Pitch Canker Disease Research Articles

Spray-induced gene silencing (SIGS) as a tool for the management of Pine Pitch Canker forest disease.

Global change is exacerbating the prevalence of plant diseases caused by pathogenic fungi in forests worldwide. The conventional use of chemical fungicides, which is commonplace in agricultural settings, is not sanctioned for application in forest ecosystems, so novel control strategies are imperative. SIGS (Spray-Induced Gene Silencing) is a promising approach that can modulate the expression of target genes in eukaryotes in response to double-stranded RNA (dsRNA) present in the environment that triggers the RNA interference (RNAi) mechanism. SIGS exhibited notable success in reducing virulence when deployed against some crop fungal pathogens, such as Fusarium graminearum, Botrytis cinerea and Sclerotinia sclerotiorum, among others. However, there is a conspicuous dearth of studies evaluating the applicability of SIGS for managing forest pathogens. This research aimed to determine whether SIGS could be used to control Fusarium circinatum, a widely impactful forest pathogen that causes Pine Pitch Canker disease. Through a bacterial synthesis, we produced dsRNA molecules to target fungal essential genes involved to vesicle trafficking (Vps51, DCTN1, and SAC1), signal transduction (Pp2a, Sit4, Ppg1, and Tap42), and cell wall biogenesis (Chs1, Chs2, Chs3b, Gls1) metabolic pathways. We confirmed that F. circinatum is able to uptake externally applied dsRNA, triggering an inhibition of the pathogen's virulence. Furthermore, this study pioneers the demonstration that recurrent applications of dsRNAs in SIGS are more effective in protecting plants than single applications. Therefore, SIGS emerges as an effective and sustainable approach for managing plant pathogens, showcasing its efficacy in controlling a globally significant forest pathogen subject to quarantine measures.

A delayed response in phytohormone signaling and production contributes to pine susceptibility to Fusarium circinatum

BackgroundFusarium circinatum is the causal agent of pine pitch canker disease, which affects Pinus species worldwide, causing significant economic and ecological losses. In Spain, two Pinus species are most affected by the pathogen; Pinus radiata is highly susceptible, while Pinus pinaster has shown moderate resistance. In F. circinatum-Pinus interactions, phytohormones are known to play a crucial role in plant defense. By comparing species with different degrees of susceptibility, we aimed to elucidate the fundamental mechanisms underlying resistance to the pathogen. For this purpose, we used an integrative approach by combining gene expression and metabolomic phytohormone analyses at 5 and 10 days post inoculation.ResultsGene expression and metabolite phytohormone contents suggested that the moderate resistance of P. pinaster to F. circinatum is determined by the induction of phytohormone signaling and hormone rearrangement beginning at 5 dpi, when symptoms are still not visible. Jasmonic acid was the hormone that showed the greatest increase by 5 dpi, together with the active gibberellic acid 4 and the cytokinin dehydrozeatin; there was also an increase in abscisic acid and salicylic acid by 10 dpi. In contrast, P. radiata hormonal changes were delayed until 10 dpi, when symptoms were already visible; however, this increase was not as high as that in P. pinaster. Indeed, in P. radiata, no differences in jasmonic acid or salicylic acid production were found. Gene expression analysis supported the hormonal data, since the activation of genes related to phytohormone synthesis was observed earlier in P. pinaster than in the susceptible P. radiata.ConclusionsWe determine that the moderate resistance of P. pinaster to F. circinatum is in part a result of early and strong activation of plant phytohormone-based defense responses before symptoms become visible. We suggest that jasmonic acid signaling and production are strongly associated with F. circinatum resistance. In contrast, P. radiata susceptibility was attributed to a delayed response to the fungus at the moment when symptoms were visible. Our results contribute to a better understanding of the phytohormone-based defense mechanism involved in the Pinus-F. circinatum interactions and provide insight into the development of new strategies for disease mitigation.

