Outbreak of Xylella fastidiosa subsp. pauca ST53 affecting wild and cultivated olive trees on the island of Mallorca, Spain

Phy opa hologia Medi e anea 63(3): 475-480, 2024 Fi enze Uni e si y P ess www. up ess.com/pm Phy opa hologia Medi e anea The in e na ional jou nal o he Medi e anean Phy opa hological Union ISSN 0031-9465 (p in ) | ISSN 1593-2095 (online) | DOI: 10.36253/phy o-15891 Sho No es Ci a ion: Mo alejo, E., Que glas, B., Mon esinos, M., Ad o e , F., Olmo, D., Nie o, A., Ped osa, A., López, M., Juan, A., Ma co-Noales, E., Na a o- He e o, I., Ba bé, S., Velasco-Amo, M.P., Oli a es-Ga cía, C., & Landa, B.B. (2024). Ou b eak o Xylella as idiosa subsp. pauca ST53 a ec ing wild and cul i a ed oli e ees on he island o Mallo ca, Spain. Phy opa hologia Med- i e anea 63(3): 475-480. doi: 10.36253/ phy o-15891 Accep ed: Decembe 27, 2024 Published: Decembe 30, 2024 © 2024 Au ho (s). This is an open access, pee - e iewed a icle published by Fi enze Uni e si y P ess (h ps://www. up ess.com) and dis ibu ed, excep whe e o he wise no ed, unde he e ms o he CC BY 4.0 License o con en and CC0 1.0 Uni e sal o me ada a. Da a A ailabili y S a emen : All el- e an da a a e wi hin he pape and i s Suppo ing In o ma ion iles. Compe ing In e es s: The Au ho (s) decla e(s) no con lic o in e es . Edi o : Anna Ma ia D’Onghia, CIHEAM/Medi e anean Ag onomic Ins i u e o Ba i, I aly. ORCID: EM: 0000-0003-4927-9367 DO: 0000-0002-8542-9089 AN: 0000-0001-5484-7088 EM-N: 0000-0001-7973-0345 SB: 0000-0002-2929-6410 MPV-A: 0000-0001-7176-0435 CO-G: 0009-0004-6511-2633 BBL: 0000-0002-9511-3731 Ou b eak o Xylella as idiosa subsp. pauca ST53 a ec ing wild and cul i a ed oli e ees on he island o Mallo ca, Spain Edua do MORALEJO1,*, Bà ba a QUETGLAS1, Ma ina MONTESINOS1, F ancisco ADROVER2, Diego OLMO2, Alicia NIETO2, Ana PEDROSA2, Ma a LÓPEZ3, And eu JUAN3, Es e MARCO-NOALES4, Inmaculada NAVARRO-HERRERO4, Sil ia BARBÉ4, Ma ía Pila VELASCO-AMO5, Concepción OLIVARES-GARCÍA5, Blanca B. LANDA5 1 T agsa, Emp esa de T ans o mación Ag a ia, Delegación de Balea es, 07005 Palma de Mallo ca, Spain 2 Ins i u de Rece ca i Fo mació Ag oalimen à ia i Pesque a de les Illes Balea s, 07009 Palma de Mallo ca, Spain 3 Se icio de Ag icul u a, Conselle ia de Medi Ambien , Ag icul u a i Pesca, 07006 Palma de Mallo ca, Spain 4 Cen o de P o ección Vege al y Bio ecnología, Ins i u o Valenciano de In es igaciones Ag a ias (IVIA), CV-315 km 10.7, 46113 Moncada, Spain 5 Ins i u e o Sus ainable Ag icul u e, Consejo Supe io de In es igaciones Cien í icas (IAS-CSIC), 14004 Có doba, Spain *Co esponding au ho : E-mail: emo alejo @gmail.com Summa y. The Balea ic Islands ha e eme ged as a ho spo o he in asi e plan pa h- ogen Xylella as idiosa (X ). Since 2016, he X subsp. as idiosa and mul iplex ha e been de ec ed causing almond lea sco ch and Pie ce’s disease on he island o Mal- lo ca, Spain, and a new sequence ype (ST), ST80, o subsp. pauca is in ec ing wild and cul i a ed oli e ees on he island o Ibiza. In addi ion, X subsp. mul