Trees In Greece Research Articles

Using Multispectral Data from UAS in Machine Learning to Detect Infestation by Xylotrechus chinensis (Chevrolat) (Coleoptera: Cerambycidae) in Mulberries

The tiger longicorn beetle, Xylotrechus chinensis Chevrolat (Coleoptera: Cerambycidae), has posed a significant threat to mulberry trees in Greece since its invasion in 2017, which may be associated with global warming. Detection typically relies on observing adult emergence holes on the bark or dried branches, indicating severe damage. Addressing pest threats linked to global warming requires efficient, targeted solutions. Remote sensing provides valuable, swift information on vegetation health, and combining these data with machine learning techniques enables early detection of pest infestations. This study utilized airborne multispectral data to detect infestations by X. chinensis in mulberry trees. Variables such as mean NDVI, mean NDRE, mean EVI, and tree crown area were calculated and used in machine learning models, alongside data on adult emergence holes and temperature. Trees were classified into two categories, infested and healthy, based on X. chinensis infestation. Evaluated models included Random Forest, Decision Tree, Gradient Boosting, Multi-Layer Perceptron, K-Nearest Neighbors, and Naïve Bayes. Random Forest proved to be the most effective predictive model, achieving the highest scores in accuracy (0.86), precision (0.84), recall (0.81), and F-score (0.82), with Gradient Boosting performing slightly lower. This study highlights the potential of combining remote sensing and machine learning for early pest detection, promoting timely interventions, and reducing environmental impacts.

First Report of Phytophthora cinnamomi causing root rot of avocado trees in Greece.

In September 2023, thirty declining 30-year-old avocado (Persea americana) trees ('Hass' grafted on 'Zutano' seedlings) were detected in a 1.5-ha orchard in the island of Crete (Chania region). Crown symptoms encompassed wilting and leaf chlorosis, advancing to defoliation and extensive dieback. Tap and feeder roots decayed and brown discoloration of root tissues was evident on heavily infected trees. The disease was severe and widespread, resulting in a 5% mortality rate among 300 trees. The pathogen was isolated with a modified soil baiting technique (Ferguson and Jeffers, 1999). Surface disinfected avocado fruits were immersed in water containing soil samples. Following a period of 2 to 8 days, tissue fragments from the resulting necrotic lesions on the fruit surface were transferred on ΡΑRP medium and subsequently incubated at 20°C (Ferguson and Jeffers, 1999). Three isolates (AV2, AV12 and AV11a) were obtained by transferring single hyphal tips to new Petri dishes containing V8 juice agar. They were grown at 20˚C and used for identification after 10 days. Isolates formed coralloid colonies with abundant clustered spherical hyphal swellings and terminal or intercalary (ratio 1:5) thick-walled chlamydospores measuring 20 to 36 μm (avg 29±0.8 μm) with characteristic thick walls (avg 1.2±0.2 μm). Sporangia, produced in non-sterile soil-extract water, were ovoid to obpyriform, persistent, non-papillate, 32 to 81 μm (avg 56±4.8 μm) long and 20 to 42 μm (avg 31±3.2 μm) wide (n=100). Isolates were heterothallic as they did not produce oospores in single cultures. Based on the morphological traits the isolates were identified as Phytophthora cinnamomi (Erwin and Ribeiro 1996). The internal transcribe spacer region (ITS) including ITS1, 5.8S rDNA region, and ITS2 as well as the cytochrome c oxidase subunit I (coxI) gene of the three representative isolates wereamplified with ITS1/ITS4 and FM83/FM84 primers, respectively (White et al. 1990; Martin and Tooley 2003), and sequenced (GenBank acc. PP506613 to PP506615 and PQ063867 to PQ063869, respectively). BLAST search revealed almost 100% identity with the sequences of P. cinnamomi ex-isotype isolate (KC478663 and KU899315 respectively). Pathogenicity tests using isolate AV2 were conducted following the soil infestation method (Jung et al. 1996) using six-year-old avocado 'Zutano' seedlings. Six non-inoculated plants treated with vermiculite-multivitamin juice mixture were used as controls. Plants (1 m tall) were grown in pots under greenhouse conditions and watered regularly. Six weeks post inoculation, all inoculated trees showed chlorosis, wilting and root rot, while control plants remained symptomless. Symptoms were similar to those observed in the field and the pathogen was re-isolated and molecularly identified as previously described. This study presents the first documented occurrence of P. cinnamomi, widely regarded as the most destructive avocado pathogen globally, on avocado crops in Greece (Rodger et al. 2019). Additionally, this marks the first recorded presence of this pathogen on the island of Crete, regardless of the host species. The accurate identification of Phytophthora species associated with avocado root rot is essential for implementing an effective disease management strategy, particularly in the selection of appropriate disease-resistant rootstocks.