From Lab to Nursery: Novel Approaches of Seed Disinfection for Managing Pine Pitch Canker Propagation

Fusarium circinatum, the causative agent of pine pitch canker disease, is a pathogenic fungus that poses a significant threat to pine forests globally. It infects various Pinus species, causing resinous cankers, needle discoloration, and tree death. The disease severely impacts forest ecosystems, necessitating cost-effective and environmentally friendly management strategies. Contaminated pine seeds and seedlings are the main pathways for introducing this fungus to disease-free areas. To mitigate this disease and prevent its spread, it is crucial to implement new processes in forest plant production systems that align with the existing conditions of forest nurseries, ensuring effective and sustainable management. With this in mind, a national collaborative study involving 14 Portuguese partners was initiated to develop new prevention and mitigation strategies. In this work, four different treatments—MennoFlorades, Captan, ethanol, and hot water—were tested for their ability to eliminate F. circinatum from contaminated Pinus seeds in vitro. The most effective treatments were selected for further in vitro assays and real-context nursery germination trials to assess their impacts on seed germination, plant production, and certification. MennoFlorades, Captan, and hot water were tested in the nursery, with hot water showing the most promising results due to its negligible impact on seedlings, eco-friendly nature, ease of implementation, and cost-effectiveness. These findings offer promising prospects for preventing pine pitch canker outbreaks in nurseries and, consequently, in forests.

Relative Expression of Genes Elicited by Clonostachys rosea in Pinus radiata Induces Systemic Resistance

Radiata pine is one of the most commonly planted tree species in Chile due to its fast growth and desirable wood and pulp properties. However, its productivity is hampered by several diseases. Pitch canker disease (PCC) caused by Fusarium circinatum, is considered the most damaging disease to the pine forest industry. Several control measures have been established, with biological control emerging as an environmentally friendly and effective way for F. circinatum control. Previous studies support the value of Clonostachys rosea in reducing PCC damage, with evidence suggesting a potential induced systemic resistance (ISR) triggered in radiata pines by this agent. Ten-month-old radiata pine plants were pre-treated with C. rosea on a substrate at 8 and 1 days before inoculation with F. circinatum on the stem tip, and expression levels were determined for DXS1, LOX, PAL, and PR3 genes 24 h later. Lesion length was 45% lower on plants pre-treated with C. rosea and infected with F. circinatum compared to non-pre-treated and infected plants. Additionally, LOX and PR3 were induced 23 and 62 times more, respectively, in comparison to untreated plants. Our results indicate that C. rosea causes an ISR response in pre-treated plants, significantly increasing the expression of resistance genes and reducing lesion length.

Genetic Control of Pitch Canker Response in Southern Pine and Southern Pine Hybrids

Fusarium circinatum causes pine pitch canker (PPC) disease and associated symptoms such as resinous lesions, mechanical weakness, and crown dieback that may lead to mortality in Pinus and Pseudotsuga spp. There are no ameliorative techniques available for the disease, and the genetic resistance among populations to support commercial plantation deployment has not been well characterized. In this study, we characterize the genetic control of PPC disease tolerance (and/or resistance) and predict the tolerance of families in existing breeding populations: open-pollinated (OP) half-sib and control-pollinated full-sib (FS) slash pine (Pinus elliottii var. elliottii Engelm.), OP loblolly pine (Pinus taeda L.), and advanced-generation OP hybrid slash × P. caribaea (Pinus elliottii var. elliottii Engelm. × Pinus caribaea var. bahamensis, caribaea, and hondurensis) using F. circinatum isolates obtained from three locations in Georgia and FL, USA. We describe a new experimental design that improves the accuracy of breeding value predictions, provides more precise genetic parameter estimates, and facilitates comparisons within and among taxa as well as comparisons among isolates. We found strong evidence for genetic control of the ratio of stem damage by F. circinatum, especially in slash pine and slash × P. caribaea hybrids. Loblolly and slash × P. caribaea hybrids exhibited less damage than slash pine. We observed a spectrum of virulence among F. circinatum isolate sources, which were not equally virulent in different pine taxa.

