Characterization of fungal pathogens (Diaporthe angelicae and D. eres) responsible for umbel browning and stem necrosis on carrot in France

A collection of 102 Diaporthe isolates was compiled from lesions on carrot, parsley and wild Apiaceae species in France from 2010 to 2014. Molecular typing based on ITS rDNA sequences resulted in the identification of 85 D. angelicae and 17 D. eres isolates. Based on sequences of the 3′ part of the IGS rDNA, intraspecific variability was analysed for 17 D. angelicae and 13 D. eres isolates from diverse plant species, locations in France, and plant tissues. The genetic diversity was greater for D. angelicae isolates than D. eres isolates. In vitro sensitivity of five D. angelicae and four D. eres isolates to each of nine fungicides was similar for isolates of both species, with a marked variation in fungicide sensitivity depending on the active ingredient. To assess the pathogenicity of D. angelicae and D. eres isolates on carrot, one isolate of each species was inoculated onto umbels in a controlled environment. Typical lesions were observed for both isolates. Carrot crop debris collected from a seed production field in France and placed in controlled conditions produced perithecia and ascospores typical of Diaporthe, that were further characterized molecularly as belonging to D. angelicae. Detection of Diaporthe species on seed lots from three carrot production fields in France was investigated. Both species were detected on seeds by conventional PCR assay, with a greater frequency for D. angelicae than D. eres (67% vs 33%, respectively). Overall, the results highlighted that umbel browning in carrot seed crops in France was mainly caused by D. angelicae.     

Identification and characterization of <Emphasis Type="Italic">Diaporthe vaccinii</Emphasis> Shear causing upright dieback and viscid rot of cranberry in Poland

During the summers of 2013–2014, symptoms similar to viscid rot and upright disease were observed on cranberries (Vaccinium macrocarpon) on one plantation in central Poland. The associated fungi were isolated from symptomatic plant tissue. On the basis of morphological and cultural characteristics and the ability of isolated fungi to elicit viscid rot symptoms on cranberry fruits, they were classified as the genus Diaporthe. Further analysis of ITS sequence data allowed for the classification of the newly obtained isolates as D. vaccinii. Additional analysis of genetic diversity using five RAPD and eight ISSR primers constituted additional confirmation of genetic distance existing between closely related D. vaccinii and D. eres species, enabling their differentiation.     

Pathogenicity of fungi associated with ash dieback towards Fraxinus excelsior

A large part of the area in Europe in which Fraxinus excelsior is native is currently affected by ash dieback, a threatening disease caused by the ascomycetous fungus Hymenoscyphus fraxineus. Fungi other than H. fraxineus also occur in large numbers on stems of the dying ash trees. To clarify their possible role in the dieback process, six fungal species common on dying stems and twigs of ash in Poland, i.e. Cytospora pruinosa, Diaporthe eres, Diplodia mutila, Fusarium avenaceum, F. lateritium and F. solani, were tested for pathogenicity using a test based on artificial wound inoculations of 6‐year‐old F. excelsior plants under field conditions, with H. fraxineus included for comparison. There were significant differences in index of pathogenicity among the fungi tested. Hymenoscyphus fraxineus (mean index 5.78) was the most pathogenic. Diplodia mutila (4.23) and C. pruinosa (4.02) were significantly less pathogenic than H. fraxineus, but significantly more than the other fungi. Diaporthe eres (2.43), F. avenaceum (1.92), F. solani (1.86) and F. lateritium (1.08) were the least pathogenic (P < 0.0001). The extent of disease symptoms caused by F. solani and F. lateritium was statistically similar to the control (P = 0.05). All tested fungi were successfully reisolated from inoculated stems. The contribution of the results to understanding the possible role of these fungi in the ash dieback process in F. excelsior, particularly in trees weakened after primary infection by H. fraxineus, is discussed.     

