Orobanche crenata is a parasitic weed that causes severe yield losses in important grain and forage legume crops. Cereals have been reported to inhibit O. crenata parasitism when grown intercropped with susceptible legumes, but the responsible metabolites have not been identified. A number of metabolites have been reported in cereals that have allelopathic properties against weeds, pests, and pathogens. We tested the effect of several allelochemicals identified in cereals on O. crenata seed germination and radicle development. We found that 2-benzoxazolinone, its derivative 6-chloroacetyl-2-benzoxazolinone, and scopoletin significantly inhibited O. crenata seed germination. Benzoxazolinones, l-tryptophan, and coumalic acid caused the stronger inhibition of radicle growth. Also, other metabolites reduced radicle length, this inhibition being dose-dependent. Only scopoletin caused cell necrotic-like darkening in the young radicles. Prospects for their application to parasitic weed management are discussed. © 2013 American Chemical Society., This research is supported by projects FP7-ARIMNet-MEDILEG and AGL2011-22524 (cofinanced by FEDER funds)., Peer Reviewed

An aggressive isolate of Ascochyta lentis obtained from lentil (Lens culinaris L.) produced various metabolites in vitro. The metabolites were isolated from the culture filtrates and characterized by spectroscopic, chemical, and optical methods. A new phytotoxic anthraquinone, named lentisone, was isolated and characterized as (1S,2S,3S)-1,2,3,8-tetrahydroxy-1,2,3,4- tetrahydro-6-methylanthraquinone together with the well-known pachybasin (1-hydroxy-3-methylanthraquinone), tyrosol, and pseurotin A. Lentisone, tyrosol, and pseurotin A were phytotoxic to lentil, with lentisone the most toxic of all. The toxicity of these compounds is light-dependent. Finally, lentisone was also found to be phytotoxic to chickpea, pea, and faba bean, with toxicity in the latter higher than in any other tested legume, including lentil. © 2013 American Chemical Society., The research was supported in part by a grant from the Italian Ministry of University and Research and MEDILEG Project ARIMNet Call 2011., Peer Reviewed

The genus Xiphinema constitutes a large group of about 260 species of plant-ectoparasitic nematodes. The group is polyphagous and distributed almost worldwide. Some of the species of this genus damage agricultural crops by direct feeding on root cells as well as by transmitting nepoviruses. Species discrimination in Xiphinema is complicated by phenotypic plasticity leading to potential misidentification. We conducted nematode surveys in cultivated and natural environments in Spain from 2009 to 2012, from which we identified 20 populations of Xiphinema species morphologically close to the virus-vector nematode species Xiphinema diversicaudatum, three apomictic populations tentatively identified as species from the complex Xiphinema aceri-pyrenaicum group, and one population morphologically different from all others that is characterized by a female tail elongate to conical and absence of uterine differentiation. We developed comparative multivariate analyses for these related species by using morphological and morphometrical features together with molecular data from nuclear ribosomal DNA genes [D2-D3 expansion segments of large ribosomal subunit 28S, internal transcribed spacer 1 (ITS1), and partial small ribosomal subunit (18S)]. The results of multivariate, molecular, and phylogenetic analysis confirmed the morphological hypotheses and allowed the delimitation and discrimination of two new species in the genus described herein as Xiphinema baetica sp. nov. and Xiphinema turdetanensis sp. nov., and ten known species: Xiphinema adenohystherum, Xiphinema belmontense, Xiphinema cohni, Xiphinema coxi europaeum, Xiphinema gersoni, Xiphinema hispidum, Xiphinema italiae, Xiphinema lupini, Xiphinema nuragicum, and Xiphinema turcicum. Multivariate analyses based on quantitative and qualitative characters and phylogenetic relationships of Xiphinema spp. based on the three molecular ribosomal markers resulted in a partial consensus of these species grouping as nematode populations were maintained for the majority of morphospecies groups (e.g. morphospecies groups 5 and 6), but not in some others (e.g. position of Xiphinema granatum), demonstrating the usefulness of these analyses for helping in the diagnosis and identification of Xiphinema spp. The clade topology of phylogenetic trees of D2-D3 and partial 18S regions in this study were congruent in supporting the polyphyletic status of some characters, such as the female tail shape and the degree of development of the genital system in species with both genital branches equally developed. This is the most complete phylogenetic study for Xiphinema non-americanum-group species. Agreement between phylogenetic trees and some morphological characters (uterine spines, pseudo-Z organ, and tail shape) was tested by reconstruction of their histories on rDNA-based trees using parsimony and Bayesian approaches. Thus, integrative taxonomy, based on the combination of multivariate, molecular analyses with morphology, constitutes a new insight into the identification of Xiphinema species., This research was supported by a grant AGL2009-06955 from ‘Ministerio de Ciencia e Innovación’ of Spain, grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), grant AGL2012-37521 from 'Ministerio de Economía y Competitividad' of Spain, grant AGR-136 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and the European Social Fund., Peer Reviewed

Background: In the last years, many olive plantations in southern Spain have been mediated by the use of self-rooted planting stocks, which have incorporated commercial AMF during the nursery period to facilitate their establishment. However, this was practised without enough knowledge on the effect of cropping practices and environment on the biodiversity of AMF in olive orchards in Spain. Methodology/Principal Findings: Two culture-independent molecular methods were used to study the AMF communities associated with olive in a wide-region analysis in southern Spain including 96 olive locations. The use of T-RFLP and pyrosequencing analysis of rDNA sequences provided the first evidence of an effect of agronomic and climatic characteristics, and soil physicochemical properties on AMF community composition associated with olive. Thus, the factors most strongly associated to AMF distribution varied according to the technique but included among the studied agronomic characteristics the cultivar genotype and age of plantation and the irrigation regimen but not the orchard management system or presence of a cover crop to prevent soil erosion. Soil physicochemical properties and climatic characteristics most strongly associated to the AMF community composition included pH, textural components and nutrient contents of soil, and average evapotranspiration, rainfall and minimum temperature of the sampled locations. Pyrosequencing analysis revealed 33 AMF OTUs belonging to five families, with Archaeospora spp., Diversispora spp. and Paraglomus spp., being first records in olive. Interestingly, two of the most frequent OTUs included a diverse group of Claroideoglomeraceae and Glomeraceae sequences, not assigned to any known AMF species commonly used as inoculants in olive during nursery propagation. Conclusions/Significance: Our data suggests that AMF can exert higher host specificity in olive than previously thought, which may have important implications for redirecting the olive nursery process in the future as well as to take into consideration the specific soils and environments where the mycorrhized olive trees will be established. © 2014 Montes-Borrego et al., This research was supported by grants from Projects AGL2008-00344/AGR from ‘Ministerio de Ciencia e Innovación’, Project P10-AGR-5908 from ‘Consejería de Economía, Innovación y Ciencia’ of Junta de Andalucía, Project AGL-2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain, and Fondo Europeo de desarrollo regional (FEDER) >Una manera de hacer Europa> from the European Union. The grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), also provided partial financial support. M. Montes-Borrego enjoyed a contract from ‘Consejería de Economía, Innovación y Ciencia’ of Junta de Andalucía, Spain., Peer Reviewed

[Background] The Lathyrus genus includes 160 species, some of which have economic importance as food, fodder and ornamental crops (mainly L. sativus, L. cicera and L. odoratus, respectively) and are cultivated in >1·5 Mha worldwide. However, in spite of their well-recognized robustness and potential as a source of calories and protein for populations in drought-prone and marginal areas, cultivation is in decline and there is a high risk of genetic erosion. [Scope] In this review, current and past taxonomic treatments of the Lathyrus genus are assessed and its current status is examined together with future prospects for germplasm conservation, characterization and utilization. A particular emphasis is placed on the importance of diversity analysis for breeding of L. sativus and L. cicera. [Conclusions] Efforts for improvement of L. sativus and L. cicera should concentrate on the development of publicly available joint core collections, and on high-resolution genotyping. This will be critical for permitting decentralized phenotyping. Such a co-ordinated international effort should result in more efficient and faster breeding approaches, which are particularly needed for these neglected, underutilized Lathyrus species. © 2014 The Author., The authors would like to acknowledge the financial support of Fundação para a Ciência e a Tecnologia, Portugal, through grants #PEst-OE/EQB/LA0004/2011 and #PTDC/AGR-GPL/103285/2008 and Research Contract by the Ciência 2008 program (to M.C.V.P.). This research was also funded by FP7-ARIMNet-MEDILEG project and by Spanish project AGL2011-22524., Peer Reviewed

Sheathoid nematodes of the genus Hemicriconemoides are migratory root-ectoparasites of many plants including various agricultural crops and fruit trees. They are generally found inhabiting warm areas of the world and presently consist of 52 valid species. In this study we provide morphological and molecular characterisation of 12 species of this genus viz.: H. alexis, H. brachyurus, H. californianus, H. chitwoodi, H. macrodorus, H. minutus, H. ortonwilliamsi, H. promissus, H. silvaticus, H. strictathecatus, H. wessoni and Hemicriconemoides sp. originating from China, Greece, Japan, Myanmar, Spain, South Africa and the USA. Morphological descriptions, measurements, light and scanning electron microscopic observations and drawings are given for several species. Phylogenetic relationships within Hemicriconemoides, as inferred from the analyses of the D2-D3 of 28S rRNA and ITS-rRNA gene sequences, resulted in trees with three major clades that corresponded with species groupings based on morphology of the lip pattern and vulval flap. PCR with species-specific primers were developed for H. californianus, H. chitwoodi and H. strictathecatus. © Koninklijke Brill NV, Leiden, 2014., Pablo Castillo acknowledges support from grant 219262 ArimNET_ERANET FP7 2012-2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), grant AGL2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain, grant AGR-136 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and the European Social Fund., Peer Reviewed

