Eutypella Citricola Research Articles

Identification and Pathogenicity of Fungal Species Associated with Branch Canker and Shoot Blight on Persimmons (Diospyros kaki) in California.

Persimmon is a relatively new crop to California agriculture. Asian persimmons (Diospyros kaki) are the dominant species commercially cultivated in the United States, primarily grown in California, covering approximately 1,153 ha of bearing trees. In the growing seasons of 2020 and 2021, unusual shoot blight and branch cankers were observed in several persimmon orchards in San Joaquin and Solano counties in California. The most prevalent symptoms were well-defined black discoloration in the cambium and streaking in the vascular tissues of green shoots. On woody branches and old pruning wounds, symptoms manifested as black wedge-shaped cankers. Isolations from affected tissues revealed the occurrence of Diaporthe species, including D. chamaeropis, D. foeniculina, and an undescribed Diaporthe sp. as the most frequent isolated pathogens, followed by Eutypella citricola and Phaeoacremonium iranianum. The isolates were identified through multilocus phylogenetic analyses using nucleotide sequences of the rDNA internal transcribed spacer, β-tubulin, and translation elongation factor 1-alpha genes. To fulfill Koch's postulates, mycelium plugs of the various fungal species identified were inserted in 2-year-old branches of mature persimmon trees after making wounds using a corkborer in field conditions. Results showed that Diaporthe spp., E. citricola, and P. iranianum are the main causal agents of branch canker and shoot dieback of persimmon trees in California, with Diaporthe spp. being the most frequently isolated pathogen.

Fungal Pathogens Associated with Almond Decline Syndrome, an Emerging Disease Complex in Intensive Almond Crops in Southern Spain.

In 2016, an almond (Prunus dulcis) decline syndrome (ADS) emerged in intensive almond plantations in the Andalusia region (southern Spain), showing branch dieback, gummosis, and general tree decline. The aim of this work was to elucidate the etiology of this disease complex. For this purpose, surveys were conducted across the Andalusia region, and a wide collection of fungi was recovered from wood samples showing gum and internal discoloration. Representative isolates were selected and identified by sequencing ITS, TEF1, TUB, ACT, LSU, and/or RPB2 genes. The following fungal species were identified to be associated with the disease: Botryosphaeria dothidea, Diplodia corticola, Di. seriata, Dothiorella iberica, Lasiodiplodia viticola, Macrophomina phaseolina, Neofusicoccum mediterraneum, N. parvum, N. vitifusiforme, Diaporthe neotheicola, Dia. rhusicola, Dia. ambigua, Eutypa lata, E. tetragona, Eutypella citricola, Eu. microtheca, Fusarium oxysporum s.l., Pleurostoma richardsiae, Phaeoacremonium iranianum, Pm. krajdenii, Pm. parasiticum, and Cytospora sp. All isolates were tested for pathogenicity by inoculating detached or attached almond shoots. Di. corticola and N. parvum were the most aggressive species, showing the largest lesions and most gummosis in attached shoots. The results suggest that the species belonging to Botryosphaeriaceae play a key role in disease development, while the remaining identified species may act as secondary pathogens or endophytes. However, further research to determine the interaction between all these fungal species and other biotic and abiotic factors in the ADS progress is needed.

Molecular detection and identification of Diatrypaceous airborne spores in Australian vineyards revealed high species diversity between regions.

The grapevine trunk disease, Eutypa dieback (ED), causes significant vine decline and yield reduction. For many years, the fungus Eutypa lata was considered the main pathogen causing ED of grapevines in Australia. Recent studies showed other Diatrypaceous fungi were also associated with vines exhibiting dieback symptoms but there is limited information on how these fungal pathogens spread in vineyards. Thus, information on the spore dispersal patterns of Diatrypaceous fungi in different wine regions will assist in identifying high-risk infection periods in vineyards. Using more than 6800 DNA samples from airborne spores collected from eight wine regions in south-eastern Australia over 8 years using a Burkard spore trap, this study investigated the diversity and abundance of Diatrypaceous species, using multi-faceted molecular tools. A multi-target quantitative PCR (qPCR) assay successfully detected and quantified Diatrypaceous spores from 30% of the total samples with spore numbers and frequency of detection varying between regions and years. The high-resolution melting analysis (HRMA) coupled with DNA sequencing identified seven species, with E. lata being present in seven regions and the most prevalent species in the Adelaide Hills, Barossa Valley and McLaren Vale. Cryptovalsa ampelina and Diatrype stigma were the predominant species in the Clare Valley and Coonawarra, respectively while Eutypella citricola and Eu. microtheca dominated in the Hunter Valley and the Riverina regions. This study represents the first report of D. stigma and Cryptosphaeria multicontinentalis in Australian vineyards. This study further showed rainfall as a primary factor that triggers spore release, however, other weather factors that may influence the spore release in different climatic regions of Australia still requires further investigation.

