Phytopathologia Mediterranea <p><em>Phytopathologia Mediterranea</em> is an international open-access, peer-reviewed journal edited by the <a title="Mediterranean phytopathological union" href="" target="_blank" rel="noopener">Mediterranean Phytopathological Union</a>. The journal deals with the main areas of plant pathology as epidemiology, control, biochemical and physiological aspects, application of molecular biology techniques, applied to fungi, bacteria, phytoplasmas, viruses, viroids, nematodes, etc. Special attention is given to phytopathological problems of the Mediterranean area. The journal includes 3 issues per year in which a review paper, original research papers, short notes and new disease reports are published. It also includes Book reviews of interest for Mediterranean phytopathologists. Papers are published in English. Phytopathologia Mediterranea is covered by CAB, BIOSIS, AGRIS, Chemical Abstracts, CSA, JSTORE.</p> <p><em>Phytopathologia Mediterranea</em> is printed with the financial support of the Ministero per i Beni Culturali, Roma, Italy.</p> Firenze University Press en-US Phytopathologia Mediterranea 0031-9465 <p>Authors retain the copyright and grant the journal right of first publication with the work simultaneously licensed under a <strong>Creative Commons Attribution 4.0 International Public License (<a href="">CC-BY-4.0</a>)</strong>&nbsp;that allows others to share the work with an acknowledgment of the work's authorship and initial publication in PHYTO</p> <p><a href="" rel="license"><img style="border-width: 0;" src="" alt="Creative Commons License"></a><br>This work is licensed under a <a href="" rel="license">Creative Commons Attribution 4.0 International License</a></p> Characterization of Pyrenophora tritici-repentis (tan spot of wheat) races in Kazakhstan <p class="p1">Tan spot, caused by <em>Pyrenophora tritici-repentis</em>, is an economically important foliar disease of wheat in Kazakhstan. Population structure of the pathogen changes every year due to climate change. This study aimed to&nbsp;characterize the race structure of&nbsp;<em>P. tritici-repentis&nbsp;</em>isolates recovered from wheat in south and north Kazakhstan, and identify tan spot resistance in host genotypes based on disease phenotypes and molecular screening. Virulence profiles were determined within 40 isolates of the pathogen from wheat crops during the 2020 growing season. Seven races, (1, 3, 4, 5, 6, 7 and 8) were identified. A collection of 80 wheat accessions, including promising lines and cultivars from Kazakhstan and Russia, were evaluated for their reactions to races 1 and 5 of the pathogen, and to Ptr ToxA and Ptr ToxB, using greenhouse assessments and molecular markers diagnostic for the&nbsp;<em>Tsn1</em>&nbsp;and&nbsp;<em>Tsc2&nbsp;</em>genes. From a practical viewpoint, 18 wheat genotypes were insensitive to the two races and the two Ptr toxins.&nbsp;This resistant germplasm can be used in breeding programmes aiming to develop wheat varieties resistant to <em>P. tritici-repentis</em>.</p> Madina KUMARBAYEVA Alma KOKHMETOVA Nadezhda KOVALENKO Makpal ATISHOVA Zhenis KEISHILOV Klara AITYMBETOVA Copyright (c) 2022 Madina KUMARBAYEVA, Alma KOKHMETOVA, Nadezhda KOVALENKO, Makpal ATISHOVA, Zhenis KEISHILOV, Klara AITYMBETOVA 2022-07-19 2022-07-19 61 2 243 257 10.36253/phyto-13178 A real time loop-mediated isothermal amplification (RealAmp) assay for rapid detection of Pleurostoma richardsiae in declining olive plants <p class="p1"><span class="s1"><em>Pleurostoma richardsiae</em> is associated with host trunk diseases, known to cause dieback, cankers and wilting of woody trees, and human infections. This fungus was isolated from wood tissues of declining olive trees and grapevines showing esca disease symptoms, in the Apulia region of Italy. Fungus detection