Occurrence of shoot blight disease, caused by Cylindrocarpon destructans (Zinss) Scholten, on cherry trees (Prunus avium) in Greece

Shoot blight is a common stone fruit disease that occurs worldwide. The disease is most commonly identified in early spring, shortly after bud break. To our knowledge, this is the first report of the fungus Cylindrocarpon destructans causing severe canker and shoot blight on cherry tree in Greece and worldwide. This study also investigated the effect of temperatures and moisture on the mycelial growth and conidial germination of C. destructans. It was found that a temperature range of 10 to 30 °C was adequate for mycelial growth and conidial germination, with 20 °C being the optimum. In addition, 6 h of continuous moisture at 20 °C was required to initiate conidial germination, and germination gradually increased as the duration of continuous moisture increased from 6 to 36 h, with no further increases observed up to 48 h. The pathogenicity and virulence of C. destructans to commercial cherry, plum, peach, apricot, apple and pear cultivars were also examined. The results of this study showed that C. destructans did not show host specificity as it was pathogenic to all peach, plum, apricot, pear and apple cultivars tested. Some differences in the level of susceptibility were found among cultivars evaluated.Based on the above results, a logistic regression model could be developed that adequately describes the effects of pre-inoculation moisture and temperature on the infection incidence in cherry trees.

Identification and complete genome sequencing of a divergent olive virus T isolate and an olive leaf yellowing-associated virus isolate naturally infecting olive trees in Greece

Several new full genome sequences of olive viruses came to light recently via high-throughput sequencing (HTS) analysis. In this study, total RNA HTS analysis of two Greek olive trees revealed the presence of an olive virus T (OlVT) isolate and an olive leaf yellowing-associated virus (OLYaV) isolate. The full viral genome of OlVT isolate (50Ch) is composed of 6862 nucleotides encoding for three proteins (replicase, movement protein, and capsid protein) with typical betaflexiviruses’ genomic features. However, both sequence and phylogenetic data analysis exhibited high levels of variability between 50Ch and the previously characterized OlVT isolates. In addition, the almost full genome of the Greek OLYaV isolate (OL2) was obtained, which is composed of 16,693 nucleotides encoding for 11 open reading frames (ORFs) and shares common genomic features with the recently characterized OLYaV isolates from Spain and Brazil. Sequence and phylogenetic analysis revealed high similarity between these three isolates. Due to problems encountered with the detection of both viruses, new nested RT-PCR assays were developed and applied. In addition, recombination events were observed in OlVT isolates (50Ch GR-168), thus highlighting the potential role of this mechanism in the evolution of the virus. This study is adding further knowledge to the limited information available about these recently characterized olive infecting viral pathogens and highlights their widespread distribution in Greece, one of the most important olive producing countries of the world.

First Report of Pseudophaeomoniella oleae Causing Wood Streaking and Decay on Olive Trees in Greece.

