Salvatore Frisullo

Fungal Planet description sheets: 214-280

Novel species of microfungi described in the present study include the following from South Africa: Cercosporella dolichandrae from Dolichandra unguiscati, Seiridium podocarpi from Podocarpus latifolius, Pseudocercospora parapseudarthriae from Pseudarthria hookeri, Neodevriesia coryneliae from Corynelia uberata on leaves of Afrocarpus falcatus, Ramichloridium eucleae from Euclea undulata and Stachybotrys aloeticola from Aloe sp. (South Africa), as novel member of the Stachybotriaceae fam. nov. Several species were also described from Zambia, and these include Chaetomella zambiensis on unknown Fabaceae, Schizoparme pseudogranati from Terminalia stuhlmannii, Diaporthe isoberliniae from Isoberlinia angolensis, Peyronellaea combreti from Combretum mossambiciensis, Zasmidium rothmanniae and Phaeococcomyces rothmanniae from Rothmannia engleriana, Diaporthe vangueriae from Vangueria infausta and Diaporthe parapterocarpi from Pterocarpus brenanii. Novel species from the Netherlands include: Stagonospora trichophoricola, Keissleriella trichophoricola and Dinemasporium trichophoricola from Trichophorum cespitosum, Phaeosphaeria poae, Keissleriella poagena, Phaeosphaeria poagena, Parastagonospora poagena and Pyrenochaetopsis poae from Poa sp., Septoriella oudemansii from Phragmites australis and Dendryphion europaeum from Hedera helix (Germany) and Heracleum sphondylium (the Netherlands). Novel species from Australia include: Anungitea eucalyptorum from Eucalyptus leaf litter, Beltraniopsis neolitseae and Acrodontium neolitseae from Neolitsea australiensis, Beltraniella endiandrae from Endiandra introrsa, Phaeophleospora parsoniae from Parsonia straminea, Penicillifer martinii from Cynodon dactylon, Ochroconis macrozamiae from Macrozamia leaf litter, Triposporium cycadicola, Circinotrichum cycadis, Cladosporium cycadicola and Acrocalymma cycadis from Cycas spp. Furthermore, Vermiculariopsiella dichapetali is described from Dichapetalum rhodesicum (Botswana), Ophiognomonia acadiensis from Picea rubens (Canada), Setophoma vernoniae from Vernonia polyanthes and Penicillium restingae from soil (Brazil), Pseudolachnella guaviyunis from Myrcianthes pungens (Uruguay) and Pseudocercospora neriicola from Nerium oleander (Italy). Novelties from Spain include: Dendryphiella eucalyptorum from Eucalyptus globulus, Conioscypha minutispora from dead wood, Diplogelasinospora moalensis and Pseudoneurospora canariensis from soil and Inocybe lanatopurpurea from reforested woodland of Pinus spp. Novelties from France include: Kellermania triseptata from Agave angustifolia, Zetiasplozna acaciae from Acacia melanoxylon, Pyrenochaeta pinicola from Pinus sp. and Pseudonectria rusci from Ruscus aculeatus. New species from China include: Dematiocladium celtidicola from Celtis bungeana, Beltrania pseudorhombica, Chaetopsina beijingensis and Toxicocladosporium pini from Pinus spp. and Setophaeosphaeria badalingensis from Hemerocallis fulva. Novel genera of Ascomycetes include Alfaria from Cyperus esculentus (Spain), Rinaldiella from a contaminated human lesion (Georgia), Hyalocladosporiella from Tectona grandis (Brazil), Pseudoacremonium from Saccharum spontaneum and Melnikomyces from leaf litter (Vietnam), Annellosympodiella from Juniperus procera (Ethiopia), Neoceratosperma from Eucalyptus leaves (Thailand), Ramopenidiella from Cycas calcicola (Australia), Cephalotrichiella from air in the Netherlands, Neocamarosporium from Mesembryanthemum sp. and Acervuloseptoria from Ziziphus mucronata (South Africa) and Setophaeosphaeria from Hemerocallis fulva (China). Several novel combinations are also introduced, namely for Phaeosphaeria setosa as Setophaeosphaeria setosa, Phoma heteroderae as Peyronellaea heteroderae and Phyllosticta maydis as Peyronellaea maydis. Morphological and culture characteristics along with ITS DNA barcodes are provided for all taxa.

