Bioassays on the role of tomato,potato and sweet pepper as sources of <Emphasis Type="Italic">Tomato chlorosis virus</Emphasis> transmitted by <Emphasis Type="Italic">Bemisia tabaci</Emphasis> MEAM1

Shot hole disease of stone fruits caused by Thyrostroma carpophilum has become a major threat to stone fruit industry of Jammu and Kashmir, India because of the failure in its management with fungicides. To understand the diversity in shot hole pathogen, a combination of conventional (morphological, cultural and pathological) and molecular (ISSR and ITS markers) approaches were employed to discern variability in 25 isolates of T. carpophilum isolated from peach, plum, apricot, almond and cherry leaves collected from Srinagar, Ganderbal, and Baramulla districts of Jammu and Kashmir, India. The studies revealed a high level of variability among the pathogen. Based on the morpho-cultural and pathological studies, the isolates were grouped into different categories based on colony growth, texture, margin and colour besides change in media colour, incubation period, leaf area infected, etc. Using ISSR markers, a high level of polymorphism in different isolates of T. carpophilum was observed which indicated that these markers are suitable for studying the genetic diversity in this pathogen. Based on dendrogram, the isolates were grouped irrespective of their geographical origin or host species. Phylogenetic analysis of the 25 sequences based on ITS region showed maximum similarity with T. carpophilum (Syn. Wilsonomyces carpophilus) sequences retrieved from NCBI and grouped them in a single clade which proved it as a powerful tool for authentic identification. The pathogen was highly variable based on morpho-cultural, pathological and molecular (ISSR) characterisation.     

<Emphasis Type="Italic">Neofusicoccum parvum</Emphasis> associated with fruit rot and shoot blight of peaches in Greece

Shoot blights and fruit rots comprise the most serious diseases of peaches in Greece. In this study, the importance of the fungus Neofusicoccum parvum as a casual agent of a fruit rot and shoot blight of peach trees in Greece was investigated. This pathogen was isolated from both immature and mature peach fruit of the cultivar “Catherine” and later on from mature fruit of the peach cultivars “Andross”, “RedHaven”, “Sun Crest” and “Sun Cloud”. In the first year of investigation, N. parvum was found causing preharvest fruit rot and shoot blights of peach trees only at the location “Ammos-Mesi-Meliki Verias” in the prefecture of Imathia (the main peach production area of Greece) at incidences of 30 and 8%, respectively. However, in 2006 N. parvum was isolated from more locations such as Diavatos, Veria, Kopanos and Agia Marina in the prefecture of Imathia, but only at less than 3% of the total surveyed rotted peach fruit and blighted shoots. The pathogen overwintered as sub-epidermal pycnidia in blighted shoots or mummified fruit that remained on peach trees. This study also showed that the optimum temperature for mycelial growth and conidial germination of N. parvum was 25 oC. Pathogenicity tests using peach fruit showed that isolates of N. parvum and Diplodia seriata (isolated from pistachio grown in the same region) showed no significant differences in their virulence. In laboratory inoculation tests using detached shoots from 25 peach and nectarine cultivars, N. parvum isolates obtained from rotted peaches caused different size cankers on these cultivars. The cultivar Big Top was the most susceptible while the cultivar Maria Bianca the least susceptible.     

Characterization of Botryosphaeriaceae species associated with diseased loquat (Eriobotrya japonica) in Spain

Loquat (Eriobotrya japonica) is an important subtropical fruit crop in Spain and other Mediterranean countries. In recent years, characteristic symptoms of branch canker and dieback have been observed in the main cultivated areas of loquat in Spain. The goal of this study was to identify and characterize the species of Botryosphaeriaceae associated with these symptoms. For this, 36 affected orchards were surveyed between 2010 and 2011 in six provinces of southeastern Spain. Eighty‐four isolates belonging to the family Botryosphaeriaceae were recovered from samples with symptoms. These isolates were characterized by means of phenotypical studies, DNA sequence analyses of the internal transcribed spacer (ITS) and part of the translation elongation factor 1‐α regions, and pathogenicity tests. Ten fungal species were identified: Diplodia malorum, Diplodia olivarum, Diplodia seriata, Diplodia pseudoseriata/Diplodia alatafructa, Diplodia sp., Dothiorella sarmentorum, Neofusicoccum mediterraneum, N. parvum, Spencermartinsia plurivora and S. viticola. In addition, Diplodia eriobotryicola and Dothiorella eriobotryae are newly described. The most frequent species isolated from cankers was D. seriata and, as far as is known, this is the first report of D. malorum, and species belonging to the complex D. pseudoseriata/D. alatafructa, in Spain. All species were pathogenic to 1‐year‐old loquat plants cv. Algerie, with Diplodia sp. and S. viticola as the most virulent.     