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated and wild olive and grapevine in Crete Island were investigated. Morphological and morphometrical studies identified five Longidorus and six Xiphinema species, with frequencies of prevalence (for wild and cultivated olives and grapevines, respectively) as follows: Longidorus closelongatus (2.0–13.3 %), L. cretensis (1.0–6.7 %), L. moesicus (13.3 % only in grapevines), L. orientalis (3.3 % only in grapevines), L. pseudoelongatus (7.0 % only in olives), Xiphinema cretense n. sp. (3.0 % only in olives), X. index (3.0–23.3 %), X. israeliae (6.3 % only in olives), X. italiae (3.3–10.0 %), X. pachtaicum (26.7–42 %) and X. simile (3.3 % only in grapevines). Xiphinema cretense n. sp. is characterized by a body size 3,872–6,135 μm long, lip region anteriorly rounded, separated from the rest of the body by a depression, odontostyle and odontophore 140.6 and 80.3 μm long respectively, vulva position at 46.0–50.5 %, female tail 31.0–38.0 μm long, nearly hemispherical with curvature essentially dorsal and with a tip completely rounded or presenting a very short bulge, c ratio (119.1–186.9), c’ ratio (0.7–0.8). Molecular characterisation using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids., 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”. E. Nasiou
and M.S. Lazanaki are employed by the ARIMNET-PESTOLIVE
project. The seventh author is a recipient of a “Juan de La Cierva”
contract from Ministerio de Economía y Competitividad of Spain., Peer Reviewed

A new mono-sexual root-lesion nematode species, Pratylenchus oleae n. sp., parasitizing roots of olive plants cv. Koroneiki in commercial fields at Ouled Chamekh (central Tunisia), and wild and cultivated olive (cv. Picual) plants in Agua Amarga (southern Spain) is described. The new species is characterised by the female having a lip region slightly offset and bearing three annuli, stylet 16.5 (14.5-17.0) μm long, with prominent rounded knobs, pharyngeal overlapping rather long (22-36) μm, lateral fields areolated and with four incisures and diagonal lines in middle band, spermatheca rounded but non-functional, tail short, conoid-rounded to subcylindrical, usually annulated terminus, males unknown, and a specific D2-D3, ITS1, 18S-rRNA, hsp90 and COI sequences. Morphologically this species is related to P. cruciferus, P. delattrei, and P. kumamotoensis. The results of the phylogenetic analysis based on sequences of the D2-D3 expansion regions of 28S, partial 18S and ITS rRNA genes confirmed the close relationship of P. oleae n. sp. with P. dunensis, P. penetrans, P. pinguicaudatus, from which was clearly separated. A PCR-based diagnostic assay was also developed for identification of P. oleae n. sp. using the species-specific primers Poleae_fw1_4 and Poleae_rv1 that amplify a 547-bp fragment in the internal transcribed spacer (ITS1) region of ribosomal DNA, which clearly separate from other root-lesion nematodes damaging olive such as P. penetrans and P. vulnus. © 2014 Koninklijke Nederlandse Planteziektenkundige Vereniging., This research was supported by a grant
219262 ArimNET_ERANET FP7 2012–2015 Project
PESTOLIVE ‘Contribution of olive history for the management
of soilborne parasites in the Mediterranean basin’ from 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”. The first author is a recipient of a “Juan de La
Cierva” contract from Ministerio de Economía y Competitividad
of Spain, Peer Reviewed

Broomrapes are weedy root parasitic plants that severely constrain faba bean (Vicia faba) production. The most widely distributed species affecting faba bean is Orobanche crenata, although O. foetida is of local importance in Tunisia. After long and extensive breeding efforts made in several countries only moderately resistant cultivars are available to farmers. In an attempt to identify new sources of resistance a germplasm collection of 483 V. faba accessions was screened for resistance to O. crenata under field conditions in Córdoba, Spain. Stability of resistance of the 37 most resistant accessions was further tested in a multi-location experiment in Egypt, Tunisia and Spain over three field seasons. Resistance to O. foetida was also tested in Tunisia.Although complete resistance was not found, and in spite of significant genotype. ×. environment interaction revealing instability of the phenotypic expression across environments, this study allowed the identification of a number of accessions showing significant levels of resistance that was stable across environments. Cultivar Baraca and accessions V-1268, V-1302, V-1301, V-268, V-231, V-319 and V-1272 were the most resistant and stable across environments, and what is most interesting, being resistant both to O. crenata and O. foetida., Financial support by the European Union FP5-EUFABA and
FP7-ARIMNet-Medileg projects and by Spanish AGL2011-22524
co-financed by FEDER is acknowledged., Peer Reviewed

This paper summarises the current knowledge concerning cryptic species of plant-parasitic nematode and briefly reviews the different methods available for their detection and characterisation. Cryptic species represent an important component of biodiversity, such speciation being common among plant-parasitic nematodes and occurring in diverse groups with different life history traits, including the spiral, virus vector, root-lesion and false root-knot nematodes. Cryptic species are important for a number of reasons, including food security, quarantine, non-chemical management technologies and species conservation, and should not be ignored. The magnitude of the phenomenon is largely unknown, but the available data on plant-parasitic nematodes demonstrate that reliance on morphology alone for species delimitation seriously underestimates the total number of taxa. Future research should focus on appropriately designed case studies using combined approaches, including large-scale, whole sample analyses by next-generation sequencing or proteomics in order to be able to answer the many questions that still remain., This research was supported by grant AGL2012-37521
from ‘Ministerio de Economía y Competitividad’ of
Spain, 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 Instituto Nacional de Investigación
y Tecnología Agraria y Alimentaria (INIA), and
grants P12-AGR 1486 and 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’., Peer Reviewed

A population of Xiphinema barense from wild olive trees in Torre Pozzella, Brindisi province, southern Italy, is described using both morphological and molecular studies and compared with the description of the type specimens. The wild olive nematode population agrees very well with all morphometrics provided in the original description. However, detailed observations of the lumen of the tubular portion of the uterus in paratypes and specimens of the new population revealed a clear pseudo-Z-organ with small granules mixed with crystalloid bodies which were previously undetected. Photomicrographs of adult paratypes, which were lacking in the original description, and of specimens of the new population from wild olive trees are provided. The results of the phylogenetic analyses based on the sequences of the D2-D3 expansion regions of the 28S rRNA gene and ITS rRNA genes confirm the species differentiation and indicate the phylogenetic position of X. barense and its relationship with closely related species., The present research was partially funded by the grant
219262 ArimNET_ERANET FP7 2012-2015 Project
PESTOLIVE “Contribution of olive history for the management
of soilborne parasites in the Mediterranean
basin”; and 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”., Peer Reviewed

Trabajo presentado en el XVIII Congreso de la Sociedad Española de Fitopatología (SEF), celebrado en Palencia del 20 al 23 de septiembre de 2016., Se han detectado endosimbiontes bacterianos en algunos grupos de nematodos
fitoparásitos y parecen tener un papel fundamental en la adaptación al medio y
supervivencia de sus especies huésped. En este trabajo se ha determinado la presencia y
la diversidad molecular de estos endosimbiontes dentro de los nematodos ectoparásitos
migratorios de la familia Longidoridae (viz Xiphinema, Longidorus y Paralongidorus).
Se han testado 124 poblaciones de nematodos (previamente identificados
morfológicamente y molecularmente) frente a la presencia de endosimbiontes
bacterianos usando el gen ribosómico 16S. Solo se detectaron endosimbiontes
bacterianos en especies de nematodos pertenecientes al género Xiphinema y
específicamente dentro del grupo Xiphinema americanum. En este estudio se ha
secuenciado el gen 16S de 57 endosimbiontes bacterianos. Uno de los grupos de
secuencias perteneció al género “Candidatus Xiphinematobacter” (19 secuencias
asociadas con 7 especies de nematodos, incluyendo 2 especies descritas y 3 especies de
endosimbiontes no descritos anteriormente). El segundo grupo de endosimbiontes (38
secuencias asociadas con 5 especies de nematodos y 5 especies de endosimbiontes no
descritos anteriormente) estuvo relacionado con la familia Burkholderiaceae que incluye
también endosimbiontes fúngicos y de plantas. El grupo de endosimbiontes
pertenecientes a la familia Burkholderiaceae son citados por primera vez en el Phylum
Nematoda. Nuestros estudios indican una elevada relación simbiótica entre las especies
de nematodos huéspedes y estos endosimbiontes. Estos resultados fueron corroborados
con una elevada congruencia filogenética entre nematodo huésped-endosimbionte en
una larga co-evolución entre ellos. Asimismo, también se estudiaron las variaciones en
el genoma entre dos endosimbiontes asociados con las especies de nematodos
transmisores de virus Xiphinema americanum y Xiphinema rivesi., Investigación financiada por P12-AGR-1486-CICE, ArimNET_ERANET-219262-INIA, y fondos FEDER (EU)., Peer Reviewed

Póster presentado en el XVIII Congreso de la Sociedad Española de Fitopatología (SEF), celebrado en Palencia del 20 al 23 de septiembre de 2016., Las infecciones de olivo por nematodos fitoparásitos se han asociado con reducciones
del crecimiento y vigor en varios países de la Cuenca Mediterránea. Los nematodos
ectoparásitos migratorios de la familia Longidoridae (Longidorus, Paralongidorus y
Xiphinema) comprenden especies polífagas, y algunas de ellas transmiten
específicamente virus vegetales (nepovirus) de importancia económica. Hasta hoy, a
nivel mundial, se han citado 9 especies de Longidorus y 16 especies de Xiphinema
asociadas con olivo. En este trabajo se han realizado prospecciones fitonematológicas
sistematizadas y extensas en olivo cultivado (O. europaea subsp. europaea) y silvestre
(O. europaea subsp. sylvestris) en un total de 453 puntos de muestreo (115 asociados a
olivo silvestre y 338 a olivo cultivado), ampliamente distribuidos en 211 localidades del
territorio andaluz. El diagnóstico integrativo (combinando estudios morfométricos y
moleculares) ha permitido identificar 13 especies del género Longidorus (6 de ellas
nuevas para la ciencia) y 32 especies del género Xiphinema, 10 especies pertenecientes
a X. americanum-group (2 de ellas nuevas para la ciencia) y 22 especies pertenecientes
a X. no-americanum-group, (8 de ellas nuevas para la ciencia). La prevalencia media fue
10 veces mayor en Xiphinema spp. que en Longidorus spp. (85,0% vs. 8,4%), siendo la
prevalencia de Longidorus spp. en olivo silvestre mayor que en olivo cultivado (16,0%
vs. 6,0%), sin embargo, no se apreciaron diferencias significativas en Xiphinema spp.
(93,9% vs. 81,7%). La distribución geográfica de Longidorus spp. sugiere un patrón
relacionado con factores edáficos y ecológicos, mientras que en Xiphinema spp.
estuvieron ampliamente distribuidas y diferencialmente asociadas con la subespecie de
olivo. Los análisis filogenéticos basados en ADN ribosómico (fragmentos D2-D3 del
gen 28S, 18S, región ITS) han demostrado una elevada diversidad críptica y han
confirmado la delimitación y discriminación de las diferentes especies encontradas.
Asimismo, la filogenia de los tres marcadores moleculares mantuvo la mayoría de los
linajes y grupos detectados en ambos géneros en estudios anteriores. En conclusión, la
extraordinaria prevalencia y diversidad de este grupo de nematodos en olivo en
Andalucía sugiere que el sur de la Península Ibérica constituye un centro evolutivo de
Longidorus spp. y Xiphinema spp., Investigación financiada por P12-AGR-1486-CICE, AGL2012-37521-MINECO, ArimNET_ERANET-219262-INIA, y fondos FEDER (EU)., Peer Reviewed