Endophytic fungi occurring in fennel, lettuce, chicory, and celery — commercial crops in southern Italy

The occurrence of endophytic fungi in fennel, lettuce, chicory, and celery crops was investigated in southern Italy. A total of 186 symptomless plants was randomly collected and sampled at the stage of commercial ripeness. Fungal species of Acremonium, Alternaria, Fusarium, and Plectosporium were detected in all four crops; Plectosporium tabacinum was the most common in all crop species and surveyed sites. The effect of eight endophytic isolates (five belonging to Plectosporium tabacinum and three to three species of Acremonium) inoculated on lettuce plants grown in gnotobiosis was assessed by recording plant height, root length and dry weight, collar diameter, root necrosis, and leaf yellowing. P. tabacinum and three species of Acremonium, inoculated on gnotobiotically grown lettuce plants, showed pathogenic activity that varied with the fungal isolate. Lettuce plants inoculated with the isolates Ak of Acremonium kiliense, Ac of Acremonium cucurbitacearum, and P35 of P. tabacinum showed an increased root growth, compared to the non-inoculated control. The high frequency of P. tabacinum isolation recorded in lettuce plants collected in Bari and Metaponto, and in fennel plants from Foggia agricultural districts, suggests a relationship not only between a crop species and P. tabacinum, but also between the occurrence of the endophyte and the crop rotation history of the soil.

Three new species of Leptographium from pine.

Leptographium species are common inhabitants of fresh conifer logs and lumber that are known for their ability to cause blue-stain and, in some cases, their association with disease. L. procerum has been associated with a root disease although controversy surrounds its role in tree death. During the course of the past two decades, a relatively large number of isolates tentatively identified as L. procerum have been collected in various parts of the world. Some of these display morphological characters unlike those of L. procerum s. str. and this has prompted us to re-examine them. Four groups of morphologically distinct isolates were identified, of which L. procerum s. str. represented one. The remaining isolates of Leptographium are newly described as L. alethinum, L. pityophilum and L. euphyes spp. nov.

Diversity Evaluation of Xylella fastidiosa from Infected Olive Trees in Apulia (Southern Italy).

Olive culture is very important in the Mediterranean Basin. A severe outbreak of Olive Quick Decline Syndrome (OQDS) caused by Xylella fastidiosa infection was first noticed in 2013 on olive trees in the southern part of Apulia region (Lecce province, southern Italy). Studies were carried out for detection and diversity evaluation of the Apulian strain of Xylella fastidiosa. The presence of the pathogen in olive samples was detected by PCR amplifying the 16S rDNA, gyrase B subunit (gyrB) and HL hypothetical protein genes and single nucleotide polymorphisms (SNPs) assessment was performed to genotype X. fastidiosa. Twelve SNPs were recorded over gyrB and six SNPs were found for HL gene. Less variations were detected on 16S rDNA gene. Only gyrB and HL provided sufficient information for dividing the Apulian X. fastidiosa olive strains into subspecies. Using HL nucleotide sequences was possible to separate X. fastidiosa into subspecies pauca and fastidiosa. Whereas, nucleotide variation present on gyrB gene allowed separation of X. fastidiosa subsp. pauca from the other subspecies multiplex and fastidiosa. The X. fastidiosa strain from Apulia region was included into the subspecies pauca based on three genes phylogenetic analyses.

BRIEF HISTORICAL ACCOUNT OF OLIVE LEAF SCORCH (“BRUSCA”) IN THE SALENTO PENINSULA OF ITALY AND STATE-OF-THE-ART OF THE OLIVE QUICK DECLINE SYNDROME

“Brusca” (leaf scorch or marginal leaf burn) is a disease of olive (Olea europaea) typically characterized by the desiccation and death of tissues at the tip and/or along the edge of the leaf blade, which can be followed by defoliation. Since the end of the 18th century this disorder has reappeared periodically, after long periods of quiescence, in the same olive- growing areas of the province of Lecce (Apulia, Salento pensinsula, southern Italy). Over time, this disease has been the object of repeated investigations, first by a couple of local physician, who were the authors of its description, then, since the beginning of the 20th century, by professional plant pathologists. These studies have established that “brusca” may have multiple origins as determined by abiotic (“brusca non parassitaria” = non parasitic scorching) or biotic (“brusca parassitaria” = parasitic scorching) causes. “Brusca non parassitaria” can be induced by any physical cause that affects water supply to the leaf margin cells (e.g. insufficient moisture in the soil, water is lost too quickly from the leaves to be replaced adequately, damaged roots), or by hot dry winds, salty winds, nutrient deficiency/toxicity. By contrast, weak foliar pathogens (i.e. the discomycete fungus Stictis panizzei) or xylem-invading fungi or bacteria that plug the water conducting vessels can, in principle, be the agents of “brusca parassitaria”. These agents may include Xylella fastidiosa, a xylem-limited bacterium, which has recently been found associated with an olive disease denoted “quick decline syndrome”. This disease occurs in some areas of the Lecce province where severe cases of “brusca” have repeatedly been observed in the past. This coincidence, and the type of symptoms (extensive scorching and desiccation of the canopy) that somewhat recall those described in the early literature, may lead one to speculate that X. fastidiosa has been sitting in the area for about 250 years, rather than being a pathogen of recent introduction. This is not the case, as discussed in the present paper.