Fungi as potential factors limiting natural regeneration of pedunculate oak (Quercus robur) in mixed-species forest stands in Poland

Dieback of young Quercus robur seedlings can limit natural regeneration in mixed-species forest stands in Poland. The aim of this study was to examine the role of fungi in the dieback of oak seedlings in central Europe. Fungi were isolated from the stems and roots of Q. robur, from both healthy seedlings and seedlings with symptoms, that were sampled from four stands in Poland. In total, 111 distinct taxa were identified. Ascomycota was dominant, representing 95.6% of the isolates. Among the taxa identified, Alternaria alternata, Colletotrichum fioriniae, Colletotrichum godetiae, Coniella quercicola, Diaporthe eres, Gnomoniopsis paraclavulata, Ilyonectria rufa, Mucor genevensis, Penicillium glandicola, Tubakia dryina, and Umbelopsis changbaiensis were most frequently isolated, and were consistently found in stems and roots of both healthy and diseased plants. The community compositions of fungi in healthy and diseased stems were similar, although Fusarium species, especially Fusarium sp. 1 (FTSC 5) and F. sporotrichioides, were found mainly in seedlings with symptoms. The pathogenicity of the most consistently isolated species from stems and roots of diseased seedlings was tested on Q. robur seedlings. F. sporotrichioides caused the largest lesions on inoculated seedlings. Six weeks after inoculation, D. eres, C. fioriniae, G. paraclavulata, T. dryina, and F. sporotrichioides killed 0%–18.8% of seedlings, while I. rufa and Ilyonectria pseudodestructans did not cause any lesions or other symptoms. This study is the first comprehensive report suggesting that massive fungal attack can lead to oak seedling dieback in mixed-species forest stands in central Europe.     

Phomopsis husk rot of macadamia in Australia and South Africa caused by novel Diaporthe species

Phomopsis husk rot (PHR) in macadamia is a disease of economic importance in major commercial production areas in Australia and South Africa. Effective control of PHR is hindered by limited knowledge about its aetiology and epidemiology. The diversity and pathogenicity of more than 50 isolates of Diaporthe associated with PHR in macadamia orchards in Australia and South Africa was assessed. Multilocus phylogenetic analyses of DNA sequences of the ITS, tef1α, and tub2 gene loci revealed four novel clades that are described as Diaporthe australiana sp. nov., D. drenthii sp. nov., D. macadamiae sp. nov., and D. searlei sp. nov. Pathogenicity tests with representative isolates found that all four species caused PHR of varying severity between and within species, as well as between the two macadamia cultivars HAES 344 and HAES 816. The Australian species, D. australiana, was the most aggressive species compared with the three South African species. This study improves our understanding of the aetiology of PHR in macadamia and paves the way for more effective disease management.     

Aetiology of branch dieback,panicle and shoot blight of pistachio associated with fungal trunk pathogens in southern Spain

Pistachio represents an emerging nut crop across the Mediterranean basin. In Spain, pistachio has been traditionally cultivated in marginal-dry areas with unfavourable climatic conditions for plant diseases. Consequently, little attention has been given to research on pistachio diseases until recently. Symptoms of branch dieback and cankers, and shoot and panicle blight have been recently observed in commercial pistachio orchards across southern Spain. In this study, 10 commercial pistachio orchards showing disease symptoms were surveyed between 2017 and 2018. Botryosphaeriaceae fungi were consistently isolated from affected shoots, among other fungal families with minor relevance. Representative isolates of each family were characterized based on colony and conidial morphology, optimum growth temperature, and the comparison of DNA sequence data (ITS, LSU, EF, TUB2, and ACT genomic regions). Detached and attached shoots, and attached panicles of pistachio cv. Kerman were inoculated with mycelial plugs or conidial suspensions to demonstrate the pathogenicity of the selected isolates. Botryosphaeria dothidea, Lasiodiplodia pseudotheobromae, Neofusicoccum mediterraneum, N. parvum, Diaporthe neotheicola, Diaporthe sp., Eutypa lata, Eutypa sp., Cytospora sp., and Phaeoacremonium minimum were identified. P. minimum had the highest optimum growth temperature (29.6 °C) and Cytospora sp. the lowest (21–22 °C). Botryosphaeriaceae isolates showed the largest lesions on inoculated shoots, with L. pseudotheobromae being the most aggressive, followed by Neofusicoccum species. Panicles inoculated with N. mediterraneum showed blight symptoms and canker formation 6 weeks after inoculation, without significant differences in aggressiveness between isolates. This work reports relevant information about this emerging disease in the novel Spanish pistachio-growing areas.     