Póster presentado en el XVIII Congreso de la Sociedad Española de Fitopatología (SEF), celebrado en Palencia del 20 al 23 de septiembre de 2016., Los nematodos de la familia Longidoridae (viz. Longidorus, Paralongidorus, y
Xiphinema) comprenden especies fitoparásitas polífagas que pueden afectar a un gran
número de plantas. El daño que causan a los diferentes huéspedes depende no sólo de su
parasitismo directo sobre células epidérmicas y corticales de la raíz, sino también por la
transmisión específica de virus vegetales (nepovirus). El genoma mitocondrial de
nematodos fitoparásitos proporciona información esencial sobre su estructura genómica,
relaciones filogenéticas e inferencias funcionales y evolutivas. Hasta la fecha, X.
americanum era el único genoma mitocondrial secuenciado de la familia Longidoridae.
En este estudio se ha secuenciado el genoma mitocondrial de 4 especies de nematodos
fitoparásitos pertenecientes a esta familia, incluyendo Xiphinema rivesi, Xiphinema
pachtaicum, Longidorus vineacola y Paralongidorus litoralis. Los cuatro genomas
tienen un tamaño estimado de 12.6, 12.5, 13.5 y 12.8 kb, respectivamente, siendo de los
más pequeños descritos en el Phylum Nematoda. Todos los genomas contienen 12 genes
codificantes de proteínas (viz. cox1-3, nad1-6, nad4L, atp6 and cob) y dos genes
ribosómicos (rrnL and rrnS), estando ausente el gen atp8. La distribución génica resultó
muy variable entre ellos, a excepción de X. rivesi y X. americanum, especies
estrechamente emparentadas. Se identificaron 22 tRNAs en L. vineacola, 21 tRNAs en
X. rivesi y P. litoralis (faltando mt-tRNAN), y 19 tRNAs en X. pachtaicum (faltando mttRNAN,
mt-tRNAA y mt-tRNAR). Las regiones codificantes fueron muy pequeñas y
estuvieron presentes en pocas posiciones dentro del genoma. Los análisis filogenéticos
mostraron dos clados dentro de Longidoridae, uno incluyendo Longidorus y
Paralongidorus y otro incluyendo especies de Xiphinema. Xiphinema americanum y X.
rivesi están claramente relacionados filogenéticamente, mientras que la posición de X.
pachtaicum no estuvo claramente definida. Estos nuevos genomas mitocondriales
pueden aportar nuevos marcadores para la identificación de especies, y futuros estudios
filogenéticos., Investigación financiada por P12-AGR-1486-CICE, ArimNET_ERANET-219262-INIA, y fondos FEDER (EU)., Peer Reviewed

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated olive (cvs. Chemlali and Chétoui) in Tunisia were investigated. Morphological and morphometrical studies identified three Longidorus and six Xiphinema species, with frequencies of prevalence as following: Longidorus africanus (23.0 %), L. euonymus (4.5 %), L. glycines (13.7 %), Xiphinema conurum (13.7 %), X. italiae (36.4 %), X. meridianum (13.7 %), X. pachtaicum (18.2 %), X. robbinsi (9.1 %), and Xiphinema sp. (4.5 %). The three Longidorus species were reported for the first time in Tunisia, in addition to two species of Xiphinema (viz. X. meridianum and X. robbinsi). Molecular characterisation using D2-D3 expansion regions of 28S rRNA and ITS1-rRNA was carried out and Bayesian inference analysis was used to reconstruct phylogenetic relationships among these species and with other longidorids. Twenty-five new D2-D3 of 28S rRNA gene sequences were obtained in the present study, seven for Longidorus and 18 for Xiphinema spp., as well as 14 new ITS1 rRNA gene sequences (seven for Longidorus and seven for Xiphinema spp.)., 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". The fourth author is a recipient of a "Juan de La Cierva" contract from Ministerio de Economía y Competitividad of Spain., Peer reviewed

The Xiphinema americanum-group constitutes a complex of about 55 species of polyphagous plant-ectoparasitic nematodes with a worldwide distribution. This group of plant-parasitic nematodes is one of the most difficult dagger nematode species complexes for diagnosis because the morphology is very conservative and morphometric characters often overlap. We conducted nematode surveys in cultivated and wild olives in southern Spain from 2012 to 2014, from which we identified 16 nematode populations of the X. americanum-group, five of which were tentatively identified as belonging to three new species and are described herein as Xiphinema plesiopachtaicum sp. nov.,Xiphinema vallense sp. nov., and Xiphinema astaregiense sp. nov., and 11 populations belonging to nine known species: Xiphinema brevisicum, Xiphinema duriense, Xiphinema incertum, Xiphinema luci, Xiphinema madeirense, Xiphinema opisthohysterum, Xiphinema pachtaicum, Xiphinema parapachydermum, and Xiphinema rivesi. A phenetic study based on multivariate factor analyses was developed to compare some of these related species by using morphometric features. In the factor analysis the first four factors accounted for 73.1% of the total variance of the selected characters, identifying body length, body length/maximum body width (a), body length/pharyngeal length (b), body length/tail length (c), and tail length/body width at anus (c′) ratios, distance from anterior end to vulva as percentage of body length (V), stylet length, oral aperture-guiding ring distance, and lip region width as key morphometric characters to differentiate a restricted set of species within the X.pachtaicum-subgroup that includes X.plesiopachtaicum sp. nov. and X.vallense sp. nov. Multivariate analysis of variance using these specific characters allowed to differentiate species in the X.pachtaicum complex or groups of them using morphometric characters (body length, a, b, c, c′, V, stylet length, lip region width, oral aperture-guiding ring distance, female tail length, and hyaline region length). Phylogenetic analyses based on nuclear ribosomal DNA genes [D2-D3 expansion segments of large ribosomal subunit 28S, and internal transcribed spacer 1 (ITS1)] and the protein-coding mitochondrial gene, cytochrome c oxidase subunit 1 (coxI) were congruent, showing two main clades separating most of the species of X.americanum-subgroup ‘sensu stricto’ from the X.pachtaicum-subgroup. Agreement between phylogenetic trees and some morphological characters (viz. total stylet length, vulva position, and a ratio) were tested by reconstruction of their histories on rRNA-based trees using parsimony and Bayesian approaches. Thus, integrative taxonomy, based on a combination of multivariate morphological and molecular analyses constitutes a new insight into the identification of X.americanum-group species., This research is part of the PhD project of the first author and was supported by grant AGL2012-37521 from Ministerio de Economía y Competitividad of Spain, grant 219262 ArimNET_ERANET FP7 2012-2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), grants P12-AGR 1486 and AGR-136 from Consejería de Economía, Innvovación y Ciencia of the Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’. A. J. Archidona-Yuste is a recipient of research contract BES-2013-063495 from Ministerio de Ciencia e Innovación of Spain. The authors thank J. Martín Barbarroja and G. León Ropero from IAS-CSIC for the excellent technical assistance., Peer reviewed

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 reviewed

Host-plant resistance is the most practical, long-term and economically efficient disease control measure for Verticillium wilt in olive caused by the soil-borne fungus Verticillium dahliae, and it is the core of the integrated disease management. In Spain, Verticillium wilt has become a main concern for the olive industry because of the widespread occurrence of a highly virulent, defoliating (D) pathotype. Available V. dahliae-resistant olive cultivars show incomplete resistance to the D pathotype and do not satisfy consumers’ demand for high yields and oil quality as well as suitability for new environments and cultural practices in modern olive production in Spain. Highly resistant rootstocks would be of paramount importance for production of agronomically-adapted and commercially-desirable olive cultivars. Nevertheless, the validity of resistant rootstocks grafted with Verticillium wilt-susceptible olive cultivars in soils highly infested with the D pathotype would be increased if the resistance is demonstrated stable in coinfection with plant-parasitic nematodes, such as root-knot nematodes. In this work, wild olive clones ‘Ac-4’, ‘Ac-13’ and ‘Ac-18’ previously developed as highly resistant to D V. dahliae were tested for infection response by the root-knot nematode Meloidogyne javanica alone and jointly with D V. dahliae under controlled conditions optimal for Verticillium wilt in olive. The three wild olive clones showed resistance to D V. dahliae under high inoculum levels in soil and also when plants were co-infected by both pathogens. However, all clones were susceptible to M. javanica, although ‘Ac-13’ and ‘Ac-18’ showed a degree of tolerance to the used inoculum level. Nevertheless, coinfected plants with M. javanica and D V. dahliae reduced the nematode reproduction rate in ‘Ac-13’ and ‘Ac-18’, but increased that in ‘Ac-4’, suggesting that different resistance mechanisms to D V. dahliae might be operating in these clones were acting also on M. javanica reproduction., Financial support was provided by Project AGR 6082 from Consejería de Innovación, Ciencia y Empresa, Junta de Andalucía, Spain, and research grant from ‘Organización Interprofesional del Aceite de Oliva Español’. Also, partial financial support was provided by grants KBBE219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA, Spain), and grants P12-AGR-1486 and 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”., Peer reviewed

