemoto et al., 1999) Disease incidence did not differ among peach cvs., but was significantly lower in 4 year old or younger peach trees than in older plants. PYLR incidence was higher in peach orchards adjacent to commercial pear orchards and the incidence decreased with distance from pears (Purcell et al., 1981). Furthermore, since there is no evidence that PYLR spreads from peach to peach, pear trees are regarded as the primary pathogen reservoir (Marcone et al., 2014) 'Ca. P. trifolii' (reference strain) Fragaria: stunting, small leaves, multiple crowns and no runners (Jomantiene et al., 1998a).
Prunus dulcis: little leaf, leaf rolling, dieback of branches, rosette and yellowing (Zirak et al., 2009a)  Prunus spp: the diseased trees develop flowers having white petals with green veins or abnormal floral structures having cupped, green petals; the affected flowers fail to set fruit . Leaf symptoms start in early summer with upward leaf curling followed by yellowing, reddening and premature fall and stunting (Paltrinieri et al., 2006).
Malus: little leaf, leaf margin involute and yellows (Li et al., 2014b) Upon grafting of symptomatic shoots on healthy jujube plants symptoms appear after 48 weeks (Ye et al., 2017). 'Ca. P. ziziphi' phytoplasma can survive in branches during winter and roots are not necessary for the multiplication of phytoplasma and the development of phytoplasma symptom (Zhao et al., 2009a) Two year old apples may show 'Ca. Phytoplasma ziziphi'-like symptoms, but the phytoplasma does not infect adult trees (Li et al., 2014b) Buckland valley grapevine yellows phytoplasma Vitis: Yellowing, downward curling of leaves, stunted shoots that do not harden off, but remain rubbery, shoot tips dye and bunches shrivel and fall

No information was found
Up to about 30% incidence was reported in Chardonnay plots . This phytoplasma affects many grapevines and is characterized by remission of symptoms, some recurrence and occurrences in previously unaffected grapevines (Constable et al., 2003)  able to acquire the phytoplasma on infected sesame plants ; Orosius argentatus is a vector of the TBB phytoplasma to tomato, several legume species, and lucerne .
Orosius cellulosa is the vector of cotton virescence phytoplasma (Desmidts et al., 1973), which is now known to be a 16SrII-C subgroup member (Marzach ı et al., 2009) Orosius lotophagorum is also a vector of sweet potato little leaf disease .