has been based on morphological and molecular features, which are time-consuming to identify and require well-trained personnel. Improvement of <em>Pl. richardsiae</em> detection in olive was achieved through development of real time loop-mediated isothermal amplification targeting the intergenic spacer (IGS) region of the fungus. Specificity of the assay was confirmed using ten <em>Pl. richardsiae</em> strains and 36 other fungus strains of species usually isolated from declining olive trees. The achieved limit of detection was 7.5 × 10<sup>-2</sup> ng μL<sup>-1</sup> of <em>Pl. richardsiae</em> genomic DNA. A preliminary validation of RealAmp was also performed using material from infected olive plants artificially inoculated in a greenhouse.</span></p> Abdraouf SADALLAH Serena Anna MINUTILLO Franco VALENTINI Maria Luisa RAIMONDO Francesco LOPS Antonia CARLUCCI Antonio IPPOLITO Anna Maria D’ONGHIA Copyright (c) 2022 Abdraouf SADALLAH, Serena Anna MINUTILLO, Franco VALENTINI, Maria Luisa RAIMONDO, Francesco LOPS, Antonia CARLUCCI, Antonio IPPOLITO, Anna Maria D’ONGHIA 2022-07-19 2022-07-19 61 2 259 267 10.36253/phyto-12748 Fusarium annulatum causes Fusarium rot of cantaloupe melons in Spain <p class="p1">During the summer of 2018, there was high incidence of fruit rots of cantaloupe melons (<em>Cucumis melo</em> var. <em>cantalupensis</em>) in Murcia province, south-eastern Spain. The fruits showed development of whitish mycelium and pulp softening. Morphological and molecular analysis of the internal transcribed spacer (ITS), translation elongation factor 1-α (<em>TEF1-α</em>) and the second largest subunit of RNA polymerase (<em>RPB2</em>) genes confirmed <em>Fusarium annulatum </em>as the causal agent of the disease. A phylogenetic study indicated that <em>F. annulatum</em> is in the <em>Fusarium</em> <em>fujikuroi</em> species complex (FFSC). Pathogenicity of the isolate was determined on healthy fruit verifying Koch’s postulates. The first symptoms of fruit rot were observed 3 d after inoculations at 28ºC. Fruit infections only occurred in artificially wounded melons, and <em>F. annulatum</em> was re-isolated from the wounds. This disease appeared after fruit harvesting, and could generate substantial economic losses mainly in fruit destined for foreign markets due to long transportation times. This is the first report of melon fruit rot caused by <em>F. annulatum</em> in Spain.</p> María Ängeles PARRA Javier GÓMEZ Fulgencio Wadi AGUILAR Juan Antonio MARTINEZ Copyright (c) 2022 María Ängeles PARRA, Javier GÓMEZ, Fulgencio Wadi AGUILAR, Juan Antonio MARTINEZ 2022-07-19 2022-07-19 61 2 269 277 10.36253/phyto-13454 Draft genome sequence of Phyllosticta ampelicida, the cause of grapevine black rot <p class="p1"><em>Phyllosticta ampelicida</em> causes grapevine black rot, a potentially damaging disease for grape production. This paper reports the draft genome sequence of <em>P. ampelicida</em> PA1 Galicia CBS 148563, which is 30.55 Mb and encodes 10,691 predicted protein-coding genes. This is the first sequence genome assembly of <em>P. ampelicida</em>, and this information is a valuable resource to support genomic attributes for determining pathogenic behaviour and comparative genomic analyses of grapevine black rot fungi.