iplex ST81 is widesp ead in sc ublands, and causes mild, sub-le hal dieback o wild oli e ees in Meno ca and Mallo ca. A new ou b eak is he e epo ed o he X subsp. pauca in he municipali y o Sencelles in he cen e o Mallo ca island. In ea ly 2024, dying pa ches we e obse ed in wild oli e ees (Olea eu opaea a . eu opaea subsp. syl es is). Sam- ples om hese ees we e X -posi i e in di e en qPCR es s, and he pa hogen was subsequen ly iden i ied as belonging o ST53 o subsp. pauca, he same gene ic a ian esponsible o oli e quick decline synd ome in Apulia, I aly. Mo e han 184 plan s o eigh hos s ha e es ed posi i e o subsp. pauca wi hin a dema ca ion zone o app ox. 1 km adius. The iden i ied hos species include 124 wild oli e ees, 40 cul i a ed oli e ees, nine Rhamnus ala e nus, six Ne ium oleande , wo La andula angus i olia, one Lau us nobilis, one La andula den a a and one Polygala my i olia. O pa icula con- ce n is de ec ion o co-in ec ions by X subsp. subsp. pauca and mul iplex on plan s om na u al se ings (wild oli es, L. den a a and R. ala e nus), posing po en ial isk o gene ic ecombina ions. In ensi e su eys a e being ca ied ou o con ain he sp ead o ST53, and in ec ed plan s ha e been des oyed in he dema ca ed zone. Keywo ds. Gene ic di e si y, oli e quick decline synd ome, in asi e pa hogens, Mul i- Locus Sequence Typing (MLST), disease ou b eak. 476 Edua do Mo alejo e alii INTRODUCTION The ec o -bo ne plan pa hogenic bac e ium Xylella as idiosa (X ) has ecen ly eme ged as a h ea o ag i- cul u e in sou he n Eu ope. Following i s i s de ec- ion in Apulia (I aly) in 2013, associa ed wi h he apid decline synd ome o oli e ees (Sapona i e al., 2013), he Eu opean Union (EU) ook decisi e ac ion wi h manda o y su eys o con ain possible sp ead o he pa hogen wi hin he EU (Reg. EU 2016/2031). The h ee main subspecies o X ha e since hen been de ec ed, and hei es ablishmen con i med, in se e al sou he n Eu o- pean coun ies, including he island o Co sica and he Balea ic Islands (Sapona i e al., 2013; Denance e al., 2017; Landa e al., 2020; Olmo e al., 2017; Ca alho-Luis e al., 2022). The Balea ic Islands ha e become a ho spo o es ablishmen o gene ic di e si y o his pa hogen, which o igina ed om No h, Cen al and Sou h Ame - ica. Since 2016, he h ee main subspecies o X , as idi- osa, mul iplex, and pauca, ha e been de ec ed, along wi h unique geno ypes such as he sequence ype (ST) 81 ( om subsp. mul iplex) on he islands o Mallo ca and Meno ca, and he ST80 ( om subsp. pauca) on he island o Ibiza (Olmo e al., 2021). Subspecies as idiosa, and in pa icula ST1, is only known on Mallo ca island, whe e i caused se e e ou b eaks o Pie ce’s disease o g ape ines and lea sco ch o almond (Mo alejo e al., 2019; Mo alejo e al., 2020). Some e idences sugges ha he geno ypes esponsible o Pie ce’s disease (ST1) and almond lea sco ch (ST1 and ST81) in Mallo ca we e likely a single in oduc ion ia in ec ed plan ma e ial om Cali o nia, in app ox. 1993 (Mo alejo e al., 2020; Velasco-Amo e al., 