Eight fungal and plant metabolites belonging to different class
es of naturally occurring compounds, a 24-oxa[14]-cytochalasan
as cytochalasin B (1), a trisubstituted isocoumarin as 6-hydroxymellein (2), a tetracyclic pimarane d
iterpene as sphaeropsidin A (3), a chalcone as cavoxin (4), a pentasubstituted benzofuranone as cyclopaldic acid (5), a bicyclic-sesquiterpene as inuloxin A (6), a epipolythiopiperazine as gliotoxin (7) and a cyclohexene epoxide as
epiepoformin (8), were tested for their effectiveness in reducing early stages of development of several major rust fungi from the genera Puccinia and Uromyces. Spore germination and appressoria formation were assessed on pre-treated detached leaves, under controlled conditions. Among the various metabolites evaluated, compounds 5 and 8 were the most effective in inhibiting fungal germination and penetration of all rust species studied at values comparable with those obtained by fungicide application, while compound 4 was phytotoxic to plant leaves at any concentration tested., The research is supported by FP7-ARIMNet-MEDILEG and AGL2014-52871 projects and Programme STAR (A.C.) (UniNa and Compagnia di San Paolo, Italy). Dr E. Barilli received a grant from the JAE-Doc program., Peer reviewed

Bacterial endosymbionts have been detected in some groups of plant-parasitic nematodes, but few cases have been reported compared to other groups in the phylum Nematoda, such as animal-parasitic or free-living nematodes. This study was performed on a wide variety of plant-parasitic nematode families and species from different host plants and nematode populations. A total of 124 nematode populations (previously identified morphologically and molecularly) were screened for the presence of potential bacterial endosymbionts using the partial 16S rRNA gene and fluorescence in situ hybridization (FISH) and confocal microscopy. Potential bacterial endosymbionts were only detected in nematode species belonging to the genus Xiphinema and specifically in the X. americanum group. Fifty-seven partial 16S rRNA sequences were obtained from bacterial endosymbionts in this study. One group of sequences was closely related to the genus ‘Candidatus Xiphinematobacter’ (19 bacterial endosymbiont sequences were associated with seven nematode host species, including two that have already been described and three unknown bacterial endosymbionts). The second bacterial endosymbiont group (38 bacterial endosymbiont sequences associated with six nematode species) was related to the family Burkholderiaceae, which includes fungal and soil–plant bacterial endosymbionts. These endosymbionts were reported for the first time in the phylum Nematoda. Our findings suggest that there is a highly specific symbiotic relationship between nematode host and bacterial endosymbionts. Overall, these results were corroborated by a phylogeny of nematode host and bacterial endosymbionts that suggested that there was a high degree of phylogenetic congruence and long-term evolutionary persistence between hosts and endosymbionts., This research was supported by grants P12-AGR 1486 and 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’, grant AGL2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain, grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA). A. J. Archidona-Yuste is a recipient of contract BES-2013-063495 from ‘Ministerio de Ciencia e Innovación’ of Spain., Peer reviewed

The genus Xiphinema constitutes a large group of ∼275 species of polyphagous, plant-ectoparasitic nematodes that are distributed almost worldwide. Some species affect agricultural crops by feeding directly on root cells as well as by transmitting nepoviruses. Despite their agricultural importance, species discrimination in Xiphinema is difficult, leading to potential misidentification. Integrative taxonomy, based on the combination of molecular analyses and morphology, constitutes a new insight into Xiphinema species identification. In this study we describe two new species of Xiphinema from the Iberian Peninsula (X. macrodora, sp. nov. and X. oleae, sp. nov.) associated with cultivated and wild olive trees. Both species have specific rRNA sequences. Morphologically, Xiphinema macrodora, sp. nov. is characterised by a very long body (7.2–8.7 mm), a very long odontostyle and odontophore (190–206 and 105–120 µm, respectively), and a well developed pseudo-Z-organ, comprising 8 to 12 sclerotised bodies. Xiphinema oleae, sp. nov. is characterised by an odontostyle and an odontophore 136–149 and 65–80 µm long, respectively, and a well developed Z-organ with refractive inclusions (3–5), variable in shape. Additionally, X. macrodora, sp. nov. has the longest body size, and the longest odontostyle and odontophore of any Xiphinema, whereas X. oleae, sp. nov. is the first species with a well developed Z-organ from the Iberian Peninsula., This research is part of the Ph.D. project of the first author. Research was supported by grants P12-AGR-1486 from ‘Consejería de Economía, Innovación y Ciencia’ of Junta de Andalucía, 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soil-borne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), and AGL-2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain, and FEDER financial support from the European Union is gratefully acknowledged. Antonio J. Archidona-Yuste is a recipient of research fellowship BES-2013-063495 from the Spanish ‘Ministerio de Economía y Competitividad’., Peer reviewed

The genus Xiphinema includes a remarkable group of invertebrates of the phylum Nematoda comprising ectoparasitic animals of many wild and cultivated plants. Damage is caused by direct feeding on root cells and by vectoring nepoviruses that cause diseases on several crops. Precise identification of Xiphinema species is critical for launching appropriate control measures. We make available the first detailed information on the diversity and distribution of Xiphinema species infesting wild and cultivated olive in a wide-region in southern Spain that included 211 locations from which 453 sampling sites were analyzed. The present study identified thirty-two Xiphinema spp. in the rhizosphere of olive trees, ten species belonging to Xiphinema americanum-group, whereas twenty-two were attributed to Xiphinema non-americanum-group. These results increase our current knowledge on the biodiversity of Xiphinema species identified in olives and include the description of four new species (Xiphinema andalusiense sp. nov., Xiphinema celtiense sp. nov., Xiphinema iznajarense sp. nov., and Xiphinema mengibarense sp. nov.), and two new records for cultivate olives (X. cadavalense and X. conurum). We also found evidence of remarkable prevalence of Xiphinema spp. in olive trees, viz. 85.0% (385 out of 453 sampling sites), and they were widely distributed in both wild and cultivated olives, with 26 and 17 Xiphinema spp., respectively. Diversity indexes (Richness, Hill´s diversity, Hill´s reciprocal of D and Hill´s evenness) were significantly affected by olive type. We also developed a comparative morphological and morphometrical study together with molecular data from three nuclear ribosomal RNA genes (D2-D3 expansion segments of 28S, ITS1, and partial 18S). Molecular characterization and phylogenetic analyses allowed the delimitation and discrimination of four new species of the genus described herein and three known species. Phylogenetic analyses of Xiphinema spp. resulted in a general consensus of these species groups. This study is the most complete phylogenetic analysis for Xiphinema non-americanum-group species to date., Financial support was received by Project P12-AGR-1486 from ‘Consejería de Economía, Innovación y Ciencia’ of Junta de Andalucía, the grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olivehistory for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Project AGL-2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain, and FEDER financial support from the European Union., Peer reviewed

A new and a known species of Rotylenchus from Crete, each collected from the rhizosphere of olive, are described and illustrated based on morphological, morphometric and molecular studies. Rotylenchus cretensis n. sp. is characterised by having a truncate lip region with irregular longitudinal striation, lateral field areolated only in pharynx region, stylet length less than 41 μm, vulva located at 53–59 %, conoid-rounded to dorsally convex-conoid female tail with 7–11 annuli and specific D2-D3 of 28S, ITS, and partial 18S rRNA. Rotylenchus cretensis n. sp. appears to be similar to R. urmiaensis, having overlapping character ranges. Rotylenchus cretensis is also similar to R. striaticeps from which has got shorter stylet length (35.5–41.5 vs 39–50 μm), female tail shape (conoid-rounded to dorsally convex-conoid vs rounded), shorter spicules length (34–40 vs 41–50 μm), and phasmid position (varying from at level to three annuli posterior to the anus vs at level to seven annuli anterior to anus). Multivariate analysis of variance performed on R. cretensis n. sp., as well as two populations of R. striaticeps and R. urmiaensis from Iran on combined morphometric characters data set revealed a significant effect by species populations, indicating that closely morphological and morphometrically species differed significantly from each other. Additionally, analysis of variance showed that morphometric characters such as lip region width, distance from phasmid to tail tip (PHS) or a ratio can be used as key morphometric characters for species differentiation in this group. The results of phylogenetic analyses based on D2-D3 region, ITS of rRNA, and the partial 18S sequences confirmed the close molecular relationships between R. cretensis n. sp., R. urmiaensis and R. striaticeps, but clearly support the proposal of R. cretensis n. sp. as a new species. Also, the recognition of this cryptic species within Rotylenchus shows that the biodiversity of these nematodes is still not fully understood and requires further studies. Additionally, R. cypriensis is described and molecularly analysed for first time in Greece., The present research was partially funded 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 was employed by the ARIMNET-PESTOLIVE project., Peer reviewed

The genus Longidorus includes a remarkable group of invertebrate animals of the phylum Nematoda comprising polyphagous root-ectoparasites of numerous plants including several agricultural crops and trees. Damage is caused by direct feeding on root cells as well as by transmitting nepoviruses that cause disease on those crops. Thus, correct identification of Longidorus species is essential to establish appropriate control measures. We provide the first detailed information on the diversity and distribution of Longidorus species infesting wild and cultivated olive soils in a wide-region in southern Spain that included 159 locations from which 449 sampling sites were analyzed. The present study doubles the known biodiversity of Longidorus species identified in olives by including six new species (Longidorus indalus sp. nov., Longidorus macrodorus sp. nov., Longidorus onubensis sp. nov., Longidorus silvestris sp. nov., Longidorus vallensis sp. nov., and Longidorus wicuolea sp. nov.), two new records for wild and cultivate olives (L. alvegus and L. vineacola), and two additional new records for wild olive (L. intermedius and L. lusitanicus). We also found evidence of some geographic species associations to western (viz. L. alvegus, L. intermedius, L. lusitanicus, L. onubensis sp. nov., L. vineacola, L. vinearum, L. wicuolea sp. nov.) and eastern distributions (viz. L. indalus sp. nov.), while only L. magnus was detected in both areas. We developed a comparative study by considering morphological and morphometrical features together with molecular data from nuclear ribosomal RNA genes (D2–D3 expansion segments of 28S, ITS1, and partial 18S). Results of molecular and phylogenetic analyses confirmed the morphological hypotheses and allowed the delimitation and discrimination of six new species of the genus described herein and four known species. Phylogenetic analyses of Longidorus spp. based on three molecular markers resulted in a general consensus of these species groups, since lineages were maintained for the majority of species. This study represents the most complete phylogenetic analysis for Longidorus species to date., Financial support was received by Projects AGL-2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain, Project P12-AGR-1486 from ‘Consejería de Economía, Innovación y Ciencia’ of Junta de Andalucía, and FEDER financial support from the European Union is gratefully acknowledged. The grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), also provided partial financial support., Peer reviewed

