Search Results (18)

English walnut (Juglans regia) is a species that is highly valued for the quality of its wood and the nutritional and nutraceutical properties of its fruit. A severe dieback of J. regia trees was observed recently in orchards located in three geographically distinct areas of Tuscany, central Italy. Symptoms included root and collar rot, necrosis of the under-bark tissue, bleeding cankers, stunted growth, and crown dieback. Four Phytophthora species were obtained from 239 isolates found on symptomatic J. regia individuals. They were identified, on the basis of macro-morphological (colony shape and texture), micro-morphometric (shape and size of oogonia, antheridia, oospores, sporangia, and chlamydospores) and molecular (ITS sequencing) characters, as P. gonapodyides, P. cactorum, P. citricola, and P. plurivora. Among these species, P. plurivora was the species isolated with overwhelming frequency from symptomatic tissue and rhizosphere soil, suggesting it to be the putative etiological agent. Pathogenicity assays were conducted on 20 cm long detached J. regia branches for a definitive establishment of disease causation. Severe symptoms (extended necroses) were exhibited by branches infected with P. plurivora, proving its pathogenicity and high virulence on this host. The other Phytophtora species produced negligible necroses around the inoculation site. P. plurivora was recovered from all the investigated orchards, providing evidence that it is quite widespread. This study highlights the growing threat posed by the polyphagous P. plurivora to walnut cultivation and the sustainable business it fuels in Europe, underscoring the need for integrated management strategies to mitigate its economic and ecological impacts. Full article

In recent years, severe decline and mortality events have been observed in holm oak (Quercus ilex L.) ecosystems in different Italian regions, including Puglia (southern Italy). Given the landscape and ecological relevance of holm oak forests in Apulia, a study was conducted to identify the causal agents related to this complex disease syndrome. The surveys, conducted in winter 2024 in three different woodlands, revealed the widespread occurrence of mature holm oak trees showing sudden death, crown thinning, shoot and branch dieback, sunken cankers, and root rot symptoms. Isolations performed from symptomatic samples collected from both stem and small roots yielded fungal and fungal-like colonies representing two distinct families: Botryosphaeriaceae and Peronosporaceae. Analysis of morphological and DNA sequence data allowed us to identify six distinct species, including Diplodia corticola and D. quercivora (Botryosphaeriaceae), Phytophthora cinnamomi, P. multivora, P. psychrophila, and P. asparagi (Peronosporaceae). For P. asparagi and P. psychrophila, isolated for the first time from declining holm oak trees in Italy, Koch’s postulates were satisfied by inoculating 1-year-old seedlings at the collar in controlled conditions. Thirty days after inoculation, all plants showed the same symptoms observed in the field. Overall, the data obtained highlights the co-occurrence of multiple Botryosphaeriaceae and Phytophthora species on declining holm oak trees and the discovery of a new haplotype of Diplodia quercivora. Full article

A severe dieback of rosemary (Rosmarinus officinalis L.) plants was observed in a medicinal/culinary herb plantation in Casole d’Elsa, Siena, central Italy. Symptoms included stunted growth, crown desiccation, root rot, collar rot and internal tissue necrosis, strongly indicative of Phytophthora root and crown rot syndrome. Morphological and molecular identification (ITS and Cox1 sequencing) of strains isolated from symptomatic stems, roots and soil revealed the occurrence of two Phytophthora species: Phytophthora pseudocryptogea, which constituted 94% of isolates obtained from the stem, root apparatus and rhizosphere; and Phytophthora megasperma, which was not recovered from plant organs or tissue, being exclusively isolated from rhizosphere soil samples at a low isolation rate (6%). The pathogenicity of the obtained strains was assessed by inoculating eighteen-month-old R. officinalis plants in a soil infestation trial. Plants inoculated with P. pseudocryptogea strains died 10 days after artificial inoculation. P. pseudocryptogea was subsequently re-isolated from the roots of inoculated, symptomatic plants, thus fulfilling Koch’s postulates. Plants inoculated with P. megasperma strains were in good vegetative condition and did not show any visible symptoms, suggesting P. megasperma to be nonpathogenic. Artificial inoculation tests thus confirmed P. pseudocryptogea to be the aetiological agent responsible for the death of R. officinalis plants in the plantation under study. This is the first report of root, collar and crown rot caused by P. pseudocryptogea on R. officinalis in Italy. There is evidence that poorly drained soils and climate constraints facilitate the spread of this oomycete. These findings highlight the critical role of nursery trade in the introduction of Phytophthora species in agroecosystems and emphasize the need for more stringent control measures. Full article