A survey was conducted in olive orchards (Olea europaea L., cv Koroneiki) showing severe decline in Milatos and Ierapetra (Lasithi, Crete, Greece) in November 2017 and January 2019, respectively. Diseased trees exhibited wilting, yellowing of leaves, twig and branch dieback, and internal discoloration of vascular tissue. Insect infestations were commonly associated with these symptoms. A yeast-like fungus was consistently isolated from discolored vessels previously surface-disinfested with 95% ethanol, on acidified potato dextrose agar (APDA). The fungus yielded several dark brown to black, globose to oval pycnidia with dimensions 120-330 × 90-300 μm (average 179.6 × 143.9 μm), after 25 days of growth on APDA. Emerging colonies were transferred to new PDA and their growth rate was 2.21 mm/day at 24 °C in the dark. The sparse aerial mycelium was initially white, turning into beige-pinkish in the centre after 21 days of growth on PDA. Microscopic observations revealed hyaline, smooth, ampulliform conidiophores, bearing conidia solitary or in slimy heads. Conidia were 1-celled, hyaline, smooth, subcylindrical with obtuse ends, 1.25-5.75 × 0.75-2.00 μm (average 3.12 × 1.16 μm) in size. Light to dark brown pycnidia, semi-immersed in PDA, with dimensions 150-490 × 90-320 μm (average 297.7 × 231.0 μm) were evident in 3-week-old cultures. Colony morphology and microscopic features were similar to Pseudophaeomoniella oleae (Crous et al. 2015). DNA from two representative isolates (EML1 and DRAGVR1) was extracted and their internal transcribed spacer region (ITS) of ribosomal DNA, actin (ACT) and translation elongation factor 1-alpha (TEF1-α) genes were amplified using the primer pairs ITS1/ITS4 (White et al. 1990), ACT-512F/ACT-783R and EF1-728F/EF1-986R (Carbone and Kohn 1999), respectively. PCR products were sequenced and deposited in GenBank (accession Nos. MZ854242-MZ854243, OK143463-OK143464 and OK143465-OK143466). BLAST search revealed high similarity to GenBank sequences of the ex-type strain of P. oleae for ITS (≥99.50%, NR_137966.1 and KP635972.2), ACT (100%, KP635974.1), and TEF-1a (98.32%, KP635968.1). Based on morphology and phylogenetic analysis of the ITS region, the fungus was identified as P. oleae. Ten 3-year-old olive trees of each of the cvs Amfissis and Koroneiki were artificially inoculated by drilling a 3-mm-diameter hole into the trunk and injecting 50 μl of a 1 × 107 ml-1 conidia suspension of the isolate DRAGVR1 (Markakis et al. 2017). Another ten trees of each cv treated similarly with sterilized distilled water served as controls. Potted trees were kept under ambient conditions. Fourteen months post inoculation, longitudinal and transverse sections of inoculated trunks revealed wood discoloration extending above and below the inoculation point in both cultivars, whereas no leaf symptoms were observed. P. oleae was steadily re-isolated from symptomatic wood tissue and identified by colony morphology. Neither symptoms nor positive isolations were observed in controls. To our knowledge, this is the first report of wood streaking and decay caused by P. oleae in olive trees in Greece. Although the fungus has been isolated previously from decayed olive trees showing vascular wilt in Italy (Crous et al. 2015), this is the first experimental evidence of the pathogenic potential of the species on olive trees worldwide. This disease could potentially be an increasing problem in olive tree growing areas and result in severe crop losses. Hence, effective management practices should be investigated and applied.

Environmental and Economic Assessment of Wood Pellet Production from Trees in Greece

Biomass is a renewable, economic and readily available resource of energy that has potential to substitute fossil fuels in many applications such as heat, electricity and biofuels. The increased use of the agricultural biomass can help the agricultural based societies in achieving energy security and creating employment without causing environmental degradation. However, the viability and feasibility of electricity generation from agricultural biomass depends upon the availability of biomass supply at a competitive cost. The present study investigates the availability of agricultural biomass for distributed power generation in Greece (Kozani). The study concludes with a discussion on significance and challenges of decentralized electricity generation for rural energy supply, including brief description about economical, social, environmental and technical aspects of bioelectricity. With the application of the life cycle analysis applied, the environmental and economic impacts that will occur in the region of Kozani in Greece, where a biomass wood pellets production workshop is operating, have been assessed. The total annual emission of CO 657.9 gr, HC 22.36 gr, PM 67.94 and NOx 8.832,2 gr was calculated. The economic evaluation estimated the payback period for the investment in this plant to be approximately 3 years.