Intraspecific diversity
Taking into account the reasoning of Section 3.1.1, intraspecific diversity is addressed up to the related strain level. Nevertheless, in most cases (with the exception of 'Ca. P. pyri'-related strain (peach yellow leaf roll, PYLR) and Buckland valley grapevine yellows phytoplasma), sequevars (groups of strains characterised by a specific DNA sequence for one or several genes) have been reported, and these are listed in Table 5. Hishimonus sellatus . Hishimonoides chinensis inoculated with phytoplasmas from infected Ziziphus mauritiana can transmit the disease, especially where Paulownia and Z. mauritiana are mixed or close together (Jin and Gao, 1984) Hishimonus lamellatus (Trivellone, 2019) Buckland valley grapevine yellows phytoplasma None reported None reported None The peanut witches' broom phytoplasma group (16SrII) includes several strains with a worldwide distribution. Twenty-one 16SrII subgroups (A-U) have been described ( 'Ca. P. australiense' (reference strain) Some evolutionary divergence in the 16SrXII-B group has been suggested. Analyses of genes (tuf, and rp operon) that are less conserved than the 16S rRNA gene can distinguish four subgroups (16SrXII-B (tuf-Australia I; rp-A), 16SrXII-B (tuf-New Zealand I; rp-B), 16SrXII-B (tuf-New Zealand II) and 16SrXII-B (rp-C). Strawberry lethal yellows 1, strawberry green petal, Australian grapevine yellows, pumpkin yellow leaf curl and cottonbush witches'broom phytoplasmas are members of the 16SrXII-B (tuf -Australia I; rp-A) subgroup. The strawberry lethal yellows 2 and cottonbush reduced yellow leaves phytoplasmas are assigned to the 16SrXII-B (tuf-New Zealand II; rp-B), subgroup. No relationship is present between these phytoplasma subgroups and collection date, location or host plant ) 'Ca. P. fraxini' (reference strain) Based on sequence homology, similarity coefficients derived from RFLP of the 16S rDNA and phylogenetic analysis, six 16S rRNA subgroups have been described (16SrVII-A to -F) (da Silva Fugita et al., 2017). The subgroup 16SrVII-A incorporates North American isolates (Griffiths et al., 1999). There is uncertainty on whether 16S rRNA other than -A should be included in the 'Ca. P. fraxini' species . Among these Phytoplasma name, reference strain/related strain name Justification subgroups, erigeron witches' broom from Brazil and Argentina (EriWB) phytoplasmas are included in VII-B subgroup (Barros et al., 2002;Meneguzzi et al., 2008), alfalfa witches'broom from Argentina (ArAWB) phytoplasma in VII-C subgroup , together with the causal agent of Crotalaria juncea shoot proliferation in Brazil . Within 16SrVII-D, a phytoplasma is reported in erigeron plants from Brazil (Flores et al., 2015), and a Chilean isolate from grapevine, previously classified as belonging to subgroup 16SrVII-A (Gajardo et al., 2009), was tentatively reclassified as a representative of subgroup 16Sr-E . Finally, a 16SrVII phytoplasma identified in Vernonia brasiliana in Brazil has been assigned to -F subgroup (da Silva Fugita et al., 2017). Strain variation in aggressiveness has been reported both in ash trees and in an experimental host (Catharanthus roseus) (Sinclair and Griffiths, 2000) 'Ca. P. hispanicum' (reference strain) Intraspecific variation exists within the 'Ca. P. hispanicum' species, as several different 16SrXIII subgroups have been described within this species: 16SrXIII-A (Mexican periwinkle virescence), 16SrXIII-B (strawberry multiplier), 16SrXIII-D (potato purple top disease), 16SrXIII-E (papaya apical curl necrosis), 16SrXIII-F (strawberry red leaf) , 16SrXIII-H (broccoli stunt) , 16SrXIII-I (strawberry green petal) , 16SrXIII-J (strawberry fruit phyllody) (Melo et al., 2018), and a novel subgroup detected in strawberry in Chile (Cui et al., 2018). Phytoplasma isolates from Melia azedarach collected in Argentina, Bolivia and Paraguay, formerly included in 'Ca. P. hispanicum' species , were then described as strains of the 'Ca. P. meliae' species  'Ca. P. pruni'related strain (North American grapevine yellows, NAGYIII) Based on their 16SrRNA gene sequence, the NAGYIII strains belong to two sequevars (NAGYIIIa and b), and they can be differentiated from 'Ca. P. pruni' by the presence of a MseI restriction site that is absent from the 'Ca. P. species' reference strain. Because NAGYIII sequevars have not been reported in X-disease, there is uncertainty about whether NAGYIII and Prunus X-disease are caused by different phytoplasma genotypes  'Ca. P. trifolii' (reference strain) Three subgroups have been classified on the basis of sequence homology and the collective RFLP patterns of amplified 16S rRNA genes. CP, AWB, BLTV, PWB and TBBc are assigned to taxonomic subgroup CP-A, Fragaria multicipita belongs to subgroup CP-B and ILEY isolates are assigned to subgroup CP-C (Hiruki and Wang, 2004). Additional variants have been described. Isolates BLL, Centaurea solstitialis virescence (CSV1), Catharanthus phyllody (CPS), and PLL are assigned to subgroups VI-D, -E, -F, and -H respectively . Heterogeneity of the 16S rRNA has been reported for the 'Fragaria multicipita', leading to assignation to -B or -G 16SrVI subgroups, depending on the sequence used for classification (Davis et al., 2003a. Sequence alignment of the 'Ca. Phytoplasma trifolii' isolates from Capsicum annum revealed a low level of genetic diversity within subgroup 16SrVI-A in Turkey (Oksal et al., 2017) and Mexico, although in Mexico a second isolate was classified into a new subgroup (16SrVI-J) .
Group/subgroup assignation of the two 16SrVI-related strains described in Araucaria heterophylla  and in Datura inoxia (Raj et al., 2009) is uncertain, as these strains share less than 97.5% sequence homology with Ca. P. trifolii, respectively 'Ca. P. ziziphi' (reference strain) The 'Ca. P. ziziphi' species includes strains from a homogenous ecological lineage , although isolates with less than 99.7% variation on their 16S rRNA sequence have been associated to similar stem fasciation of persimmon trees in China . Also, Indian16SrV-B isolates infecting peach differ from 'Ca. P. zyzyphi' reference isolates upon sequence analyses of other genomic regions (eg. ribosomal protein rpl22 and rps3 and at the secY genes) (Marcone et al., 2014). A JWB disease in northeastern China is associated with JWB-DL strain representing a new, distinct 'Ca. P. ziziphi'-related strain (Wei et al., 2007)

Detection and identification of the pest
For all the categorised phytoplasmas, molecular detection methods are available. Phytoplasmas are routinely detected by polymerase chain reaction (PCR). Universal and specific primers are available, and nested PCR protocols have been developed to overcome low pathogen titre in certain hosts and/or in the case of asymptomatic infection. However, there is a potential lack of specificity in the case of the design of nested PCR with universal primers. Diagnostics in woody host plants is sometimes difficult because of the uneven pathogen distribution, low phytoplasma loads or presence of inhibitors in the extracts to be tested. Several robust diagnostic protocols have also become available based on Real Time PCR and LAMP (loop mediated isothermal amplification) approaches. Identification of phytoplasmas is routinely achieved by sequencing of a specific 16S rRNA fragment followed by virtual restriction fragment length polymorphism (RFLP) analyses according to an available online tool (Zhao et al., 2009b;iPhyClassifier, 2019). Phytoplasma-specific symptoms may indicate phytoplasma infection, but cannot be used to identify the infecting 'Ca. P. species'.
In Table 6, the detection and identification methods for each categorised phytoplasma is summarized together with the associated uncertainty.
Are detection and identification methods available for the pest?
YES, the non-EU phytoplasmas categorised here can be detected by molecular methods. In the case of papaya, that can be infected by several phytoplasma species, ribosomal primers are available for the specific amplification of 16SrII phytoplasmas (P1/rSPLLS SUNH, ).

