Armillaria Root Rot Research Articles

In vitro co-culture system for investigating Armillaria root rot in Prunus spp. using a fiber-supported liquid approach.

In vitro co-culture techniques that allow the growth of plants and pathogens under controlled environmental conditions are being used to re-create host plant infection. These approaches reduce infection times, promote reproducibility, and enable a rapid evaluation of plant-pathogen interactions. As a result, these systems have become essential in breeding programs aimed at developing plant resistance to diseases. In this study, we developed and validated an in vitro co-culture system to investigate the Armillaria root rot (ARR) affecting Prunus spp. This disease, caused by fungi Armillaria spp. and Desarmillaria caespitosa, poses a severe threat to the stone and nut fruit industry due to the susceptibility of most commercial rootstocks to infection and the lack of effective management options for its control. The system consists of a fiber-supported liquid approach in sterile plastic vessels that allows a fast and reproducible fungal infection under controlled environmental conditions. The floor of the vessels was covered with a polyester-fiber matte and a germination paper that served as an interface between the mycelia and the plant roots. The vessels were subjected to inoculation with Armillaria mellea and D. caespitosa, and three Prunus genotypes ('Guardian®', 'MP-29', and Prunus cerasifera '14-4') were co-cultured with both fungi. Disease progression and plant and fungal biomass were monitored during co-culture. The presented in vitro co-culture approach facilitates the concurrent growth of Armillaria/Desarmillaria spp. and Prunus spp., excluding most of the limitations associated with greenhouses and field experiments. This system provides consistent and reproducible conditions for investigating a prominent plant disease affecting Prunus spp.

Armillaria Species Causing Kiwifruit Vine Decline and Root Rot in Northeastern Türkiye: A Growing Concern for Kiwifruit Health

ABSTRACTKiwifruit (Actinidia deliciosa) production in northeastern Türkiye is facing an emerging threat from Armillaria root rot. Surveys conducted in August 2021 and 2022 revealed that 10.5%–17.5% of the 200–500 vines within each of 35 kiwifruit orchards in the Perşembe, Gülyalı and Altınordu districts of Ordu province exhibited symptoms of Armillaria root rot, including complete defoliation. Diseased plants exhibited diagnostic signs on their root collars and woody roots, including white mycelial fans, extensive wood rot and rhizomorphs. A single Armillaria isolate was obtained from each of the 35 symptomatic orchards. Molecular identification, employing DNA sequencing of the large subunit (LSU) region of rDNA, the translation elongation factor subunit 1‐alpha (TEF1) gene and the second largest RNA polymerase II B‐subunit (RPB2) gene, identified 20 isolates as Armillaria gallica and 15 as A. mellea. Phylogenetic analyses, based on TEF1 alignments, further supported the species identification with a 100% bootstrap value. To examine genetic diversity, start codon targeted (SCoT) marker 13 was used, successfully differentiating the two Armillaria species and revealing three distinct clades. Clades I and II comprised 15 and 5 isolates of A. gallica, respectively, while Clade III contained all 15 A. mellea isolates. Pathogenicity testing on kiwifruit seedlings (cv. Hayward) revealed the ability of both A. gallica and A. mellea isolates to induce disease. Rhizomorph‐producing isolates in kiwifruit seedlings displayed high virulence, inducing severe leaf necrosis and seedling mortality, while isolates lacking these structures were non‐pathogenic. This study identifies the first global instance of A. gallica causing kiwifruit root rot. This finding, along with the identification of A. mellea as a causal agent in the country, highlights the emerging threat of Armillaria root rot to kiwifruit production in the region.

Using local knowledge to reconstruct climate‐mediated changes in disease dynamics and yield—A case study on Arabica coffee in its native range