Identification and pathogenicity of six fungal species causing canker and dieback disease on golden rain tree in Beijing, China

ABSTRACT Golden rain trees (Koelreuteria paniculata) are largely cultivated because of their important ornamental, medicinal, and economic value. However, they are affected by canker and dieback disease to a large extent. To determine the fungi associated with canker and dieback disease of golden rain trees, isolations were obtained from diseased branches and twigs during 2019 and 2020 in greenbelts and nurseries in Beijing, China. Isolates were identified as six species (Allocryptovalsa castaneicola, Botryosphaeria dothidea, Cytospora koelreutericola sp. nov., Dothiorella acericola, Eutypella citricola, and Peroneutypa scoparia) based on morphological features and phylogenetic analyses of ITS, act, rpb2, tef1-α, and tub2. The results of pathogenicity tests indicated that all fungi produced discoloration and Botryosphaeria dothidea was highly aggressive to golden rain tree. In conclusion, this study explored the taxonomy, phylogeny, and pathogenicity of different fungal species associated with canker and dieback disease on golden rain tree and provided fundamental knowledge to improve disease management.

The Hidden Diversity of Diatrypaceous Fungi in China.

In this study, we investigated the diversity of diatrypaceous fungi from six regions in China based on morpho-molecular analyses of combined ITS and tub2 gene regions. We accept 23 genera in Diatrypaceae with 18 genera involved in the phylogram, and the other five genera are lacking living materials with sequences data. Eleven species included in four genera (viz. Allocryptovalsa, Diatrype, Diatrypella, and Eutypella) have been isolated from seven host species, of which nine novel species (viz. Allocryptovalsa castaneae, A. castaneicola, Diatrype betulae, D. castaneicola, D. quercicola, Diatrypella betulae, Da. betulicola, Da. hubeiensis, and Da. shennongensis), a known species of Diatrypella favacea, and a new record of Eutypella citricola from the host genus Morus are included. Current results show the high diversity of Diatrypaceae which are wood-inhabiting fungi in China.

Paraeutypella guizhouensis gen. et sp. nov. and Diatrypella longiasca sp. nov. (Diatrypaceae) from China.

BackgroundIn this study, we introduce a novel genus, Paraeutypella, of the family Diatrypaceae comprising three species viz. Paraeutypella guizhouensis sp. nov. and P. citricola (basionym: Eutypella citricola) and P. vitis (basionym: Sphaeria vitis). Diatrypella longiasca sp. nov. is also introduced, which forms a distinct clade in Diatrypella sensu stricto. The discovery of this new genus will contribute to expanding the knowledge and taxonomic framework of Diatrypaceae (Xylariales).New informationGeneric delimitations in Diatrypaceae are unsettled because the phylogeny has yet to be resolved using extensive taxon sampling and sequencing of ex-type cultures. During an investigation of xylarialean fungi, we collected eutypella-like fungi which is distinct from Eutypella sensu stricto in our phylogenetic analyses (ITS and β-tubulin), thus, introduced as Paraeutypella guizhouensis gen. et sp. nov.. Paraeutypella is characterised by having 4–25 perithecia in a stroma each with 3–6 sulcate, long ostiolar necks. Paraeutypella citricola comb. nov. (basionym: Eutypella citricola) is introduced on Acer sp. from China. Diatrypella longiasca sp. nov. is introduced as a new species in Diatrypella sensu stricto. which has 2–5 ascomata per stroma and long ascospores, unusual when compared to other Diatrypella species and distinct phylogenetically.