None
'Ca. P. fraxini' (reference strain) Phytoplasma-specific primers (PI and Tint) and AshY specific primers (fBl/rASHYS) are available to amplify a portion of the phytoplasma16S rDNA (Smart et al., 1996). Amplicon sequencing may be required to confirm the identification (Feeley et al., 2001) None 'Ca. P. hispanicum' (reference strain) Molecular characterization has been successfully performed using chaperonin-60 (cpn60) and DNAdependent RNA polymerase b-subunit (rpoB) genes  None 'Ca. P. pruni'-related strain (North American grapevine yellows, NAGYIII) NAGYIII strains can be differentiated from 'Ca. P. pruni' by the presence of a MseI restriction site on the 16S rRNA gene that is absent from that of the 'Ca. P. species' reference strain  None 'Ca. P. pyri'-related strain (peach yellow leaf roll, PYLR) Nested PCR with universal primers was applied in the certification of dormant Prunus buds in the US (Waterworth and Mock, 1999). A Real Time PCR system has also been reported (Sudarshana et al., 2011) No details of the Real Time PCR protocol are provided in the reference (conference abstract only) (Sudarshana et al., 2011) 'Ca. P. trifolii' (reference strain) Sensitive and quantitative diagnostic tools to investigate mixed infections of two phytoplasma isolates of 'Ca. P. trifolii' are available, based on PCR-RFLP and micro-channel capillary electrophoresis (Wei There are shortcomings in the qPCR assay, since it also detected aster yellows (group 16SrI) and pigeon pea witches'-

Pest distribution outside the EU
The distribution outside the EU of the phytoplasmas categorised here is reported in Table 7, based on data from the EPPO GD and/or the CABI Crop Protection Compendium (CPC) (CABI, 2019), and, when not available in these sources, from extensive literature searches.
The available distribution maps from the EPPO GD (for Ca. P. australiense (reference strain), Ca. P. fraxini (reference strain) and Ca. P. trifolii (reference strain)) are provided in Appendix B.
Phytoplasma name, reference strain/ related strain name Available detection and identification methods Uncertainties et al., 2016). A qPCR protocol based on ribosomal 16 gene was developed to detect Columbia Basin potato purple top in in plants and beet leafhoppers (Crosslin et al., 2006). A specific nested PCR protocol is also available (Smart et al., 1996). broom (group 16SrIX) phytoplasmas in infected periwinkle plants (Crosslin et al., 2006) 'Ca. P. ziziphi' (reference strain) Direct and nested PCRs were performed to target amplifications of three phytoplasma genomic loci, namely, a partial ribosomal rRNA operon (rrn), a ribosomal protein gene cluster rpsS-rplV-rpsC, and an extended secY locus that encodes ribosomal protein L15, preprotein translocation subunit secY, and methionine aminopeptidase (rplO-secYmap) ).

Buckland valley grapevine yellows phytoplasma
Nested PCR with universal primers followed by RFLP was applied for pathogen detection and characterisation (Constable et al., 2002) None AMERICA: Brazil (Silva et al., 2014); Cuba ; Peru  ASIA: Israel ; Myanmar ; Pakistan ; Saudi Arabia ; Turkey (Ikten et al., 2014;Ozdemir and Cagirgan, 2015) OCEANIA: Wallis and Futuna Islands  EUROPE (non-EU): Serbia  Serbia: reported in six Picris hieracioides asymptomatic samples collected in vineyards . OCEANIA: New Caledonia  The Strawberry witches'broom mycoplasm (SYWB00) reported in Canada and Japan by the EPPO GD was detected before the development of molecular identification tools.

Pest distribution in the EU
Four of the phytoplasmas categorised here were reported in the EU (Table 8), where they can be considered to have a restricted distribution, as all of them were reported only in few plants, in up to four EU MS and mostly by a single research group. In addition, in some cases ('Ca. P. aurantifolia'related strains', 'Ca. P. trifolii'), reports from the EPPO GD are presented with 'no details'.   -Is the pest present in the EU territory? If present, is the pest widely distributed within the EU?
NO, 'Ca. P. australiense' (reference strain), 'Ca. P. hispanicum' (reference strain), 'Ca. P. pruni'-related strain (North American grapevine yellows, NAGYIII), 'Ca. P. pyri'-related strain (peach yellow leaf roll, PYLR), and the unclassified Buckland valley grapevine yellows phytoplasma are not known to be present in the EU. Greece, Portugal (EPPO GD: Present, no details), Italy Davino et al., 2007;Paltrinieri and Bertaccini, 2007;Parrella et al., 2008), UK  Reports from the EPPO GD in Greece and Portugal have no further details. The pest was reported i) in few batches of symptom-less potato plantlets obtained from two lots of seeds from different undescribed Italian locations and from unknown origins (Paltrinieri and Bertaccini, 2007); ii) in one batch (10 insects) out of 3 of field-collected Empoasca decipiens in Italy (Parrella et al., 2008); iii) in 3 field-collected Calendula arvensis plants, one Solanum nigrum plant, and one Chenopodium species ; iv) in the UK, where 50 (57%) of 88 plants showed obvious symptoms, at one location Italy (Bruni et al., 2005;Zambon et al., 2018) The reports from Italy were published by the same group but not confirmed. Vitis: there is only one report concerning 9 plants detected by nested PCR out of 171 tested ones (Zambon et al., 2018). Hypericum perforatum: it is not known on how many plants the phytoplasma was identified (Bruni et al., 2005) 3.3.

