The occurrence of nematodes of the family Longidoridae was investigated in soil samples collected from cultivated and wild olives and grapevines in Crete. The first-stage juveniles of Xiphinema cretense and X. herakliense are described for the first time. The species X. israeliae, X. cretense, X. herakliense and Longidorus pseudoelongatus, previously recorded exclusively from olives in Crete, are herein reported in the rhizosphere of grapevines. Also L. iranicus is reported for the first time in cultivated olive, while X. italiae and L. closelongatus are reported for the first time in wild olive in Crete. Data on the occurrence of phytoparasitic nematode species in cultivated olives, wild olives and grapevines are updated with those previously published.This  research  was  supported  by  grant  KBBE 219262  ArimNET-ERANET  FP7  2012-2015  Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the 
Mediterranean basin’ from Hellenic Agricultural  Organization-DEMETER  and  Instituto  Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), grant AGR-136 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, “Una manera de hacer Europa”. I.G. Birmpilis and E. Nasiou  were  employed  by  the  ARIMNET-PESTOL-IVE  project.Peer reviewe

Archidona-Yuste, Antonio

Birmpilis, I. G.

Cantalapiedra-Navarrete, C.

Castillo, Pablo

Nasiou, Eleni

Palomares Rius, Juan E.

Tzortzakakis, E. A.