</p> Ales EICHMEIER Emilia DIAZ-LOSADA Eliska HAKALOVA Jakub PECENKA Katerina STUSKOVA Sonia OJEDA David GRAMAJE Copyright (c) 2022 Ales EICHMEIER , Emilia DIAZ-LOSADA, Eliska HAKALOVA, Jakub PECENKA, Katerina STUSKOVA, Sonia OJEDA , David GRAMAJE 2022-07-19 2022-07-19 61 2 279 282 10.36253/phyto-13516 Fig rust caused by Phakopsora nishidana in South Africa <p class="p1">Fig rust, caused by <em>Cerotelium fici</em>, was first recorded in South Africa in 1927. Recent observations have revealed high incidence of rust and untimely defoliation of fig trees (<em>Ficus carica</em>) in residential gardens and commercial orchards. Using phylogenetic analysis, the causal organism of a fig rust isolate (PREM63073) collected in 2020 was confirmed as <em>Phakopsora nishidana</em>. Inoculation and microscope studies showed that mulberry plants were immune to <em>P. nishidana</em> isolate PREM63073. Infection of fig leaves occurred through stomata on the abaxial leaf surfaces. Very long germ tubes were observed for <em>P. nishidana</em>, often with no clear contact with the leaf surfaces and an apparent lack of directional growth towards stomata. Inoculated plants from 15 fig cultivars varied in their severity of leaf infection, whereas fruit of the cultivar Kadota developed reddish-brown blemishes without sporulation. Currently, <em>C. fici</em> and <em>P. nishidana</em> are recognised as occurring on <em>F. carica</em> in South Africa. This suggests a need to resolve the worldwide distribution and identity of the rust species involved.</p> Willem H.P. BOSHOFF Botma VISSER Cornel M. BENDER Alan R. WOOD Lisa ROTHMANN Keith WILSON Victor HAMILTON-ATTWELL Zacharias A. PRETORIUS Copyright (c) 2022 Willem H.P. BOSHOFF, Botma VISSER, Cornel M. BENDER, Alan R. WOOD, Lisa ROTHMANN, Keith WILSON, Victor HAMILTON-ATTWELL, Zacharias A. PRETORIUS 2022-07-19 2022-07-19 61 2 283 298 10.36253/phyto-13034 Activity of essential oils from Syzygium aromaticum and Rosmarinus officinalis against growth and ochratoxin A production by Aspergillus tubingensis and Aspergillus luchuensis from Moroccan grapes <p class="p1">Essential oils have been used since ancient times in traditional medicine and agri-food science to preserve food, and to combat human diseases. Essential oils (EOs) from clove and rosemary, obtained by hydro-distillation, were analyzed by GC/MS and evaluated for their antifungal activity against strains of <em>Aspergillus tubingensis</em> and <em>A. luchuensis </em>and their effect on ochratoxin A (OTA) production by these fungi. The major constituent of clove EO was eugenol (86.4%), and of rosemary EO was eucalyptol (35.9%). Mycelium growth inhibition assays showed that the EOs had dose-depend inhibition effects, which reached 36.6% for rosemary EO, and 100% for clove EO, at EO concentration of 200 μL L<sup>-1</sup>. These reductions were different for different <em>Aspergillus</em> strains, the essential oil type, and the strain/oil type interaction. Analyses of OTA in the culture medium extracts of <em>Aspergillus</em> strains was carried out using HPLC-FLD, and was confirmed by LC-MS/MS for positive controls of two OTA-producing strains of <em>A</em>. <em>tubingensis</em>. In general, OTA was reduced (from 45 to 100% reduction), except for two strains of <em>A. tubingensis</em>, where OTA production was stimulated.</p> Adil LAAZIZ Yousra EL HAMMOUDI Souad QJIDAA Abdelouahed HAJJAJI Hassan HAJJAJ Geert HAESAERT Amina BOUSETA Copyright (c) 2022 Adil LAAZIZ, Yousra EL HAMMOUDI, Souad QJIDAA, Abdelouahed HAJJAJI, Hassan HAJJAJ, Geert HAESAERT, Amina BOUSETA 2022-07-19 2022-07-19 61 2 299 310 10.36253/phyto-12841 First report of Serratia marcescens from oleander in Hungary <p class="p1">Oleander (<em>Nerium oleander </em>L.) is a popular woody ornamental plant, often used for decorating public areas, terraces and gardens. Many diseases may decrease in the ornamental value of these plantings. Between 2018 and 2020, plant pathogenic bacteria of oleander were examined, and many samples of infected plants were collected from different sites in Hungary. Two non-pigmented <em>Serratia marcescens</em> isolates were identified from oleander by classical and molecular methods. The isolates caused necrotic lesions on oleander leaves. <em>Serratia marcescens </em>is known as an opportunistic mammal or plant pathogen, but non-pathogenic strains are known to be useful biological control agents or plant growth-promoting bacteria. This is the first report of the plant pathogen <em>S. marcescens</em> from oleander, and the first identification of the bacterium in Hungary.