Council Directive 2000/29/EC
Non-EU phytoplasmas of the host plants are listed in Council Directive 2000/29/EC. All phytoplasma categorised here are listed in Annex IAI, under the generic definition 'Non-European viruses and viruslike organisms of Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L.'. Details are presented in Table 9.
Annex IAI also mentions peach rosette (Annex IAI 5(f)), Peach X-disease (Annex IAI 5(g)) and Peach yellows (Annex IAI 5(h)) mycoplasmas. These are all strains of the 'Ca. P. pruni' (reference strain), which has been reported several times in the EU (EFSA PLH Panel, 2020). However, 'Ca. P. pruni'-related strain (NAGYIII) is closely related to, but distinct from, the above-mentioned strains of 'Ca. Phytoplasma pruni'. Therefore, it cannot be considered regulated as Peach rosette, Peach X-disease and Peach yellows mycoplasmas.
Annex IAI also includes strawberry witches' broom mycoplasm (SYWB00), which is probably a strain of the 'Ca. P. hispanicum'. None A disease named 'peach yellow leaf roll' was reported in peach orchards from Spain, but the identified causal agent was 'Ca. P. pyri', isolate PD, based on sequence analyses of 16S rRNA, pnp, secY, imp, aceF genes (Sabate et al., 2014;Sabate et al., 2018). Therefore, the phytoplasma reported in Spain was not PYLR 'Ca. P. trifolii' (reference strain) Italy, Spain (EPPO GD: Present, no details), Austria , Czech Republic  EPPO GD: Reports from EU MS refer to few infected plants (ranging from 1 to 28). Therefore, 'Ca. P. trifolii' is not considered to be widely present in the EU 'Ca. P. ziziphi' (reference strain) Italy (Pasquini et al., 2000;Paltrinieri et al., 2006) Only a conference report from one plant from Italy with no details beside PCR-RFLP detection of mixed infections with 'Ca. P. solani' and 'Ca. P. asteris' (Paltrinieri et al., 2006). Only one full report from Italy (Pasquini et al., 2000), in the absence of any further characterization beside PCR and RFLP analyses

Buckland valley grapevine yellows phytoplasma
None None   chips, sawdust and shavings, obtained in whole or part from these plants, wood packaging material, in the form of packing cases, boxes, crates, drums and similar packings, pallets, box pallets and other load boards, pallet collars, dunnage, whether or not actually in use in the transport of objects of all kinds, except dunnage supporting consignments of wood, which is constructed from wood of the same type and quality as the wood in the consignments and which meets the same Union phytosanitary requirements as the wood in the consignment, but including that which has not kept its natural round surface, originating in Canada and the USA Official statement that the wood: (a) originates in an area free from Saperda candida Fabricius, established by the national plant protection organisation in the country of origin, in accordance with the relevant International Standards for Phytosanitary Measures, which is mentioned on the certificates referred to in Article 13(1)(ii) under the rubric 'Additional declaration', or (b) has undergone an appropriate heat treatment to achieve a minimum temperature of 56°C for a minimum duration of 30 continuous minutes throughout the entire profile of the wood, which is to be indicated on the certificates referred to in Article 13(1)(ii), or (c) has undergone an appropriate ionising radiation to achieve a minimum absorbed dose of 1 kGy throughout the wood, to be indicated on the certificates referred to in Article 13 (1) Phytosanitary Measures, which is mentioned on the certificates referred to in Article 13(1)(ii) under the rubric 'Additional declaration', or (b) has been processed into pieces of not more than 2,5 cm thickness and width, or (c) has undergone an appropriate heat treatment to achieve a minimum temperature of 56°C for a minimum duration of 30 minutes throughout the entire profile of the chips, which is to be indicated on the certificates referred to in Article 13 (1) (9) and (18), and Annex IV(A)(I) (15) and (17) (a) the plants, other than those raised from seed, have been: either officially certified under a certification scheme requiring them to be derived in direct line from material which has been maintained under appropriate conditions and subjected to official testing for at least the relevant harmful organisms using appropriate indicators or equivalent methods and has been found free, in these tests, from those harmful organisms, or derived in direct line from material which is maintained under appropriate conditions and has been subjected, within the last three complete cycles of vegetation, at least once, to official testing for at least the relevant harmful organisms using appropriate indicators or equivalent methods and has been found free, in these tests, from those farmful organisms,  (9) and (18), Annex III(B)(1) and Annex IV(A)(I) (15), (17) and (19.2), official statement that: (a) the plants have been: either officially certified under a certification scheme requiring them to be derived in direct line from material which has been maintained under appropriate conditions and subjected to official testing for at least the relevant harmful organisms using appropriate indicatos or equivalent methods and has been found free, in these thests, from those harmful organisms, or derived in direct line from material which is maintained under appropriate conditions and subjected, within the last three complete cycles of vegetation, at least once, to official testing for at least the relevant harmful organisms using appropriate indicators or equivalent methods and has been found free, in these tests, from those harmful organisms; either officially certified under a certification scheme requiring them to be derived in direct line from material which has been maintained under appropriate conditions and subjected to official testing for at least Apple proliferation mycoplasm using appropriate indicators or equivalent methods and has been found free, in these tests, from that harmful organism, or derived in direct line from material which is maintained under appropriate conditions and subjected, within the last six complete cycles of vegetation, at least once, to official testing for at least Apple proliferation mycoplasm using appropriate indicators or equivalent methods and has been found free, in these tests, from the harmful organism, (bb) no symptoms of diseases caused by Apple proliferation mycoplasm have been observed on plants at the place of production, or on susceptible plants in its immediative vicinity, (a) the plants, other than those raised from seed, have been: either officially certified under a certification scheme requiring them to be derived in direct line from material which has been maintained under appropriate conditions and subjected to official testing for, at least, Plum pox virus using appropriate indicators or equivalent methods and has been found free, in these tests, from that harmful organism, or derived in direct line from material which is maintained under appropriate conditions and has been subjected, within the last three complete cycles of vegetation, at least once, to official testing for at least Plum pox virus using appropriate indicators or equivalent methods and has been found free, in these tests, from that harmful organism; either officially certified under a certification scheme requiring them to be derived in direct line from material which has been maintained under appropriate conditions and subjected to official testing for at least the relevant harmful organisms using appropriate indicators or equivalent methods and has been found free, in these tests, from those harmful organisms, or derived in direct line from material which is maintained under appropriate conditions and has been subjected, within the last three complete cycles of vegetation, at least once, to official testing for at least the relevant harmful organisms using appropriate indicators or equivalent methods and has been found free, in these tests, from those harmful organisms, either officially certified under a certification scheme requiring them to be derived in direct line from material which has been maintained under appropriate conditions and subjected to official testing for at least the relevant harmful organisms using appropriate indicators or equivalent methods and has been found free, in these tests, from those harmful organism, or derived in direct line from material which is maintained under appropriate conditions and has been subjected, within the last three complete cycles of vegetation, at least once, to official testing for at least relevant harmful organisms using appropriate indicators for equivalent methods and has been found free, in these tests, from those harmful organism (bb) no symptoms of diseases caused by the relevant harmful organisms have been observed on plants at the place of production, or on susceptible plants in its immediate vicinity, since the beginning of the last complete cycles of vegetation  municipalities of Babtai and K _ edainiai (region of Kaunas)), P, SI (except the regions Gorenjska, Koro ska, Maribor and Notranjska, and the communes of Lendava and Ren ce-Vogrsko (south from the highway H4)), SK (except the county of Dunajsk (a) originate in the protected zones listed in the right hand column; or (b) have been subjected to an appropriate treatment to ensure freedom from Daktulosphaira vitifoliae (Fitch) according to a specification approved in accordance with the procedure referred to in Article 18(2).  Plants intended for planting, other than seeds, of the genera Abies Mill., Apium graveolens L., Argyranthemum spp., Asparagus officinalis L., Aster spp., Brassica spp., Castanea Mill., Cucumis spp., Dendranthema (DC.) Des Moul., Dianthus L. and hybrids, Exacum spp., Fragaria L., Gerbera Cass., Gypsophila L., all varieties of New Guinea hybrids of Impatiens L., Lactuca spp., Larix Mill., Leucanthemum L., Lupinus L., Pelargonium l'H erit. Ex Ait., Picea A. Dietr., Pinus L., Platanus L., Populus L., Prunus laurocerasus L., Prunus lusitanica L., Pseudotsuga Carr., Quercus L., Rubus L., Spinacia L., Tanacetum L., Tsuga Carr., Ulmus L., Verbena L. and other plants of herbaceous species, other than plants of the family Gramineae, intended for planting, and other than bulbs, corms, rhizomes, seeds and tubers.