Effect of Diaporthe RNA virus 1 (DRV1) on growth and pathogenicity of different <Emphasis Type="Italic">Diaporthe</Emphasis> species

A 4.1 kbp positive-strand RNA virus known as Diaporthe RNA virus 1 (DRV1) occurs in hypovirulent, non-sporulating isolates of the fungal pathogen Diaporthe perjuncta. A full-length cDNA clone of DRV1 was developed and RNA transcribed from the cDNA clone used to transfect different Diaporthe spp. The transfected species included three D. ambigua isolates and an unidentified Phomopsis asexual state of a Diaporthe sp. Successful transfections were confirmed using RT-PCR. Although the in vitro-transcribed positive sense single-stranded RNA used for transfection included vector sequences at both ends, the genomes of progeny virus from DRV1-transfected isolates were free of the vector sequences. Transfection resulted in morphological changes in these fungal pathogens. However, the presence of DRV1 did not reduce growth rate in two of the three D. ambigua or the Phomopsis sp. significantly. Pathogenicity studies showed that the transfected isolates have reduced aggresiveness.     

Pathogenicity of Colletotrichum species causing anthracnose of Capsicum in Asia

Anthracnose of chili is caused by a complex of Colletotrichum species, with recent surveys reporting at least 28 different species implicated. However, there have been very few studies to identify the relative pathogenicity of the various species or to optimize a bioassay to assess pathogenicity. A detached Capsicum fruit bioassay to determine the pathogenicity of a diverse geographical range of isolates of Colletotrichum scovillei showed fruit maturity, host genotype, and inoculation method all interact to affect infection and rate of lesion development. On Capsicum annuum ‘Bangchang’ fruit wounded prior to inoculation, pathogenicity was consistent regardless of fruit maturity. In contrast, without wounding there was variability in pathogenicity. On the relatively resistant host Capsicum chinense PBC932, pathogenicity was dependent on both the inoculation method and the maturity stage of the fruit. In addition, lack of correlation in pathogenicity of isolates between the two Capsicum lines indicated that there was host–isolate specialization that would make prediction of pathogenicity of isolates on host difficult. In a further study, 10 species of Colletotrichum isolated from diseased chili fruits in Asia caused anthracnose symptoms on C. annuum ‘Bangchang’ under all testing conditions, with large differences in aggressiveness. C. chinense PBC932 was generally more resistant to all the species, with smaller lesions produced in different host conditions. Colletotrichum javanense and C. scovillei were highly aggressive relative to other species, especially when inoculated on nonwounded fruit. Pathotype differences were identified within multiple isolates of C. scovillei and C. siamense, the two most frequently identified pathogenic species on chili.     

Diversity and pathogenicity of Fusarium graminearum species complex from maize stalk and ear rot strains in northeast China

The Fusarium graminearum species complex (FGSC) is an important group of pathogens distributed in maize‐producing areas worldwide. This study investigated the genetic diversity and pathogenicity of 40 FGSC isolates obtained from stalk rot and ear rot samples collected from 42 locations in northeastern China during 2013 and 2014. A phylogenetic tree of translation elongation factor (EF‐la) sequences designated the 40 isolates as F. graminearum sensu stricto (67.5%) and F. boothii (32.5%). By using inter‐simple sequence repeat analysis (ISSR), it was shown that the isolates were divided into two clades, which corresponded to the species identity of the isolates. However, the isolates from the two different diseases could not be distinguished in pathogenicity. The disease severity index of seedlings inoculated with stalk isolates was slightly higher than that of seedlings inoculated with isolates from infected ears, whereas the pathogenicity of the stalk and ear isolates were identical.     

Diaporthe sclerotioides exhibits no host specificity among cucurbit species

Plant–fungal specificity between cucurbitaceous crops and Diaporthe sclerotioides, the causal agent of black root rot, was studied using cucumbers (Cucumis sativa), melons (Cucumis melo), pumpkins (Cucurbita maxima), watermelons (Citrullus lanatus) and bottlegourd (Lagenaria siceraria var. gourda). Twelve D. sclerotioides isolates from these cucurbit species were cross‐inoculated. The virulence of the isolates was evaluated as the area under the disease progress curve (AUDPC). All cucurbit species were susceptible to each isolate, but AUDPCs were significantly different among the hosts, with the order of greatest to least being melon, cucumber, watermelon, bottlegourd and finally, pumpkin. The infectiveness of isolates was assessed as the quantity of D. sclerotioides DNA detected in the hypocotyls of seedlings 2 weeks after inoculation using a real‐time PCR protocol. The fungal DNA quantities varied among the species in the same order as the AUDPCs. Whilst there were statistically significant correlations between the virulence and infectiveness of D. sclerotioides isolates in cucumbers, melons and bottlegourds, their coefficients of determination were not high (r² < 0·6). Orthogonal contrasts indicated no specificity in either the fungal virulence or infectiveness between D. sclerotioides isolates and the cucurbit hosts from which these isolates originated. Thus, although the degree of host susceptibility to D. sclerotioides varies among cucurbit species, the absence of specificity to the host species in either virulence or infectiveness suggests the pathogen may spread via various cucurbit crops, irrespective of their original host species.     