Temporal and Spatial Variation in the Population Structure of Spanish Fusarium circinatum Infecting Pine Stands

Fusarium circinatum is an introduced fungal pathogen extended to the northern regions of Spain that causes Pine Pitch Canker (PPC) disease. In this work, we analyzed the pathogen's genetic diversity to study changes over time and space since the first outbreak occurred in Spain. Using six polymorphic SSR markers, 15 MLGs were identified in 66 isolates, and only three haplotypes were found with frequencies higher than one. In general, genotypic diversity was low and decreased shortly over time in the northwestern regions while maintained at País Vasco, where only one haplotype (MLG32) was detected 10 years. This population also included isolates of a single mating type (MAT-2) and VCGs identified in only two groups, while isolates from NW regions were of both mating types and VCGs represented in 11 groups. The existence of haplotype MLG32 maintained on time and widely distributed suggests its good adaptation to the environment and the host. Results showed that the pathogen in País Vasco remains clearly differentiated from other northwestern populations. This fact was supported with no evidence of migration among regions. Results are explained by the asexual reproduction, but also selfing at least to a lesser extent that leads to identification of two new haplotypes.

Co-Infections by Fusarium circinatum and Phytophthora spp. on Pinus radiata: Complex Phenotypic and Molecular Interactions.

This study investigated the complex phenotypic and genetic response of Monterey pine (Pinus radiata) seedlings to co-infections by F. circinatum, the causal agent of pine pitch canker disease, and the oomycetes Phytophthora xcambivora and P. parvispora. Monterey pine seedlings were wound-inoculated with each single pathogen and with the combinations F. circinatum/P. xcambivora and F. circinatum/P. parvispora. Initially, seedlings inoculated only with F. circinatum showed less severe symptoms than seedlings co-inoculated or inoculated only with P. xcambivora or P. parvispora. However, 30 days post-inoculation (dpi), all inoculated seedlings, including those inoculated only with F. circinatum, showed severe symptoms with no significant differences among treatments. The transcriptomic profiles of three genes encoding pathogenesis-related proteins, i.e., chitinase (PR3), thaumatin-like protein (PR5), phenylalanine ammonia-lyase (PAL), and the pyruvate decarboxylase (PDC)-encoding gene were analyzed at various time intervals after inoculation. In seedlings inoculated with single pathogens, F. circinatum stimulated the up-regulation of all genes, while between the two oomycetes, only P. xcambivora induced significant up-regulations. In seedlings co-inoculated with F. circinatum and P. xcambivora or P. parvispora none of the genes showed a significant over-expression 4 dpi. In contrast, at 11 dpi, significant up-regulation was observed for PR5 in the combination F. circinatum/P. xcambivora and PDC in the combination F. circinatum/P. parvispora, thus suggesting a possible synergism of multiple infections in triggering this plant defense mechanism.

Ras2 is important for growth and pathogenicity in Fusarium circinatum

In this study, we investigated to possible role of Ras2 in Fusarium circinatum- a fungus that causes pine pitch canker disease on many different pine species and has a wide geographic distribution. This protein is encoded by the RAS2 gene and has been shown to control growth and pathogenicity in a number of fungi in a mitogen-activated protein kinase- and/or cyclic adenosyl monophosphate pathway-dependent manner. The aim was therefore to characterize the phenotypes of RAS2 gene knockout and complementation mutants of F. circinatum. These mutants were generated by transforming protoplasts of the fungus with suitable split-marker constructs. The mutant strains, together with the wild type strain, were used in growth studies as well as pathogenicity assays on Pinus patula seedlings. Results showed that the knockout mutant strain produced significantly smaller lesions compared to the complementation mutant and wild type strains. Growth studies also showed significantly smaller colonies and delayed conidial germination in the knockout mutant strain compared to the complement mutant and wild type strains. Interestingly, the knockout mutant strain produced more macroconidia than the wild type strain. Collectively, these results showed that Ras2 plays an important role in both growth and pathogenicity of F. circinatum. Future studies will seek to determine the pathway(s) through which Ras2 controls these traits in F. circinatum.