Temperature and incubation period affect Septoria pistaciarum conidium germination: disease forecasting and validation

Septoria leaf spot is an important disease of pistachio trees in Greece. This study aimed to determine effects of temperature and the incubation period on germination of conidia of Septoria pistaciarum, and to evaluate a generic model to forecast pistachio leaf spot under the field conditions of Aegina Island, Greece. The optimum temperature for conidium germination was 23°C, and germination was inhibited at 35 and 4°C. At constant temperature of 23°C, conidia commenced germination after 9 h. The predictive model indexed disease risk close to 100 at 10 May at two locations (Rachi Moschona and Vigla) in 2017, and first leaf spot symptoms were observed on 17 May. Moderate to high disease severity (>25% leaves infected) were observed in unsprayed trees at the end of May. In 2018, the model indexed risk close to 100 on 9 May at Rachi Moschona, and first symptoms were observed on 18 May. Moderate to high disease severity (>25% leaves infected) were observed in unsprayed trees on 25th of May. This study has shown that the forecasting model can be used in Aegina Island, Greece, to predict the severity of Septoria leaf spot of pistachio.

Comparative study on the chemical composition of laurel (Laurus nobilis L.) leaves from Greece and Georgia and the antibacterial activity of their essential oil

Laurel (Laurus nobilis L.) is a plant species from Lauraceae family, and is native to the Mediterranean region. The goal of this study was to compare chemical composition of laurel leaves and antibacterial activity of its essential oil (EO) from wild-grown trees in Greece and Georgia. The laurel leaves from the two native habitats had dissimilar concentrations of phenolic acids. Of the conjugated flavonols and flavons, kaempferol (1981.3 μg/g) and apigenin (1433.6 μg/g) were the major representatives in the leaves from Greece, while luteolin (839.1 μg/g) and kaempferol (688.1 μg/g) were the major ones in the leaves from Georgia, respectively. The EO content was 1.42% and 4.54% in the leaves from Greece and Georgia, respectively. The main EO constituents of the Greek laurel plants were 1,8-cineole (30.8%), α-terpinyl acetate (14.9%), α-terpineol (8.0%), sabinene (7.9%), and terpinen-4-ol (6.0%). The main EO constituents of the Georgian laurel plants were 1,8-cineole (29.2%), α-terpinyl acetate (22.6%), sabinene (12.2%), and methyleugenol (8.1%). The EO antimicrobial activities against 20 microorganisms were determined. Among the Gram-positive bacteria, the Enterococcus faecalis strain was the most sensitive, followed by Staphylococcus aureus ATCC 6538. Among the Candida species, C. albicans ATCC 10231 was the most sensitive to the laurel leaf EOs.