2022). METHODS Ea ly in 2024, plan heal h inspec o s in he Balea ic Islands no iced an unusually se e e decline in a s and o wild oli e ees (Olea eu opaea subsp. eu opaea a . syl- es is) (Figu e 1). Samples om di e en ee b anches we e collec ed, we e pooled, and hen sen o he Balea - ic Islands O icial Plan Heal h Labo a o y (LOSVIB) o analyses. Sample ex ac s we e p epa ed by homogeniz- ing 0.5 g o lea pe ioles in an ex ac ion bag (BIORE- BA®) and g inding hem in 5 mL (1:10 weigh : olume) o phosha e bu e ed saline (PBS) using a semi-au oma ed homogenize (Homex 7; BIOREBA®). To al DNA ex ac ion was ca ied ou using 200 µL o each sample ex ac , and he EZNA HP Plan Mini ki (Omega-Bio ek), which employs a CTAB-based me hod, as desc ibed in he EPPO p o ocol (EPPO, 2023). The DNA ex ac s we e subsequen ly es ed o he p esence o X by quan i a i e eal- ime PCR (qPCR), acco ding o Ha pe e al. (2010). Aliquo s o DNA om he X posi i e samples om he wild oli e ee we e sen o he Ins i u e o Sus ainable Ag icul u e (IAS-CSIC), Có do- ba, Spain, o ST de e mina ion, which was ca ied ou by con en ional Mul iLocus Sequence Typing (MLST) analysis (Yuan e al., 2010). A nes ed-MLST (Cesb on e al., 2020) analysis was used when no enough ampli ica- ion p oduc o di ec sequencing was ob ained when using he con en ional MLST app oach. RESULTS AND DISCUSSION MLST analyses iden i ied he p esence o alleles leuA-7, pe C-6, malF-16, cysG-24, holC-10, nuoL-16 and gl T-14 associa ed wi h X subsp. pauca ST53 in he a ec ed wild oli e ee. Addi ionally, he qPCR p o o- col o Dupas e al. (2019) alida ed he diagnosis as X subsp. pauca. A e con i ming he diagnosis and ecognizing he po en ial de as a ing impac i his subspecies we e o sp ead, he phy osani a y au ho i y o he Balea ic Go - e nmen implemen ed a a ge ed ac ion plan. This includ- ed inc easing su eys and in ensi e sampling in he a ea o he new ou b eak. Addi ional measu es we e also es ab- lished o hose al eady in place om he cu en Regula- ion (EU) 2016/2031. These included analyses o de e - mine he subspecies p esen in all he X -posi i e samples, in collabo a ion wi h he Valencian Ins i u e o Ag icul- u al Resea ch (IVIA), using he qPCR p o ocols de el- oped by Dupas e al. (2019) and Hodge s e al. (2021). Since he i s de ec ion in Janua y 2024 and up o Decembe 2024, a o al o 1,328 samples om wi hin he disease ou b eak a ea ha e been analyzed. This has shown ha 184 plan s we e in ec ed by X subsp. pauca, Figu e 1. A g oup o wild oli e ees wi h se e e symp oms o quick decline synd ome. This pho og aph was aken a ocus 0 nea he municipali y o Sencelles, Mallo ca Island, Spain, whe e Xylella as id- iosa subsp. pauca ST53 was ound o he i s ime in Janua y 2024. 