The transcriptome of Pinus pinaster under Fusarium circinatum challenge

BackgroundFusarium circinatum, the causal agent of pitch canker disease, poses a serious threat to several Pinus species affecting plantations and nurseries. Although Pinus pinaster has shown moderate resistance to F. circinatum, the molecular mechanisms of defense in this host are still unknown. Phytohormones produced by the plant and by the pathogen are known to play a crucial role in determining the outcome of plant-pathogen interactions. Therefore, the aim of this study was to determine the role of phytohormones in F. circinatum virulence, that compromise host resistance.ResultsA high quality P. pinaster de novo transcriptome assembly was generated, represented by 24,375 sequences from which 17,593 were full length genes, and utilized to determine the expression profiles of both organisms during the infection process at 3, 5 and 10 days post-inoculation using a dual RNA-sequencing approach. The moderate resistance shown by Pinus pinaster at the early time points may be explained by the expression profiles pertaining to early recognition of the pathogen, the induction of pathogenesis-related proteins and the activation of complex phytohormone signaling pathways that involves crosstalk between salicylic acid, jasmonic acid, ethylene and possibly auxins. Moreover, the expression of F. circinatum genes related to hormone biosynthesis suggests manipulation of the host phytohormone balance to its own benefit.ConclusionsWe hypothesize three key steps of host manipulation: perturbing ethylene homeostasis by fungal expression of genes related to ethylene biosynthesis, blocking jasmonic acid signaling by coronatine insensitive 1 (COI1) suppression, and preventing salicylic acid biosynthesis from the chorismate pathway by the synthesis of isochorismatase family hydrolase (ICSH) genes. These results warrant further testing in F. circinatum mutants to confirm the mechanism behind perturbing host phytohormone homeostasis.

Pine Pitch Canker (PPC): Pathways of Pathogen Spread and Preventive Measures

Fusarium circinatum (Nirenberg and O’ Donnell) is the causal agent of pine pitch canker (PPC) disease, one of the most devastating forest diseases worldwide. Long-distance spread occurs mainly through the movement of infected seeds whereas at regional level, the movement of seedlings, substrates, or containers may play an important role in fungal dispersal. Invasion of nurseries takes place via infected seeds and further spread can occur by planting contaminated seedlings, especially due to the possibility of infected plants remaining symptomless. Once established, F. circinatum spreads by rain, wind, and insects. The natural spread of the pathogen is limited due to the short dispersal distances of the spores and the fairly short flight distances of disseminating insects. In this review, we summarize the currently known dispersal pathways of the pathogen, discussing both natural and human-assisted processes. With the purpose of understanding how to best intervene in the disease’s development in nurseries and forests, we outline the epidemiology of the pathogen describing the key factors influencing its spread. Preventive measures to control the spread of F. circinatum locally and globally are described with special emphasis on the challenges in implementing them.

Pine Pitch Canker and Insects: Relationships and Implications for Disease Spread in Europe

The fungal pathogen Fusarium circinatum (Nirenberg and O’ Donnell) is the causal agent of pine pitch canker (PPC) disease, which seriously affects conifer species in forests and nurseries worldwide. In Europe, PPC is only established in the Iberian Peninsula; however, it is presumed that its range could expand through the continent in the near future. Infection caused by this fungus requires open wounds on the tree, including physical damage caused by insects. Therefore, a relationship probably occurs between PPC and a wide variety of insects. The aim of this review is to outline the taxonomic and ecological diversity of insect species with high potential association with F. circinatum in Europe and elsewhere. The insects were classified as vectors, carriers and wounding agents according to the association level with the PPC disease. In addition, we discuss the insect-mediated spreading of PPC disease in relation to the different phases of forest stand development, from seeds and seedlings in nurseries to mature stands. Lastly, to improve our predictive capacities and to design appropriate intervention measures and strategies for controlling disease dissemination by insects, variables such as geographic location, time of the year and host species should be considered. Our review provides a framework of the multiple factors that regulate the insect–host interactions and determine the success of the infection.

