Alien scale insects (Hemiptera Coccoidea) in European and Mediterranean countries: the fate of new and old introductions

This contribution focuses on recent interceptions and introductions of alien scale insects and their current distribution in European and Mediterranean countries. Data and collections were gathered in markets, nurseries, and botanical gardens, mostly in Italy, either indoors or outdoors. New or recent records of the following alien species are presented: Exallomochlus hispidus (Morrison); Ferrisia virgata (Cockerell) (Pseudococcidae); Coccus viridis (Green); Milviscutulus mangiferae (Green) (Coccidae); Aonidiella orientalis (Newstead); Aspidiotus destructor Signoret; Aulacaspis tubercularis Newstead; Fiorinia fioriniae Targioni Tozzetti; Lepidosaphes pinnaeformis (Bouché); Pseudaulacaspis brimblecombei Williams (Diaspididae). New data and pest status of Phoenicococcus marlatti Cockerell (Phoenicococcidae) and Trabutina mannipara (Hemprich & Ehrenberg) (Pseudococcidae) are also reported. The possible repeated introductions of the latter from North Africa to south Italy by trans-Mediterranean winds, is hypothesized.     

Plant ferredoxin-like protein enhances resistance to bacterial soft rot disease through PAMP-triggered immunity in Arabidopsis thaliana

The protection of plants against bacterial disease is one of the important issues that need to be studied in agricultural applications. The application of a transgene, such as a gene that encodes plant ferredoxin-like protein (PFLP), to generate resistant plants is one possible strategy. Our previous reports have demonstrated that transgenic plants that express extracellular PFLP (ESF plants) are more resistant to bacterial pathogens. This protein intensifies the hypersensitive response (HR) in plants when they are infiltrated by a pathogen-associated molecular pattern (PAMP), harpin (HrpZ), from Pseudomonas syringae. In addition, this intensified HR is associated with the expression of membrane-bound NADPH oxidase. Thus, we attempted to determine the involvement of PFLP in intensifying PAMP-triggered immunity (PTI) to enhance disease resistance. First, we showed that transgenic Arabidopsis plants with the pflp gene were resistant to bacterial soft rot caused by Pectobacterium carotovorum subsp. carotovorum (Pcc). Then, the fliC gene which encoded flagellin from Pcc was cloned and expressed. The FliC protein was used in the functional study with PFLP in Arabidopsis Col-0 plants. The reactive oxygen species (ROS) generation and HR ratio were induced by the treatment with both PFLP and FliC together, but they were not induced by treatment with PFLP or FliC alone. Similar results were confirmed in ESF plants, where FliC elicited rapid ROS accumulation and callose deposition. Moreover, we demonstrated that the PFLP-intensified ROS generation and HR were related to Ca²⁺ influx and activation of NADPH oxidase. We concluded that the PFLP-intensified disease resistance is associated with the intensification of PAMP-triggered immunity.     

Analysis of genetic and virulence variability of Stemphylium lycopersici associated with leaf spot of vegetable crops

Stemphylium lycopersici (Enjoji) W. Yamam was initially described from tomato and has been reported to infect different hosts worldwide. Sequence analyses of the internal transcribed spacer (ITS) regions 1 and 2, including 5.8S rDNA (ITS-5.8S rDNA) and glyceraldehyde-3-phosphate dehydrogenase (gpd) gene, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), as well as virulence studies were conducted to analyze 46?S. lycopersici isolates. Stemphylium lycopersici isolates used in this study were obtained from diseased tomato (Solanum lycopersicum L.), eggplant (Solanum melongena L.), pepper (Capsicum annuum L.) and lettuce (Lactuca sativa L.) from major vegetable growing regions of Malaysia, including the three states of Pahang, Johor and Selangor between 2011 and 2012. Phylogenetic analysis of a combined dataset of the ITS-5.8S rDNA and gpd regions indicated that all isolates were clustered in the sub-cluster that comprised S. lycopersici, and were distinguished from other Stemphylium species. Cluster analyses using the UPGMA method for both RAPD and ISSR markers grouped S. lycopersici isolates into three main clusters with similarity index values of 67 and 68 %. The genetic diversity data confirmed that isolates of S. lycopersici are in concordance to host plants, and not geographical origin of the isolates. All S. lycopersici isolates were pathogenic on their original host plants and showed leaf spot symptoms; however, virulence variability was observed among the isolates. In cross-inoculation assays, the representative isolates were able to cause leaf spot symptoms on eggplant, pepper, lettuce and tomato, but not on cabbage.     