477 Xylella as idiosa subsp. pauca ST53 on oli e ees on he island o Mallo ca, Spain and eigh di e en hos s es ed posi i e o X subsp. pauca, om wi hin a dema ca ed zone (app ox. 1 km adius) a ound he i s disease de ec ion. The in ec ed hos species include 124 wild oli e ees, 40 cul i a ed oli e ees, nine Rhamnus ala e nus plan s, six o Ne i- um oleande , wo o La andula angus i olia, and one each o Lau us nobilis, La andula den a a and Polygala my i olia (Table 1). Despi e he nume ous cases o X subsp. pauca in ec ions de ec ed (Table 1), all cases analyzed by MLST, up o p esen , and wi hin he ini ial ocus o he ou b eak belong o he ST53 (Figu e 2). Howe e , gi en ha X subsp. as idiosa and X subsp. mul iplex we e al eady es ablished in he a ea, i was no su p ising o ind some mixed in ec ions by hese wo subspecies in some indi idual plan s. Ele en pe cen o X -posi i e samples we e o mixed in ec ions, wi h 26 cases o sub- spp. pauca/mul iplex, one o subspp. pauca/ as idiosa and wo o subspp. mul iplex/ as idiosa (Table 1). The mul iple in ec ions p obably may inc ease isk o gene ic ecombina ions in he pa hogen and he o ma ion o new sequence ypes wi h di e en i ulence and/o hos anges (Po nis e al., 2019). Al hough i is p ema u e o d aw de ini i e conclu- sions, he ST53 s ain iden i ied in Mallo ca exhibi ed a le el o i ulence compa able o ha associa ed wi h he oli e quick decline synd ome obse ed in Apulia, based on he obse ed highe se e i y o symp oms in bo h wild and cul i a ed oli e ees, compa ed o hose induced by X subsp. mul iplex ST81. E o s a e cu en - ly unde way (Decembe 2024) o isola e he bac e ium, o enable comple e genome sequencing and ca y ou compa a i e analysis o he pa hogen’s compe e genome wi h hose o ST53 s ains om Apulia. A new p ocedu e has been es ablished based on a - ge ed sequence cap u e en ichmen o X in combina ion wi h high- h oughpu sequencing using he Illumina pla o m. This is being implemen ed o p o ide addi ion- al genomic in o ma ion (Velasco-Amo e al., 2021). Ini- ial esul s based on app ox. 126 genes ha e shown close gene ic ela edness o he ST53 s ain in ec ing he wild oli e in Mallo ca wi h all o he ST53 s ains isola ed om Cos a Rica and I aly (M.P. Velasco-Amo and B.B. Landa; unpublished esul s). A complica ing ac o in con ainmen o ST53 s ains is he la ge popula ion o wild oli e ees h oughou Mallo ca, which may acili a e ansmission o he pa hogen o cul i a ed oli e ees. Tigh wild oli e ee popula ion ne wo ks can be dense ese oi s o X , and p o ide e ec i e dispe sal pa hways o comme cial oli e plan a ions. I is also unclea whe he p e ious in ec ions o wild oli e ees by ST81 o X . subsp. mul- iplex will ha e in luence subsequen in ec ions by ST53. I is es ima ed ha mo e han 50% o wild oli e ees a e in ec ed wi h ST81 (Olmo e al., 2021), so any in e ac ion be ween ST53 and ST81 in co-in ec ions, ega dless o i s in ensi y, could ha e impo an epidemiological implica- ions (Jege and B aga d, 2019). Epidemiological models ha e shown ha he mos e ec i e s a egy o p e en ing sp ead o X is o decel- Table 1. Hos plan species, numbe s o plan s analysed, and numbe s in ec ed by di e en subspecies Xylella as idiosa (X ), in single o in mixed in ec ions, in he ou b eak o subspecies pauca a ound he locali y o Sencelles (Mallo ca island, Spain) Hos