Societal Impact StatementAdapting agriculture to climate change requires an understanding of the long‐term relationship between climate, disease dynamics, and yield. While some countries have monitored major crop diseases for decades or centuries, comparable data is scarce or non‐existent for many countries that are most vulnerable to climate change. For this, a novel approach was developed to reconstruct climate‐mediated changes in disease dynamics and yield. Here, a case study on Arabica coffee in its area of origin demonstrates how to combine local knowledge, climate data, and spatial field surveys to reconstruct disease and yield time series and to postulate and test hypotheses for climate–disease–yield relationships.Summary While some countries have monitored crop diseases for several decades or centuries, other countries have very limited historical time series. In such areas, we lack data on long‐term patterns and drivers of disease dynamics, which is important for developing climate‐resilient disease management strategies. We adopted a novel approach, combining local knowledge, climate data, and spatial field surveys to understand long‐term climate‐mediated changes in disease dynamics in coffee agroforestry systems. For this, we worked with 58 smallholder farmers in southwestern Ethiopia, the area of origin of Arabica coffee. The majority of farmers perceived an increase in coffee leaf rust and a decrease in coffee berry disease, whereas perceptions of changes in coffee wilt disease and Armillaria root rot were highly variable among farmers. Climate data supported farmers' understanding of the climatic drivers (increased temperature, less rainy days) of these changes. Temporal disease‐climate relationships were matched by spatial disease‐climate relationships, as expected with space‐for‐time substitution. Understanding long‐term disease dynamics and yield is crucial to adapt disease management to climate change. Our study demonstrates how to combine local knowledge, climate data and spatial field surveys to reconstruct disease time series and postulate hypotheses for disease‐climate relationships in areas where few long‐term time series exist.

АРМИЛЛЯРИОЗНАЯ КОРНЕВАЯ ГНИЛЬ АБРИКОСОВЫХ ДЕРЕВЬЕВ В УСЛОВИЯХ ЗАПАДНОЙ ЧАСТИ АЗЕРБАЙДЖАНА

АРМИЛЛЯРИОЗНАЯ КОРНЕВАЯ ГНИЛЬ АБРИКОСОВЫХ ДЕРЕВЬЕВ В УСЛОВИЯХ ЗАПАДНОЙ ЧАСТИ АЗЕРБАЙДЖАНА

Rapid In Vitro Screening of Prunus Genotypes for Resistance to Armillaria Root Rot Using Roots of Young Rootstocks

Armillaria root rot (ARR), caused by Armillaria species and Desarmillaria tabescens, is a severe disease that affects stone fruit trees in the United States. One strategy to mitigate the impact of this disease is to develop ARR-resistant rootstocks. However, current techniques to screen Prunus species for resistance to ARR are time-consuming, labor-intensive, and may not fully replicate field conditions. To address these limitations, we developed a new rapid in vitro screening assay, which uses roots of 2-year-old Prunus rootstock genotypes. We screened 12 Prunus genotypes against Armillaria mellea, Armillaria solidipes, and Desarmillaria tabescens in vitro. Freshly excavated root segments were placed next to or on top of fungal cultures. After 21 days, the circumferential percentage and horizontal length of the fungal colonization and the ability of the fungus to enter through root periderm were evaluated. The root tissue surrounding the infection was also evaluated to assess any response reactions against the ARR pathogens. Our results showed that inoculated root tissues displayed signs of fungal infection, and infection and host responses varied among the Prunus genotypes. Host responses similar to those observed in the field, such as compartmentalization of infected tissue with barrier zones, necrophylactic periderm formation, and callus formation on root surfaces, were observed and were more evident in less susceptible genotypes. In conclusion, our newly developed assay, which uses freshly excavated roots from 2-year-old rootstocks, can rapidly screen Prunus genotypes for resistance to ARR.

Belarusian Oak Forests Afflicted with Armillaria Root Rot during Their Mass Desiccation

During the period of the oak forests’ mass desiccation, Armillaria root rot became widespread, exacerbating the state of depression. The infection of stands by white sapwood rot caused by fungi of the genus Armillaria was more often detected in Bug-Polesia and Berezina-Pre-Polesia forest growth areas. The incidence of the disease in the oak forests of Belarus increases with the raising of the stands age and the decrease in their density. More often, the affliction with Armillaria root rot occurs in floodplain oak forests, as well as in the dry lands forest types with the most fertile soils. The oak’s proportion in the stand’s composition does not significantly affect the occurrence of the infection. Under the conditions of massive weakening of the Belarussian oak forests, facultative parasites from the Armillaria genus can occur as dangerous secondary pathogens that accelerate the death of weakened oak trees. Their pathogenicity persists in the north of the republic, where the negative role of other pathological factors in oak forests decreases. After the end of the depression period, the transition of Armillaria from a parasitic to a predominantly saprotrophic strategy was recorded. The affliction of oak forests by Armillaria root rot can act as one of the oak formation’s condition indicators, marking the depression phase in oak stands. Therefore, the spread of foci of Armillaria root rot should be monitored while conducting forest pathology surveys and monitoring the condition of oak forests.