Hellenic Plant Protection Journal

Digital.CSIC

© Benaki Phytopathological InstituteHellenic Plant Protection Journal 9: 73-77, 2016DOI 10.1515/hppj-2016-00091 Department of Viticulture, Vegetable Crops, Flori-culture and Plant Protection, Institute of Olive Tree, Subtropical Crops and Viticulture, N.AG.RE.F., Hellen-ic Agricultural Organization-DEMETER, PO BOX 2228, 71003 Heraklion, Crete, Greece2 Institute for Sustainable Agriculture (IAS), Spanish National Research Council (CSIC), Avenida Menéndez Pidal s/n, 14004 Córdoba, Campus de Excelencia In-ternacional Agroalimentario, ceiA3, Spain* Corresponding author: etzortza@her.forthnet.grThe presence of Xiphinema and Longido-rus nematodes on cultivated and wild olive and grapevines in Crete was investigated by Tzortzakakis et al. (2014, 2015). The soil sam-ples were collected from:a) 101 olive orchards in Heraklion and La-sithi provinces,b) 22 individual wild olive trees in Heraklion province andc) 30 vineyards in Heraklion province and assigned to the Nematology Laboratory (affi  liation of the 1st author) by farmers for nematode diagnosis. Five known Xiphinema species (viz. X. in-dex, X. israeliae, X. italiae, X. pachtaicum and X. simile), fi ve known Longidorus species (viz. L. closelongatus, L. cretensis, L. iranicus (syn-onym of L. moesicus), L. orientalis, and L. pseudoelongatus) and two newly described (for the fi rst time) Xiphinema species (viz. X. cretense and X. herakliense) were found.The current work presents supplementa-ry data on the occurrence of Xiphinema and Longidorus nematodes on olive trees and grapevines in Crete based on additional soil samples, which were collected from:a) the topotype locality (cultivated olive) of SHORT COMMUNICATIONDescription of the fi rst-stage juveniles of Xiphinema cretense and X. herakliense - Distribution of Xiphinema and Longidorus species in olive orchards and grapevines in Crete, GreeceE.A. Tzortzakakis1*, A. Archidona-Yuste2, C. Cantalapiedra-Navarrete2, I.G. Birmpilis1, E. Nasiou1, J.E. Palomares-Rius2 and P. Castillo2Summary   The occurrence of nematodes of the family Longidoridae was investigated in soil sam-ples collected from cultivated and wild olives and grapevines in Crete. The fi rst-stage juveniles of Xi-phinema cretense and X. herakliense are described for the fi rst time. The species X. israeliae, X. cretense, X. herakliense and Longidorus pseudoelongatus, previously recorded exclusively from olives in Crete, are herein reported in the rhizosphere of grapevines. Also L. iranicus is reported for the fi rst time in culti-vated olive, while X. italiae and L. closelongatus are reported for the fi rst time in wild olive in Crete. Data on the occurrence of phytoparasitic nematode species in cultivated olives, wild olives and grapevines are updated with those previously published.Additional keywords:  Longidorus closelongatus, L. cretensis, L. iranicus, L. pseudoelongatus, X. israeliae Olive tree and grapevine are the most im-portant crops on the island of Crete, occu-pying 177,000 and 25,500 hectares, respec-tively. These represent about 22% for olive trees and 20% for grapevines of the total corresponding cultivated areas in Greece. In addition, wild olive trees are also located in some south coastal areas of the island. Dagger and needle nematodes of the genera Xiphinema and Longidorus, respec-tively, include a number of large plant ec-toparasitic nematode species with long life cycles.  They cause damage to a wide range of fruit and vegetable crops as well as wild plants by their direct feeding on root cells and transmission of nepoviruses (Decrae-mer and Robbins, 2007; Taylor and Brown, 1997). Brought to you by | CSIC - Unidad Coordinacion BibliotecasAuthenticatedDownload Date | 11/27/17 1:38 PM© Benaki Phytopathological InstituteTzortzakakis et al.74X. cretense (Tzortzakakis et al., 2014) and an olive orchard where X. herakliense had been found (Tzortzakakis et al., 2015), aiming at the detection of fi rst-stage ju-veniles (J1) which could not be detected after repeated samplings at the original description of these species. This life-stage may have a practical signifi cance when distinguishing species closely re-lated (Hunt, 1995),b) 45 olive orchards in Heraklion, Rethym-no and Lasithi provinces and 14 individ-ual wild olive trees in Heraklion province, diff erent to the ones in