Cross-pathogenicity of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> between potato and sunflower

This study examined cross-pathogenicity of the soilborne pathogen Verticillium dahliae between potato and sunflower. Four week-old potato and sunflower seedlings were inoculated with ten isolates from each of the two host species. Potato cultivars (Kennebec, susceptible, and Ranger Russet, moderately resistant) and sunflower hybrids (IS8048, susceptible, and 6946, moderately resistant) were assessed for disease severity and percent infection at 2 weeks, 3 weeks, 4 weeks, 5 weeks, and 6 weeks after inoculation (w.a.i), and for vascular discolouration at 6 w.a.i., using visual scales developed for each host species. The experiments were conducted in 2006 and repeated in 2007. Based on percent infection and disease severity, most V. dahliae isolates were highly aggressive on both host species. The tested isolates caused higher disease levels in the susceptible than in the moderately resistant phenotypes. They also caused more vascular discolouration in their original than in the alternative host. However, the isolates originating from sunflower caused less infection and disease severity on both hosts, compared to their potato counterparts. Cluster analysis based on all of the criteria used to assess pathogenicity led to three groups of isolates: (i) most V. dahliae potato isolates, which ranged with the highly aggressive control isolates, (ii) one V. dahliae sunflower isolate, which showed a similar pathogenicity level to the weakly-aggressive V. albo-atrum sub-group II control isolate, with no more symptoms than in the non-inoculated plants, and (iii) most V. dahliae sunflower isolates with mildly- to weakly-aggressive levels. Based on these results, V. dahliae cross-pathogenicity is very effective between potato and sunflower. Therefore, rotations involving these species should be avoided, especially where sunflower follows potato.     

Death of endemic Virgilia oroboides trees in South Africa caused by Diaporthe virgiliae sp. nov.

Numerous dead and dying individuals of the Western Cape endemic tree Virgilia oroboides (Fabaceae) were recently observed within a South African national botanical garden. Root‐rot fungi and fungi symbiotic with bark beetles (Curculionidae; Scolytinae) from diseased trees were assessed for their respective roles in V. oroboides mortality. Disease progression was also monitored over 1 year. Fungi were isolated from surface sterilized bark and root samples from diseased trees and provisionally identified using data from the internal transcribed spacer regions (ITS1, ITS2), including the 5·8S rRNA gene (ITS). Pathogenicity of selected fungi towards V. oroboides was tested under field conditions. The pathogenicity of various bark beetle‐associated Geosmithia (Hypocreales: Hypocreomycetidae) spp. from V. oroboides were similarly assessed. The only fungus consistently isolated from lesions on the roots and bark of declining V. oroboides, and never from healthy individuals, represented an undescribed Diaporthe (Diaporthales, Diaporthaceae) species that was characterized using molecular (using data from the ITS marker and part of the β‐tubulin gene, TUB), cultural and morphological characters. It is an aggressive pathogen of V. oroboides, newly described here as Diaporthe virgiliae sp. nov. Trees of all ages are susceptible to this pathogen with subsequent bark beetle attack of mature trees only. All Geosmithia spp. from beetles and/or infected trees were nonpathogenic towards V. oroboides. Diaporthe virgiliae caused a severe decline in the health of the monitored V. oroboides population over a period of only 1 year and should be considered as a significant threat to these trees.     