Transcriptomics reveal the genetic coordination of early defense to Armillaria root rot (ARR) in Prunus spp.

Armillaria root rot (ARR) poses a significant threat to the long-term productivity of stone-fruit and nut crops in the predominant production area of the United States. To mitigate this issue, the development of ARR-resistant and horticulturally-acceptable rootstocks is a crucial step towards the maintenance of production sustainability. To date, genetic resistance to ARR has been found in exotic plum germplasm and a peach/plum hybrid rootstock, 'MP-29'. However, the widely-used peach rootstock Guardian® is susceptible to the pathogen. To understand the molecular defense mechanisms involved in ARR resistance in Prunus rootstocks, transcriptomic analyses of one susceptible and two resistant Prunus spp. were performed using two causal agents of ARR, including Armillaria mellea and Desarmillaria tabescens. The results of in vitro co-culture experiments revealed that the two resistant genotypes showed different temporal response dynamics and fungus-specific responses, as seen in the genetic response. Gene expression analysis over time indicated an enrichment of defense-related ontologies, including glucosyltransferase activity, monooxygenase activity, glutathione transferase activity, and peroxidase activity. Differential gene expression and co-expression network analysis highlighted key hub genes involved in the sensing and enzymatic degradation of chitin, GSTs, oxidoreductases, transcription factors, and biochemical pathways likely involved in Armillaria resistance. These data provide valuable resources for the improvement of ARR resistance in Prunus rootstocks through breeding.

Chemical sensitivity phenome of an Armillaria mellea isolate determined using phenotype microarrays and potential use of caffeine-rich wastes for disease control

Armillaria fungi are frequently documented as economically and ecologically significant plant pathogens, recognised as the causal agents of Armillaria root rot disease. Armillaria mellea is one of the most aggressive pathogens in the Armillaria genus. None of the chemicals tested against Armillaria have been effective in fully eradicating an established A. mellea mycelium from an infection site and/or preventing plant mortality, which makes studies about the chemical sensitivity of A. mellea essential. In this work, the inhibitory effects of 120 different chemical agents on the growth rate of A. mellea were examined using the Biolog Phenotype MicroArray system of chemical sensitivity panels. Among the tested substances, aromatic and membrane function compounds showed the highest inhibitory activity against A. mellea. Interestingly, our results demonstrated promising potential for application of caffeine as an A. mellea-oriented fungicide. Further studies were conducted to explore the antifungal activity of a low-cost and locally available caffeine-rich waste, i.e. spent coffee grounds (SCG) against A. mellea. It is noteworthy that the hyphal growth of A. mellea was significantly inhibited when cultivated on malt extract agar supplemented with SCG. Current findings uncovered, for the first time, the potential use of caffeine-rich wastes for designing management strategies to practically control the spread of A. mellea.

Armillaria root and crown rot disease in pistachio orchards

Armillaria root and crown rot disease in pistachio orchards

Endophytic Trichoderma spp. can protect strawberry and privet plants from infection by the fungus Armillaria mellea.

Armillaria mellea is an important fungal pathogen worldwide, affecting a large number of hosts in the horticulture and forestry industries. Controlling A. mellea infection is expensive, labour intensive and time-consuming, so a new, environmentally friendly management solution is required. To this effect, endophytic Trichoderma species were studied as a potential protective agent for Armillaria root rot (ARR) in strawberry and privet plants. A collection of forty endophytic Trichoderma isolates were inoculated into strawberry (Fragaria × ananassa) plants and plant growth was monitored for two months, during which time Trichoderma treatment had no apparent effect. Trichoderma-colonised strawberry plants were then inoculated with A. mellea and after three months plants were assessed for A. mellea infection. There was considerable variation in ARR disease levels between plants inoculated with different Trichoderma spp. isolates, but seven isolates reduced ARR below the level of positive controls. These isolates were further tested for protective potential in Trichoderma-colonized privet (Ligustrum vulgare) plants where five Trichoderma spp. isolates, including two highly effective Trichoderma atrobrunneum isolates, were able to significantly reduce levels of disease. This study highlights the potential of plants pre-colonised with T. atrobrunneum for effective protection against A. mellea in two hosts from different plant families.