Projecting Monterey Pine (Pinus radiata) Populations over Time in the Presence of Various Representations of Pitch Canker Disease, Caused by Fusarium circinatum

Monterey pine (Pinus radiata D. Don) is native to California and widely planted in Mediterranean climates around the world. Pitch canker, a disease caused by Fusarium circinatum Nirenberg and O’Donnell, is a serious threat to P. radiata in native forests and in plantations. Because of its economic importance worldwide, conservation of P. radiata native populations is a high priority. We developed a demographic matrix projection model to simulate dynamics of naturally occurring P. radiata populations in California in the presence of the disease. Tree demography data for the model were collected from seven sites in a native forest on the Monterey Peninsula. Height and stem diameter were recorded for all trees, which were divided into five size classes based on these data. Transition probabilities were calculated for each size class; cone production, seed release, and seed transition probability were estimated using an iterative search process. In the model, pitch canker influences state-specific fecundity and survival probabilities. Five different approaches to include these effects were compared. Populations were projected over 50 decades from three specified initial population sizes. Elasticity analysis indicated that facilitating survival of all size classes is vital to maintaining the structure of the forest.

Factors Affecting the Distribution of Pine Pitch Canker in Northern Spain

Fusarium circinatum is the causal agent of pine pitch canker disease (PPC), affecting Pinus species and other conifers (i.e., Pseudotsuga menziesii (Mirb.) Franco.), forming resinous cankers on the main stem and branches and causing dieback in the terminal guide. This pathogen is spreading worldwide, causing economic losses by converting plantations into standing timber without any potential for future production. The disease was recently detected in Northern Spain in plantations of Pinus radiata and forest nurseries. The aim of the work reported here was to study the role of climatic and topographic variables, soil properties, and stand characteristics on PPC. For this purpose, we surveyed 50 pine stands in Cantabria and quantified the percentage of trees showing three symptoms in each stand: canker, defoliation, and dieback. We investigated the predictive power of 30 variables using generalized linear models and hierarchical partitioning. Both approaches yielded similar results. We found that the three symptoms correlated with different explanatory variables. In addition, more trees exhibited cankers in the proximity of the coast and the Basque Country. Additionally, our results showed that low canopy cover is related to a high level of the dieback symptom. Overall, this study highlights the important variables affecting the distribution of PPC in Cantabria.

Pine defenses against the pitch canker disease are modulated by a native insect newly associated with the invasive fungus

Tomicus piniperda (Coleptera, Curculionidae) is a native insect in Europe and now potential enabler of the invasive fungal pathogen of pines, Fusarium circinatum. The co-occurence of T. piniperda and F. circunatum could increase disease incidence via (1) insect wounding when they bore galleries into shoots of healthy trees, and (2) by acting as vectors that introduce fungal propagules directly. The extent of damage by F. circinatum is likely to be influenced by tree defenses, which can depend upon pine species, environmental conditions, and responses to other biotic and abiotic challenges. Indirect interactions among plant enemies can arise when the attack by one enemy alters the shared host plant in a way that affects a second enemy. We tested for indirect interactions between T. piniperda and F. circinatum mediated by plant defenses. Our study employed 100 potted, identically treated, seedlings of Pinus pinaster and P. radiata. Plants were randomly allocated to one of five different treatments: inoculation with T. piniperda alone, inoculation with F. circinatum alone, inoculation with T. piniperda and F. circinatum together, a mock treatment with sterilized water, and true controls. We assessed plant mortality, and measured plant height and diameter, crown size, resin flow after T. piniperda attack, the size of lesions from fungal infection, and phloem chemistry (terpenes and nitrogen). There were conspicuous differences between pine species in their defensive responses to inoculations: P. pinaster showed increases in numerous terpenes following attack by T. piniperda but was unresponsive to F. circinatum, while P. radiata increased terpene concentrations with exposure to either F. circinatum or T. piniperda. In P. radiata, fungal growth within the plant (as reflected by lesion size) was reduced by coincident insect attack. This suggests that plant defensive responses against the insect also functioned as deterrents to a fungal enemy. Thus the combination of T. piniperda and F. circinatum should not be as dangerous to pine stands as if the insect disabled defences against the fungus. Nonetheless, the insect can only increase fungal introductions by increasing wounds and moving propagules. There is a need for field studies of damage from F. circinatum in different pine species with low vs. high abundance of T. piniperda. Better understanding of pine defenses against F. circinatum, and genetic variation in defense responses, would contribute to the development of safe, effective, and sustainable tactics for managing damage from F. circinatum to pine forests in Spain and elsewhere.