II.
Plants, plant products and other objects which are potential carriers of harmful organisms of relevance for certain protected zones, and which must be accompanied by a plant passport valid for the appropriate zone when introduced into or moved within that zone 1.2 Plants intended for planting, other than seeds, of Beta vulgaris L., Platanus L., Populus L., Prunus L. and Quercus spp., other than Quercus suber and Ulmus L.

Part B
Plants, plant products and other objects originating in territories, other than those territories referred to in Part A I. Plants, plant products and other objects which are potential carriers of harmful organisms of relevance for the entire Community 1.

3.4.
Entry, establishment and spread in the EU

Entry
As of September 2019, there were no records of interception of non-EU phytoplasmas of the host plants in the Europhyt database.
All the phytoplasmas of the host plants categorised here can be transmitted by vegetative propagation material. Therefore, plants for planting of the host species are the most important entry pathway. Moreover, some of these phytoplasmas have additional natural hosts that also are vegetatively propagated, thus providing additional entry pathways. No pollen and seed transmissions have been reported for the phytoplasmas categorised here.
The legislation shows discrepancies between the nomenclature of phytoplasmas used in Annex IAI ("Non-European (. . .) virus-like organisms" and mycoplasms) and the one used in the present pest categorisation ('Candidatus Phytoplasma species'). This discrepancy may generate confusion at entry points.
The current legislation prohibits entry in the EU of plants for planting of Cydonia, Malus, Prunus, and Pyrus, from non-EU countries (Annex IIIAI 9), but introduction of dormant plants of these genera and of Fragaria is permitted from Mediterranean countries, Australia, New Zealand, Canada and the continental states of the USA (Annex IIIAI 18). This means that the entry pathway regarding plants for planting is only partially regulated for those phytoplasmas present in the above-mentioned countries (Mediterranean countries, Australia, New Zealand, Canada, the continental states of the USA and Switzerland). The legislation prohibits entry in the EU of commercial plants of Vitis with the exception of plants coming from Switzerland (Annex IIIAI 15).
In the case of Ribes and Rubus, the current legislation does not prohibit entry in the EU from non-EU countries of plants for planting of these genera. Although in Annex IVAI 19.2 an "official statement that no symptoms of diseases caused by the relevant harmful organisms" (e.g. non-European viruses and virus-like organisms) "have been observed on the plants at the place of production since the beginning of last complete cycle of vegetation" is requested. This measure is considered to have limited impact in preventing import of virus-infected plants. Indeed, symptoms in the infected plants might not be obvious.
Annex VA lists all the potential hosts which must be checked and accompanied by a plant passport for movement within EU MS. This measure may impair the spread of phytoplasmas on listed genera and/or plant species, but has no effect on the dissemination of phytoplasmas on unregulated host plants.
Annexes VBI 1, 2 and VBII 3, 4 determine that plants for plantings of several host species (Cydonia, Malus, Pyrus, Prunus, Rosa and Rubus) must be accompanied by a valid phytosanitary certificate in order to be introduced in the EU. Although this measure may prevent the introduction of the phytoplasmas explicitly mentioned in Annex IAI (Peach rosette, Peach X-disease and Peach yellows mycoplasmas and Strawberry witches' broom mycoplasm), it might not be effective for the phytoplasmas categorised here, which are not explicitly mentioned, and are only covered by the general definition of "Non-European (. . .) virus-like organisms".
All phytoplasmas categorised here can also be transmitted by vectors (Table 4). Vectors may thus act as entry pathway. Information on vector transmission is limited for some of the categorised phytoplasmas. The risk of introducing insects that have not yet been reported as competent vectors for these pathogens generates uncertainties on the entry pathways.