FIRST REPORT OF TWO PHOMOPSIS SPECIES ON OLIVE TREES IN ITALY

During a survey (2002-2012) on olive groves in Apulia (southern Italy), many trees of cv. Coratina were found, exhibiting cankers on twigs and branch dieback. Phomopsis spp. were isolated (15-65%) from symptomatic twigs. Genomic DNA was extracted from 43 fungal isolates. The ITS gene region was amplified with primers ITS1/ITS4 (White et al., 1990) and sequenced. Blast analysis of ITS sequences of 38 isolates showed 100% similarity with Diaporthe neotheicola A.J.L. Phillips et J.M. Santos (GenBank accession No. EU814480) whereas five isolates had 100% similarity with D. ambigua Nitschke (EU814478). ITS sequences of D. ambigua and D. neotheicola isolates from olive were deposited in GenBank under the accession Nos. HM575419, HM575420 and HM575421, HM575422, respectively. Microscopic features of D. neotheicola from Apulian olive trees and matched those described by Phillips and Santos (2009) while features of D. ambigua were identical with those previously described for this species. Pathogenicity tests were conducted in May 2012 and 2013 using two isolates/species on 1-year-old twigs of cv. Coratina by inoculating them with each isolate or with sterile agar (controls). Symptoms were observed after 90 days and Diaporthe species were re-isolated thus, fulfilling Koch’s postulates. Control plants were symptomless. The two species of Phomopsis from olive could be a potential inoculum source for grapevines. Indeed, Mostert et al. (2001) reported D. ambigua and Phomopsis sp. 1, the latter referred to as D. neotheicola, from Vitis vinifera. To our knowledge, this is the first report of D. neotheicola and D. ambigua on olive.

First report of wilt caused by Verticillium dahliae on cosmos (Cosmos bipinnatus) in Italy.

Cosmos (Cosmos bipinnatus Cav., Asteraceae) is an herbaceous plant that is grown for landscape use. During August and September of 2008 in five public and three private gardens located in Monopoli (Apulia, southern Italy), 3 to 8% of the plants showed severe symptoms of vine decline, stunting, gradual yellowing and wilting of the leaves, and final collapse of the whole plant. External symptoms were associated with brown or black streaking of the vascular tissue of roots, collar, and stem. Dead plants had numerous microsclerotia embedded in the xylem of plant tissues. Stem, collar, and root sections (0.5 cm long) from symptomatic plants collected in five gardens were surface disinfested in 5% NaOCl for 1 min and transferred to petri dishes containing potato dextrose agar (PDA) amended with 100 μg ml-1 of streptomycin sulfate and 10 μg ml-1 of neomycin. After 10 days of incubation, at 25°C in the dark, hyaline hyphae with dark microsclerotia (37 to 112 μm) and verticillate conidiophores were produced. Conidia were single celled and hyaline with dimensions of 3.3 to 7.8 × 1.8 to 3.3 μm (mean dimensions 4.2 × 2.5 μm). According to morphological characteristics, the fungus was identified as Verticillium dahliae Kleb. (1) (isolates no. Vd1818, Vd1819, and Vd1820 stored in a collection at the Department DiSACD, University of Foggia). Molecular analyses were performed on the basis of nucleotide sequences of the internal transcribed spacer region (ITS1-5.8S-ITS2) of ribosomal DNA. ITS sequences of this fungus, compared with sequences found in GenBank and attributed to V. dahliae (no. GQ130129, GQ130130, GQ130131), showed 98 to 99% sequence similarity. Healthy 40-day-old plants of C. bipinnatus (garden cosmos) cv. Sonata Pink Blusk and C. sulphurous (yellow cosmos) cv. Bilbo, obtained from seeds previously disinfested for 1 min in 3% NaOCl and ascertained to be healthy by isolation on PDA medium, were used for pathogenicity tests. Plants were grown in 3-liter pots in a steam-disinfested peat, sand, and soil mixture (2:1:1) in the greenhouse at 23 to 26°C. Ten plants of each cultivar were inoculated by root dipping into a conidial suspension of each fungal isolate (1.5 × 106 CFU ml-1). Six noninoculated cosmos plants of each cultivar served as controls. The experiment was repeated three times. First symptoms of wilting were observed on all inoculated plants of each cultivar 20 days after the inoculation; at 40 days, symptom severity ratings on plants were taken, in which 1 = asymptomatic, 2 = stunted, 3 = wilting, and 4 = dead. All three isolates caused vascular discoloration, stunting, wilting, and plant death. The mean disease rating was 3.2 and did not differ significantly among isolates. The pathogen was consistently reisolated from infected plants, fulfilling Kochs postulates. Noninoculated plants remained healthy. To our knowledge, this is the first report of Verticillium wilt on cosmos in Italy. The finding is important since other ornamental plants that are susceptible to Verticillium wilt are also grown in landscapes in the region. The disease was previously reported in Turkey (2). References: (1) G. F. Pegg and B. L. Brandy. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002. (2) E. Sezgin et al. Turk. Phytopathol. 14:43, 1985.