First Report of Apple Luteovirus 1 in Apple Trees in Greece

In June 2018, shoots were collected from five apple trees from different orchards in Zagora (Magnesia prefecture, east-central Greece) exhibiting various virus-like disease symptoms such as leaf mosaic and bark scaling or suffering from apple small fruit disorder. To identify the agents associated with the symptomatology, total RNAs were extracted following a CTAB protocol (Gambino et al. 2008), and after DNase treatment, equimolar quantities from each of the five extracts were mixed in a pooled sample and subjected to high-throughput sequencing (HTS) after ribosomal RNA was removed using a Ribo-Zero rRNA Removal Kit (Plant Leaf, Illumina). Library preparation followed by sequencing on an Illumina NextSeq Sequencing System (Biomedical Research Foundation Academy of Athens, Greece) resulted in 55 million 75-bp single-end RNA sequencing reads. The analysis of the HTS data with Geneious 11.1.5 software revealed the presence of viruses frequently found in apple cultivation in Greece (apple chlorotic leaf spot virus, apple mosaic virus, apple stem pitting virus, apple stem grooving virus), but also the presence of the recently discovered apple luteovirus 1 (ALV1) (Liu et al. 2018). In total, 316,984 reads were de novo assembled in one viral contig annotated by BLAST as ALV1 sequence, covering 99% (devoid of untranslated regions) of the genome of the reference isolate PA8 (GenBank accession no. NC_040680) and sharing 92.3% nucleotide identity. Based on the HTS analysis, primers were designed (LUTEO-Forward, 5′-GCCAGCTGAGGTCTTCACTT-3′; LUTEO-Reverse, 5′-GAAGTCGAATACCAACGCGC-3′) to amplify part (674 bp) of the ORF3 to ORF5 genomic regions of the virus. Subsequently, the five original trees and 12 more with similar symptoms from the same region were tested with reverse transcription PCR using the new pair of primers. PCR amplicons of the expected size were obtained in four of the five trees subjected to HTS, and in 11 of the other 12 trees examined. Two of the PCR products were subjected to Sanger sequencing (100% identity with each other, GenBank accession no. MT068445), confirming the presence of ALV1 (98.4% nucleotide identity with isolate PA8). The virus was first described in apple trees suffering from “rapid apple decline” in the Northeastern United States (Liu et al. 2018), but it was not found in declining trees in Washington State (Wright et al. 2020). It was also detected in apple rootstocks showing yellowing and mottling symptoms in Korea (Lim et al. 2019). In all cases, including the present study, the virus was found in mixed infections with other viruses, making difficult its association with a specific symptomatology. Nevertheless, our results suggest that this recently discovered virus is widespread in Zagora, one historical apple-producing area in mountainous Greece with many old orchards, a fact that can increase our knowledge on the diversity and possible pathogenicity of ALV1. Moreover, due to its proposed association with rapid apple decline, it constitutes a possible threat for apple cultivation in Greece. To the best of our knowledge, this is the first report of ALV1 in apple trees in Greece and generally in Europe.

Novel species of Huntiella from naturally-occurring forest trees in Greece and South Africa.

Huntiella species are wood-infecting, filamentous ascomycetes that occur in fresh wounds on a wide variety of tree species. These fungi are mainly known as saprobes although some have been associated with disease symptoms. Six fungal isolates with typical culture characteristics of Huntiella spp. were collected from wounds on native forest trees in Greece and South Africa. The aim of this study was to identify these isolates, using morphological characters and multigene phylogenies of the rRNA internal transcribed spacer (ITS) region, portions of the β-tubulin (BT1) and translation elongation factor 1α (TEF-1α) genes. The mating strategies of these fungi were also determined through PCR amplification of mating type genes. The study revealed two new species; one from Platanus orientalis in Greece and one from Colophospermum mopane and Senegalia nigrescens in South Africa. These novel taxa have been provided with the names, H. hellenicasp. nov. and H. krugerisp. nov., respectively. The former species was found to have a homothallic and the latter a heterothallic mating system.

Registration and promotion of monumental olive trees in Greece.

The history of the olive tree, its cultivation and its products is known for centuries. Some olive tree have survived over millennia and their history dates back to antiquity. In many cases, it is related to mythology and religion. The olive tree is associated with folk tradition, people's everyday life, and customs. In Greece, monumental olive trees are found in the Peloponnese, Crete, Euboea, Chios, Pelion and Attica. This paper explores and describes the particular morphological features such as shape, size, wood, cavities and age, as well as the cultural characteristics such as historical or religious events, myths and traditions that define an olive tree and characterize it as monumental. The main aim of our research is to examine the key position that monumental olive trees and their materialistic and symbolic manifestations consist a natural and cultural heritage as well. In this framework the study focuses on various key issues related to monumental olives trees and their natural, historical, social and cultural value.

Symptomatic Citrus trees reveal a new pathogenic lineage in Fusarium and two new Neocosmospora species.

