Armillaria root rot spreading into a natural woody ecosystem in South Africa

Signs and symptoms of a disease similar to those of armillaria root rot have recently been observed on various native woody plants on the foothills of Table Mountain in South Africa, one of the most botanically diverse natural environments globally. This is of concern because the root rot fungus Armillaria mellea has previously been shown to be an alien pathogen of European origin in planted gardens in the City of Cape Town. An aim of this study was to identify the cause of the root rot disease on infected plants. Based on DNA‐sequence phylogeny, it was shown that isolates collected from at least 16 native tree and woody shrub species represented the non‐native A. mellea. Microsatellite markers were then used to determine the genetic diversity and population structure of the A. mellea isolates from Table Mountain and two planted gardens where the pathogen has previously been found. Population genetic analyses revealed low levels of gene diversity and no population differentiation amongst the three populations. The results provide the first firm evidence that A. mellea has escaped the planted environment and invaded a sensitive and ecologically important natural woody environment in South Africa. This is only the second definitive case of a non‐native tree pathogen invading a natural ecosystem in the country.     

A semiaxenic phototrophic system to study interactions between arbuscular mycorrhizal and pathogenic fungi in woody plants

Among other benefits, arbuscular mycorrhizal (AM) fungi may increase plant tolerance to root diseases. The research on the underlying mechanisms requires growth conditions that are both controlled and realistic. To study these interactions, a semiaxenic phototrophic system was developed in which the roots grow in a controlled environment and can be inoculated with both pathogenic and symbiotic fungi. Micropropagated fig plantlets were grown in containers having shoots in the outside and roots in a growth medium without sugar, inoculated or not with the AM fungus Rhizophagus irregularis and the pathogenic fungus Armillaria mellea. Dual inoculated plants developed the mycorrhizal association and pathogen infection symptoms. Mycorrhizal inoculation lowered disease index and increased plant growth. Colonization of A. mellea in fig roots was quantified by real-time PCR, showing that R. irregularis did not significantly lower the quantity of Armillaria, suggesting that other mechanisms were involved in increased tolerance to the pathogen. The results show that the system proposed is suitable to study the triple interaction involving plant, AM and root pathogenic fungi.     

Fumigation with methyl bromide II. Equipment and methods for sampling and analysing deep field soil atmospheres

Procedures were developed for the rapid sampling of deep soil atmospheres in the field and analysing them by gas chromatography. The methods described here have been used in studies on the diffusion of methyl bromide in relation to the control of Armillaria mellea.     

Root rot pathogens in field soil,roots and seeds in relation to common bean (Phaseolus vulgaris), disease and seed production

The interrelationships among bean productivity, prevalence of pathogens in roots, seeds and soil, and root rot disease were described at the pod maturity stage in 13 commercial fields. The soil population and frequency of pathogens isolated from seeds varied by pathogen species and field location. Fusarium solani was the most prevalent fungus isolated from bean seeds and field soil compared to Rhizoctonia solani, Macrophomina phaseolina and F. oxysporum. Principal component analysis revealed that the first component explaining 32% of the total variance was correlated with the root rot index. PC1 was more strongly linked to root and seed infections in comparison with soil populations of pathogens. Based on a correlation between PC2 (accounting for 23% of the total variance) and the number of seeds per bean plant, charcoal, Fusarium and Rhizoctonia root rots were recognized as more important determinants of seed losses to root rot disease. There were correlations among the major pathogens infecting either roots or seeds of beans. These findings provide useful information for future experimental plans to optimize management strategies for bean root rots.     

New South Wales Department of Agriculture,Entomology Branch,Annual Report 1966-7 (extracts)

Abstract

Damage to tea plants due to a root infection was observed on an estate in southern Malawi. Initial symptoms on plants in the infected areas varied from yellowing of leaves on individual branches followed by dieback to a more frequent rapid wilting of the leaves of entire bushes. The bark surface of superficial roots and collar region was covered by a loose mat of coarse fungal mycelium varying from white or cream to pale or bright yellow in colour. Infection was well developed on a large proportion of roots of affected plants before foliage symptoms appeared. The causal agent was identified as Pseudophaeolus baudonii (Pat.) Ryv.; this is the first record of the fungus in Malawi and the first record of its occurrence anywhere on tea. The fungus is widely distributed in Africa on a range of hosts. It is not yet known how the fungus is transmitted and definite control methods cannot be recommended although it is suggested that entire tea crops in diseased areas be removed and the land used for tobacco production. It is unlikely thatP. baudonii will have serious economic effects on tea crops in the area.     