In Chile and worldwide, walnut (Juglans regia L.) production faces significant losses due to crown and root rot caused by the phytopathogen Phytophthora cinnamomi. Currently, control methods have proven insufficient or unfavorable for the environment, increasing the need for sustainable alternatives. This research evaluates nanoemulsions based on extracts of medicinal plants endemic to Chile to control P. cinnamomi in walnut crops. The methodology included an in vitro test to determine the effective inhibitory concentrations of three nanoemulsions (N80, N90, and N100) on the mycelial growth of the phytopathogen, a test on walnut plants under controlled conditions, and two field tests using concentrations between 300 and 500 ppm. The in vitro results showed that the nanoemulsions could inhibit 90% of mycelial growth at 80 to 100 ppm concentrations. In the field, the N90 nanoemulsion at 500 ppm significantly reduced disease symptoms preventively and post-inoculation, compared with the control. This research is the first to study the use of nanoemulsions from native Chilean plants to control P. cinnamomi, showing potential to reduce the use of synthetic fungicides, contributing to safer and more ecological phytosanitary management. Full article

Crown gall disease (Agrobacterium tumefaciens), crown/root rot disease (Phytophthora spp.), root lesion disease (Pratylenchus vulnus) and tree vigor are key traits affecting the productivity and quality of walnuts in California. Unchallenged hybrid rootstocks were analyzed by RNA-seq to examine pre-formed factors affecting these traits. Enrichment analysis of the differentially expressed genes revealed that the increased expression of cell wall biogenesis-related genes plays a key role in susceptibility to A. tumefaciens, susceptibility to Phytophthora spp. and increased vigor. Analysis of the predicted subcellular loci of the encoded proteins revealed that many gene products associated with vigor and susceptibility were targeted to the plasma membrane and extracellular space, connecting these traits to sustaining barrier function. We observed that RNA processing and splicing, along with predicted nuclear targeting, were associated with resistance to A. tumefaciens, resistance to Phytophthora spp. and low vigor. Four genes within the J. microcarpa QTL region for resistance to A. tumefaciens and Phytophthora spp. were represented among our transcripts, with two of the genes being differentially expressed in association with resistance to A. tumefaciens and decreased vigor. No differential expression related to Phytophthora spp. or P. vulnus resistance was observed in this region. Additionally, the J. microcarpa haplotype expressed more transcripts associated with resistance to A. tumefaciens, Phytophthora spp. and low vigor, but not P. vulnus, than the J. regia haplotype. We also report unique and shared hormone and defense responses associated with each trait. This research suggests a link between cell wall biogenesis, vigor and critical root diseases of walnut. Full article

Phytophthora capsici incites foliar blight, root, fruit, and crown rot in squash (Cucurbita spp.) and limits production worldwide. Resistance to crown rot in C. moschata breeding line #394-1-27-12 is conferred by three dominant genes, but the molecular mechanisms underlying this resistance are poorly understood. In the current study, RNA sequencing was used to investigate transcriptional changes in #394-1-27-12 (resistant) and Butterbush (susceptible) following infection by P. capsici at 12, 24, 48, 72, and 120 h post inoculation (hpi). Overall, the number of differentially expressed genes (DEGs) in Butterbush (2648) exceeded those in #394-1-27-12 (1729), but in both genotypes, the highest number of DEGs was observed at 72 hpi and least at 24 hpi. Our gene ontology (GO) analysis revealed a downregulation of the genes involved in polysaccharide and lignin metabolism in Butterbush but as an upregulation of those associated with regulation of peptidase activity. However, in #394-1-27-12, the downregulated genes were primarily associated with response to stimuli, whereas those upregulated were involved in oxidation–reduction and response to stress. The upregulated genes in #394-1-27-12 included defensin-like proteins, respiratory-burst oxidases, ethylene-responsive transcription factors, cytochrome P450 proteins, and peroxidases. These findings provide a framework for the functional validation of the molecular mechanisms underlying resistance to P. capsici in cucurbits. Full article