Fungal pathogens associated with branch and trunk cankers of nut crops in Iran

Branch and trunk canker diseases have become prevalent on nut crops in Iran. During 2015 to 2018, extensive field surveys were conducted on 58 almond, 43 pistachio and 80 walnut orchards in Iran to study fungal pathogens associated with symptomatic trees. One hundred and fifty-six representative fungal isolates were selected and identified based on morphological characteristics and by phylogenetic comparison of DNA sequence data. Fungal species found were Collophorina hispanica, Pleurostoma richardsiae, nine species of Phaeoacremonium (namely P. angustius, P. cinereum, P. italicum, P. fraxinopennsylvanicum, P. minimum, P. parasiticum, P. scolyti, P. tuscanum and P. viticola), 11 species of Botryosphaeriaceae (namely Botryosphaeria dothidea, Diplodia gallae, D. mutila, D. seriata, Dothiorella plurivora, Do. sarmentorum, Do. viticola, Lasiodiplodia citricola, L. mahajangana, L. theobromae and Neofusicoccum parvum), four species of Diatrypaceae (namely Cryptosphaeria pullmanensis, Diatrype whitmanensis, Eutypella citricola and E. vitis) and two non-identified Eutypella spp. (Eutypella sp. 1 and Eutypella sp. 2). Some of these species represent new reports in Iran and/or are reported for the first time in their respective hosts. Pathogenicity tests demonstrated that most of these fungi were pathogenic to inoculated almond, pistachio and walnut shoots. Therefore, more importance should be given to fungal trunk pathogens in Iran, and specific management strategies should be included within the nut crops IPM management programs, with the aim of improving their sustainability.

Hongos asociados con la muerte regresiva de los cítricos en Nuevo León y Tamaulipas, México


 El cultivo de cítricos es afectado por varios hongos fitopatógenos, los cuales pueden causar enfermedades y reducir su producción. El hongo Lasiodiplodia spp. causa muerte regresiva y otros síntomas en cítricos y en otros cultivos en diversos países, incluyendo México. El objetivo de este trabajo fue identificar cepas fúngicas aisladas de árboles de cítricos con síntomas de muerte regresiva y evaluar su patogenicidad en condiciones de invernadero. Los hongos aislados fueron identificados con base en sus características morfológicas como Lasiodiplodia theobromae, Fomitopsis meliae y Eutypella citricola, confirmados con la amplificación por PCR y secuenciación de la región ITS al comparar con las secuencias del GenBank. En el bioensayo de invernadero se comprobó que L. theobromae y F. meliae provocan síntomas de marchitez, muerte descendente y lesiones necróticas en las áreas inoculadas a partir del cuarto día posterior a la inoculación en plantas de naranjo dulce variedad Valencia. F. meliae + L. theobromae causaron daños más severos, provocando lesiones necróticas de 22 a 27 cm de longitud, marchitez y muerte regresiva y ambos fueron reaislados a partir de las lesiones producidas. E. citricola sólo produjo necrosis alrededor del área inoculada. De acuerdo con los resultados de la prueba de patogenicidad, los hongos L. theobromae y F. meliae causan muerte regresiva de los cítricos.

Diatrypaceae species overlap between vineyards and natural ecosystems in South Africa

Abstract Diatrypaceae species collected from 33 woody hosts occurring near vineyards were characterised. Plants showed symptoms of dieback, necrosis and cankers. Phylogenetic analyses based on ITS-rDNA and the β-tubulin gene separated the 265 isolates obtained into 14 species. The five most frequently encountered species, Cryptovalsa ampelina, Eutypa cremea, Eutypa lata, Eutypella citricola and Eutypella microtheca, were also found on grapevine in a previous study in South Africa. Several plant species, therefore, serve as hosts for these pathogens. First reports in South Africa include Cryptosphaeria ligniota, Eutypella leprosa and Eutypella australiensis. Host ranges of Cryptosphaeria multicontinentalis, Eu. australiensis and Eu. microtheca were also expanded. Diatrypaceae species caused brown discolouration when inoculated onto wounded grapevine tissues, which suggests that cross-infections are possible between grapevine and other woody hosts. Campelina., Eu. microtheca and Eu. citricola were the most virulent species and their virulence was comparable to that of Eu. lata, a known pathogen causing Eutypa dieback.

Eutypa, Eutypella, and Cryptovalsa Species (Diatrypaceae) Associated with Prunus Species in South Africa.

Stone fruit trees (Prunus spp.) are economically important fruit trees cultivated in South Africa. These trees are often grown in close proximity to vineyards and are to a large extent affected by the same trunk disease pathogens as grapevines. The aim of the present study was to determine whether stone fruit trees are inhabited by Diatrypaceae species known from grapevines and whether these trees could act as alternative hosts for these fungal species. Isolations were carried out from symptomatic wood of Prunus species (almond, apricot, cherry, nectarine, peach, and plum) in stone fruit growing areas in South Africa. Identification of isolates was based on phylogenetic analyses of the internal transcribed spacer region and β-tubulin gene. Forty-six Diatrypaceae isolates were obtained from a total of 380 wood samples, from which five species were identified. All five species have also been associated with dieback of grapevine. The highest number of isolates was found on apricot followed by plum. No Diatrypaceae species were isolated from peach and nectarine. Eutypa lata was the dominant species isolated (26 isolates), followed by Cryptovalsa ampelina (7), Eutypa cremea (5), Eutypella citricola (5), and Eutypella microtheca (3). First reports from Prunus spp. are E. cremea, E. citricola, and E. microtheca. Pathogenicity tests conducted on apricot and plum revealed that all these species are pathogenic to these hosts, causing red-brown necrotic lesions like those typical of Eutypa dieback on apricot.