The effect of Cryphonectria parasitica on water relations of American chestnut

Chestnut blight has commonly been regarded as a phloem disease due to conspicuous stem cankers that result from infection by the fungal pathogen Cryphonectria (Endothia) parasitica. Stomatal conductance (g_s), transpiration rate (E) and leaf water potential were measured throughout the day on leaves distal to naturally-occurring virulent (sunken bark with abundant stromata) or hypovirulent (swollen bark lacking stromata) cankers and cankers induced by inoculation with virulent or hypovirulent strains of C. parasitica. Relative to control stems, hydraulic conductivity (K_h), g_s and E were reduced significantly (α = 0·05) for both natural virulent cankers, and cankers that were induced by a virulent strain. These effects were less pronounced for both natural and induced hypovirulent cankers. Isolation experiments revealed that the percentage of xylem tissue chips yielding C. parasitica was greater for virulent than for hypovirulent cankers. The data provide evidence that the localized presence of C. parasitica in cankers of American chestnut results in stomatal closure, possibly as a direct result of xylem dysfunction.     

Significant in vitro antagonism of the laurel wilt pathogen by endophytic fungi from the xylem of avocado does not predict their ability to control the disease

The ascomycete Raffaelea lauricola causes laurel wilt, a lethal vascular disease of avocado, Persea americana, and other members of the Lauraceae plant family. Few effective control measures for laurel wilt exist and new measures are needed. In this study, biological control of the disease with endophytic fungi from avocado was examined. Thirty‐two endophytes (24 operational taxonomic units or OTUs) isolated from the xylem of healthy trees (the infection court of R. lauricola) were evaluated against R. lauricola with in vitro dual‐culture assays. Nine OTUs that showed strong in vitro antagonism of the pathogen were tested in planta against laurel wilt. In three greenhouse experiments, grafted avocado plants of Simmonds or Russell cultivars, which are both susceptible to laurel wilt, were inoculated with endophytes and, after 10–16 days, inoculated with the same isolate of R. lauricola that was used in the in vitro assays. Within 14 days of inoculation with R. lauricola, laurel wilt developed in plants that were not treated with endophytes (positive controls) but also developed in endophyte‐treated plants to the extent observed in the positive controls (P = 0.05). The pathogen colonized plants rapidly and systemically, but endophytes generally did not colonize xylem more than 2 cm above the point at which plants were inoculated. Although the tested endophytes strongly antagonized the pathogen in vitro, this did not translate to an ability to reduce development of laurel wilt. The management of laurel wilt and other plant diseases with fungal endophytes is discussed.     

Ability of aLeptosphaeria maculans isolate to form stem cankers on Indian mustard (Brassica juncea) segregates as a single locus

Australian isolates of the blackleg fungusLeptosphaeria maculans, that form cankers on two Indian mustard (Brassica juncea) varieties (Stoke and Zaria) are described. This ability to form cankers on var. Stoke segregates as a single locus in both F₁ and backcross progeny from a cross between twoL. maculans isolates.     

Genetic diversity and pathogenicity of Monilinia polystroma – the new pathogen of cherries

Brown rot caused by fungi belonging to the genus Monilinia is one of the major limiting factors of sour and sweet cherry production. Up to now, three species, M. fructigena, M. laxa and M. fructicola, have been identified as causal agents of brown rot on cherries worldwide. From 2010 to 2013, during the monitoring of cherry orchards in different areas of Poland, a fourth species, M. polystroma, was isolated from brown rot symptoms on sour and sweet cherry fruits. To the best of the authors’ knowledge, this is the first time M. polystroma has been reported as the causal agent of brown rot on cherries. The genetic diversity of M. polystroma isolates from cherries and other hosts was analysed using PCR MP, ISSR and RAPD techniques and showed its clear distinctness from other Monilinia spp. tested. The cluster analysis of fingerprinting data revealed a high similarity of M. polystroma isolates from Poland and their close relationship with the reference strain from Japan, indicating that this species is a recently introduced pathogen. The highest genetic distance between the examined isolates and the highest number of different genotypes was observed in an ISSR assay. Detailed genetic diversity characteristics revealed that M. polystroma isolates from cherries did not create a distinct group but were intermingled with M. polystroma isolates from other hosts. The results of the pathogenicity test conducted on different fruit species indicated a lack of host specificity for M. polystroma isolates.     