s To al analyzed X posi i ea% Xylella as idiosa subspeciesb pauca mu liplex as idiosa pauca/ mul iplex pauca/ as idiosa mul iplex/ as idiosa Lau us nobilis 6 2 33 1 1     La andula angus i olia 8 8 100 2 6     La andula den a a 6 5 83  4  1   Ne ium oleande 22 6 27 6      Olea eu opaea a . eu opaea subsp. eu opaea 57 41 72 37 1  3   Olea eu opaea a . eu opaea subsp. syl es is 929 155 17 106 29 2 18   Polygala my i olia 1 1 100    1   P unus dulcis 8 1 13  1     Rhamnus ala e nus 190 26 14 5 8 6 3 1 2 Sal ia osma inus 23 19 83 19     Thi een o he plan species 78 -        To als 1328 264 19.88 157 69 8 26 1 2 a Diagnoses pe o med using he qPCR es o Ha pe e al. (2010). b Subspecies assigna ion based on he qPCR es s as desc ibed by Dupas e al. (2019) and Hodge s e al. (2021). 478 Edua do Mo alejo e alii e a e ansmission chains (Whi e e al., 2020; Giménez- Rome o e al., 2023). Acco dingly, in Mallo ca, he deci- sion has been aken o e adica e in ec ed plan s whe e e easible. Al hough his is a di icul and a guably una - ainable objec i e, he a emp can ha e signi ican long- e m impac s. I is an icipa ed ha popula ions o he p incipal insec ec o o X , Philaenus spuma ius, will p og essi ely decline in Mallo ca in esponse o ising empe a u es associa ed wi h clima e change (Giménez- Rome o e al., 2024). Unsui able condi ions o he ec- o may become signi ican in a pe iod o app ox. 15 o 20 yea s. Clima e change could be bene icial o con ain- ing he ou b eak, p o ided ha exponen ial in ec ion a es can be educed. App op ia e implemen a ion o hese ac ions will alle ia e he economic consequences o X o he Mallo can oli e p oduc ion sec o in he medium e m. FUNDING This esea ch was unded by P ojec s E-RTA2017- 00004-C06-02 om AEI-INIA Spain, and BeXyl (Beyond Xylella, In eg a ed Managemen S a egies o Mi iga ing Xylella as idiosa impac in Eu ope; G an ID No. 101060593), om he Eu opean Union Ho izon Ac ion P og amme ‘Food, Bioeconomy Na u al Resou c- es, Ag icul u e and En i onmen ’. ACKNOWLEDGEMENTS The au ho s o his pape hank he owne s o he s ud- ied o cha ds, and especially Ani a an de We , who has always been kind and willing o collabo a e wi h he inspec- o s and echnicians o he Mallo can plan heal h se ices. Figu e 2. Map o he ou b eak ocus o Xylella as idiosa subsp.paucaST53 in he municipali y o Sencelles, nea he cen e o Mallo ca Island, Spain. The X pauca-posi i e plan s we e ound wi hin a adius o < 1 km. Bo h squa es wi h ampli ied iews show X -posi i e plan s wi hin he na u al ege a ion, o med mainly by wild oli e ees nea a ms whe e o he X subsp. pauca ST53 posi i e plan s we e ound wi hin oli e o cha ds. The posi i e samples o X subsp. pauca ST53 plan s we e mixed wi h o he in ec ed wi h subsp. mul iplex, and o a lesse ex en wi h subsp. as idiosa. In a ew cases, co-in ec ions by X subsp. pauca ST53 and subsp. mul iplex we e ound in he same plan (Table 1). 