Tzortzakakis et al. (2014, 2015) study andc) 100 vineyards in Heraklion and Chania provinces and assigned to the Nematolo-gy Laboratory (affi  liation of the 1st author) by farmers for nematode diagnosis.Soil sampling (for olive trees), nematode extraction, fi xing and identifi cation proto-cols were carried out as by Tzortzakakis et al. (2014, 2015). Description of J1s of Xiphinema cretense and Xiphinema heraklienseIn both species, J1s were positively identifi ed by the position of the replacement odonto-style, which lies mostly within the odonto-phore, with the anterior tip near the base of the functional odontostyle (Figures 1, 2) (Hunt, 1995).Xiphinema cretense (J1)Measurements (n = 5): L = 1103 ± 24.4 (1083-1128) μm; a = 51.7 ± 1.8 (50.4-53.7); b = 4.3 ± 0.3 (4.0-4.6); c = 22.9 ± 1.3 (22.1-24.4); c’ = 3.2 ± 0.1 (3.1-3.3); odontostyle length = 55.3 ± 0.5 (56.0-57.0) μm; replacement odontostyle length = 73.2 ± 1.2 (73.0-75.0) μm; odonto-phore length = 41.9 ± 1.0 (41.0-43.0) μm; lip region width = 7.7 ± 0.5 (7.5-8.5) μm; oral aperture-guiding ring distance = 38.7 ± 4.0 (34.0-41.5) μm; tail length = 48.6 ± 2.5 (46.0-51.0) μm; hyaline region at tail tip = 11.5 ± 0.6 (11.0-12.0) μm. Description: Morphologically similar to adult specimens described by Tzortzaka-kis et al. (2014), apart from developed re-productive system, shorter body length, tail shape and presence of replacement odonto-style (Figure 1A). Anterior part characterized by position of replacement odontostyle just posterior to functional odontostyle, its tip touching or very close to base of function-al odontostyle (Figure 1B). Bluntly conoid tail shape well curved dorso-ventrally with a slight dorsal depression at hyaline region level (Figures 1C-E). Xiphinema herakliense (J1)Measurements (n = 1): L = 1183 μm; a = 42.3; b = 5.0; c = 27.5; c’ = 3.0; odontostyle length = 63.0 μm; replacement odontostyle length = 81.5 μm; odontophore length = 39.0 μm; lip region width = 8.0 μm; oral aperture-guiding ring distance = 43.0 μm; tail length = 43.0 μm; hyaline region at tail tip = 20.0 μm.Description: Only one specimen was found. General morphology agrees closely to adults specimens described by Tzortzakakis et al. (2015), except for its developed repro-ductive system, shorter body length in open C-shape, tail shape and presence of replace-ment odontostyle (Figure 2A). Anterior part characterized by position of replacement odontostyle into odontophore, just posteri-or to base of functional odontostyle (Figure 2B). Bluntly conoid tail shape well curved dorso-ventrally with a strong dorsal depres-sion at hyaline region level (Figure 2C). Distribution of Xiphinema and Longido-rus species in cultivated olive, oleaster and grapevineIn the soil samples of 45 olive orchards, three Xiphinema (viz. X. israeliae, X. italiae, and X. pachtaicum) and two Longidorus (viz. L. irani-cus, and L. pseudoelongatus) species were found. In three out of the 14 wild olive tree sampling points, two Xiphinema (viz. X. her-akliense and X. italiae), and one Longidorus species (viz. L. closelongatus) were found. In the grapevine samples, the data on the presence of X. index, X. italiae and X. pachta-icum were not considered in this study, as these are quite common nematode species found on grapevine in Crete. However, in 12 out of the 100 examined grapevine soil Brought to you by | CSIC - Unidad Coordinacion BibliotecasAuthenticatedDownload Date | 11/27/17 1:38 PM© Benaki Phytopathological InstituteXiphinema and Longidorus species on olive and grapevine in Crete 75Figure 1. Light micrographs of fi rst-stage juvenile of Xiphin-ema cretensis Tzortzakakis et al., 2014. A) Whole body. B) An-terior region showing odontostyle (odt), replacement odonto-style (rodt), and guiding-ring (gr). C-E) Tail regions showing anus (a). (Scale bars: A = 100 μm; B-E = 20 μm).Figure 2. Light micrographs of fi rst-stage juvenile of Xiphine-ma herakliense Tzortzakakis et al., 2015. A) Whole body. B) An-terior region showing odontostyle (odt), replacement odonto-style (rodt), and guiding-ring (gr). C) Tail region showing anus (a). (Scale bars: A = 100 μm; B = 20 μm; C = 10 μm).Table 1. Longidoridae species found in cultivated olive (OLI), wild olive (OLE) and grapevine (GRA) in Crete.Nematode species Plant, locality Sample codeLongidorus closelongatus Stoyanov 1964 Grapevine, Xamoudochori GRA36Longidorus