Colonization of the vegetative stage of rice plants by the false smut fungus Villosiclava virens,as revealed by a combination of species‐specific detection methods

Rice false smut disease caused by the ascomycete fungus Villosiclava virens (Clavicipitaceae) reduces rice yield worldwide. It invades rice panicles and forms dark‐green false smut balls composed of thick‐walled conidia. Although the infection process during the booting stage is well studied, its infection route before this is unclear. It was hypothesized that the thick‐walled conidia in soil penetrate rice roots, and the fungus latently colonizes roots and tiller buds at the vegetative stage. This hypothesis was tested using species‐specific detection methods. First, real‐time PCR with species‐specific primers and probe was used to estimate thick‐walled conidial number in the paddy field soil. Secondly, nested PCR with species‐specific primers showed that fungal DNA was detected in roots and shoot apices of rice plants in the vegetative stage. Thirdly, colourimetric in situ hybridization with a species‐specific oligonucleotide probe targeting 18S rRNA suggested that sparse mycelia or tightly condensed mycelia were present on the external surface of tiller buds enveloped by juvenile leaf sheaths at the vegetative stage. Thin hyphae were found around leaf axils at the surface of elongated stems at the heading stage, and the fungal hyphae grew in the rice root tissues. In addition, it was demonstrated that eGFP‐tagged transformants of the fungus invaded rice roots and colonized the surface of roots and leaf sheaths under artificial conditions.     

Suppression of clubroot by Clonostachys rosea via antibiosis and induced host resistance

The mechanism of the biofungicide Prestop^® (Clonostachys rosea) was investigated for control of clubroot (Plasmodiophora brassicae) on canola. The key product components were partitioned and assessed for their effect on pathogen resting spores, root hair infection (RHI) and disease development using light microscopy, quantitative PCR and different application treatments during infection. The whole product of Prestop was consistently more effective than the C. rosea conidial suspension or product filtrate alone in reducing RHI and clubroot development. This biofungicide showed little effect on germination or viability of resting spores. Two‐application treatments at seeding and 7–14 days after seeding achieved greater clubroot control than a single application of the biofungicide at either seeding or post‐seeding stage. This may indicate the need to maintain a high biofungicide dose in the soil during primary and secondary infection. This biocontrol fungus colonized the rhizosphere and interior of canola roots extensively, and possibly induced plant resistance based on up‐regulation of the genes that are involved in jasmonic acid (BnOPR2), ethylene (BnACO) and phenylpropanoid (BnOPCL, BnCCR) biosynthetic pathways. It is concluded that the biofungicide Prestop suppressed clubroot on canola at least via root colonization and induced systemic resistance (ISR), and the latter may be through the modulation of phenylpropanoid and jasmonic acid/ethylene metabolic pathways elicited by the fungus.     

A Root Box Method to Estimate Virulence of Helicobasidium mompa Using Carrots and Its Comparison with the Conventional Method Using Apple Stocks

A root box method with carrots was developed to estimate virulence of the violet root rot fungus, Helicobasidium mompa, to facilitate short-term screening of many isolates during a year. The root box consisted of two transparent acrylic plates and a plastic bag of vermiculite in which two taproots of carrot were growing and inoculated with the fungus growing on fragments of mulberry twigs. The boxes were kept in a greenhouse at 25°C, and the surface of carrots was observed weekly up to 14 weeks. The virulence of each isolate was determined based on the number of weeks after inoculation required for the fungus to develop infection cushions on the surface of carrots. Results were compared with those from the conventional inoculation method using apple stocks. Two-year-old 456 apple stocks were planted with or without fungal inoculum in 30-cm diam. plastic pots containing commercial soil and placed outdoors in April 1999. Symptoms on plant tops were observed weekly, and the first stocks were killed 14 weeks after inoculation. At the end of trial 1 (6 months) and trial 2 (14 months), apple stocks were dug up to rate disease index (DI) based on hyphal growth and infection cushion formation on the stem base. There was variability in disease severity among replicates as well as isolate variability ; however, the results were similar in both trails. The level of virulence estimated by both methods was almost parallel for a total of 23 isolates from five plant species, except for two isolates from sweet potato that formed no obvious infection cushion on apple roots but on carrot were the most virulent. Received 11 December 2000/ Accepted in revised form 23 March 2001     