A dieback was observed on three-year-old pot-grown plants of Cycas revoluta in Sicily (Italy). Symptoms, including stunting, yellowing and blight of the leaf crown, root rot and internal browning and decay of the basal stem, closely resembled the Phytophthora root and crown rot syndrome, common in other ornamentals. Isolations from rotten stem and roots, using a selective medium, and from rhizosphere soil of symptomatic plants, using leaf baiting, yielded three Phytophthora species, P. multivora, P. nicotianae and P. pseudocryptogea, were obtained. Isolates were identified by both morphological characters and DNA barcoding analysis, using three gene regions: ITS, β-tub and COI. Phytophthora pseudocryptogea was the sole species isolated directly from the stem and roots. The pathogenicity of the isolates of the three Phytophthora species was tested on one-year-old potted plants of C. revoluta, using both stem inoculation by wounding, and root inoculation through infested soil. Phytophthora pseudocryptogea was the most virulent and, like P. nicotianae, reproduced all the symptoms of natural infections, while P. multivora was the least virulent and induced solely very mild symptoms. Phytophthora pseudocryptogea was identified as the causal agent of the decline of C. revoluta, as it was re-isolated from both the roots and stems of artificially infected symptomatic plants, thus fulfilling Koch’s postulates. Full article

Cinnamon is a tree introduced to the lowlands of Mexico in the mid-16th century, but it spread to other places at the beginning of the 20th century due to its important commercial value as an aromatic spice. In the state of Veracruz, symptoms of dieback have been observed in 12-year-old cinnamon plantations cultivated in an agroforestry system, causing concern among producers. For this reason, the present investigation was carried out to determine the causal agent of these symptoms observed in cinnamon trees. Fifty symptomatic plants were recovered from established plantations. One hundred cinnamon root fragments showing dieback were selected and separated; isolates were made from tissue showing crown and root rot on clarified juice V-8 agar medium. After eight days, the growth of whitish coralloid mycelium with characteristics similar to the Phytophthora oomycete was consistently observed. Subsequently, the identity corresponding to P. cinnamomi was confirmed by morphological, taxonomic studies and Bayesian inference of the rDNA internal transcribed spacer. The pathogenicity test was performed on 20 6-month-old cinnamon plants grown in pots by inoculating 2.5 × 10⁴/mL of zoospores around the roots. Control plants were inoculated with sterile distilled water and kept in a greenhouse under conditions controlled. After five weeks, symptoms of root rot were observed in the inoculated plants; however, the control group plants remained healthy. The results showed that P. cinnamomi subclade 7c was responsible for the symptoms observed in lowland cinnamon plantations in Mexico. Our findings suggest that this phytopathogen is a new threat for cinnamon growers; likewise, it is recommended that growers implement management strategies to avoid its introduction into nurseries or new plantations that could be susceptible to this pathogen. Full article

Holm oak (Quercus ilex subsp. ballota (Desf.) Samp.) bark is a commonly used remedy to treat gastrointestinal disorders, throat and skin infections, hemorrhages, and dysentery. It has also been previously reported that its methanol extracts possess antibacterial activity, which can be related to the richness of Quercus spp. extracts in phenolic compounds, such as flavonoids and tannins. However, there is no information on the antifungal (including oomycete) properties of the bark from Q. ilex or its subspecies (ilex and ballota). In this work, we report the characterization of the aqueous ammonia extract of its bark by FTIR and GC-MS and the results of in vitro and ex situ inhibition tests against three phytopathogens. The main phytochemical components identified were inositols (19.5%), trans-squalene (13%), 4-butoxy-1-butanol (11.4%), gulopyranose (9.6%), lyxose (6.5%), 2,4-dimethyl-benzo[H]quinoline (5.1%), catechol (4.5%), and methoxyphenols (4.2%). The efficacy of the extract in controlling forest phytopathogens was tested in vitro against Fusarium circinatum (responsible for pitch canker of Pinus spp.), Cryphonectria parasitica (which causes chestnut blight), and Phytophthora cinnamomi (which causes ‘root and crown rot’ in a variety of hosts, including Castanea, conifers, Eucalyptus, Fagus, Juglans, Quercus, etc.), obtaining EC₉₀ values of 322, 295, and 75 μg·mL⁻¹, respectively, much lower than those attained for a commercial strobilurin fungicide (azoxystrobin). The extract was further tested ex situ against P. cinnamomi on artificially inoculated, excised stems of ‘Garnem’ almond rootstock, attaining complete protection at a dose of 782 μg·mL⁻¹. The results suggest that holm oak bark extract may be a promising source of bioactive compounds against invasive forest pathogens, including the oomycete that is causing its decline, the so-called ‘seca’ in Spain. Full article