PCR–RFLP and Sequence Data Delineate Three Diaporthe Species Associated with Stone and Pome Fruit Trees in South Africa

Diaporthe species include canker pathogens on a wide variety of hosts. In South Africa, Diaporthe canker of apple, pear and plum rootstocks has been attributed to Diaporthe ambigua. Recently, we recognized that isolates of D. ambigua exhibited different morphological features and thus questioned the identity of these isolates. A small set of isolates was thus chosen for comparison using DNA-based methods. Polymerase chain reaction amplification of ribosomal DNA, Restriction Fragment Length Polymorphisms and DNA sequencing revealed that isolates which had been regarded as D. ambigua in the past were three distinct species. These are D. ambigua, D. perjuncta and an unknown Phomopsis sp. This discovery has special relevance to research done on a dsRNA virus previously thought to occur in D. ambigua and now shown to infect D. perjuncta.     

Evaluating aggressiveness and host range of Alternaria dauci in a controlled environment

The aggressiveness of Alternaria dauci isolates was investigated in greenhouse conditions. Twenty‐seven isolates were pre‐selected from a large collection to represent high diversity according to geographic or host origins and intergenic spacer (IGS) polymorphism. IGS sequence analysis revealed that isolates were grouped within three different clusters. Eleven isolates were selected and inoculated on a susceptible carrot cultivar. Three criteria (mean lesion number, mean necrotic leaf area and mean disease index) were used to assess the aggressiveness of isolates. Continuous variation in aggressiveness was shown and no clear division into isolate classes was evident. For the host range study, two isolates were inoculated under greenhouse conditions onto nine cultivated Apiaceae species, two wild Daucus species and six cultivated non‐Apiaceae species representing six botanical families. Lesions varying in severity were observed on all dicot species (Apiaceae and non‐Apiaceae), but no symptoms developed on the two monocots studied (leek and sweetcorn). Plant species were also differentiated on the basis of expanding lesions (cultivated and wild carrot, dill and fennel) or non‐expanding lesions (other dicot species). Typical A. dauci conidia were observed after in vitro incubation of leaves with symptoms. Fungal structures were isolated from lesions and A. dauci was confirmed on the basis of conidial morphology and specific conventional PCR results. Genotyping of individual isolates performed with microsatellite markers confirmed the presence of the inoculated isolate. The results clearly showed that, in controlled conditions, the host range of A. dauci is not restricted to carrot.     

First report of blueberry red ringspot disease caused by <Emphasis Type="Italic">Blueberry</Emphasis><Emphasis Type="Italic">red</Emphasis><Emphasis Type="Italic">ringspot</Emphasis><Emphasis Type="Italic">virus</Emphasis> in Japan

Virus-like symptoms—red ringspots on stems and leaves, circular blotches or pale spots on fruit—were found on commercial highbush blueberry (Vaccinium corymbosum) cultivars Blueray, Weymouth, Duke and Sierra in Japan. In PCR testing, single DNA fragments were amplified from total nucleic acid samples of the diseased blueberry bushes using primers specific to Blueberry red ringspot virus (BRRV). Sequencing analysis of the amplified products revealed 95.7–97.7% nucleotide sequence identity with the BRRV genome. This paper is the first report of blueberry red ringspot disease caused by BRRV in Japan. The nucleotide sequence data reported in this paper are available in the GenBank/EMBL/DDBJ database as accessions AB469884 to AB469893 for BRRV isolates from Japan.     

Genotypic and phenotypic characterization of Streptomyces species associated with potato crops in the central part of Colombia

In Colombia, Streptomyces scabiei (syn. S. scabies) is commonly believed to be the causal organism of scab disease in local potato crops. However, very little is known about this organism and about the diversity and pathogenicity of the Streptomyces species associated with potato crops in Colombia. This study, therefore, aimed to elucidate aspects regarding the diversity of these bacteria associated with potato crops in a particular region of Colombia and evaluate their pathogenicity. We obtained 33 isolates of Streptomyces from netted, superficial and deep-pitted potato scab lesions from two main potato-producing regions in Colombia. Of these, 17 were pathogenic based on in vitro and in planta assays. None of these isolates carried the txtA, txtB, or nec1 genes, commonly associated with pathogenicity in Streptomyces, and characteristic of the pathogenicity island (PAI). We also characterized all isolates based on phenotypic characteristics and analysed their phylogenetic relationships using the 16S rRNA, atpD, recA, rpoB, and trpB genes. The isolates were highly diverse, placed in nine clades with 15 different phenotypes. The 17 pathogenic isolates were placed into three clades, namely S. pratensis, S. xiamenensis, and unknown species. This study is a preliminary investigation towards understanding scab disease in Colombia through the study of both pathogenic and nonpathogenic species present in scab disease lesions in potatoes. Also, this is the first report of Streptomyces species associated with potato tubers in Colombia.     