Diversity of Botryosphaeriaceae species associated with conifers in Portugal

Botryosphaeriaceous fungi were isolated from conifers showing disease symptoms such as diebacks, blights, and cankers. The isolates were grouped based on morphology and ERIC-PCR fingerprinting patterns and representatives of each group were identified by sequencing of the internal transcribed spacer (ITS) region. Nine species from four different genera in Botryosphaeriaceae were identified within the isolates. Many new fungus-host associations were established and several species of Botryosphaeriaceae are reported from conifers for the first time. Most of these species also represent new reports from Portugal. The genus Neofusicoccum that was thought to be mainly restricted to angiosperms was the most frequent within the collection of isolates, followed by Diplodia. Dothiorella and Botryosphaeria represented a minor fraction of the isolates. Interestingly, the most common species was N. luteum, which had never been reported from coniferous hosts. Our results indicate that Neofusicoccum species may be more important as pathogens of conifers than it was previously recognised.     

Identification,potential inoculum sources and pathogenicity of botryosphaeriaceous species associated with grapevine dieback disease in New Zealand

This study investigated the prevalence and identity of botryosphaeriaceous dieback pathogens in necrotic grapevines tissues in New Zealand vineyards, and other woody hosts growing nearby. The presumptive identities of the isolates by conidial and cultural morphology were confirmed with ITS sequence data as Neofusicoccum australe, N. luteum, N. parvum and Diplodia seriata. They were isolated predominantly from necrotic stems of grapevine and other hosts, but also from leaves, flowers and wood debris of grapevines. Inoculation with conidia and mycelium of multiple isolates of each species onto excised and attached green shoots and trunks of five grapevine varieties, Cabernet sauvignon, Chardonnay, Pinot noir, Riesling, and Sauvignon blanc, showed that all varieties became infected to a similar extent. All species except D. seriata were pathogenic, irrespective of the host source, with N. luteum being the most and D. mutila the least pathogenic (P < 0.05). On trunks, N. parvum caused cankers and the other pathogenic species caused die-back when the inoculated vines became winter-dormant. Conidia were produced from green shoot lesions and die-back wood, which indicates potential inoculum sources for vineyard infection.     

Incidence of grapevine trunk diseases is influenced by soil,topology and vineyard age,but not by <Emphasis Type="Italic">Diplodia seriata</Emphasis> infection rate in the Tokaj Wine Region,Hungary

Grapevine Trunk Diseases (GTD) are of great importance worldwide, including Hungary, a Center European country with long wine producing history. Several GTD pathogens have been described till now in Europe, but only a few from Hungary. The presence of a GTD pathogen in the vine does not necessarily result in the immediate appearance of disease symptoms, and information on the importance of environmental factors related to disease incidence are still limited. The aim of this research was to assess the occurrence of GTD in the Tokaj Wine Region, and to determine the biotic and abiotic factors influencing disease incidence. Five vineyards within 15 km radius – each with different topology, soil types, varieties and age – were studied for three consecutive years (2013 – 2015). The incidence of GTD-infection was determined every year for each vineyard. Diplodia seriata was isolated with incidence ranging from 50 to 100%, while Diaporthe spp. were the only other – minor – GTD pathogen found. Topology and soil type appeared to be major abiotic factors affecting incidence of GTD symptom. Disease incidence was also positively correlating with the age of the vineyards, and it was in fact found to be the definitive biotic factor regarding incidence. In contrast, D. seriata infection rate appeared unrelated to disease incidence or to any of the biotic or abiotic factors investigated.     

引起甜樱桃叶斑病的葡萄座腔菌的鉴定

欧洲甜樱桃(Prunus avium L.)又名大樱桃,其色鲜味美,成熟期早,具有较高的经济价值.近年来我国甜樱桃种植面积逐年增加,伴随的病害问题也愈加凸显[1,2].叶斑病是樱桃上发生普遍,且最严重的病害之一,直接影响叶片的光合作用[3,4],削弱树势,致使樱桃产量和品质降低,经济损失严重.国际上报道的樱桃叶斑病的病...     