closelongatus Grapevine, Ag Paraskies GRA37Longidorus closelongatus Grapevine, Akrotiri GRA39Longidorus closelongatus Wild olive, Agiofaraggo OLE  34Longidorus cretensis Tzortzakakis et al., 2001 Grapevine, P. Elias GRA34Longidorus iranicus Sturhan and Barooti, 1983 Olive, Arkalochori OLI131Longidorus iranicus Grapevine, Archanes GRA33Longidorus iranicus Grapevine, Ag. Thomas GRA35Longidorus iranicus Grapevine, Roukani GRA38Longidorus iranicus Grapevine, Lousakies GRA31Longidorus pseudoelongatus Altherr, 1976 Olive, Arkalochori OLI132Longidorus pseudoelongatus Olive, Faneromeni OLI136Longidorus pseudoelongatus Grapevine, Gazi GRA 32Longidorus pseudoelongatus Grapevine, P. Elias GRA40Xiphinema cretense Tzortzakakis et al., 2014 Grapevine, P. Elias GRA40Xiphinema herakliense Tzortzakakis et al., 2015 Wild olive, Agiofaraggo OLE 33, 34, 36Xiphinema herakliense Grapevine, P. Elias GRA41Xiphinema herakliense Grapevine, P. Elias GRA42Xiphinema israeliae Luc et al., 1982 Olive, Roufas OLI14Xiphinema israeliae Olive, Neapoli OLI115Xiphinema israeliae Olive, Pyrgiotisa OLI138Xiphinema israeliae Olive, Dermatos OLI145Xiphinema israeliae Olive, Dermatos OLI146Xiphinema israeliae Grapevine, Akrotiri GRA39Xiphinema italiae Meyl, 1953 Wild olive, Agiofaraggo OLE 34Xiphinema italiae Olive, Episkopi OLI 104Xiphinema pachtaicum (Tulaganov, 1938) Kirjanova 1951 Olive, 12 samples **Sample codes = 101, 102, 103, 105, 107, 110, 123, 129, 130, 137, 139, 140Brought to you by | CSIC - Unidad Coordinacion BibliotecasAuthenticatedDownload Date | 11/27/17 1:38 PM© Benaki Phytopathological InstituteTzortzakakis et al.76samples, three Xiphinema (viz. X. cretense, X. herakliense and X. israeliae) and four Longi-dorus species (viz. L. closelongatus, L. creten-sis, L. iranicus, and L. pseudoelongatus) were found. Although L. moesicus was previously reported on grapevine in Crete (Tzortzaka-kis et al., 2014), after recent studies (Maafi et al., 2015), this species has been synon-ymized with Longidorus iranicus. All these data are presented in Table 1, which supple-ments previously published results (Tzort-zakakis et al., 2014, 2015) for cultivated and wild olive trees and grapevines from Crete. Thus the updated records, considering also the previous studies, on the percentage of occurrence for the detected nematode spe-cies is as follows: a) In 146 soil samples from olive orchards: L. closelongatus 1.4%, L. cretensis 0.7%, L. iranicus 0.7%,  L. pseudoelongatus 6.2%, X. cretense 2%, X. herakliense 0.7%, X. index 2% , X. israeliae 7.5%, X. italiae 7.5% and X. pachtaicum 39%, b) In 36 soil samples from wild olive trees: L. closelongatus 2.8%, X. herakliense 36.1%, X. israeliae 2.8%, X. italiae 2.8% and. X. pachtaicum 5.6%,c) In 130 samples from vineyards (exclud-ing X. index, X. italiae and X. pachtaicum): L. closelongatus 5.4%, L. cretensis 2.3%, L. iranicus 6.2%, L. orientalis 0.8%, L. pseu-doelongatus 1.5%, X. cretense 0.8%, X. her-akliense 1.5%, X. israeliae 0.8% and X. sim-ile 0.8%.In conclusion, the data presented here-in, indicate some new information for the presence of Longidoridae in Crete. Xiphine-ma israeliae, X. cretense, X. herakliense and L. pseudoelongatus were found on grapevine, while until now they had been found exclu-sively on olive trees; L. iranicus was found on cultivated olive trees, whereas until now it had been found only on grapevines; X. itali-ae and L. closelongatus are reported for fi rst time on wild olive trees. This research was supported by grant KBBE 219262 ArimNET-ERANET FP7 2012-2015 Proj-ect PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Hellenic Agricultur-al Organization-DEMETER and Instituto Na-cional de Investigación y Tecnología Agraria y Alimentaria (INIA), grant AGR-136 from ‘Conse-jería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and Union Europea, Fon-do Europeo de Desarrollo regional, “Una man-era de hacer Europa”. I.G. Birmpilis and E. Na-siou were employed by the ARIMNET-PESTOL-IVE project. The authors thank J. Martín Bar-barroja and G. León Ropero from IAS-CSIC for the excellent technical assistance.Literature citedAltherr, E. 1976. La faune des eaux profondes inter-stitielles de la région de Wiesbaden. Bulletin de la Société Vaudoise des Sciences Naturelles, 73: 97–116.Decraemer, W. and Robbins, R.T. 2007. The who, what and where of Longidoridae and