The overwintering mode ofBemisia tabaci and its parasitoids in Israel

Wild and cultivated plants in the vicinity of Kibbutz Nahshon and a few additional locations in Israel were sampled for the presence ofBemisia tabaci Genna-dius (Homoptera: Aleyrodidae). The whiteflies, together with their parasites,Eretmocerus mundus andEncarsia lutea, were found to develop on numerous host species throughout the winter. Especially high levels were reached onLan-tana camara, Abutilon grandifolium andIpomoea batatas. During late winter and spring the population on these hosts declined. From April onwards the populations increased on potatoes and sunflowers.     

Development of conventional and real-time PCR protocols for specific and sensitive detection of Colletotrichum capsici in chilli (Capsicum annuum L.)

Anthracnose, caused by Colletotrichum capsici, is a major disease of chilli (Capsicum annuum L.) affecting both fruit and seed quality. The pathogen is both internally and externally seedborne. However, a rapid and sensitive method for detection of this pathogen in seeds is currently limited. In this study, a polymerase chain reaction (PCR) method based on sequence characterized amplified region (SCAR) marker was developed for specific and sensitive detection of C. capsici in chilli seeds and fruits. The developed SCAR primers were highly specific to C. capsici and resulted in the amplification of an expected 250-bp fragment from genomic DNA of all seven of the C. capsici isolates tested. No amplification occurred when the SCAR primers were tested with genomic DNA from three other fungal isolates and four other Colletotrichum species. The SCAR primers successfully amplified similar sized fragments from DNA derived from C. capsici-infected chilli fruits. The molecular detection sensitivity of C. capsici was 1 pg of purified C. capsici DNA template and 25 ng of DNA from C. capsici-infected chilli fruits. A real-time PCR assay was also developed using SYBR Green chemistry for detection of C. capsici in chilli fruits and seeds. The standard curve obtained showed a linear correlation between copy number of the cloned target DNA sequence of C. capsici and cycle threshold (Ct) values, with R² of 0.98. These PCR-based assays may be highly useful in detection of this important pathogen in chilli seeds and fruits in plant quarantine laboratories.     

Control of phytophthora leaf blight of taro (Colocasia esculenta) by fungicides and roguing

Six fungicides were evaluated under laboratory and field conditions for control of Phytophthora leaf blight of taro,Coloeasia esculenta, incited byPhytophthora colocasiae. Inin vitro tests Deraosan 65W was the most effective in inhibiting the mycelial growth of the test pathogen followed by Difolatan 80W, Fytolan (copper oxychloride), Apron 35F, Topsin-M 50W and Dithane Z-78 75W. Excellent control was obtained with Demosan 65W and Difolatan 80W, good control with Apron 35F, fair control with Fytolan, and poor control with Topsin-M 50W and Dithane Z-78 75W. Results ofin vivo tests were correlated with those of thein vitro tests. Roguing of infected leaves did not eradicate the pathogen but can only delay epiphytotics.     

Identification and species status of the mango biotype of Colletotrichum gloeosporioides in Ghana

Research work was carried out to identify and ascertain the species status of the mango biotype of Colletotrichum gloeosporioides infecting mangoes in Ghana. Forty five isolates of Colletotrichum species were collected from 12 districts in Ghana while five each were obtained from mango fruits from Florida, Mexico and Puerto Rico. The entire internal transcribed spacer region, partial beta-tubulin gene and partial glyceraldehyde-3-phosphate dehydrogenase gene of isolates were sequenced and used in phylogenetic studies. The results of the sequence analysis of the first ribosomal transcribed spacer (ITS 1) region showed that 35 % of the isolates from Ghana and all the five isolates from Mexico were the mango biotype of C. gloeosporioides, while the others were not. Phylogenetic studies showed that the mango biotype of the pathogen was Colletotrichum asianum but not C. gloeosporioides as previously thought. However, the other isolates that were not the mango biotype were identified as Colletotrichum siamense and Colletotrichum species which had probably cross-infected mango from other fruit crops in the field.     