A Greenhouse Test for Screening Sugar Beet (Beta Vulgaris) for Resistance to Rhizoctonia Solani

Rhizoctonia solani Kühn is a serious plant pathogenic fungus, causing various types of damage to sugar beet (Beta vulgaris L.). In Europe, the disease is spreading and becoming a threat for the growing of this crop. Plant resistance seems to be the most practical and economical way to control the disease. Experiments were carried out to optimise a greenhouse procedure to screen plants of sugar beet for resistance to R. solani. In the first experiment, two susceptible accessions were evaluated for root and leaf symptoms, after being grown in seven different soil mixtures and inoculated with R. solani. The fungus infected all plants. It was concluded that leaf symptoms were not reliable for the rating of disease severity. Statistically significant differences between the soil mixtures were observed, and there were no significant differences between the two accessions. The two soil mixtures, showing the most severe disease symptoms, were selected for a second experiment, including both resistant and susceptible accessions. As in the first experiment, root symptoms were recorded using a 1–7 scale, and a significant expression of resistance was observed. The average severity of the disease in the greenhouse experiment generally was comparable with the infection in field experiments, and the ranking of the accessions was the same in the two types of experiments. It was concluded that evaluation procedures in the greenhouse could be used as a rapid assay to screen sugar beet plants for resistance to R. solani.     

Morphophysiological response of young olive trees to verticillium wilt under different surface drip irrigation regimes

The effects of irrigation on verticillium wilt in olive, in terms of morphological, biomass and physiological parameters were evaluated on pot‐grown trees maintained in the field for 3 years. Plants inoculated and noninoculated with Verticillium dahliae were irrigated to high and low range of soil water content (HR and LR) at daily (DF; about 2 days/event), weekly (WF) and daily‐weekly (DWF) drip‐irrigation frequency. Morphological parameters, relative biomass and biomass water‐use efficiency were higher at LR than at HR (with few exceptions) and at DF than at other frequencies in noninoculated and inoculated plants, but the fungus reduced those parameters by 17.0–38.5%. Lower root weight ratio, relative biomass and shoot length as area originated at HR in noninoculated plants, could be favourable to the accumulation of root infections and the amount of fungus per tissue length in inoculated plants because higher infection was known at HR. Moreover, higher aerial biomass and length promoted by irrigation at DF could prevent the more severe expression of symptoms, which occurred at WF and DWF in the presence of Verticillium. Negative correlations were found between indicated parameters and disease. Lower water stress (S_Ψ), and higher stomatal conductance (g_s) and net photosynthesis at DF in noninoculated plants could limit the disease by improving water status, as S_Ψ was increased by the fungus only at WF and DWF, and g_s and disease were negatively correlated. LR‐DF treatment minimized the disease and kept the growth, water‐use efficiency and physiological parameters in inoculated plants to levels close to noninoculated plants.     

Armillaria Species Infesting Vineyards in Northwestern Spain

Twelve vineyards in northwestern Spain were studied to assess the incidence of white root rot during 1995 and 1997. In both years, diseased plant material was collected and the Armillaria species responsible was identified on the basis of compatibility testing. Subsequently, restriction fragment length polymorphism (RFLP) analysis of the intergenic spacer region of the nuclear ribosomal RNA gene cluster was used to identify Armillaria species in another 45 symptomatic samples submitted for diagnosis from this area. During 1995, 9 of the 12 vineyards showed white root rot, although in eight cases less than 2% of plants were affected; in the remaining vineyard 17% of plants were diseased. During 1997, 10 of the 12 vineyards were affected and three showed a marked increase in the number of plants with white root rot (to 43%, in the vineyard in which 17% were affected in 1995). The compatibility method detected Armillaria mellea in samples from 10 of the 12 vineyards, and Armillaria gallica in samples from two vineyards. RFLP–PCR analysis detected four restriction patterns corresponding to A. mellea, A. gallica and Armillaria cepistipes: patterns mel 1 (A. mellea, 71% of samples), mel 2 (A. mellea, 18%), gal 1 (A. gallica, 9%), and cep 1 (A. cepistipes, 2%, i.e. a single sample). This is the first report of A. gallica and A. cepistipes infecting Vitis spp. The presence of these two Armillaria species may be related to the fact that the vineyards from which they were isolated were located on cleared forestry sites.     