Antagonistic Effects of Trichoderma Species in Biocontrol of Armillaria Root Rot Disease In Vitro Conditions

Aim of study: Armillaria root rot is a disease of fruit trees, some herbaceous plants and forest trees. Armillaria spp. which cause this disease, are among the most destructive fungal pathogens of trees. In this study, antagonistic effects of Trichoderma species in biocontrol of Armillaria were investigated in vitro conditions Area of study: The study area was in the Düzce University, Konuralp Campus. Material and methods: Trichoderma harzianum was provided from Düzce University, Faculty of Forestry, Department of Forestry Industrial Engineering (FS19) and Trichoderma viride was provided from Düzce University, Konuralp Campus. Armillaria mellea and Desarmillaria tabescens were isolated from the infected roots of hazelnut and oak trees in Konuralp Campus and cultured in potato dextrose agar. Trichoderma species were inoculated to confront with the pathogens in cultures of Armillaria and Desarmillaria to determine antagonistic effects for 15 days. Penetration of Trichoderma hyphae in rhizomorphs and sporulation of Trichoderma species in the surface of rhizomorphs were investigated by scanning electron microscopy. Main results: The statistically significant decrease was observed for A. mellea and Desarmillaria species caused by Trichoderma. One day after Trichoderma inoculation, Trichoderma hyphae grew approximately twice as much towards the host rhizomorphs. Three days after colonization of Armillaria and Desarmillaria rhizomorphs by Trichoderma, growth of Armillaria and Desarmillaria seemed to completely stopped. Highlights: Trichoderma species have been studied as the most widely used antagonists in biological control against many plant-pathogenic fungi worldwide. To develop an effective biological method for plant protection in the future, biological control studies are highly important in oak forest under natural stand conditions.

Armillaria root rot fungi host single-stranded RNA viruses

Species of Armillaria are distributed globally and include some of the most important pathogens of forest and ornamental trees. Some of them form large long-living clones that are considered as one of the largest organisms on earth and are capable of long-range spore-mediated transfer as well as vegetative spread by drought-resistant hyphal cords called rhizomorphs. However, the virus community infecting these species has remained unknown. In this study we used dsRNA screening and high-throughput sequencing to search for possible virus infections in a collection of Armillaria isolates representing three different species: Armillaria mellea from South Africa, A. borealis from Finland and Russia (Siberia) and A. cepistipes from Finland. Our analysis revealed the presence of both negative-sense RNA viruses and positive-sense RNA viruses, while no dsRNA viruses were detected. The viruses included putative new members of virus families Mymonaviridae, Botourmiaviridae and Virgaviridae and members of a recently discovered virus group tentatively named “ambiviruses” with ambisense bicistronic genomic organization. We demonstrated that Armillaria isolates can be cured of viruses by thermal treatment, which enables the examination of virus effects on host growth and phenotype using isogenic virus-infected and virus-free strains.

Evaluation of chemical, genetic and biological control strategies for Armillaria root rot of peach

Evaluation of chemical, genetic and biological control strategies for Armillaria root rot of peach

Identification of native endophytic Trichoderma spp. for investigation of in vitro antagonism towards Armillaria mellea using synthetic- and plant-based substrates.

To isolate endophytic Trichoderma species and investigate the potential for biological control of the root rot pathogen Armillaria mellea. In all, 40 Trichoderma isolates were obtained from a range of host plants and identities were confirmed by ITS, rpb2 and tef1 sequence. When tested in dual culture assays for antagonism against A. mellea, Trichoderma isolates overgrew the A. mellea colonies within four days and by eightdays 38 Trichoderma isolates significantly reduced A. mellea colony size. Armillaria mellea was unable to be recovered from five of eight co-cultivations tested, suggesting Trichoderma had killed the A. mellea in these cases. Pre-colonized hazel disks were used to determine what happens in a more heterogeneous situation with A. mellea and a refined set of eight Trichoderma isolates. Similar to plate-based assays, Trichoderma quickly covered A. mellea stopping any further growth and two Trichoderma isolates were able to eradicate A. mellea. Of the Trichoderma spp. tested, endophytic isolates of Trichoderma virens and T. hamatum offered the greatest antagonism towards A. mellea. Using pre-colonized hazel disks was of great importance for this work to demonstrate the fungal interactions in plant material. Controlling Armillaria root rot is difficult with chemical treatments, thus an environmentally benign and cost-effective alternative is required. This study highlights the prospect of biological control as an effective, environmentally friendly alternative to chemicals.