First report of Cadophora malorum on Asparagus officinalis in Italy.

In this note Cadophora malorum infections on asparagus plants cultivated in different localities in the Foggia area (southern Italy) are reported. Pathogenicity tests demostrated that C. malorum showed the capacity to infect healthy plants of asparagus even if this capacity is less pronounced than that of Fusarium oxysporum f. sp. asparagi and F. proliferatum.

The fungus Nectria aurantiicola Berk. et Br. (Fusarium larvarum Fuckel) a biocontrol agent of the armored scale Suturaspis archangelskyae (Lindinger) in Apulia (Southern Italy)

Suturaspis archangelskyae [(Lindinger) 1929] (Homoptera, Diaspididae), an armored scale insect collected in Apulia (Italy) on Prunus dulcis (Miller) D. A. Webb (=almond) and Prunus webbii (Spach) Vierh., is reported to be infected by Fusarium larvarum Fuckel. The cycle of the fungus, as studied on female lineage of S. archangelskyae, shows mycelium in August and September, orange sporodochia from November to March, while mature red perithe- cia of the teleomorph Nectria aurantiicola Berk. et Br. are observed since middle January up to the first half of February. The fungus survives during the hot dry season as mycelium in the dead body of the host. The fungus is able to kill about 50% of the scales and spreads among demes by conidia carried on crawlers. Both varieties, the creamy var. larvarum Fuckel and the carmine-red var. rubrum Gerlach, are found on natural infected scales. The male progeny of S. archangelskyae seems not to be affected by the fungus. Ascospores are observed as well, but their role in pathogenesis remains unknown. Fusarium larvarum seems to be the main “natural” biocontrol agent (sensu Rosen et DeBach, 1990) of S. archangelskyae in Apulia. Key words: semi-arid (mediterranean) habitat, entomopathogens, pathogenic symbiosis, armo- red scale, coccids, Cheyletidae (Acari preying crawlers), biology, Puglia. NECTRIA AURANTIICOLA BERK. ET BR. (FUSARIUM LARVARUM FUCKEL) ANTAGONISTA DI SUTURASPIS ARCHANGEL- SKYAE (LINDINGER) IN PUGLIA (ITALIA MERIDIONALE) Gli autori descrivono la simbiosi fra un fungo entomopatogeno, Fusarium larvarum Fuckel (teleomorfo Nectria aurantiicola Berk. et Br.), ed il diaspino Suturaspis archangelskyae (Lindinger, 1929) su Prunus dulcis (Miller) D. A. Webb e P. webbii (Spach) Vierh. presso “Castel del Monte” (Andria, BA) ed in numerose localita della Puglia. Sono state osservate le modalita di disseminazione del fungo ed il procedere dell’infezione, che culmina con l’uccisione delle cocciniglie prima che queste possano riprodursi. Durante i mesi invernali il fungo produce sporodochi reniformi di colore arancio, ricchi di conidi falciformi, e periteci globoso-piriformi di colore rosso arancio, contenenti le ascospore. Il fungo risulta essere il principale fattore biotico limitante del diaspino del quale uccide le femmine adulte. Esso sembra suscettibile di utilizzo come agente di controllo biologico per inoculazione od incremento, sempre che vengano assicurati a fondo i fattori, ecologici che regolano l’infezione. Aspetto favorevole all’uso di F. larvarum nelle regioni temperate e la sua predilezione per le temperature moderate. Non e stato possibile chiarire il ruolo delle ascospore nella diffusione del fungo. Parole chiave: mandorlo, habitat semi-aridi (mediterranei), coccoidei, Cheyletidae (Acari predatori di neanidi).