Pest categorisation of non-EU fruit phytoplasmas
The majority of the phytoplasmas categorised here ('Ca. P. aurantifolia'-related strains, 'Ca. P. australiense', 'Ca. P. fraxini', 'Ca. P. pyri'-related strain, 'Ca. P. trifolii', 'Ca. P. ziziphi') are transmitted by different species of Hemiptera insects which are not regulated, thus providing additional entry pathways. Bactericera cockerelli, vector of 'Ca. P. hispanicum', is listed in Annex IAI, which bans the entry of this vector. Table 12 provides an overview of the main potential entry pathways for the non-EU phytoplasmas categorised here. In summary, the current legislation: À regulates the plants for planting entry pathway for some of the phytoplasmas categorised here (Cydonia, Fragaria, Malus, Prunus, and Pyrus) if coming from specific countries (excluding Mediterranean countries, Australia, New Zealand, Canada, the continental states of the USA), À closes the Vitis plants pathway, À requires only visual inspection and an official declaration of absence of symptoms in the case of Ribes and Rubus.
For phytoplasmas with a wide host range, the plants for planting pathway is only partially regulated because these organisms may also enter in the EU through plants for planting of unregulated host species. The vector pathway is mainly not regulated. The analysis of entry pathways is affected by uncertainties due to the transmission biology, host range and geographical distribution of the non-EU phytoplasmas. Based on the above data and considerations, the entry pathways of the phytoplasmas categorised here are summarized as follows: • entry pathway involving plants for planting of the host plants: this pathway is closed for: 'Ca.
• entry pathway involving other hosts: this pathway is partially regulated for other hosts of: 'Ca.
• entry pathway involving infectious vectors: the pathway is open for 'Ca. P. aurantifolia'-related strains, 'Ca. P. australiense', 'Ca. P. fraxini', 'Ca. P. pyri'-related strain (PYLR), 'Ca. P. trifolii' and 'Ca. P. ziziphi' due to the unregulated status of their competent vectors. This pathway is possibly open for 'Ca. P. pruni'-related strain (NAGYIII) and Buckland Valley grapevine yellows phytoplasma because of the possible existence of unknown competent vectors. This pathway is regulated for 'Ca. P. hispanicum'. The host plants widely occur in the EU as commercial crops as well as wild plants. Details on the area of their production in individual EU MSs are provided in Table 13, as mean values for the period 2014-2018. Are the pests able to become established in the EU territory?
YES, the host plants of the phytoplasmas under categorisation are widespread in the EU and climatic conditions are not limiting for phytoplasma development as long as they are suitable for host growth. The absence of vectors for some phytoplasmas may affect their establishment (see Section 3.4.4.).

Climatic conditions affecting establishment
Phytoplasma multiplication rates may be influenced in opposite ways by temperature in vectors and plants (Galetto et al., 2011;Salar et al., 2013). Moreover, climate affects vector and host plant biological parameters (eg. synchronicity between egg hatching and inoculum availability). Therefore, foreseeing the influence of climate on the establishment of phytoplasma diseases is difficult, with a lack of data in the literature.
Nevertheless, it is expected that the phytoplasmas categorised here would be able to establish wherever their host plants are grown, unless the absence of vectors prevents their establishment. The host plants are widely cultivated in the EU. The Panel therefore considers that climatic conditions will not impair the ability of the phytoplasmas addressed here to establish in the EU. At the same time, symptom expression and severity may be affected by climatic conditions (e.g. temperature and light) (see Section 3.5).

Vectors and their distribution in the EU
The presence and the geographic distribution in the EU of competent vectors of the non-EU phytoplasmas categorised here are reported in Table 14. Competent vectors of 'Ca. P. aurantifolia'related strains, 'Ca. P. pyri'-related strain (peach yellow leafroll, PYLR), and 'Ca. P. trifolii' are described and known to be present in the EU, according to the EPPO GD, Fauna Europaea, Catalogue of Life and a WoS literature search. Competent vectors of 'Ca. P. australiense', 'Ca. P hispanicum', 'Ca. P. fraxini', and 'Ca. P. ziziphi' are described, but not reported to be present in the EU. No competent vectors are known for 'Ca. P. pruni'-related strain (NAGYIII) and Buckland Valley grapevine yellows phytoplasmas.
The possible unreported presence in the EU of known competent vectors of the phytoplasmas categorised here and the possibility that European phloem feeder insects may act as vectors of newly introduced phytoplasmas are sources of uncertainty in predicting the spread of non-EU phytoplasmas.