Identification of Colletotrichum siamense causing litchi pepper spot disease in mainland China

Litchi (Litchi chinensis) pepper spot disease results in black spotting symptoms on litchi fruits. This disease was first observed on litchi cultivar Guiwei, in Guangzhou, China, in 2009, and then found widespread in other litchi-growing regions of China. Colletotrichum isolates were consistently recovered from typical black spot lesions of diseased fruits with frequency ranging from 83% to 100%. Three representative Colletotrichum isolates from Maoming, Guangzhou and Shenzhen were selected for identification and pathogenicity testing in the field. Based on morphology and phylogenetic analysis using the ribosomal internal transcribed spacer region (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), calmodulin (CAL), actin (ACT), β-tubulin (TUB2) and glutamine synthetase (GS) gene sequences, the three isolates were identified as C. siamense. In the pathogenicity experiments, typical symptoms appeared on the inoculated litchi fruits, including black spots and green patches around these black spots. These symptoms were consistent with the symptoms originally observed in the field. Colletotrichum siamense was successfully reisolated from the typical black spot lesions of the inoculated litchi fruits. To the authors' knowledge, this is the first report on characterization of C. siamense as the causal agent of litchi pepper spot disease in mainland China by successful inoculation on fruits under field conditions.     

Virulence affected by assay parameters during grapevine pathogenicity studies with Botryosphaeriaceae nursery isolates

This study assessed the symptoms that developed when 114 Botryosphaeriaceae isolates from grapevine nursery plant materials were used to inoculate excised green shoots and 1‐year‐old rooted canes of Sauvignon blanc. The experiments showed that all isolates and species were able to produce lesions. Overall, the Neofusicoccum species were shown to be highly pathogenic in both tissue types while the Diplodia species were highly pathogenic on canes but not on green shoots. Isolates of the most prevalent species, N. luteum and N. parvum, showed varying lesion lengths on excised green shoots and canes. An evaluation of the factors associated with lengths of lesions showed that they were significantly affected by experimental batch which reflected inherent host and environmental factors over time. Reisolation from inoculated canes also indicated that most isolates of all species except D. seriata were able to spread internally beyond the lesions. Genetic variability analysis using UP‐PCR showed that N. luteum isolates were genetically diverse but no association was observed between the phylogenetic group and degree of pathogenicity caused by the isolates. This study demonstrated that all Botryosphaeriaceae species from grapevine nurseries were pathogenic to grapevines and that the lesion lengths varied between species, among isolates within a species and among nursery sources, and was affected by the test method.     

Species of the Colletotrichum gloeosporioides and C. boninense complexes associated with olive anthracnose

The taxonomic status of Colletotrichum gloeosporioides sensu lato (s.l.) associated with olive anthracnose is still undetermined and the pathogenic ability of this species complex is controversial. In the present study, isolates obtained from olive and provisionally identified as C. gloeosporioides s.l. on the basis of morphological and cultural features were reclassified using ITS and TUB2 as DNA barcode markers and referred to seven distinct species, recently separated within C. gloeosporioides (C. aenigma, C. gloeosporioides sensu stricto (s.s.), C. kahawae, C. queenslandicum, C. siamense and C. theobromicola) and C. boninense (C. karstii) species complexes. Furthermore, isolates of C. kahawae were ascribed to the subspecies ciggaro by analysing the GS gene. A single isolate, not in either of these two species complexes, was not identified at the species level. In pathogenicity tests on detached olive drupes some of these species, including C. aenigma, C. kahawae subsp. ciggaro, C. queenslandicum, C. siamense and C. karstii, were shown to be weakly pathogenic. Moreover, they were found very sporadically on olive. In contrast, some isolates of C. gloeosporioides s.s. and isolates of C. theobromicola proved to be virulent on both green and ripening olives. This study gives a better insight into both the aetiology and the epidemiology of olive anthracnose and might have implications for biosecurity and quarantine because C. theobromicola has never been reported in major European olive‐producing countries.     

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.