The diversity of fusaria in symptomatic Citrus trees in Greece, Italy and Spain was evaluated using morphological and molecular multi-locus analyses based on fragments of the calmodulin (CAM), intergenic spacer region of the rDNA (IGS), internal transcribed spacer region of the rDNA (ITS), large subunit of the rDNA (LSU), RNA polymerase largest subunit (RPB1), RNA polymerase second largest subunit (RPB2), translation elongation factor 1-alpha (EF-1α) and beta-tubulin (TUB) genes. A total of 11 species (six Fusarium spp., and five Neocosmospora spp.) were isolated from dry root rot, crown, trunk or twig canker or twig dieback of citrus trees. The most commonly isolated species were Fusarium sarcochroum, F. oxysporum and Neocosmospora solani. Three new Fusarium species are described, i.e., F. citricola and F. salinense belonging to the newly described F. citricola species complex; and F. siculi belonging to the F. fujikuroi species complex. Results of pathogenicity tests showed this new complex to include prominent canker causing agents affecting several Citrus spp. In addition, two new species are described in Neocosmospora, named N. croci and N. macrospora, the latter species being clearly differentiated from most members of this genus by producing large, up to nine-septate sporodochial conidia.

Description of Hystripalpus chalkidicus new species from olive trees in Greece (Acari: Tenuipalpidae)

The female and deutonyph of Hystripalpus chalkidicus, new species, are described and illustrated. The species was found on an olive tree in Chalkidiki, Greece.

Cryphonectria parasitica, the chestnut blight fungus, causes cankers on Quercus frainetto in Greece

In 2011, cankers were observed on Hungarian oak trees (Quercus frainetto) in three localities in Greece. The pathogen was identified as Cryphonectria parasitica on the basis of morphology and sequence analysis of the internal transcribed region (ITS), part of the elongation factor 1a (EF-1a) and part of the beta-tubulin region (βT). A pathogenicity test was performed and Koch’s postulates were confirmed by re-isolation of the fungus C. parasitica from artificially inoculated detached twigs. This is the first report of C. parasitica as the cause of cankers on Hungarian oak trees in Greece.

Platypus cylindrus, a vector of Ceratocystis platani in Platanus orientalis stands in Greece

SummaryThe ambrosia beetle Platypus cylindrus was very common in stands of Platanus orientalis trees in Greece, infesting trees already infected by Ceratocystis platani. The fungus C. platani was isolated from 59% of adult beetles of P. cylindrus collected on the wing in a heavily infected stand of P. orientalis. Transmission of C. platani was demonstrated in 80% of P. orientalis seedlings challenged with P. cylindrus beetles, which were previously fed in cultures of the fungus. In most of the cases, the beetles had bored tunnels in stems, where abundant perithecia of C. platani were observed. Platypus cylindrus adults regularly visited artificially wounded P. orientalis trees in a natural stand. Although this ambrosia beetle normally infests stressed or dead trees, it appears to play a role as a vector of C. platani, especially by visiting wounded trees.

First Report of Verticillium Wilt Caused by Verticillium dahliae on Avocado Trees in Greece.