Fusarium pseudograminearum causes crown rot in wheat. This study aimed to assess the effects of the bacterial strain QTH8 isolated from Cotinus coggygria rhizosphere soil against F. pseudograminearum. Bacterial strain QTH8 was identified as Bacillus halotolerans in accordance with the phenotypic traits and the phylogenetic analysis of 16S rDNA and gyrB gene sequence. Culture filtrates of bacterial strain QTH8 inhibited the mycelial growth of F. pseudograminearum and resulted in mycelial malformation such as tumor formation, protoplast condensation, and mycelial fracture. In addition, bacterial strain QTH8 also inhibited the mycelial growth of Hainesia lythri, Pestalotiopsis sp., Botrytis cinerea, Curvularia lunata, Phyllosticta theaefolia, Fusarium graminearum, Phytophthora nicotianae, and Sclerotinia sclerotiorum. The active compounds produced by bacterial strain QTH8 were resistant to pH, ultraviolet irradiation, and low temperature, and were relatively sensitive to high temperature. After 4 h exposure, culture filtrates of bacterial strain QTH8—when applied at 5%, 10%, 15%, 20%, 25%, and 30%—significantly reduced conidial germination of F. pseudograminearum. The coleoptile infection assay proved that bacterial strain QTH8 reduced the disease index of wheat crown rot. In vivo application of QTH8 to wheat seedlings decreased the disease index of wheat crown rot and increased root length, plant height, and fresh weight. Iturin, surfactin, and fengycin were detected in the culture extract of bacterial strain QTH8 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Bacterial strain QTH8 was identified for the presence of the ituC, bacA, bmyB, spaS, srfAB, fend, and srfAA genes using the specific polymerase chain reaction primers. B. halotolerans QTH8 has a vital potential for the sustainable biocontrol of wheat crown rot. Full article

From 2018 to 2020, surveys of oomycetes associated with root and crown rot of almond (Prunus dulcis) were conducted on diseased young almond trees in commercial orchards and nurseries in six provinces of Spain. A total of 104 oomycete isolates were obtained from plant and soil samples, which h were identified by sequencing the internal transcribed spacer (ITS) region of the ribosomal DNA. Diverse species belonging to the genera Globisporangium, Phytophthora, Phytopythium and Pythium were found, Phytopythium vexans and Phytophthora niederhauserii being the most frequent. The pathogenicity of these two species to one-year-old almond seedlings of ‘Garnem’ (P. dulcis × P. persica) rootstock was studied. All seedlings inoculated with Pp. vexans and Ph. niederhauserii isolates showed severe symptoms at the late stage of the pathogenicity test (defoliation, wilting and dieback) and several plants died. Some isolates of Ph. niederhauserii significantly reduced the dry weight of the roots compared with the control, but this effect was not observed in seedlings inoculated with Pp. vexans. These results provide new information about the oomycete species present in almond crops in Spain and highlight the importance of carrying out frequent phytosanitary surveys for a better knowledge of potential risks posed by these soil-borne pathogens. Full article

Paulownia species are fast growing trees native to China, which are being grown in managed plantings in several European countries for the production of wood and biomasses. In 2018, wilting, stunting, leaf yellowing, and collapse, as a consequence of root and crown rot, were observed in around 40% of trees of a 2-year-old planting of Paulownia elongata × P. fortunei in Calabria (Southern Italy). Two species of Phytophthora were consistently recovered from roots, basal stem bark, and rhizosphere soil of symptomatic trees and were identified as Ph. nicotianae and Ph. palmivora on the basis of both morphological characteristics and phylogenetic analysis of rDNA ITS sequences. Koch’s postulates were fulfilled by reproducing the symptoms on potted paulownia saplings transplanted into infested soil or stem-inoculated by wounding. Both Phytophthora species were pathogenic and caused root rot and stem cankers. Even though P. palmivora was the only species recovered from roots of naturally infected plants, in pathogenicity tests through infested soil P. nicotianae was more virulent. This is the first report of Phytophthora root and crown rot of a Paulownia species in Europe. Strategies to prevent this emerging disease include the use of healthy nursery plants, choice of well-drained soils for new plantations, and proper irrigation management. Full article