Survey on viroids infecting grapevine in Italy: identification and characterization of Australian grapevine viroid and Grapevine yellow speckle viroid 2

Five viroid species have been reported from grapevine. Hop stunt viroid (HSVd) and Grapevine yellow speckle viroid 1 (GYSVd-1) are distributed worldwide, whereas Grapevine yellow speckle viroid 2 (GYSVd-2), Australian grapevine viroid (AGVd) and Citrus exocortis viroid (CEVd) are found only sporadically. However, the presence of AGVd and GYSVd-2 in several countries, including China, Turkey and Tunisia, suggests a wider dissemination, possibly also in Europe, where AGVd has never been found and GYSVd-2 has been occasionally identified in Italy. Taking advantage of a multiplex RT-PCR assay recently developed for detecting simultaneously these five viroids, vines growing in Italy in commercial vineyards and germplasm collections were surveyed. Besides confirming the widespread presence of HSVd and GYSVd-1 in the field, GYSVd-2 and/or AGVd were identified in two grapevine table cultivars (Sultanina Bianca and Red Globe) from germplasm collections. Tests extended to vines cultivated in southern Italy confirmed the presence of both viroids, which were further characterized. No major sequence divergences between the AGVd and GYSVd-2 variants from Italy and those previously described from other countries were observed. Phylogenetic analysis supported the close relationships among AGVd variants from Italy, Tunisia and Australia. To our knowledge this is the first report of AGVd in Europe and the first molecular characterization of GYSVd-2 isolates from a European country.     

Effect of avermectin B1 on the development and fecundity of the california red scale,Aonidiella auranth,and on two of its natural enemies:Chilocorus bipustulatus andAphytis holoxanthus

Various immature stages of male and female California red scale,Aonidiella aurantii (Maskell) (Homoptera: Diaspididae), were treated with avermectin B₁ (MK-936) at concentrations ranging from 10 to 100 ppm. Treatments at 10 ppm completely arrested crawler development and 20 and 100 ppm caused 100% mortality of 48-h and 7-day-old male and female 1st instar nymphs, respectively. The development of the 2nd instar was slightly affected by MK-936 at up to 40 ppm. Sublethal doses of the chemical applied to immature stages caused no dramatic reduction in the fecundity of the subsequent, mature females. MK-936 applied to male prepupal and pupal stages and mated females did not prevent male emergence, nor did it affect the ability of males to copulate and inseminate female scales, or the female fecundity. The development and adult emergence of treatedAphytis holoxanthus DeBach (Hymenoptera: Aphelinidae) larvae and pupae parasitizingChrysomphalus aonidum L. scales were not influenced by a 9 ppm MK-936 treatment; leaves sprayed with this concentration had residual toxicity onA. holoxanthus adults for only 24 h post-treatment. Feeding byChilocorus bipustulatus L. (Coleoptera: Coc-cinellidae) on 0.9 ppm MK-936-treated Florida red scales resulted in 100% mortality of 2nd instars and induced sterility in the females.     

Response ofSpodoptera littoralis (Boisd.) andEarias insulana (Boisd.) larvae to azadirachtin and salannin

The effect of azadirachtin and salannin, two triterpenoids isolated from seeds of neem (Azadirachta indica A. Juss), on the feeding response ofSpodoptera littoralis (Boisd.) andEarias insulana (Boisd.) larvae, was investigated. Styropor (foamed polystyrene) lamellae were painted on both sides with a mixture of 5% sucrose with different concentrations of either azadirachtin or salannin dissoled in methanol-water (3∶7). Azadirachtin strongly suppressed feeding inS. littoralis larvae even at 0.001%, whereas salannin showed some antifeedant activity at 0.005% and above. Larvae ofE. insulana were deterred from feeding on azadirachtin-treated lamellae even at 0.005%, whereas salannin was effective only at 0.01% and above. Azadirachtin applied on cotton leaves deterred larvae ofS. littoralis from feeding at all concentrations ranging between 0.001 and 0.02%.     