Avocado (Persea americana) is an important crop for Chania, Crete, Greece, and is grown on more than 800 ha. In November 2013, 4-year-old trees in a new avocado grove of cv. Hass grafted onto the rootstock 'Bacon,' previously planted in citrus trees, showed symptoms of yellowing, leaf fall, twig and branch dieback and vascular tissue discoloration. Disease incidence was estimated at 2.3% (12 out of 530 trees affected). A fungus was consistently and readily isolated from symptomatic vascular tissue, previously surface-disinfested with 95% ethanol, on acidified potato dextrose agar (APDA). After 7 days, slow-growing colonies were transferred to PDA and the growth rate of the fungus was 2.9 mm/day at 24°C in the dark. Microscopic observations revealed hyaline hyphae with many irregular, dark microsclerotia measuring 40 to 200 × 30 to 75 μm (average 94.5 × 50.3 μm) developing after 21 days of growth. Hyaline, elliptical, single-celled conidia measuring 2.8 to 7.5 × 2.5 to 4.3 μm (average 4.8 × 3.1 μm) developed on verticillate conidiophores. For molecular characterization, Verticillium dahliae specific primer pair ITS1-F/ITS2-R that amplifies the rRNA internal transcribed spacer (ITS) region was used (2). Band of expected size was amplified, sequenced, and deposited in GenBank (Accession No. KJ818294). On the basis of morphological characteristics (3) and a BLAST search with 100% identity to the published ITS sequence of a V. dahliae isolate in GenBank (KC834733.1), the fungus was identified as V. dahliae. Five 1-year-old avocado plants of cv. Hass, grafted onto the rootstock 'Bacon,' were used for pathogenicity tests. Artificial inoculation was performed by making a 5.0 × 3.5 mm hole in the rootstock trunk, injecting approximately 40 μl of a 2.8 × 107 conidia/ml suspension into the vessels (spores were introduced passively), sealing with Vaseline, and covering with adhesive paper tape. Five control plants were mock inoculated with sterilized distilled water. Disease symptoms that appeared 18 days post artificial inoculation were similar to those observed under natural infection conditions. Thirty-five days post artificial inoculation, disease incidence was 80%, whereas the percentage of positive V. dahliae re-isolations from infected tissues was 95% (96.7 and 93.3% from rootstock and graft, respectively). The extent of vascular tissue discoloration from the point of inoculation ranged from 11 to 62 cm, whereas V. dahliae was successfully re-isolated even from the end of the graft (approximately 60 cm above the initial inoculation point), thus confirming Koch's postulates. Neither symptoms nor positive isolations were observed in control plants. The pathogenicity test was repeated twice with similar results. Verticillium wilt of avocado has been observed in several countries including Argentina, Chile, Ecuador, Israel, Mexico, Morocco, Spain, and the United States (1). To the best of our knowledge, this is the first report of Verticillium wilt on avocado in Greece. This disease could potentially be an increasing problem in areas where young avocado trees are established on land previously planted in vegetable crops.

Partitiviruses of a fungal forest pathogen have species-specific quantities of genome segments and transcripts

Heterobasidion partitiviruses infect forest pathogenic fungi of the genus Heterobasidion. We have studied the amounts of genomes and transcripts of four partitiviruses isolated from four different Heterobasidion strains infecting different host trees in Greece, Poland, Finland, and China. Heterobasidion partitiviruses have bisegmented genomes encoding coat protein and RNA-dependent RNA polymerase. Our results show that the coat protein genome segment is generally more abundant in infected mycelia than the RNA-dependent RNA polymerase segment and that this bias persists also at transcript levels. The different virus species all have unique ratios of the genome segments and the ratio is generally stable over different temperatures and hosts. The amounts of transcripts of each virus respond to host growth temperatures in a distinctive and consistent manner. The Heterobasidion partitiviruses studied here affect only rarely the growth of their natural hosts but do influence the growth of a new host more frequently.

Antecolic reconstruction during pancreatoduodenectomy

Heterobasidion partitiviruses infect forest pathogenic fungi of the genus Heterobasidion. We have studied the amounts of genomes and transcripts of four partitiviruses isolated from four different Heterobasidion strains infecting different host trees in Greece, Poland, Finland, and China. Heterobasidion partitiviruses have bisegmented genomes encoding coat protein and RNA-dependent RNA polymerase. Our results show that the coat protein genome segment is generally more abundant in infected mycelia than the RNA-dependent RNA polymerase segment and that this bias persists also at transcript levels. The different virus species all have unique ratios of the genome segments and the ratio is generally stable over different temperatures and hosts. The amounts of transcripts of each virus respond to host growth temperatures in a distinctive and consistent manner. The Heterobasidion partitiviruses studied here affect only rarely the growth of their natural hosts but do influence the growth of a new host more frequently.

First Report of Hop stunt viroid Infecting Japanese Plum, Cherry Plum, and Peach in Greece.