<span class="Apple-converted-space">&nbsp;</span></p> Attila FODOR Laszlo PALKOVICS Anita VÉGH Copyright (c) 2022 Attila FODOR, Laszlo PALKOVICS, Anita VÉGH 2022-07-19 2022-07-19 61 2 311 317 10.36253/phyto-13354 Abstracts of oral and poster presentations given at the 12th International Workshop on Grapevine Trunk Diseases, Mikulov, Czech Republic, 10–14 July 2022 <p class="p1"><span class="s1">The 12th International Workshop on Grapevine Trunk Diseases (12th IWGTD) was held in Mikulov, Czech Republic, from 10 to 14 July, 2022. The Workshop was chaired by Dr Aleš Eichmeier and organized by Mendel University in Brno and Svaz Vinařů České Republiky/Union of Winemakers of the Czech Republic. The ICGTD Council meeting took place on 10 July, and that evening the welcome reception was opened by Rostislav Koštial (Mayor of Mikulov, Senator and viticulturist), Dr Martin Chlad (President of Svaz Vinařů České Republiky/Union of Winemakers of the Czech Republic), and Dr Svatopluk Kapounek (Vice-Rector, Mendel University, Brno).<br></span>The scientific programme was opened by Dr Alena Salašová (Dean, Faculty of Horticulture, Mendel University, Brno). The Workshop was attended by 133 researchers from 25 countries, presenting 61 oral and 49 poster papers. These presentations were in five sessions, including; Pathogen Characterization and Identification, Epidemiology, Plant-pathogen Interactions, Microbial Ecology, and Disease Management in Nurseries and Vineyards. The Disease Management session aimed to provide grape growers with an overview of practical options for trunk disease control. This session included contributions from researchers who have completed field trials on trunk disease management. David Gramaje (Instituto de Ciencias de la Vid y el Vino (ICVV), Logroño, La Rioja, Spain, outlined an evaluation of treatments for protection of grapevine pruning wounds from natural pathogen infections; Francois Halleen (University of Stellenbosch, South Africa) presented detailed results of efficiency of hot water treatments in nurseries; and Josè Úrbez-Torres (Agriculture and Agri-Food Canada SuRDC) outlined research on biological control of Botryosphaeria dieback in grapevines.<br>A field trip (13 July) visited vineyards in the South Moravia. Delegates were presented with an overview of grapevine production in South Moravia, followed by discussions on the main diseases (including GTDs) affecting grapevines in the Czech Republic. Vineyards affected by GTD pathogens were also visited, where the growers outlined their viewpoints. The field trip focused on presenting the current situation of GTDs in South Moravia, and the management strategies being adopted.<span class="Apple-converted-space">&nbsp;</span></p> <p class="p1">Student competitions for best oral and poster presentations included papers presented by 29 postgraduate students.<span class="Apple-converted-space">&nbsp;<br></span>For best posters:<br>1st place went to Catarina Leal (University of Reims Champagne-Ardenne, France, and Polytechnic University of Valencia, Spain), “Evaluation of <em>Trichoderma atroviride</em> SC1 and <em>Bacillus subtilis</em> PTA-271 combination against grapevine trunk diseases pathogens in nursery propagation process”;<br>2nd place went to Marcelo Bustamante (University of California, Davis, United States of America), “Investigating the role of <em>Fusarium</em> spp. in young vine decline in California”;</p> <p class="p2">3rd place went to María Julia Carbone (Universidad de la República, Montevideo, Uruguay), “Interactive effects of <em>Dactylonectria macrodidyma</em> inoculation on the rhizosphere and root microbiome of grapevine”.