This work has studied for the first time the structure and diversity of plant-parasitic nematodes (PPNs) infesting olive orchard soils in a wide-region in Spain that included 92 locations. It aims at determining which agronomical or environmental factors associated to the olive orchards are the main drivers of the PPNs community structure and diversity. Classical morphological and morphometric identification methods were used to determine the frequency and densities of PPNs. Thirteen families, 34 genera and 77 species of PPNs were identified. The highest diversity was found in Helicotylenchus genus, with six species previously reported in Spain and with H. oleae being a first report. Neodolichorhynchus microphasmis and Diptenchus sp., Diphtherophora sp., and Discotylenchus sp., usually considered fungal feeders, were also reported for the first time associated to olive rhizosphere. PPNs abundance ranged from 66 to 16,288 individuals/500-cm3 of soil with Helicotylenchus digonicus being the most prevalent species, followed by Filenchus sp., Merlinius brevidens and Xiphinema pachtaicum. Nematode abundance and diversity indexes were influenced by olive cultivar, and orchard and soil management practices; while olive variety and soil texture were the main factors driving PPN community composition. Soil physicochemical properties and climatic characteristics most strongly associated to the PPN community composition included pH, sand content and exchangeable K, and maximum and minimum average temperature of the sampled locations. Our data suggests that there is a high diversity of PPNs associated to olive in Southern Spain that can exert different damage to olive roots depending on the olive variety and their abundance. Further analysis to determine the resistance levels of most common olive varieties to the prevalent PPNs in Spain will help to choose the most appropriate ones for the establishment of new plantations. This choice will take into consideration the specific soils and environments where those olive varieties will be established., Financial support was received by Projects AGL2008-00344/AGR from ‘Ministerio de Ciencia e Innovación’, Project AGL-2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain and Projects P10-AGR-5908 and P12-AGR-1486 from ‘Consejería de Economía, Innovación y Ciencia’ of Junta de Andalucía, and FEDER financial support from the European Union is gratefully acknowledged. The grant 219262 ArimNET_ERANET FP7 2012-2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), and the ‘Internacional del Aceite de Oliva Español-Citoliva’ also provided partial financial support., Peer reviewed

Xiphinema herakliense n. sp. from wild olive and cultivated trees in Vathy Rema and other localities, Heraklion province of Crete, Greece, is described based on morphological and molecular characters. The new species is characterised by having differentiation in the tubular part of the uterus, body length 4.1 (3.3-5.5) mm, odontostyle 149 (135-163) μm long, odontophore 85 (71-98) μm long, female tail dorsally convex-conoid with subdigitate peg, and presence of functional males. The polytomous identification code of the new species is: A4, B2 + 3, C5a, D6(5), E5, F4(5), G3, H2, I3, J5, K5, L2. The new species belongs to morphospecies Xiphinema non-americanum Group 5 characterised by having two equally developed female genital branches with uterine differentiation, presence of pseudo-Z-organ with small granular bodies plus small spines (in low number) and crystalloid bodies along uterus, and a short, rounded tail ending in a peg. Xiphinema herakliense n. sp. appears closely related to X. barense, X. horvatovicae, X. lanceolatum, X. loteni, X. sphaerocephalum and X. thorneanum, from which it may be differentiated morphologically and/or molecularly. The results of the phylogenetic analyses based on the sequences of the D2-D3 expansion regions of the 28S rRNA gene and ITS1 rRNA genes are also supportive of its specific status., The present research was partially funded 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’., Peer reviewed

Low density of an unknown root-knot nematode was found on wild olive soils at Cape Spartel near Tanger city in northern Morocco. Morphometry, esterase and malate dehydrogenase electrophoretic phenotypes, as well as ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) sequences demonstrated that this nematode species differs clearly from other previously described root-knot nematodes. The species is herein described, illustrated and named as Meloidogyne spartelensis n. sp. This new root-knot nematode can be morphologically distinguished from other Meloidogyne spp. by: (i) roundish perineal pattern, dorsal arch low, with fine, sinuous cuticle striae, lateral field faintly visible; (ii) female excretory pore posterior to stylet knobs, EP/ST ratio 1.4-2.0; (iii) second-stage juveniles with hemizonid located 1 to 2 annuli anterior to excretory pore and long, sub-digitate tail; and (iv) males with lateral field composed of four incisures, with areolated outer bands. Phylogenetic trees based on 18S, ITS1-5.8S-ITS2, D2-D3 of 28S rDNA, and partial coxII-16S rRNA and coxI gene of mtDNA showed that M. spartelensis n. sp. belongs to an undescribed root-knot nematode lineage that is clearly separated from other species with resemblance in morphology, such as M. dunensis, M. kralli, and M. sewelli., This research was supported by grant KBBE219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Agence Nationale de la Recherche (ANR, France) and Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA, Spain), grant Long-Term Mission Program from Institut de Recherche pour le Développement (IRD, France), PhD grant from Tishreen University (Syrian Arab Republic), grant ANR-10-LABX-41 from LABEX TULIP (France), 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”., Peer reviewed

Stunt nematodes are characterized by phenotypic plasticity, with overlapping morphology and morphometry leading to potential misidentification. Consequently, the application of integrative taxonomic approaches is useful to species delimitation based on a combination of different perspectives, e.g. morphology and DNA sequences. We conducted nematode surveys in cultivated and natural environments in Spain and the USA, from which we identified 18 known species of the family Telotylenchidae and two new taxa within the studied samples. These species were morphologically, morphometrically, and molecularly characterized. The results of light and scanning electron microscopic observations, and molecular and phylogenetic analysis also allowed two new species to be distinguished, described herein as Bitylenchus hispaniensis sp. nov. and Tylenchorhynchus mediterraneus sp. nov. The phylogenetic analysis was carried out using molecular data from nuclear ribosomal DNA genes [D2–D3 expansion segments of the large ribosomal subunit (28S), internal transcribed spacer (ITS), and partial small ribosomal subunit (18S)]. We also provide here a test of alternative hypotheses that confirms the monophyly of both Tylenchorhynchus and Bitylenchus sensu Siddiqi's classification but does not support Fortuner & Luc's conceptual view of Tylenchorhynchus as a large genus. Ancestral state reconstructions of several diagnostic morphological characters using a maximum parsimony approach showed congruence in morphological and molecular evolution for stylet knob inclination and tail tip annulation. Our analysis emphasizes some of the problems related to the taxonomy and phylogeny of nematodes of Telotylenchinae. © 2014 The Linnean Society of London, The last author acknowledges support from 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 Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), grant AGL2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain, grants P12-AGR 1486 and AGR-136 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, grant AGL2009-06955 from ‘Ministerio de Ciencia e Innovación’ of Spain, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’. The fifth author acknowledges the support from the Russian Foundation of Basic Research, project number 14-04-00953 A., Peer reviewed

Subbotin, Sergei A. et al., The genus Hemicycliophora (Nematoda: Hemicycliophoridae) contains 132 valid species of plant-parasitic nematodes, collectively known as ‘sheath nematodes’. Hemicycliophora spp. are characterized morphologically by a long stylet with rounded basal knobs and a cuticular sheath, present in juvenile and adult stages. Populations of 20 valid and 14 putative species of Hemicycliophora and Loofia from several countries were characterized morphologically using light (LM) and scanning electron microscopy (SEM) and molecularly using the D2-D3 segments of 28S rRNA and internal transcribed spacer (ITS) rRNA gene sequences. LM and SEM observations provided new details on the morphology of these species. PCR-restriction fragment length polymorphisms (PCR-RFLPs) of the D2-D3 of 28S rDNA were proposed for identification of the species. Phylogenetic relationships within populations of 36 species of the genus Hemicycliophora using 102 D2-D3 of 28S rDNA and 97 ITS rRNA gene sequences as inferred from Bayesian analysis are reconstructed and discussed. Ancestral state reconstructions of diagnostic characters (body and stylet length, number of body annuli, shape of vulval lip and tail), using maximum parsimony and Bayesian inference, revealed that none of the traits are individually reliable characters for classifying the studied sheath nematode. The Shimodaira–Hasegawa test rejected the validity of the genus Loofia. This is the most complete phylogenetic analysis of Hemicycliophora species conducted so far. © 2014 The Linnean Society of London, P. Castillo also acknowledges support from grant AGL2009-06955 from ‘Ministerio de Ciencia e Innovación’ of Spain, grant AGR-136 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, Union Europea, Fondo Europeo de Desarrollo regional, Una manera de hacer Europa, and grant 219262 ARIMnet_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)., Peer reviewed

