Sasanelli, N., Vovlas, N., Trisciuzzi, N., Cantalapiedra-Navarrete, C, Palomares-Rius, J. E., and Castillo, P. 2013. Pathogenicity and host-parasite relationships oí Heterodera cruciferae in cabbage. Plant Dis. 97:333-338.Stunted cabbage {'Lupini') associated with severe soil infestations by a cyst-forming nematode were observed in large patches of open fields in Castellaneta, province of Taranto, southern Italy. Morphological traits based on mature cysts, males, and second-stage juveniles (J2s) and molecular analysis of ribosomal DNA (D2 to D3 expansion segments of 28S and internal transcribed spacer [ITS]1 region) were used to identify the species. ITSl sequence information supported the identity of Heterodera cruciferae, also showing a high degree of similarity to other species of the Heterodera Goettingiana group, including H. goettingiana, H. carotae, and H. urticae. Nematodes successfully established permanent feeding sites in cabbage roots which caused cellular alterations in the root cortex, endodermis, pericycle, and vascular cylinder by inducing typical multinucleate syncytia. Syncytial cytoplasm was granular and dense, with variously sized vacuoles and hypertrophied nuclei with nucleoli. Cabbage plant growth was also reduced in pathogenicity tests. The relationship between the initial nematode population density in soil and shoot plant weight was well described by the Seinhorst's equation. Tolerance limits with respect to shoot plant weight of cabbage to H. cruciferae was estimated as 1.50 units of eggs plus J2s/cm^ of soil. The minimum relative value (m) for plant height was 0.71 at an initial nematode population density of (Pi) > 64 units of eggs plus J2s/cm^ of soil. The maximum nematode reproduction rate {Pf/Pi) was 4.6 times that of the initial population density of 8 units of eggs plus J2s/cm^ of soil.

Sanitary selection and certification of olive cultivars require sensitive diagnostic methods and effective sanitation protocols. Although much attention has been paid in the past few years to the development of diagnostic tools for reliable virus identification, the need to define a common and standardized diagnostic protocol led to the implementation of a ring test among nine Italian diagnostic laboratories. A one‐step RT‐PCR protocol and different primer sets, targeting the most common olive viruses covered by phytosanitary rules, were tested in each laboratory, using the same batch of positive and healthy controls as well as the same amplification conditions and reaction components. The one‐step RT‐PCR, performed using several specific primer sets, was able efficiently to detect the target viruses in all laboratories. Furthermore, a one‐step RT‐PCR protocol was used successfully for the first time for detection of Tobacco necrosis virus (TNV) and Olive mild mosaic virus (OMMV). Results showed that all target viruses were not uniformly distributed in the canopy, and that at least two subsets of samples must be collected from each plant. This standardized protocol is now being used to produce nuclear stocks for 70 different Italian olive cultivars, in the framework of the national project OLVIVA, which involves 25 national research institutions.

The non-cyst forming heteroderid nematode Cryphodera brinkmani was detected in Italy parasitizing roots of Japanese white pine bonsai (Pinus parviflora) trees imported from Japan. Morphology and morphometrical traits of the intercepted population on this new host for C. brinkmani were in agreement with the original description, except for some minor differences on male morphology. Integrative molecular data for this species were obtained using D2-D3 expansion regions of 28S rDNA, ITS1-rDNA, the partial 18S rDNA, and the protein-coding mitochondrial gene, cytochrome oxidase c subunit I (COI). The phylogenetic relationships of this species with other representatives of non-cyst and cyst-forming Heteroderidae using ITS1 are presented and indicated that C. brinkmani clustered together with other Cryphodera spp. and with Meloidodera alni suggesting a monophyletic origin of non-cyst forming nematodes (Heteroderinae sensu Luc et al. 1988), which have been considered close to the ancestor of most species of Heteroderidae. Histological observations of P. parviflora feeder roots infected by C. brinkmani indicated that nematode females induce similar anatomical alterations to those reported for C. kalesari and consisting of formation of a single uninucleate giant cell (nurse cell) with hypertrophied nucleus, prominenet nucleolus, thickened cell wall and expanding into the stele and in contact of xylem, vacuum cambium and phloem. These findings are in agreement with the results of the phylogenetic analysis and indicate a close relationship in the plant responses induced by Cryphodera nematode females with those caused by the genetically related Meloidodera spp., which also induce formation of a uninucletate giant cell.

A survey of nematodes associated with the red palm weevil Rhynchophorus ferrugineus was conducted in southern Italy in 2015 and 2016 in order to create a species inventory and obtain data about nematode biodiversity. A total of 70 insect samples (pupae and adults) were collected from infested Phoenix canariensis, Phoenix dactylifera, and Chamaerops humilis palms in three Italian Regions: sampling took place at 11 locations in Apulia, 1 in Basilicata, and 1 in Sardinia regions. Individual insects were dissected to determine nematode presence, and different nematode species were also recovered from red palm weevil cocoons collected at the sites in Apulia. Individual nematodes were molecularly identified by sequencing the ITS, D2‐D3 expansion domains of the 28SrRNA gene and the mitochondrial COI and inferring the phylogenetic relationships. The insect‐associated nematofauna identified belonged to the families Rhabditidae, Cephalobidae, and Diplogastridae. Just two nematode species, Teratorhabditis synpapillata and Mononchoides macrospiculum, were always found in association with adult insects and cocoons taken from all sampling sites. This paper reports on the biodiversity of the nematodes associated with R. ferrugineus and on current knowledge of the specific habitat of specialized and divergent entomophilic nematodes.

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