Trichodor-idae. Journal of Nematology, 39: 295-297.Hunt, D.J. 1995. Aphelenchida, Longidoridae and Trichodoridae: Their Systematics and Bionomics. CAB International, Wallingford, U.K., p. 352.Kirjanova, E.S. 1951. Soil nematodes found in cot-ton fi elds and in virgin soil of Golodnaya Steppe (Uzbekistan). Trudy Zoologie Institute Akademie Nauk SSSR, 9: 625–657.Luc, M., Brown, D.J.F. and Cohn, E. 1982. Xiphinema israeliae n. sp. (Nematoda: Dorylaimoidea). Re-vue de Nématologie, 5: 233-239.Maafi , Z.S., Subbotin, S.A., Sturhan, D., Barooti, S.and Taheri, Z.M. 2015. Characterisation of Longido-rus iranicus Sturhan & Barooti, 1983 (Nematoda: Longidoridae) from Iran and synonymisation of L. moesicus Lamberti, Choleva & Agostinelli, 1983. Russian Journal of Nematology, 23: 21-28.Meyl, A.H. 1953. Beitrage zur Kenntnis der Nem-atodenfauna vulkanisch erhitzter biotope. 1. Mitt., die terrikolen nematoden in bereich von fumarolen auf der insel ischia.Zeitschrift fuer Morphologie und Okologie der Tiere, 42: 67–116.Stoyanov, D. 1964. A contribution to the nematofau-na of the grapevine. Rastitelna Zashtita Sofi a, 12: 16-24.Sturhan, D. and Barooti, S.H. 1983. Longidorus irani-cus n. sp. (Nematoda: Dorylaimida). Systematic Parasitology, 5: 21-24.Taylor, C.E. and Brown, D.J.F. 1997.Nematode vectors of plant viruses. CABI, Wallingford, UK, p.296.Tulaganov, A. T. 1938. The fauna of nematodes of cotton and surrounding soil in Katta-Kurgan district of the Uzbeck SSE. Tr Uzbekskistan Gos. Brought to you by | CSIC - Unidad Coordinacion BibliotecasAuthenticatedDownload Date | 11/27/17 1:38 PM© Benaki Phytopathological InstituteXiphinema and Longidorus species on olive and grapevine in Crete 77University, 12: 1-25.Tzortzakakis, E.A., Peneva, V., Terzakis, M., Neilson, R. and Brown, D.J.F. 2001. Longidorus cretensis n. sp. (Nematoda: Longidoridae) from a vineyard infected with a foliar ‘yellow mosaic’ on Crete, Greece. Systematic Parasitology, 48: 131-139.Tzortzakakis,E.A., Archidona-Yuste, A., Cantalapie-dra-Navarrete, C., Nasiou, E., Lazanaki, M.S., Ka-bourakis, E.M., Palomares-Rius, J.E. and Castil-lo, P. 2014. Integrative diagnosis and molecular phylogeny of dagger and needle nematodes of olives and grapevines in the island of Crete, Greece, with description of Xiphinema cretense ΣΥΝΤΟΜΗ ΑΝΑΚΟΙΝΩΣΗΠεριγραφή της προνύμφης πρώτου σταδίου των νηματωδών Xiphinema cretense και X. herakliense. Διασπορά των νηματωδών Xiphinema και Longidorus σε ελιές, αγριελιές και αμπέλια στην Κρήτη Ε.Α. Τζωρτζακάκης, A. Archidona-Yuste, C. Cantalapiedra-Navarrete, Ι.Γ. Μπιρμπίλης, Ε. Νάσιου, J.E. Palomares-Rius and P. CastilloΠερίληψη   Η παρουσία νηματωδών της οικογένειας Longidoridae διερευνήθηκε σε δείγματα εδά-φους προερχόμενα από καλλιεργούμενες ελιές, αγριελιές και αμπέλια στην Κρήτη. Έγινε περιγραφή της προνύμφης πρώτου σταδίου των ειδών Xiphinema cretense και X. herakliense. Τα είδη X. israeliae, X. cretense, X. herakliense και Longidorus pseudoelongatus, τα οποία μέχρι τώρα είχαν αναφερθεί μόνο σε ελιά στην Κρήτη, βρέθηκαν στην ριζόσφαιρα αμπελιών. Επιπλέον, το είδος L. iranicus αναφέρεται για πρώτη φορά σε καλλιεργούμενη ελιά, ενώ τα είδη X. italiae και L. closelongatus αναφέρονται για πρώτη φορά σε αγριελιά στην Κρήτη. Η παρουσία  φυτοπαρασιτικών ειδών νηματωδών σε καλλιεργούμενη ελιά, αγριελιά και αμπέλι συνοψίζεται λαμβάνοντας υπόψη προηγούμενες δημοσιεύσεις. Hellenic Plant Protection Journal 9: 73-77, 2016n. sp. (Nematoda, Longidoridae). European Jour-nal of Plant Pathology,140: 563-590.Tzortzakakis,E.A., Archidona-Yuste, A., Cantalapie-dra-Navarrete, C., Nasiou, Palomares-Rius, J.E. and Castillo, P. 2015. Description and molecular characterisation of Xiphinema herakliense n. sp. (Nematoda: Longidoridae) from wild and culti-vated olives in Crete. Nematology,17: 231-245.Received: 11 January 2016; Accepted: 20 May 2016Brought to you by | CSIC - Unidad Coordinacion BibliotecasAuthenticatedDownload Date | 11/27/17 1:38 PM

Description of the first-stage juveniles of Xiphinema cretense and X. herakliense - Distribution of Xiphinema and Longidorus species in olive orchards and grapevines in Crete, Greece

Description of the first-stage juveniles of Xiphinema cretense and X. herakliense - Distribution of Xiphinema and Longidorus species in olive orchards and grapevines in Crete, Greece

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