Pseudomonas syringae pv. avii (pv. nov.), the Causal Agent of Bacterial Canker of Wild Cherries (Prunus avium) in France

Bacterial strains isolated from cankers of wild cherry trees (Prunus avium) in France were characterized using numerical taxonomy of biochemical tests, DNA–DNA hybridization, repeat sequence primed-PCR (rep-PCR) based on REP, ERIC and BOX sequences, heteroduplex mobility assay (HMA) of internal transcribed spacer (ITS) as well as pathogenicity on wild cherry trees and other species of Prunus. They were compared to reference strains of Pseudomonas syringae pathovars isolated from wild and sweet cherry and various host plants. Wild cherry strains were closely related to P. syringae (sensu lato) in LOPAT group Ia (+ - - - +). Wild cherry strains were pathogenic to wild cherry trees and produced symptoms similar to those observed in orchards. They were pathogenic also, but at a lesser extent, to sweet cherry trees (cv. Napoléon). The wild cherry strains were collected from five different areas in France and appeared to constitute a very homogeneous group. They showed an homogenous profile of a biochemical and physiological characteristics. They were closely related by DNA–DNA hybridization and belonged to genomospecies 3 `tomato'. Rep-PCR showed that wild cherry strains constitute a tight group distinct from P. s. pv. morsprunorum races 1 and 2 and from other P. syringae pathovars. HMA profiles indicated that the ITS of all wild cherry strains were identical but different from P. s. pv. persicae strains since the two heteroduplex bands with reduced mobility were generated by hybridization with the P. s. pv. persicae pathotype strain CFBP 1573. The 8 genomospecies of Gardan et al. (1999) have not been converted into formal species as they cannot be differentiated by biochemical tests. Therefore, the pathovar system within P. syringae was currently used. P. syringae pv. avii is proposed for this bacterium causing a wild cherry bacterial canker and strain CFBP 3846 (NCPPB 4290, ICMP 14479) is designated as the pathotype.     

Transmission of Phytophthora ramorum in Mixed-Evergreen Forest in California

ABSTRACT During 2001 to 2003, the transmission biology of Phytophthora ramorum, the causal agent of sudden oak death, was studied in mixedevergreen forest, a common forest type in northern, coastal California. Investigation of the sources of spore production focused on coast live oak (Quercus agrifolia) and bay laurel (Umbellularia californica), dominant hosts that comprised 39.7 and 46.2% of the individuals at the study site, respectively. All tests for inoculum production from the surface of infected coast live oak bark or exudates from cankers were negative. In contrast, sporangia and chlamydospores were produced on the surface of infected bay laurel leaves. Mean number of zoospores produced from infected bay laurel leaves under natural field conditions during rainstorms was 1,173.0 +/- SE 301.48, and ranged as high as 5,200 spores/leaf. P. ramorum was recovered from rainwater, soil, litter, and streamwater during the mid- to late rainy season in all 3 years of the study. P. ramorum was not recovered from sporadic summer rains or soil and litter during the hot, dry summer months. Concentrations of inoculum in rainwater varied significantly from year to year and increased as the rainy season progressed for the two complete seasons that were studied. Potential dispersal distances were investigated for rainwater, soil, and streamwater. In rainwater, inoculum moved 5 and 10 m from the inoculum source. For soil, transmission of inoculum was demonstrated from infested soil to bay laurel green leaf litter, and from bay laurel green leaf litter to aerial leaves of bay laurel seedlings. One-third to one-half of the hikers tested at the study site during the rainy season also were carrying infested soil on their shoes. In streamwater, P. ramorum was recovered from an unforested site in pasture 1 km downstream of forest with inoculum sources. In total, these studies provide details on the production and spread of P. ramorum inoculum in mixed-evergreen forest to aid forecasting and managing disease transmission of this environmentally destructive pathogen.     

Molecular characterization of Pseudomonas syringae isolates from fruit trees and raspberry in Serbia

Infection of fruit trees by Pseudomonas syringae is a potentially serious problem that may limit the establishment and sustained productivity of pome and stone fruit orchards in Serbia. To estimate possible diversity of Pseudomonas syringae fruit trees strains, we collected a set of strains in several areas of Serbia. The samples were taken from infected orchards with raspberry, plum, cherry, sour cherry, peach, pear and apple trees. Genetic diversity of P. syringae strains isolated from fruit trees was determined by using SpeI macrorestriction analysis of genomic DNAs by pulsed-field gel electrophoresis (PFGE) and REP-PCR. Molecular analysis showed that most of isolates had unique profiles, with the exception of isolates from plum and cherry that displayed profiles identical to each other and similar to P. syringae pv. morsprunorum. The study presented here clearly demonstrates the discriminative power of molecular techniques in enabling a detailed analysis of the genetic variations between strains of P. syringae from different pome and stone fruit hosts in Serbia.     