Impacts
Reported impacts caused by the phytoplasmas categorised here on the host plants are reported in Table 15, those on other hosts are listed in Appendix D. These phytoplasmas cause damage to the host plants in countries with environmental conditions similar to those present in the EU. They can be spread by infected plants for planting, therefore introduction of these pests in the EU is likely to impact the production of the host plants. Given that some of the pests also affect economically important crops beside the target host plants, their introduction in the EU may cause a broader damage. The susceptibility of Fragaria, Malus, and Vitis to 'Ca. P. aurantifolia'-related strains, of Prunus to 'Ca. P. australiense', and of Prunus and Vitis to'Ca. P. trifolii', as well as the actual presence of 'Ca. P. pyri'related strain (PYLR) in Pyrus are uncertain. Based on 2012-2015 surveys in some orchards in Faraghe (Iran), 12.5% of the studied apricot trees showed symptoms (Rasoulpour et al., 2019).
Disease severity observed in Australian pear orchard ranged from reduced bud viability to numerous dead limbs  The susceptibility of Fragaria (Streten et al., 2005c), Malus 'Ca. P. australiense' infection has been consistently reported on Fragaria, Rubus, and Vitis plants. Impacts on strawberry plants with little leaf and lethal yellows disease range from stunting to reduced leaf size, yellowing of younger leaves and occasional plant death. Impacts on grape range from irregular chlorosis or yellowing of leaves.
There is a single report of infection of Prunus (peach) in Bolivia  The susceptibility of Prunus needs to be confirmed (only one disease note without details on number of infected plants)  'Ca. P. fraxini' (reference strain) 'Ca. P. fraxini' phytoplasma strains have been reported in Vitis vinifera in Italy (Zambon et al., 2018), Chile (Gajardo et al., 2009) and Iran (Ghayeb Zamharir et al., 2017).
Among the host plants, there are two reports in Prunus spp in China  and Iran (Zunnoon- , and one in Fragaria in Argentina .
Infected Fragaria and Prunus show reduced fruit production, with lower quality -Would the pests' introduction have an economic or environmental impact on the EU territory?
YES, the introduction in the EU of the phytoplasmas categorised here would have an economic impact.  (Wolf, 2015) The susceptibility of other EU cvs is not known 'Ca. P. pyri'related strain (peach yellow leaf roll, PYLR) Peach is the known host of the phytoplasma and well present in the EU. PYLR-infected trees are severely and rapidly damaged. Yields of PYLR-affected peach trees are drastically reduced by premature fruit drop (Marcone et al., 2014). Apricot, European plum and sweet cherry have failed to become infected by grafting with PYLR infected scions (Marcone et al., 2014). Marianna 2624 rootstock is probably resistant to the PYLR-MLO (Uyemoto et al., 1992) The presence of PYLR in Pyrus has not been definitively assessed (Marcone et al., 2014) 'Ca. P. trifolii' (reference strain) Fragaria virginiana and F. chiloensis can be infected by 'Ca. P. trifolii', and this phytoplasma can presumably infect the commercial hybrid F. x ananassa (Jomantiene et al., 1998a). Impacts range from stunted to distorted and chlorotic leaves Susceptibility of Prunus and Vitis is uncertain (fewer than 5 plants from one single report for each species) (Zirak et al., 2010); (Zambon et al., 2018) 'Ca. P. ziziphi' (reference strain) A month following the first appearance of the virescence symptoms on sweet cherry plants (Prunus avium), the diseased trees became wilted and eventually died . The plants in a peach orchard in Northern Italy (identification made by RFLP and detected mixed infection with 'Ca. P. solani' and 'Ca. P. asteris') dried up and died in one to two weeks and sometimes sprouting of rootstock 'GF 677' was also observed; percentages of affected plants varied with cultivars and years: Cultivar 'Silver King' shows a stable symptom percentage of about 3%, cultivar 'Spring Crest' had 9% infected plants in 2001 and 2002, while in 2003 the percentage was 3%, cultivar 'May Crest' in 2001 had 3% infected plants but in the following years the disease presence was erratic (less than 1%) (Paltrinieri et al., 2006).
The visual survey incidence of the disease on apple trees in China was about 3% (Li et al., 2014b) -

Buckland valley grapevine yellows phytoplasma
Infection of grapevines with this phytoplasma causes weakening of the plants, lack of winter hardening of the new shoots, shoot tip death and bunches shriveling and falling, hampering grape production and reducing the longevity of the plants  There is a gap of knowledge on the vectors of this phytoplasma and on the susceptibility of grapevine cvs other than Chardonnay (pers. À explicitly list in the legislation the categorised pests as "phytoplasmas", replacing "virus-like organisms of the host plants" in Annex IAI; À extension of phytosanitary measures to specifically include hosts other than Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L. for the phytoplasmas categorised here, that may be introduced as plants for planting; À banning import of host plants for planting from the third countries where the phytoplasmas categorised here are reported; À extension of certification schemes, testing requirements and phytosanitary certificates to natural hosts other than Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L. for the phytoplasmas categorised here, that may be introduced as plants for planting.

Additional control measures
Potential additional control measures are listed in Table 16. Are there measures available to prevent the entry into, establishment within or spread of the pest within the EU such that the risk becomes mitigated?
YES, measures are already in place (see Section 3.3) and potential additional measures for further regulating the identified pathways to limit entry, establishment, spread or impacts are listed in Section 3.6.1.

Additional supporting measures
Potential additional supporting measures are listed in Table 17. • The asymptomatic phase of phytoplasma infection hampers visual detection; • The low concentration and uneven distribution in the woody hosts impairs reliable detection; • There is a wide host range for some phytoplasmas ('Ca. P. aurantifolia'-related strains, 'Ca. P. australiense', 'Ca. P. hispanicum', 'Ca. P. fraxini', 'Ca. P. trifolii', and 'Ca. P. ziziphi); • There is a lack of information on competent vectors for some agents (NAGYIII and Buckland valley grapevine yellows phytoplasmas). ISPM 5 defines a buffer zone as "an area surrounding or adjacent to an area officially delimited for phytosanitary purposes in order to minimize the probability of spread of the target pest into or out of the delimited area, and subject to phytosanitary or other control measures, if appropriate" (ISPM 5). The objectives for delimiting a buffer zone can be to prevent spread from the outbreak area and to maintain a pest free production place, site or area.
If the presence of the pathogen is restricted, a buffer zone (based on the flight capability of the vector) may help reduce the risk of contamination of infected entry material Spread All phytoplasmas categorised here 3.6.1.4. Biological or technical factors limiting the ability to prevent the presence of the pest on plants for planting Although Annex IV AI, at point 19.2, requires an"official statement that no symptoms of diseases caused by the relevant harmful organisms" (in our case non-European virus-like organisms, here non-EU phytoplasmas) "have been observed on the plants at the place of production since the beginning of the last complete cycle of vegetation", this measure is considered to have limited impact in preventing import of infected plants of host plants intended for planting. This is because symptoms in the infected woody plants often appear one year after inoculation by the insect vector.