Trabajo presentado en el XVII Congreso de la Sociedad Española de Fitopatología, celebrado en Lleida del 7 al 10 de octubre de 2014., Las infecciones de olivo por nematodos fitoparásitos se han asociado con reducciones del
crecimiento y vigor de la planta en varios países de la Cuenca Mediterránea . Sin embargo, no
existen estudios sistematizados sobre la importancia y ecología de nematodos fitoparásitos que
infectan/infestan olivar en Andalucía, así como sobre los factores climáticos, agronómicos o del
cultivo, que determinan su distribución e interacción con la Verticilosis .
En este trabajo se han realizado prospecciones fitonematológicas sistematizadas en 60 de las 69
zonas biológicas de las ocho provincias Andaluzas establecidas por la Red de Información y Alerta
Fitosanitaria del olivar de Andalucía, que delimita áreas de cultivo de olivar con características
homogéneas de clima, orografía, hidrología, paisaje, etc ., así como zonas de vegetación natural de
olivo silvestre. Se han identificado infestaciones por más de un centenar de especies de nematodos
fitoparásitos pertenecientes a diferentes grupos tróficos de los cuales, uno de los grupos con mayor
extensión y biodiversidad fue el de los ectoparásitos migratorios del grupo de longidóridos que incluye
los géneros
Longidorus
(10 especies) y
Xiphinema
(24 especies) . La integración de caracterizaciones
convencionales (morfológicas y morfométricas) junto con análisis moleculares y filogenéticos
basados en ADN ribosómico (fragmentos D2-D3 del gen 28S, 18S, región ITS), y con análisis de
datos multivariantes han demostrado una elevada diversidad críptica (especies morfológicamente
indistinguibles pero filogenéticamente distantes) en el complejo del grupo
Xiphinema americanum
que
comprende alrededor de 60 especies cosmopolitas y de difícil identificación. El diagnóstico preciso en
este grupo es particularmente importante por incluir especies de cuarentena (grupos EPPO A1 y A2) y
vectores de virus vegetales de importancia económica .
Este trabajo demuestra la importancia, distribución y diversidad de nematodos fitoparásitos en
olivo en Andalucía así como la necesidad de integración de diferentes abordajes para el diagnóstico
rápido y preciso de los mismos . La información generada permitirá determinar los mecanismos
potenciales de dispersión de estos agentes y los factores que facilitan y/o determinan sus ataques
en olivar, lo que facilitará el desarrollo y aplicación de estrategias eficientes para su manejo
integrado., Investigación financiada por AGL2012-37521-MINECO, ArimNET_ERANET-219262-INIA, P12-AGR 1486-CICE, y fondos FEDER (EU)., No

Orobanche and Phelipanche species (the broomrapes) are root parasitic plants, some of which represent serious weed problems causing heavy yield losses on important crops. Current control relies on the use of certain agronomic practices, resistant crop varieties, and herbicides, albeit success has been marginal. Agronomic practices such as the use of allelopathic species in intercropping or cover crops, or the use of direct seedling over residues of allelopathic species incorporate the principle of allelopathy exerted by molecules exuded from roots or released by crop residues to control broomrapes. In addition, the isolation of natural substances from root exudates of plants with potential to inhibit broomrape development opens the door to the design of new herbicides based on natural and benign sources. Ryecyanatines A and B and ryecarbonitrilines A and B, the first new substituted cyanatophenol, substituted cyanatobenzo[1,3]dioxole, and the latter two new substituted benzo[1,3]dioxolecarbonitriles were isolated from rye (Secale cereale L.) root exudates. They were characterized as 4-cyanato-2-methoxyphenol, 2-cyanato-benzo[1,3]dioxole, 2-methoxybenzo[1,3]dioxole-5-carbonitrile and benzo[1,3]dioxole-2-carbonitrile by spectroscopic (essentially NMR and HRESI MS spectra) methods. These compounds were investigated for allelopathic activity on Orobanche germination and development. Ryecarbonitriline A induced germination of Orobanche cumana seeds, and this germination can be considered as suicidal because O. cumana does not parasite rye roots and cannot survive without host resources beyond germination stage. In addition, ryecyanatine A promotes a rapid cessation of O. cumana, Orobanche crenata and Orobanche minor radicle growth with the promotion of a layer of papillae at the radicle tip in O. cumana and O. crenata hampering the contact of the parasite to the host. Ryecarbonitriline B also displayed the same activity although being less active than ryecyanatine A and mainly restricted to O. cumana., This research was carried out in part in the frame of Programme STAR, financially supported by UNINA – Italy and Compagnia di San Paolo – Italy and in part by project FP7-ARIMNet-MEDILEG. This work was carried out within the COST ACTION FA1206 Strigolactones: biological roles and applications.

Mónica Fernández-Aparicio has received the support of the Japan Society for the Promotion of Science Postdoctoral Fellowship ID noP 13390 and from the European Union, in the framework of the Marie-Curie FP7 COFUND People Programme, through the award of an AgreenSkills’ fellowship (under grant agreement noP COFUND-GA-2010-267196)., Peer reviewed

Trabajo presentado en la 5th OLIVEBIOTEQ Conference (International Conference for OliveTrees and Olive Products), celebrada en Ammam (Jordania) del 3 al 6 de noviemnre de 2014.-- Mateille, T. et al.., PESTOLIVE (Contribution of olive history for the management of soil-borne
parasites in the Mediterranean Basin) is a project funded by ARIMNet, an ERANET
action supported by the 7th European Framework Programme and by non-European
Mediterranean countries.
PESTOLIVE (www1.montpellier.inra.fr/CBGP/pestolive) aims at producing knowledge
and tools for a new and efficient management of plant-parasitic nematodes (PPN) and
plant-pathogenic fungi (PPF) in olive (Olea europaea L.) cropping systems and
nurseries, while reducing the use of pesticides. Because of the anthropic continuum
from Olea post-glacial refuges to oleasters (domestication) and then to olive-trees
(breeding and cropping), the fragmentation of the PPN and PPF communities and of
their natural enemies could explain the scattered diversity of the control techniques
(especially resistance rootstocks, biocontrol, cropping strategies) developed and applied
all around the Mediterranean Basin. The novelty of PESTOLIVE is based on i) the
analysis and the management of the parasite diversity (ecology of communities) instead
of controlling emblematic species (population approach) and ii) the involvement of
knowledge about the historical co-adaptation of soil-borne parasite and natural enemies
communities to olive-tree domestication (origins and past assemblages) and breeding
that follows the history of O. europaea around the Mediterranean Basin., ARIMNet is coordinated by Dr. F. Jacquet and M. Ollagnon (INRA, France) and PESTOLIVE is co-funded by ANR (France), GDAR (Turkey), INIA (Spain), IRESA (Tunisia), MENESF CRS (Morocco) and NAGREF (Greece)., No

Mediterranean environments are of most interest to study pea adaptability to terminal drought conditions especially in the current context of global climate change. In our work we have tested nine pea cultivars in five South European and North African locations, characterised by different agro climatic conditions within the Mediterranean climate. Data were processed through the additive main effects and multiplicative interaction analysis. Grain yield, aboveground biomass and flowering date were assessed within each mega-environment with parametric and non-parametric methods, establishing ranks for the genotypes within each condition attending to their stability parameters and mean values. The field analysis revealed HR1 as a wide-adapted genotype, whereas others such as Desso showed the best adaptation in South Mediterranean areas. Our results also highlighted the potential interest of these genotypes and others (i.e. Messire and ZP108) in breeding programs and further studies on drought tolerance., Financial support by Spanish project AGL2014-52871, Andalusian project AGR-2883, Serbian project TR-31024 and the FP7 ERANet-ARIMNET-Medileg is acknowledged., Peer reviewed

Trabajo presentado en el 15th Congress of the Mediterranean Phytopathological Union (Plant health sistaining Mediterranean ecosystems), celebrado en Córdoba (España) del 20 al 23 de junio de 2017., Foliar diseases caused by biotrophic pathogens, such
as rusts and powdery mildews, are major limiting
factors in legume and cereal production worldwide.
Crop protection is largely based on chemical control,
although there is renewed interest in the discovery
of natural products as alternatives to synthetic fungicides., This research was supported by FP7-ARIMNet-MEDILEG
and AGL2014-52871 projects and Programme STAR (A.C.)
financially supported by UniNa and Compagnia di San
Paolo, Italia., Peer reviewed

Trabajo presentado en el 15th Congress of the Mediterranean Phytopathological Union (Plant health sistaining Mediterranean ecosystems), celebrado en Córdoba (España) del 20 al 23 de junio de 2017., Traditional identification of plant-parasitic nematode
species by morphology and morphometric methods
is difficult because of the high morphological variability
which can lead to considerable overlapping of
many characters and ambiguous nematode identification.
It is essential to use several approaches to give
accurate species identification (integrative taxonomy)., This research was financially supported by grants P12-AGR 1486 and AGR-136 from ‘Consejeria de Economia, Innvovacion y Ciencia’ from Junta de Andalucia, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’, grant 219262 ArimNET_ERANET FP7 2012-2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in
the Mediterranean basin’ from Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA), and Project AGL-2012-37521 from `Ministerio de Economía y Competitividad’ of Spain., Peer reviewed

Dagger and needle nematodes included in the family Longidoridae (viz. Longidorus, Paralongidorus, and Xiphinema) are highly polyphagous plant-parasitic nematodes in wild and cultivated plants and some of them are plant-virus vectors (nepovirus). The mitochondrial (mt) genomes of the dagger and needle nematodes, Xiphinema rivesi, Xiphinema pachtaicum, Longidorus vineacola and Paralongidorus litoralis were sequenced in this study. The four circular mt genomes have an estimated size of 12.6, 12.5, 13.5 and 12.7 kb, respectively. Up to date, the mt genome of X. pachtaicum is the smallest genome found in Nematoda. The four mt genomes contain 12 protein-coding genes (viz. cox1-3, nad1-6, nad4L, atp6 and cob) and two ribosomal RNA genes (rrnL and rrnS), but the atp8 gene was not detected. These mt genomes showed a gene arrangement very different within the Longidoridae species sequenced, with the exception of very closely related species (X. americanum and X. rivesi). The sizes of non-coding regions in the Longidoridae nematodes were very small and were present in a few places in the mt genome. Phylogenetic analysis of all coding genes showed a closer relationship between Longidorus and Paralongidorus and different phylogenetic possibilities for the three Xiphinema species., This research was supported by grants P12-AGR 1486 and AGR-136 from ‘Consejeria de Economia, Innvovacion y Ciencia’ from Junta de Andalucia, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’, and grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA). The James Hutton Institute receives financial support from the Scottish Government, Rural and Environment Science and Analyical Services Division., Peer reviewed