Characteristics and pathogenicity of six Phytophthora isolates from pot plants

From several greenhouse plants showing foot and root rot symptomsPhytophthora isolates were gathered. Isolates fromPeperomia, Saintpaulia andSinningia were identified asP. nicotianae van Breda de Haan var.nicotianae and an isolate fromBegonia asP. cryptogea Pethybr. & Laff. Cardinal temperatures for the various isolates were determined. The specific and non-specific pathogenicity of the isolates was studied by inoculating the different crops with the different isolates, including aP. cryptogea isolate fromGerbera and aP. nicotianae var.nicotianae isolate from carnation. TheP. nicotianae var.nicotianae isolates appeared to be morphologically identical. Some of the isolates were similar in host range, but others exhibited differences in host specificity at 20°C as well as 25, 30 or 35°C. The same applies for the twoP. cryptogea isolates.     

Arthropod fauna and main insect pests of plane trees in Israel

Fifty-four insect and four mite species are included in a list of the entomofauna of plane trees in Israel. Only two species are monophagous:Phyllonorycter platani (Stgr.) (Lepidoptera: Gracillariidae) andEdwardsiana iranicola Zachv. (Heteroptera: Cicadellidae). Four species are noxious:P. platani, the main insect pest of the plane trees in Israel;Zeuzera pyrina L. (Lepidoptera: Cossidae);Kalotermes flavicollis F. (Isoptera: Kalotermitidae); andE. iranicola. Of much lesser importance areTargionia vitis Sign. (Homoptera: Diaspididae),Heliothrips haemorrhoidalis Bché. andRetithrips syriacus May. (Thysanoptera: Thripidae). About half of the listed species are natural enemies and many are parasites ofP. platani. Details are given on the noxious species, together with recommendations for prevention and control.     

Integrative diagnosis and molecular phylogeny of dagger and needle nematodes of olives and grapevines in the island of Crete,Greece, with description of Xiphinema cretense n. sp. (Nematoda,Longidoridae)

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.     

Phenotypic and genotypic structure of Phytophthora infestans populations on tomato and potato in the North of Thailand in 2000–2002

A total of 80 single–lesion isolates of Phytophthora infestans were collected from tomatoes and potatoes in several locations in Chiang Mai and Tak provinces in 2000–2002. These isolates were analyzed for mating type, metalaxyl sensitivity, mitochondrial DNA (mtDNA) haplotype RFLP pattern as determined by probe RG57, and for microsatellite markers. All isolates were A1 mating type. Isolates from tomato were usually sensitive to metalaxyl, but isolates from potato were usually resistant to metalaxyl. With one exception, all tomato isolates were related to the US-1 clonal lineage. With two exceptions, all potato isolates were related to two European lineages. In these two provinces, the populations of P. infestans on tomatoes are clearly different from those on potatoes.     

Detection and estimation of Citrus Tristeza Virus infection rates based on Elisa Assays of packing house fruit samples

Enzyme-linked immunosorbent assay (ELISA) proved to be a sensitive detector for citrus tristeza virus (CTV) in orange fruits (Citrus sinensis (L.) Osbeck). Samples of five fruits were taken from 350-kg packing house containers and tested by ELISA to predict the infection rate of CTV in two infected orange groves. The predicted infection rates, 1% and 11%, were in reasonable agreement with the observed rates of 1% (15/1400) and 16% (324/2053), respectively. The 360 test samples from reputedly uninfected groves all tested negative. These results suggest that the ELISA procedure may provide a general method of detecting viral or other systemic pathogenic infections using the fruit as the test material in place of tree tissue. Fruit samples can be collected routinely at the packing house to reduce test costs.     

Serological recognition of Streptomyces species causing scab on potato tubers

Antisera were prepared against extracts of two tyrosinase-positiveStreptomyces spp., one of which caused a “deep” and the other a “russet” scab. Tyrosinase-positiveStreptomyces isolates not reacting with either of these antisera proved to be nonpathogenic to potato tubers, with few exceptions only. Not all isolates reacting with one or both antisera, however, were pathogenic and so all the serological positive ones had to be tested for pathogenicity to potato tubers. To obtain this relative specificity the antisera had to be absorbed with an extract of a non-pathogenic tyrosinase-positive isolate.     