Dapple plum and peach fruit is a widely distributed disorder of plum and peach resulting in significant economic losses (4). During a survey for the presence of Hop stunt viroid (HSVd) on stone fruit trees in Greece, samples from 30 European plums (Prunus domestica L., cvs. President, Tuleu Grass), 45 Japanese plums (Prunus salicina Lindl., cvs. Angeleno, Diamond, Santa Rosa), 12 cherry plums (Prunus domestica L. var. insititia (L.) Fiori & Paoletti of unknown cultivar), and 107 peaches (Prunus persica (L.) Batsch, cvs. Red Haven, Elberta, June Gold, Spring Crest, Lemonato) were collected in several orchards around Greece. Their fruit skin symptomatology indicated viroid infection (reddish dappling blotches and cracks in European and Japanese plum, green dappling in cherry plum, and light colored blotches and lines in peach). Samples were screened with tissue-print hybridization (TPH) for HSVd using a full length DIG-labelled riboprobe deriving from in vitro transcription of the positive control, a citrus isolate of HSVd (G. Vidalakis, CCPP, University of California, Riverside). In total, 44 out of the 194 trees surveyed were HSVd-positive with TPH. For a small number (40) of TPH-positive field samples, TNA phenol extraction from fruit skin, leaves, and bark and one-tube two-step reverse transcription (RT)-PCR assays followed, using a standardized protocol (3) with two different primer pairs, one new primer pair (this study) and a previously reported primer pair (2). RT-PCR analysis showed the presence of HSVd in peach and Japanese plum in prefectures Pella (Central Macedonia), Achaia, and Korinthia (Peloponnesus) and in cherry plum in Achaia (Peloponnesus). Six of 11 Japanese plums (cvs. Angeleno, Santa Rosa), 2 of 12 cherry plums, and 8 of 12 peaches (cvs. Spring Crest, Red Haven) examined were found HSVd-infected, but none of the five European plums were. Nucleotide sequence analyses of purified and cloned amplicons from peaches and Japanese and cherry plums revealed sizes of 297 to 308 nt and similarity to sequence variants of other HSVd isolates previously characterized: 95 to 97% identity with the Moroccan isolates apr.9, apr.10, apr.11, and apr.12 and the Spanish isolate apr.4 from apricot (1) (GenBank Accession Nos. AJ297825 to AJ297828 and Y09346, respectively). For confirmation of HSVd presence in field trees, 10 Japanese plums cv. Angeleno, 10 peaches cv. June Gold, and 10 peaches cv. Spring Crest, HSVd-negative (TPH), were bud- or chip-grafted from two of the aforementioned Japanese plums cv. Angeleno and two of the aforementioned peaches cv. Red Haven. Two years later, five Japanese plum trees (cv. Angeleno) and five peach trees (three cv. Spring Crest and two cv. June Gold) were found HSVd-positive with TPH; no fruits were observed to produce fruit symptoms as the grafted trees were kept in an insect-proof greenhouse (no bees for cross-pollination). To our knowledge, our investigation reports for the first time the occurrence of HSVd infecting Japanese plum, cherry plum, and peach in Greece, emphasizing the need for a certification program for the prevention of spreading stone fruit tree viroids in this country.

Phyllocnistis citrella and its parasitoids in three citrus species in Greece

The relative abundance as well as the percentage of parasitism of Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae) and its parasitoids were studied in a citrus orchard with orange, mandarin and lemon trees in Greece. Infestation of P. citrella on each citrus host, as expressed by the number of mines per leaf, was significantly higher on lemon than on mandarin. The P. citrella parasitoid complex included the native species Neochrysocharis formosa and Pnigalio pectinicornis, as well as the introduced Citrostichus phyllocnistoides. The most abundant of those was N. formosa in all of the citrus species. The average percentage of parasitism was 13.1%, 13.8% and 11.7% on orange, mandarin and lemon, respectively. No significant differences in parasitism rate by each of the three parasitoid species separately were recorded among the three citrus species.