Response to Temperature and Virulence Assessment of Fusarium circinatum Isolates in the Context of Climate Change

With future global temperatures predicted to increase, the relationship between a host, pathogen, and environment, becomes less predictable and epidemics may pose a greater risk to forests worldwide. Resistance breeding is an important disease management tool, but because tree species require long breeding times, it is necessary to develop techniques for testing current pathogen isolates against their hosts. Pitch canker disease of pines, caused by the pathogen Fusarium circinatum, is no exception and represents a threat to pine forests and commercial plantations worldwide, as it thrives at warm temperatures and high humidity. We tested growth of 15 F. circinatum isolates in culture at three temperatures: 25, 27, and 31 °C. We also evaluated the sporulation and pathogenicity of eight of the isolates on two susceptible Pinus elliotti (slash pine) open-pollinated families and one tolerant open-pollinated Pinus taeda (loblolly pine) family. Our results showed significant differences among isolates in the temperature and pathogenicity tests. All isolates showed a significant decrease in growth at 31 °C, although some showed similar growth at 25 and 27 °C. Several of the new isolates tested were more pathogenic than the isolates that the USDA Forest Service Resistance Screening Center (RSC) had been using. The new isolates have now been incorporated into their operational screening program.

A Low-Cost Spore Trap Allows Collection and Real-Time PCR Quantification of Airborne Fusarium circinatum Spores

A variety of commercial instruments are available for sampling and quantifying microscopic airborne organisms from the environment. Although most samplers are highly sensitive, they are also expensive, costing thousands of dollars per unit, a price that is out of reach for many researchers, especially those looking to design experiments with replication. While looking at options to monitor pine stands for the presence of Fusarium circinatum, the causal agent of pitch canker disease, on multiple sites with several units per site, we developed a simple, low-cost spore trap that allows surveying spore abundance in outdoor environments. The trap consists of a rotating motor that holds a metal rod and two petroleum jelly-coated microscope slides. As the motor rotates, the slides collect airborne particles. To test whether the traps allowed detection of F. circinatum spores, we placed six traps on three sites: an actively-managed slash pine commercial stand located in Lake Butler, FL, a semi-managed loblolly and slash pine stand near Gainesville, FL, and a site with little perturbance at Goethe State Forest, FL, consisting of mainly slash pine trees. The slides were replaced weekly, and F. circinatum was detected by quantitative PCR using species-specific primers. Results show detection of low levels ( X ¯ = 1.7–77.1 picograms ± SE = 0.3–39.7) of the pathogen spores with high reproducibility. These traps offer a low-cost solution to spore, pollen, or small insect trapping experiments for initial or general assessment of a pathogen or species population. Their low cost has the added benefit that multiple traps can be deployed per experiment, thus increasing statistical power by using multiple replications.