Detection and partial molecular characterization of Plum pox virus on almond trees in Turkey

Almond (Prunus dulcis) is one of the well known stone fruit species grown for its unripe fruits and delicious seeds in Turkey. In the Trakya region, however, some prevailing virus infections have reduced almond yields and quality. In ten districts of Trakya, 260 leaf samples were collected from affected almond trees in June 2010. DAS–ELISA assays and RT-PCR tests were employed for the identification of viruses. As a result of these detection studies, five of the 260 leaf samples gathered from symptomatic almond trees had Plum pox virus (PPV), 81 of them had Prunus necrotic ringspot virus (PNRSV), and 11 samples contained Prune dwarf virus (PDV). Only four out of 260 samples had a mixture of these viruses. Partial nucleotide sequences of five almond isolates of PPV were determined and compared with 17 other PPV isolates in databases. Computer analysis of obtained and published nucleotide sequences showed identity ranged from 75.72% to 96.87%. Of the five PPV almond isolates obtained, however, there was a close nucleotide identity of 95.82–96.61% to Turkish isolates. Phylogenetic analysis of nucleotides and amino acids showed that five PPV isolates of almond from the Trakya Region of Turkey were clustered in the same subgroup with PPV-T Turkish isolates in GenBank. Therefore we can consider almond isolates of PPV as PPV-T strain, like the two other isolates from apricot trees in Turkey.     

Synthetic pyrethroids and avermectin for controlling the grapevine pestsLobesia botrana,Cryptoblabes gnidiella andDrosophila melanogaster

Field treatments in a vineyard with 0.015 or 0.01% a.i. of cypermethrin, fenvalerate, fenpropathrin or AC-222,705 were more efficient in controlling the grape-berry moth (Lobesia botrana Schiff.) and the honeydew moth (Cryptoblabes gnidiella Mill.) than four standard treatments consisting of two with 0.05% a.i. fenitrothion and two with 0.075% a.i. diazinon. In pyrethroid-treated plots, infestation at the end of the trials ranged between 2.5 and 12%, compared with 21% in the standard treatment plots and 34% in the untreated plots. Cypermethrin, fenpropathrin and AC-222,705 exhibited similar field activity, while that of fenvalerate was somewhat lower. Under laboratory conditions, cypermethrin at 0.005 and 0.01% a.i. was significantly more potent than fenvalerate, fenpropathrin and AC-222,705; at a higher concentration, 0.015% a.i., all pyrethroids were highly active, with mortality ranging between 75 and 95%. Under laboratory conditions the vinegar fly (Drosophila melanogaster Meig.) was in general more susceptible to pyrethroids than was the grape-berry moth. Cypermethrin and AC-222,705 at 0.005% a.i. and avermectin at 0.0035% a.i. were potent compounds against the vinegar fly and more active than fenvalerate and fenpropathrin.     

Braconidae (Hymenoptera) associated with forest and ornamental trees and shrubs in Israel

Data are presented on the occurrence of Braconidae (Hymenoptera parasitica) parasitizing insects associated with forest and ornamental trees and shrubs in Israel. Fifty-five genera of plants are listed, the richest in braconid fauna being tamarisk (9 species); acacia, pistachio and poplar (8 species each); carob and oak (7 species each). Of the 95 species of insect hosts mentioned, 53 are Lepidoptera, mostly Gelechiidae (7 species), Pyralidae (6 species), Noctuidae (5 species), Gracillariidae, Tortricidae, Geometridae, Lymantriidae and Lycaenidae (3 species each); 44 are Coleoptera, mostly Cerambycidae (13 species), Scolytidae (12 species), and Bostrichidae (9 species); three are Diptera. Of the 92 species of braconids listed, of which only 65 have been fully named, 56 develop in Lepidoptera, mostly Noctuidae (15 species), Gelechiidae (11 species) and Pyralidae (9 species); 33 species develop in Coleoptera, mostly Cerambycidae (12 species, Bostrichidae (10 species) and Scolytidae (5 species); and three species develop in Diptera. Thirty-eight species are new to the fauna of Israel; at least three of them are new to science,viz., Gnaptodon, Gildoria andDendrosotinus titubatus Papp.