Diversity of Diatrypaceae Species Associated with Dieback of Grapevines in South Africa, with the Description of Eutypa cremea sp. nov.

Recent studies in grape-growing areas including Australia, California, and Spain have revealed an extensive diversity of Diatrypaceae species on grapevines showing dieback symptoms and cankers. However, in South Africa, little is known regarding the diversity of these species in vineyards. The aim of this study was, therefore, to identify and characterize Diatrypaceae species associated with dieback symptoms of grapevine in South Africa. Isolates were collected from dying spurs of grapevines aged 4 to 8 years old, grapevine wood showing wedge-shaped necrosis when cut in cross section as well as from perithecia on dead grapevine wood. The collected isolates were identified based on morphological characters and phylogenetic analyses of the internal transcribed spacer region (ITS) and β-tubulin gene. Seven Diatrypaceae species were identified on grapevine, namely Cryptovalsa ampelina, C. rabenhorstii, Eutypa consobrina, E. lata, E. cremea sp. nov., Eutypella citricola, and E. microtheca. The dying spurs yielded the highest diversity of species when compared with the wedge-shaped necrosis and/or perithecia. C. ampelina was the dominant species in the dying spurs, followed by E. citricola, whereas E. lata was the dominant species isolated from the wedge-shaped necroses and perithecia. These results confirm E. lata as an important grapevine canker pathogen in South Africa, but the frequent association of C. ampelina with spur dieback suggests that this pathogen plays a more prominent role in dieback than previously assumed. In some cases, more than one species were isolated from a single symptom, which suggests that interactions may be occurring leading to decline of grapevines. C. rabenhorstii, E. consobrina, E. citricola, E. microtheca, and E. cremea are reported for the first time on grapevine in South Africa.

Identification, Distribution, and Pathogenicity of Diatrypaceae and Botryosphaeriaceae Associated with Citrus Branch Canker in the Southern California Desert.

Several members of the families Botryosphaeriaceae and Diatrypaceae are known as canker and dieback pathogens of a number of woody hosts. Because desert citrus production in California can occur in proximity to table grape production, it was suspected that fungi associated with grapevine cankers might also be associated with citrus branch canker and dieback decline. To determine the fungi associated with branch canker and dieback disease of citrus in the southern California desert regions, surveys were conducted from 2011 to 2013 in the major citrus-growing regions of Riverside, Imperial, and San Diego Counties. Cankered tissues were collected from branches showing symptoms typical of branch canker and dieback. Various fungal species were recovered from necrotic tissues and species were identified morphologically and by phylogenetic comparison of partial sequences of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2), β-tubulin gene, and elongation factor 1-α genes with those of other species in GenBank. Four fungi, including Neoscytalidium hyalinum, Eutypella citricola, E. microtheca, and an unnamed Eutypella sp., were associated with branch canker. N. hyalinum was the most frequently recovered fungus from symptomatic tissues (31%) followed by E. citricola (10%), E. microtheca (4%), and the Eutypella sp. (2%). In pathogenicity tests, all fungi caused lesions when inoculated on 'Lisbon' lemon (citrus) branches. Lesions caused by the Eutypella sp. were significantly longer than those of the other Eutypella spp.; however, they did not differ significantly from those produced by N. hyalinum. The most-parsimonious unrooted trees based on the combined data of ITS and partial β-tubulin gene region sequences showed three distinct clades of Eutypella spp. (E. citricola, E. microtheca, and an unidentified Eutypella sp.). Similarly, ITS and partial translation elongation factor 1-α gene region sequences differentiated two species of Neoscytalidium, N. hyalinum and N. novaehollandiae. Identifying the diversity, distribution, and occurrence of these fungal pathogens is useful for the management of citrus branch canker and dieback disease in the desert citrus-growing regions of California.

Trunk Disease Fungi Associated With Diospyros kaki in South Africa.