479 Xylella as idiosa subsp. pauca ST53 on oli e ees on he island o Mallo ca, Spain LITERATURE CITED Ca alho-Luis C., Rod igues J.M., Ma ins L.M., 2022. Dispe sion o he bac e ium Xylella as idiosa in Po - ugal. Jou nal o Ag icul u al Science and Technol- ogy. A 12(1): 35–41. h ps://doi.o g/10.17265/2161- 6256/2022.01.005 Cesb on S., Dupas E., Beau epè e Q., B iand M., Mon es- Bo ego M., Velasco-Amo MdP., Landa B.B., Jacques M-A., 2020. De elopmen o a nes ed-Mul iLocus Sequence Typing app oach o a highly sensi i e and speci ic iden i ica ion o Xylella as idiosa subspecies di ec ly om plan samples.Ag onomy 10(8): 1099. h ps://doi.o g/10.3390/ag onomy10081099 Denancé N., Legend e B., B iand M., Oli ie V., de Bois- seson C., … Jacques M.A., 2017. Se e al subspecies and sequence ypes a e associa ed wi h he eme - gence o Xylella Fas idiosa in na u al se ings in F ance. Plan Pa hology 66(7): 1054–1064. h ps://doi. o g/10.1111/ppa.12695 Dupas E., B iand M., Jacques M.A., Cesb on S., 2019. No el e aplex quan i a i e PCR assays o simul a- neous de ec ion and iden i ica ion o Xylella as idiosa subspecies in plan issues. F on ie s in Plan Science 10: 1732. h ps://doi.o g/10.3389/ pls.2019.01732 EPPO (Eu opean Plan P o ec ion O ganiza ion), 2023. PM 7/24 (5) Xylella as idiosa. EPPO/OEPP Bulle in 53(2): 205–276. h ps://doi.o g/10.1111/epp.12923 Eu opean Commission, 2016. Regula ion (EU) 2016/2031 o he Eu opean Pa liamen o he Council o 26 Oc obe 2016 on p o ec i e measu es agains pes s o plan s, amending Regula ions (EU) No 228/2013, (EU) No 652/2014 and (EU) No 1143/2014 o he Eu opean Pa liamen and o he Council and epeal- ing Council Di ec i es 69/464/EEC, 74/647/EEC, 93/85/EEC, 98/57/EC, 2000/29/EC, 2006/91/EC and 2007/33/EC. O icial Jou nal o he Eu opean Union L 317: 4–104. h p://da a.eu opa.eu/eli/ eg/2016/2031/ oj/eng Giménez-Rome o À., Mo alejo E., Ma ías M.A., 2023. A compa men al model o Xylella as idiosa diseases wi h explici ec o seasonal dynamics. Phy opa hol- ogy 113(9): 1686–1696. h ps://doi.o g/10.1094/PHY- TO-11-22-0428-V Giménez-Rome o À., I u bide M., Mo alejo E., 2024. Global wa ming signi ican ly inc eases he isk o Pie ce’s disease epidemics in Eu opean ineya ds. Scien i ic Repo s 14: 9648. h ps://doi.o g/10.1038/ s41598-024-59947-y Ha pe S.J., Wa d L.I., Clo e G.R.G., 2010. De elopmen o LAMP and eal-Time PCR me hods o he apid de ec ion o Xylella as idiosa o qua an ine and ield Applica ions. Phy opa hology 100(12): 1282–1288. h ps://doi.o g/10.1094/PHYTO-06-10-0168 Hodge s J., Glo e R., Cole J., Hall J., Boonham N., 2021. Genomics in o med design o a sui e o eal‐ ime PCR assays o he speci ic de ec ion o each Xylella as idiosa subspecies. Jou nal o Applied Mic obiology 131(2): 855–872. h ps://doi.o g/10.1111/jam.14903 Jege M., B aga d C., 2019. The epidemiology o Xylella as idiosa; A pe spec i e on cu en knowledge and amewo k o in es iga e plan hos – ec o –pa hogen in e ac ions. Phy opa hology 109: 200–209. h ps:// doi.o g/10.1094/PHYTO-07-18-0239-FI Landa B.B., Cas illo A.I., Giampe uzzi A., Kahn A., Román-Écija M, …Almeida R-P.P., 2020. Eme gence o a plan pa hogen in Eu ope