The crown condition of Norway spruce and occurrence of symptoms caused by Armillaria spp. in mixed stands

Mixed stands are currently supported as effective management options to reduce forest vulnerability to climate change. However, our issues about benefits of mixtures, mechanisms of their incidence and conditions for their effectiveness have yet been only partly clarified. We assessed the crown condition of Norway spruce (Picea abies /L./ Karst) and occurrence of symptoms caused by Armillaria spp. on spruce in a small area in the Drahanská vrchovina Highlands (Czech Republic) in differently mixed stands in 2002 and 2019. We found that although the broadleaf abundance affected the total defoliation of Norway spruce, the effect was unstable in time. The observed effect of altitudinal zones and Norway spruce abundance on the occurrence of symptoms of Armillaria root rot in spruce was also unstable. The drought during recent four years and the implemented forestry measures (salvage and intermediate felling) can be reasons for the relationship instability. We did not identify any statistically significant relationship between tree species diversity (Simpson’s Index) and defoliation.

Evaluation of the susceptibility of Prunus rootstock genotypes to Armillaria and Desarmillaria species

Variation in susceptibility of 28 Prunus rootstock genotypes to the causal agents of Armillaria root rot, Armillaria mellea, A. solidipes, and Desarmillaria tabescens, were studied by conducting in vitro root screening assays. Root segments with wounded and intact periderm were placed next to and on the top of the fungal cultures. At day 21, the percent success of fungal penetration and the circumferential and longitudinal lengths of fungal growth were measured. A parallel investigation using the inoculated root segments was carried out to characterize the active host defense mechanisms involved, including anatomical responses in bark and wood. Overall, the success of penetration and the longitudinal and circumferential spread of Armillaria spp. and D. tabescens were significantly different among various Prunus rootstock genotypes. None of the tested rootstock genotypes were completely resistant to infection. However, plum and plum derived rootstocks, and some of the cherry genotypes were less susceptible to infection compared to the peach genotypes. The host’s ability to limit infection by Armillaria spp. and D. tabescens was not limited to a single mechanism but appeared to be regulated by several collective nonspecific host responses acting together. Differential levels of a series of non-specific coordinated events were triggered in Prunus genotypes such as the formation of new callus tissue on the root surface, a colored reaction zone, necrophylactic periderm, new cells, and new vascular cambium to compartmentalize the pathogen. The host responses were elevated in the genotypes with low level of infection as compared to highly infected genotypes.

Armillaria root rot threatens Cameroon’s Penja pepper (Piper nigrum L.)

Penja pepper (Piper nigrum) produced in Cameroon has long been recognized for its exceptional organoleptic quality. The pepper vine is grown using a support tree (Spondias mombin) in Cameroon. A root disease is associated with plant deaths in both plants. The disease symptoms are characterized by collar cracking and gummosis and the disease was tentatively identified as Armillaria root rot. In this work the extent of the problem was characterized by surveying 35 farms in Cameroon. Samples were taken from diseased support trees and pepper vines. Support trees exhibiting typical symptoms were found in approximately one third of the surveyed farms. In these farms, disease incidence and Pepper vine mortality ranged from 1.3 to 50% and 1.2 to 87.7%, respectively. Analysis of the ribosomal internal transcribed spacer region (ITS) locus suggested that the isolates collected from S. mombin and P. nigrum are most likely, which is a first for both hosts, represented by Armillaria camerunensis (Henn.) Volk & Burdsall, a fungus previously associated with declining cacao trees in Cameroon. Given the increasing demand for Penja pepper and the great damaging potential of this root rot, research on management strategies should be prioritized.