Persimmon trees with dieback symptoms and cankers were observed in three production areas in Western Cape Province in South Africa. Isolations were made from diseased branches, cankers, and pruning wounds as well as fungal fruiting bodies on dead branches and old pruning wounds. Several trunk disease pathogens were identified based on morphological characteristics and by molecular methods, including Diaporthe eres, D. infecunda, Eutypella citricola, E. microtheca, Phaeoacremonium parasiticum, P. scolyti, P. australiense, P. minimum, Fomitiporia capensis, Fomitiporia sp., Fomitiporella sp., and Inocutis sp., which were isolated from persimmon for the first time in the world. Other first reports from persimmon in South Africa include D. foeniculina, D. ambigua, D. mutila, Diaporthe sp., Neofusicoccum australe, N. parvum, Diplodia seriata, and Eutypa lata. Pathogenicity tests conducted with all species, except the basidiomycetes, confirmed their status as possible persimmon pathogens. This is the first study to determine and identify fungi associated with diseased persimmon in South Africa. The knowledge gained in this study forms the basis for further research to determine the impact of these fungi on persimmon productivity.

Diatrypella macrospora sp. nov. and new records of diatrypaceous fungi from Iran

Descriptions and illustrations are provided for six diatrypaceous species from Iran. Diatrypella macrospora is introduced as new to science. It differs from all other Diatrypella species by its larger ascospores. New records for the Iranian mycota include Cryptovalsa rabenhorstii, Eutypella citricola, Peroneutypa scoparia and Quaternaria quaternata. Descriptions and illustrations of microscopic characters, ITS sequence data and taxonomic notes are provided for these newly reported taxa.

Pathogenicity of Diatrypaceous Fungi on Grapevines in Australia.

In addition to Eutypa lata, which causes Eutypa dieback, numerous other fungi in the Diatrypaceae family have been isolated from diseased grapevines (Vitis vinifera) and other woody hosts. Pathogenicity trials comprising 70 strains of diatrypaceous fungi representing nine species in six genera were conducted to determine whether these fungi, collected in Australia, were pathogenic to grapevines. When inoculated into wounded trunks of 'Cabernet Sauvignon', eight species, including E. lata, E. leptoplaca, Cryptovalsa ampelina, C. rabenhorstii, Eutypella citricola, E. microtheca, Diatrypella vulgaris, and a Diatrype sp. produced necrotic lesions significantly longer than on controls. In addition, all nine species (including a Cryptosphaeria sp.) were reisolated from the margins of developing lesions and at varying distances above and below the point of inoculation. Diatrypaceous fungi were frequently isolated from asymptomatic or otherwise healthy tissue several centimeters ahead of the disease margin. Methods to control diseases associated with diatrypaceous fungi must take into account their propensity to colonize woody tissues ahead of or in the absence of visible symptoms. Current recommendations for the management of Eutypa dieback using remedial surgery and pruning wound protection appear sufficient for the control of the other diatrypaceous fungi included in this study.

Species of Diatrypaceae associated with grapevine trunk diseases in Eastern Spain

This research was supported by the projects RTA2007-00023-C04 and RTA2010-00009-C03, with matching funds from the Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA), Spain, and the European Regional Development Fund (ERDF). The authors thank Dr. T. Garnatje (Institut Botanic de Barcelona - CSIC, Barcelona, Spain) for her valuable help on the phylogenetic analyses.

Efficacy of fungicides on mycelial growth of diatrypaceous fungi associated with grapevine trunk disease

Several species of Diatrypaceae have been recently isolated from the wood of cankered grapevines in several regions of the world and shown to be pathogenic with varying degrees of virulence when inoculated in stems of potted vines. Existing management strategies have focussed on the development of procedures and products to prevent or reduce Eutypa lata infection. The best method to prevent infection is by applying fungicides to wounds, but there are few registered chemicals for any of the diatrypaceous fungi in Australia. Six selected fungicides were evaluated in vitro for their efficacy in reducing mycelial growth of Eutypa lata, Cryptovalsa ampelina, Diatrypella vulgaris, Eutypa leptoplaca, Eutypella citricola and Eutypella microtheca. Carbendazim, fluazinam, tebuconazole, and prothioconazole + tebuconazole were effective at inhibiting mycelial growth of all Diatrypaceae spp. tested. Pyraclostrobin reduced colony diameter of most of the fungal species by 50% or more. Pyrimethanil was ineffective at reducing mycelial growth of these pathogens. Five of the six fungicides evaluated in this study have shown efficacy in vitro against Diatrypaceae spp. and require further evaluation in the field. This study represents the first approach for fungicide evaluation against mycelial growth of diatrypaceous fungi, other than Ea. lata. It contributes to the development of integrated management strategies for grapevine trunk diseases.