Phytopathogenic microorganisms belonging to the genus Phytophthora have been recognized many times as causal agents of diseases that lower the yield of many plants important for agriculture. Meanwhile, Phytophthora cactorum causes crown rot and leather rot of berry fruits, mainly strawberries. However, widely-applied culture-based methods used for the detection of pathogens are time-consuming and often inaccurate. What is more, molecular techniques require costly equipment. Here we show a rapid and effective detection method for the aforementioned targets, deploying a simple molecular biology technique, Loop-Mediated Isothermal Amplification (LAMP). We optimized assays to amplify the translation elongation factor 1-α (EF1a) gene for two targets: Phytophthora sp. And Phytophthora cactorum. We optimized the LAMP on pure strains of the pathogens, isolated from organic plantations of strawberry, and successfully validated the assay on biological material from the environment including soil samples, rhizosphere, shoots and roots of strawberry, and with SYBR Green. Our results demonstrate that a simple and reliable molecular detection method, that requires only a thermoblock and simple DNA isolation kit, can be successfully applied to detect pathogens that are difficult to separate from the field. We anticipate our findings to be a starting point for developing easier and faster modifications of the isothermal detection methods and which can be applied directly in the plantation, in particular with the use of freeze-dried reagents and chemistry, allowing observation of the results with the naked eye. Full article

Field surveys conducted on nine farms over a 2-year period showed the widespread presence of Phytophthora-related diseases on globe artichoke plants in the main growing area in Sardinia (Italy). Characteristic symptoms included wilting and necrosis of the outermost leaves and dark brown discoloration of stem tissues, as well as root rot. A total of 18 Phytophthora colonies belonging to three species were isolated and characterized. Based on morphological features and ITS sequence data, Phytophthora isolates were identified as P. crassamura (eight isolates) and P. cactorum (four isolates). Six isolates could not be assigned to any formally described species of Phytophthora and are therefore described here as Phytophthora marrasii sp. nov. The ITS phylogeny places P. marrasii in a terminal clade basal to the sister taxa (P. foliorum, P. hibernalis, P. lateralis, and P. ramorum) of the clade 8c. In particular, P. marrasii is phylogenetically related to P. foliorum, a species from which it differs in 62 nucleotides in the ITS region. At the same time, it can easily be distinguished morphologically from P. foliorum mainly because of the low minimum temperature for growth, the bigger and persistent non-papillate sporangia, and smaller oogonia. Pathogenicity tests confirmed that all three Phytophthora species are pathogenic on globe artichokes, which represent a new host for these pathogens. Full article

In this study, the early activation of plant-defense-related genes during a three-way plant–antagonist–pathogen interaction in a tomato–Trichoderma–Phytophthora nicotianae model system was evaluated. Thirty-day-old tomato seedlings were treated at the root systems with a suspension of germinated conidia of two selected strains of T. asperellum and T. atroviride and then inoculated with zoospores of P. nicotianae. The defense mechanisms activated by tomato plants upon the simultaneous colonization of the root systems by Trichoderma spp. and P. nicotianae were evaluated 72 h post-inoculation by analyzing the transcriptomic profiles of genes involved in the pathways of salicylic acid (i.e., pathogenesis-related proteins—PR1b1 andPR-P2-encoding genes), jasmonic acid (i.e., lipoxygenases enzymes—TomLoxC- and TomLoxA-encoding genes), and the tomato plant defensin protein (i.e., SlyDF2-encoding gene). The results showed that PR1b1 was more strongly up-regulated in the three-way system including T. asperellum, while the gene PR-P2 was up-regulated in the system including T. atroviride. TomLoxA was significantly up-regulated only in the three-way system including T. asperellum, while TomLoxC was significantly up-regulated in neither of the analyzed three-way systems. Finally, the gene SlyDF2 was significantly up-regulated in tomato seedlings in both three-way systems. Full article