Inoculation of Mature Pedunculate Oaks (Quercus robur) with the Root Rot Fungus Collybia fusipes: Relationships with Tree Vigour and Soil Factors

The severity of natural infections induced by the root rot fungus Collybia fusipes depends on soil factors. Severely infected trees usually show reduced vigour, as evidenced by poor growth. However, it is not known whether reduced vigour could be a cause of the severe infection. The objective was to clarify the relationships between soil factors, tree vigour and susceptibility of Quercus robur to C. fusipes by artificially inoculating mature trees. Two experiments compared oak trees of different ages and dominance class and oak trees growing on different type of soils. The inoculum survival and the infection success were poor in both experiments when waterlogging was severe. Inoculum survival was better in soil with increasing sand/clay ratio, carbon/nitrogen ratio and decreasing pH and calcium, magnesium and phosphorus availability. The relationships between oak vigour and success of infection or surface area of lesions were inconsistent, with a slightly higher infection success on co-dominant/suppressed oaks in one experiment and a higher surface area of successful infection on dominant oaks in the second.     

Epidemiology of root rot caused by <Emphasis Type="Italic">Leptosphaeria maculans</Emphasis> in <Emphasis Type="Italic">Brassica napus</Emphasis> crops

The infection of above-ground tissues of Brassica napus by Leptosphaeria maculans is well understood. However, root infection (root rot) under field conditions, the development of root rot over time and its relationship to other disease symptoms caused by L. maculans has not been described. A survey of B. napus crops was conducted in Australia to investigate the incidence and severity of root rot. Additionally, the pathway of root infection was examined in field experiments. Root rot was present in 95% of the 127 crops surveyed. The severity and incidence of root rot was significantly correlated with that of crown canker; however, the strength of this relationship was dependent on the season. Root rot symptoms appeared before flowering and increased in severity during flowering and at maturity, a pattern similar to crown canker suggesting that the infection of the root is an extension of the crown canker phase of the L. maculans lifecycle. All isolates of L. maculans tested in glasshouse experiments caused root rot and crown canker in B. napus and Brassica juncea. In the field, the main pathway of root infection is via invasion of cotyledons or leaves by airborne ascospores, rather than from inoculum in the soil. Root rot was present in crops in fields that had never been sown to B. napus previously, in plants grown in fumigated fields, and in glasshouse-grown plants inoculated in the hypocotyl with L. maculans.     

Root‐knot nematodes do not affect black root rot disease severity on watermelon and bottle gourd

The effects of root‐knot nematodes on black root rot of watermelon and bottle gourd were studied using field surveys and co‐inoculation tests with Meloidogyne incognita (southern root‐knot nematode) and Diaporthe sclerotioides. The results of the field survey suggested that root‐knot nematodes had little effect on either the severity of black root rot or infection with D. sclerotioides. None of the three fields showed a significant positive correlation between disease severity and nematode gall index, with low correlation coefficients. Co‐inoculation experiments under controlled conditions found no significant effect of root‐knot nematodes on black root rot of watermelon and bottle gourd based on area under disease progress curves (AUDPC). These results were supported by the quantities of DNA of the two agents in root tissues because no significant difference was found between dual‐ and single‐inoculation treatments with M. incognita and/or D. sclerotioides. These findings suggest that root‐knot nematodes probably do not affect the infection of watermelon or bottle gourd roots by D. sclerotioides or the incidence of black root rot in these crops caused by this fungus.     