<br><br>For best oral presentations:<br>1st place went to Catarina Leal, (University of Reims Champagne-Ardenne, France, and Polytechnic University of Valencia, Spain), “Beneficial effects of <em>Bacillus subtilis</em> PTA-271 and <em>Trichoderma atroviride</em> SC1 against the Botryosphaeria-dieback pathogen <em>Neofusicoccum parvum</em> may vary with grapevine cultivar”;<br>2nd place went to Isidora Silva-Valderama (University of British Columbia, Vancouver, Canada), “Predicting pathogens virulence: linking host breadth and pathogenicity of the <em>Botryosphaeriaceae</em> fungal family in wine grapes (<em>Vitis vinifera</em>)”;<br>3rd place went to Colin Todd (University of California, Riverside, United States of America), “Tracking the fungal pathobiome associated with young grapevine decline in California nurseries”.<br><br>The 13th IWGTD will be held in Ensenada, Mexico, in 2025.</p> AA. VV. Copyright (c) 2022 2022-08-02 2022-08-02 61 2 319 369 10.36253/phyto-13818 Pseudomonas syringae pv. syringae causes bacterial canker on Japanese quince (Chaenomeles japonica) <p class="p1">Japanese quince trees are grown as ornamental plants in Iran, in parks and in orchards close to stone fruit and pome fruit trees. Shoots of Japanese quince (<em>Chaenomeles japonica</em>) showing sunken brown canker symptoms were observed and collected near Sari, the center of Mazandaran province in the North of Iran, during the 2016 growing season. Gram negative bacteria isolated from symptomatic tissues were similar to <em>Pseudomonas syringae </em>pv. <em>syringae </em>(<em>Pss</em>) were pathogenic on Japanese quince and on quince (<em>Cydonia oblonga</em>) seedlings after artificial inoculation, and were re-isolated from diseased hosts. Phylogenetic tree construction using partial sequences of ITS and <em>rpo</em>D genes showed that the Japanese quince isolates were in the same clade as <em>Pss </em>strains. The isolates had ice nucleation activity, and the <em>Ina</em>K gene was amplified successfully. According to the results of phenotypic and genotypic characteristics, genomic DNA fingerprinting using REP-PCR, BOX-PCR and IS50-PCR and isolation of total cell proteins, we conclude that <em>Pss </em>is the causal agent of canker of the Japanese quince trees. Therefore, Japanese quince is a new host for <em>Pss </em>causing bacterial canker on many different host plants.</p> Touhid ALLAHVERDIPOUR Mina RASTGOU Heshmatollah RAHIMIAN Emanuela NORIS Slavica Matić Copyright (c) 2022 Touhid ALLAHVERDIPOUR, Mina RASTGOU, Heshmatollah RAHIMIAN, Emanuela NORIS, Slavica Matić 2022-09-15 2022-09-15 61 2 371 382 10.36253/phyto-13106 Alternaria species and related mycotoxin detection in Lebanese durum wheat grain <p class="p1"><em>Alternaria</em> is a ubiquitous genus that may infect wheat in many countries, causing the disease black point. The present study aimed to assess contamination by fungi, of durum wheat kernels from Lebanon, and identify the main <em>Alternaria </em>species contaminants. <em>Alternaria</em> was detected in the majority (97%) of the inspected fields. Contamination by <em>Alternaria</em> differed among the samples according to their geographical origins. The greatest contamination was detected in the West Bekaa area (average 59%), followed by Akkar (55%), and lowest was observed in Baalbeck (2%). HPLC-DAD analyses performed on grain samples showed that altenuene, alternariol, alternariol monomethyl ether, and tenuazonic acid were not detected in any sample. Phylogenetic analyses, based on DNA sequences of <em>β-tubulin</em>, <em>glyceraldehyde-3-phosphate dehydrogenase</em> and <em>calmodulin</em> gene fragments, showed that <em>Alternaria</em> field strains belonged to two major sections: <em>Alternaria</em> (51%) and <em>Infectoriae</em> (40%). The remaining strains were in separate clades in sections <em>Ulocladioides </em>(3%),<em> Chalastospora</em> (3%) and<em> Pseudoalternaria </em>(3%). Although this study revealed no contamination of wheat kernels by <em>Alternaria</em> mycotoxins, the potential risk of mycotoxin accumulation remains high due to the widespread occurrence of toxigenic <em>Alternaria</em> species on kernels.