associa ed wi h mul i- ple in e con inen al in oduc ions. Applied and En i- onmen al Mic obiology 86: e01521-19. h ps://doi. o g/10.1128/AEM.01521-19 Mo alejo E., Bo às D., Gomila M., Mon esinos M., Ad o e F., … Landa B. B., 2019. Insigh s in o he epidemiology o Pie ce’s disease in ineya ds o Mal- lo ca, Spain. Plan Pa holology 68: 1458–1471. h ps:// doi.o g/10.1111/ppa.13076 Mo alejo E., Gomila M., Mon esinos M., Bo às D., Pas- cual A., … Olmo D., 2020. Phylogene ic in e ence enables econs uc ion o a long-o e looked ou b eak o almond lea sco ch disease (Xylella as idiosa) in Eu ope. Communica ions Biology 3: 560. h ps://doi. o g/10.1038/s42003-020-01284-7 Olmo D., Nie o A., Ad o e F., U bano A., Beidas O., … Landa B.B., 2017. Fi s de ec ion o Xylella as idiosa in ec ing che y (P unus a ium) and Polygala my i- olia plan s, in Mallo ca Island, Spain. Plan Disease 101(10): 1820–1820. h ps://doi.o g/10.1094/PDIS- 04-17-0590-PDN Olmo D., Nie o A., Bo às D., Mon esinos M., Ad o e F., … Landa B.B., 2021. Landscape epidemiology o Xylella as idiosa in he Balea ic Islands. Ag on- omy 11(3): 473. h ps://doi.o g/10.3390/ag ono- my11030473 Po nis N., Kandel P.P., Me a M.V., Re chless A.C., Pa k- e J.K., … De La Fuen e L., 2019. Pa e ns o in e - and in asubspeci ic homologous ecombina ion in o m eco-e olu iona y dynamics o Xylella as idi- osa. The ISME jou nal 13(9): 2319–2333. h ps://doi. o g/10.1038/s41396-019-0423-y Sapona i M., Boscia D., Nig o F., Ma elli G.P., 2013. Iden i ica ion o DNA sequences ela ed o Xylella as idiosa in oleande , almond and oli e ees exhibi - ing lea sco ch symp oms in Apulia (sou he n I aly). Jou nal o Plan Pa hology 95(3): 668. h ps://doi. o g/10.4454/JPP.V95I3.035 480 Edua do Mo alejo e alii Velasco-Amo M.P., A ias-Gi aldo L.F., Ma ín Sanz J.A., Fe nández So ia V.M., Impe ial J., Landa B.B., 2021. Assessing genome-wide di e si y in Xylella as idiosa h ough a ge en ichmen ia hyb idiza ion-based cap u e om na u al hos plan and insec samples. In: Book o Abs ac s o he 3 d Eu opean con e ence on Xylella as idiosa and XF-ACTORS inal mee ing, 61, Ap il 26-30, On-line e en , 2021. Velasco-Amo M.P., A ias-Gi aldo L.F., Oli a es-Ga cía C., Denancé N., Jacques M.-A., Landa, B.B., 2022. Use o aC Gene o ype he incidence and dis i- bu ion o pXFAS_5235 plasmid-bea ing s ains o Xylella as idiosa subsp. as idiosa ST1 in Spain. Plan s (Basel) 11(12):1562. h ps://www.mdpi.com/2223- 7747/11/12/1562. E a um in: Plan s (Basel). 2024. 13(2): 200. h ps://doi.o g/10.3390/plan s13020200. Whi e S.M., Na as‐Co és J.A., Bullock J.M., Boscia D., Chapman D.S., 2020. Es ima ing he epidemiology o eme ging Xylella as idiosa ou b eaks in oli es. Plan Pa hology 69(8): 1403–1413. h ps://doi.o g/10.1111/ ppa.13238 Yuan X., Mo ano L., B omley R., Sp ing-Pea son S., S ou hame R., Nunney, L., 2010. Mul ilocus Sequence Typing o Xylella as idiosa causing Pie ce’s disease and oleande lea sco ch in he Uni ed S a es. Phy opa hology 100(6): 601–611. h ps://doi. o g/10.1094/PHYTO-100-6-0601