Patterns and drivers of fungal disease communities on Arabica coffee along a management gradient

Plants, including those managed by humans, are often attacked by multiple diseases. Yet, most studies focus on single diseases, even if the disease dynamics of multiple species is more interesting from a farmers’ perspective. Moreover, most studies are from single management systems, although it is valuable to understand how diseases are distributed across broad management gradients, especially in cases where less intensive management also provides biodiversity values in the landscape. To understand the spatial dynamics and drivers of diseases across such a broad management gradient, we assessed four major fungal diseases on Arabica coffee (Coffea arabica) at 60 sites in southwestern Ethiopia along a gradient from only little managed wild coffee in the forest understory to intensively managed coffee plantations. We found that environmental and management factors related to disease incidence and severity differed strongly among the four fungal diseases. Coffee leaf rust (Hemileia vastatrix) and Armillaria root rot (Armillaria mellea) were more severe in intensively managed sites, whereas coffee berry disease (Colletotrichum kahawae) and wilt disease (Gibberella xylarioides) were more severe in less managed sites. Among sites, incidence and severity of the four fungal diseases poorly correlated with each other. Within sites, however, shrubs that were severely attacked by coffee leaf rust also had high levels of berry disease symptoms. A better understanding of disease dynamics is important for providing management recommendations that benefit smallholder farmers, but also to evaluate possibilities for maintaining biodiversity values in the landscape related to shade cover complexity and wild coffee genetic variation.

Long-term effects of stump removal and tree species composition on the diversity and structure of soil fungal communities.

Stump removal is a common forest management practice used to reduce the mortality of trees affected by the fungal pathogen-mediated root disease, Armillaria root rot, but the impact of stumping on soil fungal community structure is not well understood. This study analyzed the long-term impact of stumping and tree species composition on the abundance, diversity and taxonomic composition of soil fungal communities using internal transcribed spacer (ITS) marker-based DNA metabarcoding in a 48-year-old trial at Skimikin, British Columbia. A total of 108 samples were collected from FH (fermented and humus layers), and soil mineral horizons (A and B) from stumped and unstumped plots of six tree species treatments (pure stands and admixtures of Douglas-fir, western red-cedar and paper birch). Fungal α-diversity in the A horizon significantly increased with stumping regardless of tree species composition, while β-diversity was significantly affected by stumping in all the horizons. We also observed that the relative abundance of the saprotrophic fungal community declined while that of the ectomycorrhizal fungal community increased with stumping. In conclusion, increase in ectomycorrhizal fungal associations, which are positively associated with tree productivity, suggests that stumping can be considered a good management practice for mitigating root disease and promoting tree regeneration.

Preventative Root-Collar Excavation Reduces Peach Tree Mortality Caused By Armillaria Root Rot On Replant Sites.

In the southeastern United States, Armillaria root rot (ARR) is caused by Desarmillaria tabescens and has become the primary cause of premature mortality in peach orchards. Most rootstocks used in commercial orchards are susceptible and management options are limited. A postinfection practice known as root-collar excavation (RCE), which involves permanent removal of the soil from the base of the trunk, has been shown to improve yields and prolong the productive life of symptomatic trees. However, symptomatic trees already have an advanced infection at the base of the trunk. This study evaluated the efficacy of preventative RCE on the progression of tree mortality in two orchards that were planted in infested replant sites. To provide convincing data for growers, the study was carried out in a commercial orchard and an experimental orchard for 8 years. Furthermore, representative enterprise budgets and net present value (NPV) analysis were utilized to compare the profitability of the two approaches. Trees were planted shallow on berms (45 by 90 cm) to facilitate RCE with hoes and AirSpade 2 years later. Tree mortality in the RCE treatment of the experimental orchard was first observed in year 6 and increased 8% on average per year thereafter. In contrast, tree mortality in the "Grower Standard" treatment was first observed in year 4 and increased 12.7% on average per year thereafter. At the commercial orchard, tree mortality in the RCE treatment was first observed in year 7 and increased 1.9% on average thereafter, while tree mortality in the Grower Standard treatment first appeared in year 5 and increased 4.3% on average thereafter. The delayed onset of ARR-associated tree mortality and the lower annual tree mortality rate in the RCE treatment led to higher NPVs in both locations. There were no negative effects on yield or fruit quality. However, the new planting system can create horticultural challenges, including the formation of a proper berm, uneven ground around the tree interfering with tree care and harvest, increased erosion due to channeling of rainwater, and increased rootstock suckering. The RCE is a valid option for southeastern growers needing to manage high ARR disease pressure on replant sites or on sites only recently cleared from ARR-infected forest land.