The traditional identification of plant-parasitic nematode species by morphology and morphometric studies is very difficult because of high morphological variability that can lead to considerable overlap of many characteristics and their ambiguous interpretation. For this reason, it is essential to implement approaches to ensure accurate species identification. DNA barcoding aids in identification and advances species discovery. This study sought to unravel the use of the mitochondrial marker cytochrome c oxidase subunit 1 (coxI) as barcode for Longidoridae species identification, and as a phylogenetic marker. The results showed that mitochondrial and ribosomal markers could be used as barcoding markers, except for some species from the Xiphinema americanum group. The ITS1 region showed a promising role in barcoding for species identification because of the clear molecular variability among species. Some species presented important molecular variability in coxI. The analysis of the newly provided sequences and the sequences deposited in GenBank showed plausible misidentifications, and the use of voucher species and topotype specimens is a priority for this group of nematodes. The use of coxI and D2 and D3 expansion segments of the 28S rRNA gene did not clarify the phylogeny at the genus level., This research was supported by grants P12-AGR 1486 and AGR-136 from the ‘Consejeria de Economia, Innovacion y Ciencia’ of the Junta de Andalucia, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’, grant 201740E042, “Análisis de diversidad molecular, barcoding, y relaciones filogenéticas de nematodos fitoparásitos en cultivos mediterráneos” from Spanish National Research Council (CSIC), grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soil-borne parasites in the Mediterranean basin’ from the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), and Project AGL-2012-37521 from the ‘Ministerio de Economía y Competitividad’ of Spain., Peer reviewed

The genus Longidorus constitutes a large group of approximately 170 species of plant-ectoparasitic nematodes that are polyphagous and distributed almost worldwide. Some of the species of this genus are vectors of plant viruses. Species discrimination in Longidorus is difficult because the morphology is very conservative, and morphometric characters often overlap, leading to potential misidentification. Integrative taxonomy, based on the combination of molecular analyses with morphology, is a useful and necessary approach in Longidorus species identification. In Spain from 2014 to 2017, we conducted nematode surveys among cultivated and wild plants, from which we identified 13 populations of Longidorus, two of which appeared to represent new species and are described herein as L. iliturgiensis sp. nov. and L. pacensis sp. nov., and 11 populations belonging to eight known species: L. africanus, L. baeticus, L. carpetanensis, L. fasciatus, L. nevesi, L. cf. olegi, L. pini, and L. vallensis. Three species are new geographical records for Spain (L. nevesi, L. cf. olegi, and L. africanus). We report molecular data for L. nevesi, L. cf. olegi, L. carpetanensis and L. pini for the first time. Additionally, we describe the males of L. pini and the juveniles of L. cf. olegi., Our research was supported by grants P12-AGR-1486 from ‘Consejería de Economía, Innovación y Ciencia’ of Junta de Andalucía, 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), and FEDER financial support from the European Union is gratefully acknowledged. Antonio J. Archidona-Yuste is a recipient of the research fellowship BES-2013-063495 from the Spanish ‘Ministerio de Economía y Competitividad’., Peer reviewed

Root-knot nematodes of the genus Meloidogyne are recognised worldwide as a major production constraint in crops of primary economic importance. Knowledge of their diversity and prevalence, as well as the major environmental and agronomical cues for understanding their distribution in specific areas is of vital importance for designing control measures to reduce significant damage. We provide the first detailed information on the diversity, distribution and levels of Meloidogyne species infecting wild and cultivated olive soils in a wide-region in southern Spain that included 499 sampling sites. Overall Meloidogyne spp. were found in 6.6% of sampled olive plants, with 6.6% and 6.5% for cultivated and wild olive, respectively. We identified five previously described Meloidogyne spp. (Meloidogyne arenaria, M. baetica, M. hapla, M. incognita, M. javanica) and one new species (Meloidogyne oleae sp. nov.) which, characterized using integrative taxonomy, increases the known biodiversity of Meloidogyne spp. affecting olive. Meloidogyne arenaria and M. incognita were only found infecting cultivated olive varieties, while, M. baetica was only found infecting wild olive. Three major parameters drive the distribution of Meloidogyne spp. in cultivated olives in southern Spain, cover vegetation on alley, irrigation and soil texture, but different species respond differently to them. In particular the presence of M. incognita is highly correlated with sandy loamy soils, the presence of M. javanica with irrigated soils and cover vegetation, while the presence of M. arenaria is correlated with the absence of cover vegetation on alley and absence of irrigation. These parameters likely influence the selection of each particular Meloidogyne species from a major dispersal source, such as the rooted plantlets used to establish the orchards., This work was supported by Project P12-AGR-1486 from the `Consejería de Economía, Innovación y Ciencia' of Junta de Andalucía to PC, the grant 219262 ArimNET_ERANET FP7 2012-2015 Project PESTOLIVE `Contribution of olive history for the management of soilborne parasites in the Mediterranean basin' from the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) to PC, Project AGL-2012-37521 from the `Ministerio de Economía y Competitividad' of Spain, FEDER financial support from the European Union, and grant 201740E042 “Análisis de diversidad molecular, barcoding, y relaciones filogenéticas de nematodos fitoparásitos en cultivos mediterráneos” from the Spanish National Research Council (CSIC) to PC., Peer reviewed

The genus Paratrichodorus currently comprises 26 species of polyphagous ectoparasitic plant nematodes that are distributed worldwide and include a few tobravirus vector species. Paratrichodorus is one of the most difficult genera for species identification because it is phenotypically conserved, with morphometric characters that often overlap between species. From 2003 to 2016, nematode surveys were conducted in southern Spain, and the present study addresses 22 nematode populations of the Paratrichodorus hispanus group. Among them, two new species, Paratrichodorus almadenensis sp. nov. and Paratrichodorus ramblensis sp. nov., are described here morphologically and molecularly and compared with topotype specimens of Paratrichodorus hispanus. Furthermore, this analysis was complemented with the first molecular data for this species complex and for the genus Monotrichodorus. The phylogeny of family Trichodoridae showed the relationship of Monotrichodorus with Trichodorus, both of which are phylogenetically associated with the P. hispanus species group, including the two newly described species. Several phylogenetic hypotheses about the monophyly of the genera with molecular data composing the family Trichodoridae were tested using Shimodaira–Hasegawa and approximately unbiased tests. This work also includes the first observations of copulatory plugs in Paratrichodorus, which clearly differ from the different types of mating plugs described in Trichodorus species., This research was supported by grants P12-AGR 1486 and 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’, grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), grant AGL2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain, and grant 201740E042, ‘Análisis de diversidad molecular, barcoding, y relaciones filogenéticas de nematodos fitoparásitos en cultivos mediterráneos’ from the Spanish National Research Council (CSIC). A. J. Archidona-Yuste is a recipient of research contract BES-2013-063495 from ‘Ministerio de Ciencia e Innovación’ of Spain. C. Gutiérrez-Gutiérrez gratefully acknowledges the support of the Prometeo Project from SENESCYT, Ecuadorian Government, and from the Portuguese Foundation for Science and Technology (FCT) through a postdoctoral fellowship (SFRH/ BPD/95315/2013)., No

The spiral nematode Helicotylenchus oleae is an ectoparasite of olive roots and is distributed in some countries in the Mediterranean Basin. In this study, we provided morphological and molecular characterisation of topotypes from southern Italy, as well as of several other populations from Crete (Greece) and Spain. Correct identification of plant-parasitic nematode species is essential to establish appropriate control strategies and for preventing their spread to other areas. Helicotylenchus oleae is reported for the first time in Greece (Crete). Integrative morphometric and molecular data for H. oleae populations using D2-D3 expansion segments of 28S rDNA, ITS-rDNA, and the cytochrome c oxidase subunit 1 (coxI), were in agreement with the original descriptions of the species, except for some minor differences, which may be a result of intraspecific variability. The phylogenetic relationships of this species with other representatives of Helicotylenchus spp. using D2-D3 expansion segments and the ITS-rRNA region was studied showing a low intraspecific diversity for H. oleae species. For the first time, coxI molecular data is obtained for the genus Helicotylenchus., This research was supported by grant P12-AGR 1486 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’, grant 201740E042, "Análisis de diversidad molecular, barcoding, y relaciones filogenéticas de nematodos fitoparásitos en cultivos mediterráneos" from Spanish National Research Council (CSIC), 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), and AGL-2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain., Peer reviewed

Nematode samplings in cultivated and wild olive in Crete, Greece, yielded the presence of Bitylenchus hispaniensis, Helicotylenchus microlobus, Helicotylenchus vulgaris, Merlinius brevidens, and Pratylenchoides alkani. With the exception of H. microlobus and M. brevidens, reports of these plant-parasitic nematode species constitute new records for Greece. Bitylenchus hispaniensis is also reported for first time in a country outside of Spain, where it was originally described. Pratylenchoides alkani is herein reported for the second time in the Mediterranean area and for the first time in association with olive. Two further populations of H. microlobus and H. vulgaris, from walnut and goji berry from Greece, were identified. Molecular data for all of these nematode species are provided, resulting in the first integrative identification of these Greek populations., This research was partially funded by grant KBBE 219262 ArimNET-ERANET FP7 2012 to 2015 Project PESTOLIVE “Contribution of olive history for the management of soilborne parasites in the Mediterranean basin” from the 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.” Maria S. Lazanaki was employed by the ARIMNET-PESTOLIVE project., Peer reviewed

The reniform nematodes of the genus Rotylenchulus are semi-endoparasites of numerous herbaceous and woody plant roots and distributed in regions with Mediterranean, subtropical and tropical climates. In this study, we provide morphological and molecular characterisation of three out of 11 valid species of the genus Rotylenchulus: R. macrodoratus, R. macrosoma, and R. reniformis from Greece (Crete), Italy and Spain. The overall prevalence of reniform nematodes in wild and cultivated olives in Greece, Italy, and Spain was 11.5%, 19.0% and 0.6%, respectively. In Greece, R. macrodoratus and R. macrosoma were detected in cultivated olive with a prevalence of 8.2% and 6.2%, respectively, but none of them were found in wild olive. This is the first report of R. macrosoma in Greece. Only one reniform nematode species was detected in olive from Italy and Spain, viz. R. macrodoratus and R. macrosoma, respectively. The parasitism of R. macrosoma on hazelnut in northern Spain was also confirmed for the first time. This study demonstrates that R. macrodoratus and R. macrosoma have two distinct rRNA gene types in their genomes, specifically the two types of D2-D3 for R. macrosoma and R. macrodoratus, the two types of ITS for R. macrodoratus and the testing of the ITS variability in other R. macrosoma populations in different countries. Rotylenchulus macrosoma from Greece and Spain showed differences in nucleotide sequences in the ITS region and D2-D3 of 28S rRNA gene., This research was supported by grant P12-AGR 1486 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’, 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), and AGL-2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain. I. G. Birmpilis was employed by the ARIMNET-PESTOLIVE project., Peer reviewed