Vertical Transmission of Fusarium circinatum Mitoviruses FcMV1 and FcMV2-2 via Microconidia

Pine Pitch Canker disease, caused by the pathogenic fungus Fusarium circinatum, affects conifer species worldwide. However, the virulence of the pathogen may be affected by the presence of mycoviruses. The aim of this laboratory-based study was to investigate the probability and rate of transmission of F. circinatum mitoviruses FcMV1 and FcMV2-2 via microconidia. Ten isolates of mitovirus-infected F. circinatum were subcultured to produce a total of 100 single-spore colonies (ten replicates per isolate). The total RNA and cDNA obtained from each spore isolate (monosporic culture) were amplified by PCR with specific primers for detection of F. circinatum mitoviruses FcMV1 and FcMV2-2. The mitoviruses were detected in a high percentage of the individual spore isolates (between 60% and 100% depending on the fungal isolate). However, the probability of transmission was not statistically significantly associated with either the F. circinatum isolate or the viral strain. A high proportion of transmission via microconidia is critical for development of a biological control program against Pine Pitch Canker (PPC) disease in forests. However, further studies are needed to establish the effect of these mitoviruses on the virulence of F. circinatum.

Root Infection of Canker Pathogens, Fusarium circinatum and Diplodia sapinea, in Asymptomatic Trees in Pinus radiata and Pinus pinaster Plantations

The existence of a latent stage within host tissue of the pine pathogens Fusarium circinatum and Diplodia sapinea, the causal agents of pitch canker and shoot blight disease respectively, has previously been cited. However, studies on this cryptic phase in each disease lifecycle has only been focused on the host aerial parts but not on the roots. Therefore, our objective was to analyze the presence of both pathogens in roots of non-symptomatic mature trees in plantations where the pathogens are known to be causing canker symptoms. For that, we sampled roots from ten non-symptomatic and ten symptomatic trees in three Pinus radiata and one Pinus pinaster plantations in Basque Country, Spain. Both pathogens were isolated from roots of non-symptomatic trees in a higher frequency than from roots of symptomatic trees, 23.3% and 6.6% respectively for D. sapinea and 16.6% and 3.3% respectively for F. circinatum. Neither pathogens was detected in the P. pinaster plantation. The two pathogens were never isolated from the same tree. A high molecular variability was observed for D. sapinea isolates with six different haplotypes and two mating types for the eleven characterized isolates, but only one haplotype and mating type was found for F. circinatum, with all isolates of both fungi being proved pathogenic. These results evidence the importance root infection may have in the disease lifecycle and, therefore, disease management.

In silico annotation of five candidate genes associated with pathogenicity in Fusarium circinatum

SummaryThe Pine Pitch Canker disease caused by the pathogenic fungus Fusarium circinatum is one of the most devastating diseases in pine forests, afforestation and nurseries around the world. Despite the importance of this phytopathogen, only a little is known about the genes that drive the infection traits and the virulence factors. In this work, five candidate genes (i.e. Fcfga1, Fcfgb1, Fcac, Fcrho1 and FcpacC) were in silico annotated using the whole genome of F. circinatum as reference. The similarity of these proposed genes at nucleotide and protein levels with genes previously described in other Fusarium species was > 90% of identity and > 90% query coverage in all cases. In addition, the gene ontology of each candidate gene was also investigated.

Herbaceous plants in the understory of a pitch canker-affected Pinus radiata plantation are endophytically infected with Fusarium circinatum

Abstract Fusarium circinatum was recently detected as an endophyte in grasses causing no apparent damage. Our goal was to describe the endophytic colonization of herbaceous host plants growing in a plantation of Pinus radiata with symptoms of pitch canker disease, which may act as a reservoir of inoculum. We detected the fungus in five species of dicot families (Asteraceae, Lamiaceae, Rosaceae), in addition to two species in the Poaceae. The fungus was found in the aerial part of non-symptomatic hosts, so we describe F. circinatum as an endophyte that is mainly transmitted by spores through the air. It was also detected in Hypochaeris radicata seeds, suggesting the potential occurrence of vertical transmission. An analysis of microsatellite markers showed a unique haplotype regardless of whether the isolates' origin was pine cankers or non-symptomatic herbaceous plants. Thus, the same genotype can adopt a pathogenic or endophytic lifestyle. We conclude that non-symptomatic plants can act as reservoirs of inoculum; pine seedlings can be infected from senescent tissue of non-symptomatic hosts colonized by the fungus.