Uncertainty
For each phytoplasma, the specific uncertainties are reported in the conclusion tables below. Uncertainties affecting all the phytoplasmas characterised here are: À The categorised pests are referred to in Council Directive 2000/29/EC as "virus-like organisms of the host plant" and not as "phytoplasmas", which could have led to inconsistencies in disease naming in official databases and legislation; À Disease names are based on symptomatology on the host where the disease was first described, but phytoplasma symptoms are not species-specific, therefore issues in the classification could have led to inconsistencies in disease naming in official databases and legislation; À Lack of biological information (competent vectors, host range, epidemiological details); À Volume of imported plants for planting of the unregulated hosts; À Distribution of the pests both in and outside the EU.

Conclusions
The Panel conclusions on this pest categorisation of non-EU phytoplasmas of Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L. are: À All the phytoplasmas categorised here meet all the criteria evaluated by EFSA to qualify as potential Union quarantine pests. À All the phytoplasmas categorised here do not meet the criteria evaluated by EFSA to qualify as potential RNQPs because they are non-EU phytoplasmas.
These conclusions are associated with uncertainty for phytoplasmas for which information on distribution, biology and epidemiology is limited. As a consequence, the categorisation presented here might change for some phytoplasmas as new data become available. However, the following general conclusions can be drawn: À The identity of all the phytoplasmas categorised here is established and diagnostic techniques are available. À All these phytoplasmas could enter the EU, especially by movement of infected plants for planting. Were this to happen, they could become established, spread and lead to impacts on the host plants, but often also on other hosts.
'Ca. P. aurantifolia'-related strains are a non-EU phytoplasma and thus do not meet all the EFSA criteria to qualify as a potential Union RNQP.
Aspects of assessment to focus on / scenarios to address in future if appropriate The main knowledge gaps are listed in this table.
Given the limited information available, the development of a full PRA would not allow solving the uncertainties of the present categorisation until more data become available The susceptibility of Prunus needs to be confirmed.
The host range is not fully known.
The potential vector ability of EU phloem feeder insects is uncertain.

4.3.
'Candidatus Phytoplasma fraxini' (reference strain) Given the limited information available, the development of a full PRA would not allow solving the uncertainties of the present categorisation until more data become available The presence of the phytoplasma in Canada and Japan is uncertain.
The host range is not fully known.
There is uncertainty on the vector ability of B. cockerelli.
The potential vector ability of EU phloem feeder insects is unclear Potential for consequences in the EU territory (Section 3.5) 'Candidatus Phytoplasma pruni'-related strain (North American grapevine yellows, NAGYIII) The geographical distribution and the host range are not fully known.
The potential vector ability of EU phloem feeder insects is unclear.
There is uncertainty about the presence of other unknown vectors Potential for consequences in the EU territory (Section 3.5) The introduction and spread of NAGYIII would have a negative impact on Vitis industry The presence of the NAGYIII on plants for planting would have a negative impact on their intended use. 'Candidatus Phytoplasma pyri'-related strain (peach yellow leaf roll, PYLR) The host range is not fully known.
The potential vector ability of EU phloem feeder insects is uncertain, as well as the existence of other unknown vectors. Potential for consequences in the EU territory (Section 3.5) The introduction and spread of PYLR would have a negative impact on Prunus industry. PYLR meets all the criteria evaluated by EFSA to qualify as a potential Union quarantine pest.
PYLR is a non-EU phytoplasma and thus does not meet all the EFSA criteria to qualify as a potential Union RNQP.
Aspects of assessment to focus on/ scenarios to address in future if appropriate The main knowledge gaps are listed in this table.
Given the limited information available, the development of a full PRA would not allow solving the uncertainties of the present categorisation until more data become available Pest categorisation of non-EU fruit phytoplasmas www.efsa.europa.eu/efsajournal 4.7.
'Candidatus Phytoplasma trifolii' (reference strain)  Unclassified Buckland valley grapevine yellows phytoplasma  The impact of the pest on the crop output and quality and on the environment in the occupied spatial units Introduction (of a pest) The entry of a pest resulting in its establishment (FAO, 2017) Measures Control (of a pest) is defined in ISPM 5 (FAO 2017) as "Suppression, containment or eradication of a pest population" (FAO, 1995 Any means that allows the entry or spread of a pest (FAO, 2017) Phytosanitary measures Any legislation, regulation or official procedure having the purpose to prevent the introduction or spread of quarantine pests, or to limit the economic impact of regulated non-quarantine pests (FAO, 2017) Protected zones (PZ) A Protected zone is an area recognised at EU level to be free from a harmful organism, which is established in one or more other parts of the Union. Quarantine pest A pest of potential economic importance to the area endangered thereby and not yet present there, or present but not widely distributed and being officially controlled (FAO, 2017) Regulated non-quarantine pest A non-quarantine pest whose presence in plants for planting affects the intended use of those plants with an economically unacceptable impact and which is therefore regulated within the territory of the importing contracting party (FAO, 2017) Risk reduction option (RRO) A measure acting on pest introduction and/or pest spread and/or the magnitude of the biological impact of the pest should the pest be present. An RRO may become a phytosanitary measure, action or procedure according to the decision of the risk manager Spread (of a pest) Expansion of the geographical distribution of a pest within an area (FAO, 2017) Abbreviations