Olive (Olea europaea L.) is one of the most important fruit crops in the Mediterranean Basin, because it occupies significant acreage in these countries and often has important cultural heritage and landscape value. This crop can be infected by several Meloidogyne species (M. javanica, M. arenaria, and M. incognita, among others), and only a few cultivars with some level of resistance to these nematodes have been found. Innovations in intensive olive growing using high planting densities, irrigation, and substantial amounts of fertilizers could increase the nematode population to further damaging levels. To further understand the interactions involved between olive and pathogenic nematodes and in the hope of finding solutions to the agricultural risks, this research aimed to determine the reaction of important olive cultivars in Spain and wild olives to M. javanica infection, including genotypes of the same and other O. europaea subspecies. All olive cultivars tested were good hosts for M. javanica, but high levels of nematode reproduction found in three cultivars (Gordal Sevillana, Hojiblanca, and Manzanilla de Sevilla) were substantially different. In the wild accessions, O. europaea subsp. cerasiformis (genotype W147) and O. europaea subsp. europaea var. sylvestris (genotype W224) were resistant to M. javanica at different levels, with strong resistance in W147 (reproduction factor [Rf] = 0.0003) and moderate resistance in W224 (Rf = 0.79). The defense reaction of W147 to M. javanica showed a strong increase of phenolic compounds but no hypersensitive reaction., This research was supported by grants P12-AGR 1486 and AGR-136 from the ‘Consejeria de Economia, Innovacion y Ciencia’ of the Junta de Andalucia, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’, grant 201740E042, “Análisis de diversidad molecular, barcoding, y relaciones filogenéticas de nematodos fitoparásitos en cultivos mediterráneos” from Spanish National Research Council (CSIC), grant 219262 ArimNET_ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soil-borne parasites in the Mediterranean basin’ from the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)., No

Datasets presented here were employed in the main work “Spatial structure and soil properties shape local community structure of plant-parasitic nematodes in cultivated olive trees in southern Spain” Archidona-Yuste et al., 2020. In this research, we aimed to unravel the diversity of plant-parasitic nematodes (PPN) associated with cultivated olive (Olea europaea subsp. europaea var. europaea) in southern Spain, Andalusia. The olive growing area of Andalusia is of high agriculture and socio-economic importance with an extensive distribution of this crop. To this end, we conducted a systematic survey comprising 376 commercial olive orchards covering the diversity of cropping systems applied. Data showed 128 species of PPN belonging to 38 genera and to 13 families. In addition, an extensive data set regarding to potential factors in structuring the community patterns of PPN found in the 376 commercial olive orchards sampled is provided. Three variables data set were compiled including above-ground environment, soil and agronomic management. Overall, 48 explanatory variables were selected as determinist processes on shaping the diversity of PPN. Finally, data also showed the values regarding to the partition of beta diversity into contributions of single sites to overall beta diversity (LCBD) and intro contributions of individual species to overall beta diversity (SCBD). Data may serve as benchmarks for other groups working in the field of PPN diversity associated with crops and of belowground communities and ecosystems., This research is part of the PhD project of the first author and was supported by grant AGL2012-37521 from Ministerio de Economía y Competitividad of Spain, grant 219262 ArimNET-ERANET FP7 2012–2015 Project PESTOLIVE and grant P12AGR 1486 Consejería de Economía, Innvovación y Ciencia of the Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’. A. Archidona-Yuste was a recipient of research contract BES-2013063495 from Ministerio de Economía y Competitividad, Spain; and at present is a recipient of Humboldt Research Fellowship for Postdoctoral Researchers at Helmholtz Centre for Environmental Research-UFZ, Leipzig, Germany., Peer reviewed

This article belongs to the Special Issue Recent Advances in Genomics and Genetics of Fruit Trees., Prunus species are important crops in temperate regions. In these regions, drought periods are predicted to occur more frequently due to climate change. In this sense, to reduce the impact of climate warming, obtaining new tolerant/resistant cultivars and rootstocks is a mandatory goal in Prunus breeding. Therefore, the current study assembled three Prunus species including almond, (P. dulcis Mill D.A. Webb), apricot (P. armeniaca L.) and peach (P. persica L.) to model the temporal effects of drought. A hybrid peach × almond and a wild almond-relative species Prunus webbii were also included in the study. Physiological traits associated with photosynthetic activity, leaf water status, and chlorophyll content were assessed under three watering treatments. Results showed that effects of time, genotype, and treatment interact significantly in all traits. In addition, results confirmed that P. webbii have a greater tolerance to drought than commercial rootstocks. However, “Real Fino” apricot showed the fastest recovery after re-irrigation while being one of the most affected cultivars. In addition, from the better response to these watering treatments by the almond genotypes, two different trends were observed after re-irrigation treatment that clearly differentiate the response of the almond cultivar “Garrigue” from the rest of Prunus genotypes. A better characterization of the short-term drought response in Prunus, an accurate and more efficient evaluation of the genotype effect was obtained from the use of mixed models considering appropriate variance–covariance structures. Although the advantages of these approaches are rarely used in Prunus breeding, these methodologies should be undertaken in the future by breeders to increase efficiency in developing new breeding materials., Peer reviewed, This research was supported by the project “Selection and characterization of drought resistant almond cultivars from the Mediterranean basin with high nutraceutical values” (Nut4Drought) of the ARIMNET-2 European Program.

Numerous studies have documented the distribution of plant and animal communities with respect to spatial structure; however, relatively little is known about the involvement of spatial structures in the diversity of soil organisms such as plant-parasitic nematodes (PPN). Host plants such as olive trees are of particular interest because they host a large number of PPN and have a high economic and cultural importance. In this study, we investigated how different aspects of the environment (i.e. the factors soil, above-ground environment, and agricultural management) and spatial structure shaped the variation of species composition (expressed as beta diversity) and species richness of plant-parasitic nematodes infesting the soil rhizosphere in 376 commercial olive orchards in the south of Spain. We used variation partitioning to assess the relative importance of the unique and shared contributions of the factors describing the environment and spatial structure. To identify sites and species of particular interest, we partitioned beta diversity into local and species contributions.

Contrary to our expectation that soil and agricultural management would largely determine the community structure of PPN, more than two-thirds of the variation remained unexplained. Spatial structure and soil were the most important factors shaping species richness and beta diversity. Surprisingly, the effects of agricultural management on species richness were lower than expected, and null [or nonexistent, or nonsignificant] on beta diversity. We found relatively high levels of shared contributions of the different factors, especially in combination with spatial structure, indicating the presence of spatial gradients of the variables describing the environmental factors.

Species contributions to beta diversity (SCBD) were positively correlated with nematode prevalence and density range; thus, SCBD could be related to the niche position as reported in other ecosystems. Local contributions to beta diversity (LCBD) were mainly related with habitat filtering mechanisms (e.g. soil physiochemical and agronomic management predictors), suggesting a relationship between nematode total biomass and ecological gradients. Overall, we revealed novel insights into the spatial structure of PPN communities and showed that its beta diversity is less structured by spatial and environmental factors compared to other organism types., This research was supported by grant AGL2012-37521 from Ministerio de Economía y Competitividad of Spain, grant 219262 ArimNET-ERANET FP7 2012-2015 Project PESTOLIVE and grant P12AGR 1486 Consejería de Economía, Innvovación y Ciencia of the Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’. A. Archidona-Yuste was a recipient of research contract BES-2013063495 from Ministerio de Economía y Competitividad, Spain; and at present is a recipient of Humboldt Research Fellowship for Postdoctoral Researchers at Helmholtz Centre for Environmental Research-UFZ, Leipzig, Germany., Peer reviewed

The present study is part of a survey for the identification of plant-parasitic nematodes in the rhizosphere of cultivated and wild olive trees in Crete, Greece. Sixteen species corresponding to 13 genera are added to 20 species belonging to 8 genera, previously reported in the survey. Seven nematode species, Filenchus ditissimus, Filenchus vulgaris, Ogma civellae, Pratylenchoides crenicauda, Psilenchus hilarulus, Tylenchus elegans, and Zygotylenchus guevarai, are recorded for the first time in Greece., The present research was funded 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’ conducted by the 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’.

The stubby-root nematode genus Paratrichodorus comprises species of plant polyphagous ectoparasitic nematodes distributed worldwide and a few of them are plant virus-vectors (tobravirus). This is one of the most difficult nematode genera in terms of species identification, since it is phenotypically conserved with closely related species and is highly sensitive to the specimen fixation and preservation processes. Using an integrative and multidisciplinary taxonomy framework, we determined species boundaries among the Paratrichodorus populations from a P. allius species-complex sampled in cultivated and natural ecosystems across Spain. We support the hypothesis that nematode diversity of the P. allius species-complex remains underestimated and describe here a total of eight new species on the basis of detailed morphological and molecular characterization of 16 Paratrichodorus populations: Paratrichodorus asidonaensis sp. nov., Paratrichodorus benalupensis sp. nov., Paratrichodorus hinogerus sp. nov., Paratrichodorus litoralis sp. nov., Paratrichodorus paraallius sp. nov., Paratrichodorus rocianerus sp. nov., Paratrichodorus tarifaensis sp. nov., and Paratrichodorus uliaensis sp. nov. This study emphasizes the importance of properly applying a broad taxonomic framework in plant-parasitic nematodes. It is also relevant for plant health, since it correctly delimits species morphologically closed to the virus vector nematode, P. allius, and gives proper molecular markers for its unequivocal identification., This research was supported by grant P12-AGR 1486 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’, grant 219262 ArimNET_ERANET FP7 2012-2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), and grant 201740E042, ‘Análisis de diversidad molecular, barcoding, y relaciones filogenéticas de nematodos fitoparásitos en cultivos mediterráneos’ from Spanish National Research Council (CSIC)., Peer reviewed