</p> Mario MASIELLO Romy EL GHORAYEB Stefania SOMMA Carine SAAB Giuseppe MECA Antonio F. LOGRIECO Wassim HABIB Antonio MORETTI Copyright (c) 2022 Mario MASIELLO, Romy EL GHORAYEB, Stefania SOMMA, Carine SAAB, Giuseppe MECA, Antonio F. LOGRIECO, Wassim HABIB, Antonio MORETTI 2022-09-15 2022-09-15 61 2 383 393 10.36253/phyto-13396 Occurrence of cherry viruses in South Tyrol (Italy) by comparing growth periods in two consecutive years <p class="p1">Sweet cherries (<em>Prunus avium</em> L.) are important as fruit crops, and can be affected by numerous viruses. An investigation on the occurrence of the three most common viruses of sweet cherry was carried out in commercially managed orchards in South Tyrol (Italy). The incidence of apple chlorotic leaf spot virus (ACLSV), Prunus necrotic ringspot virus (PNRSV) and prune dwarf virus (PDV) was investigated using enzyme-linked immunosorbent assays (ELISA) and the reverse transcriptase-polymerase chain reaction technique (RT-PCR) in spring 2018 and 2020, and during the summer and autumn of 2020. All three viruses were detected in the surveyed orchards. Comparative analyses showed that detection was more effective with RT-PCR than with ELISA, especially for detecting PNRSV and PDV. Mixed infections were detected in all the surveyed orchards. The results also showed clear differences between and during host growth periods, likely due to a variable virus concentration in the host trees.</p> Evi DELTEDESCO Magdalena NIEDRIST Sabine OETTL Copyright (c) 2022 Evi DELTEDESCO, Magdalena NIEDRIST, Sabine OETTL 2022-09-15 2022-09-15 61 2 395 404 10.36253/phyto-12807 A SYBR Green qPCR assay for specific detection of Colletotrichum ocimi, which causes black spot of basil <p class="p1"><em>Colletotrichum ocimi</em> causes black spot of basil (<em>Ocimum basilicum</em>) and is a serious threat to basil cultivation as it compromises leaf production. The pathogen also infects seeds, which could become primary sources of inoculum for spread of black spot. A SYBR Green real-time PCR assay was developed to detect <em>Colletotrichum ocimi </em>in basil leaves and seeds, based on the partial <em>β-tubulin</em> (<em>tub2</em>) gene sequence. Two primer sets were designed and tested. The selected primer pairs produced amplicons of 130 bp. The real-time PCR assay was validated for analytical specificity, sensitivity, selectivity, repeatability and reproducibility. The assay was specific for <em>C. ocimi </em>with respect to ten <em>Colletotrichum </em>spp. and to another 12 pathogens of basil plants. Sensitivity was 1 pg µL<sup>-1 </sup>of genomic fungal DNA and amplification analyses were not influenced by basil genomic DNA. The assay detected and quantified <em>C. ocimi</em> in artificially inoculated basil leaves. This is the first specific primer set for <em>C. ocimi</em>, which allows rapid detection and quantification of the pathogen is a useful tool for diagnostics in plants. Detection in seeds would also be possible, but will require an optimized extraction method. The qPCR detection of <em>C. ocimi in planta </em>can contribute to adoption of effective preventive disease management strategies.</p> Ilaria MARTINO Pedro Willem CROUS Angelo GARIBALDI Maria Lodovica GULLINO Vladimiro GUARNACCIA Copyright (c) 2022 Ilaria MARTINO, Pedro Willem CROUS, Angelo GARIBALDI, Maria Lodovica GULLINO, Vladimiro GUARNACCIA 2022-09-15 2022-09-15 61 2 405 413 10.36253/phyto-13606