Host switching between native and non‐native trees in a population of the canker pathogen Chrysoporthe cubensis from Colombia

The purpose of this study was to test the hypothesis that Chrysoporthe cubensis on native trees in South America could be the source of the pathogen that causes severe stem cankers and often mortality in commercially propagated Eucalyptus trees. This was done by investigating populations originating from two adjacent Eucalyptus (Myrtaceae) plantations in Colombia, and wild Miconia rubiginosa trees (Melastomataceae) growing alongside these stands. Polymorphic microsatellite markers were used to quantify allele sizes in 20 and 39 isolates from the two Eucalyptus stands and 32 isolates from adjacent M. rubiginosa trees. Gene and genotypic diversities were calculated from these data, and population differentiation and assignment tests were performed to ascertain whether the populations were genetically different. Results showed that there were no differences between any of the populations using these techniques, and that they can be treated as a single population. Therefore, the results support the hypothesis that host switching has occurred in C. cubensis in Colombia.     

Analysis of the natural infection of European horse chestnut (Aesculus hippocastanum) by Pseudomonas syringae pv. aesculi

Pseudomonas syringae pv. aesculi (Psa) is an emerging bacterial pathogen responsible for a recent epidemic of bleeding canker of European horse chestnut (Aesculus hippocastanum) in northwest Europe. Very little is known about the infection biology of this pathogen, which can cause lethal cankers in the branches and stem of its host. In this study, branches and whole trees of European horse chestnut naturally infected with Psa were subjected to detailed morphological and histological examination to identify the primary infection sites, the time of infection, and the patterns of subsequent lesion expansion within the host. Lesions developed during the host dormant season on the 2003–2009 extension growth increments and were centred mainly on lenticels, leaf scars and nodes. The oldest lesion developed in the 2004/2005 dormant season and the number of new lesions increased in each subsequent year. The lesions developed in the cortex and phloem and extended into the cambium to cause cankers, but there was no evidence of necrosis in the xylem. All lesions on the branches were discrete and apparently contained by a necrophylactic periderm, although there was evidence that Psa could survive within such periderms and subsequently breach them. Examination of two whole 30‐year‐old trees revealed extensive, continuous cankers in the phloem and cambium which had formed within a single growing season. Thus, the success of Psa as a tree pathogen and the causal agent of a large‐scale epidemic may in part reflect an ability to infect the aerial woody parts of its host directly.     

Brenneria quercina and Serratia spp. isolated from Spanish oak trees: molecular characterization and development of PCR primers

Brenneria quercina has been reported as one of the causal agents of oak decline in Spain. To investigate the bacterial variability of this pathogen from different Spanish oak forests, a collection of 38 bacterial isolates from seven geographic locations and from different oak species was analysed by sequencing 16S rDNA and rep-PCR fingerprinting. All Spanish isolates of B. quercina were grouped by rep-PCR into a homogenous cluster that differed significantly from B. quercina reference strains from California. 16S rDNA analysis revealed that 34 out of 38 isolates were Brenneria . However, four isolates belonged to the genus Serratia , suggesting that this bacterium could cause cankers in oak trees. The information obtained by rep-PCR fingerprint analysis was used to develop PCR primers for the sensitive and specific detection of B. quercina from infected plant tissues. Pathogenicity tests performed with Brenneria and Serratia isolates showed that both were able to grow and cause cankers in oak trees.     

Identification and Discrimination of Pseudomonas syringae Isolates from Wild Cherry in England

A survey of wild cherry (Prunus avium) woodland plantations and nurseries was carried out in 2000/01. Trees with symptoms of bacterial canker were found in 20 of the 24 plantations visited and in three of seven nurseries. Fifty-four Pseudomonas syringae isolates from wild cherry together with 22 representative isolates from sweet cherry and 13 isolates from other Prunus spp., pear and lilac were characterised by physiological, biochemical, serological and pathogenicity tests. Isolates from wild cherry were predominantly P. syringae pv. syringae (Pss), but P. syringae pv. morsprunorum (Psm) races 1 and 2 were also found. Physiological and biochemical tests discriminated Psm races 1 and 2 from other P. syringae isolates. Agglutination and indirect-enzyme-linked immunosorbent assay tests with three different antisera showed that Psm race 1 and race 2 were very uniform and indicated high variability amongst other P. syringae isolates. However, pathogenic Pss isolates could not be distinguished from non-pathogenic isolates of P. syringae on the basis of physiological, biochemical or serological tests. Pathogenicity tests on rooted lilac plants and on micropropagated plantlets of lilac and two wild cherry clones differentiated Pss and Psm isolates and demonstrated a range of aggressiveness amongst Pss isolates. Serological tests could be used as an alternative to the classical physiological and biochemical tests to increase the speed of detection and discrimination of isolates, but pathogenicity tests are still necessary to discriminate the pathogenic Pss isolates.