Hot water treatment of Japanese pear trees is effective against white root rot caused by <Emphasis Type="Italic">Rosellinia necatrix</Emphasis> Prillieux

Hot water was dripped into the rhizosphere of Japanese pear trees (Pyrus serotina Rehd. grafted on P. betulifolia Bunge.) infested with the white root rot fungus Rosellinia necatrix Prillieux, to destroy the fungus. Isolates of R. necatrix from diseased roots of Japanese pear were vulnerable to water at temperatures above 35°C, and the fungus was eradicated from the colonized substrate when water at 35°C was provided for 3 days. The time required to eradicate R. necatrix decreased exponentially with increasing temperature. Japanese pear trees tolerated a temperature of 45°C without reduction in vigor. Field experiments demonstrated the practical use of hot water drip irrigation (HWD). HWD at 50°C completely destroyed white root rot mycelia on diseased roots, and many rootlets grew after the treatment. HWD at this temperature caused no injury to the trees. HWD of diseased orchard trees was assessed in Takamori and Iida in southern Nagano, Japan. The fungus recurred in two of four trees 28 months after treatment in Takamori and in two of ten trees 16 months after treatment in Iida. The new mycelia emerged on thick roots deep within the soil. Although there is a possibility of recurrence, HWD treatment is a practical control measure for white root rot.     

Seasonal dynamics of the pink root fungus (Setophoma terrestris) in rhizosphere soil: Effect of crop species and rotation

Setophoma terrestris, a ubiquitous inhabitant of soil, causes pink root rot in various crops. In the present study, the density of S. terrestris was estimated by quantitative real-time PCR in onion and non-onion fields of Hokkaido, the northernmost island of Japan. Three-year observations in monoculture and rotation fields demonstrated that the fungus grew significantly from the third year onwards, and declined in fields planted with poor hosts (e.g., sugar beet and soybean) that produced few or no chlamydospores of S. terrestris. Seasonal analysis revealed that the population of S. terrestris consistently increased when the tops of onions fell over in summer, which is when root activity declines. However, the soil inoculum potential estimated by a seedling bioassay showed distinct seasonal patterns, which rose from post-harvest in winter and remained high until the subsequent planting in spring. Detailed surveys on depth distribution in an onion field detected a high population of S. terrestris in the effective layer (10–30 cm deep) but not below the hardpan (40 cm), implying that the fungus is intimately associated with roots. These results indicate that the proliferation of the fungus is closely related to root senescence and that over-wintered propagules play an important role in primary infections, affecting disease severity. The present study shows that the temporal dynamics of S. terrestris depend exclusively on the activity of infecting roots and provides circumstantial evidence on the deleterious impact of monoculture on crop production.     

Dumontinia root rot of liver leaf caused by <Emphasis Type="Italic">Dumontinia tuberosa</Emphasis>

Foliar wilt as well as crown and root rot with sclerotia formation has affected potted liver leaf (Hepatica nobilis var. japonica f. magna) in Ojiya, Niigata Prefecture, Japan, since 2006. Apothecia developed from the sclerotia on soil surface of pots with the diseased plants in March. A fungus forming the apothecia was identified as Dumontinia tuberosa (Sclerotiniaceae) based on its morphology and demonstrated to cause the disease. We coined the name “Dumontinia root rot (Dumontinia-negusare-byo in Japanese) of liver leaf” for the new disease.     

Potato wart disease in Newfoundland: a review of current research studies1

Research includes studies on germination of resting spores, substances affecting infection, eradication, bioassay for Synchytrium endobioticum, and sub-surface vectoring of the fungus. The sequence of events in germination has been exposed and the mechanisms of germination are currently being studied. Chitin and in particular crabshell induce zero infection, and there is promise in using these materials as eradicants. Using the bioassay so far developed, it is possible to assay for 0.1 spore per g of soil. There is evidence that the fungus can migrate in soil through the agency of vector animals. Problems long posed by this disease now seem soluble.     

Growth,population dynamics,and diversity of <Emphasis Type="Italic">Fusarium equiseti</Emphasis> in ginseng fields

Fusarium equiseti is prevalent in ginseng soil, straw mulch and in ginseng root tissues and is the cause of a root surface discolouration on ginseng grown in British Columbia. Population levels of the fungus in ginseng fields ranged from 3.8 × 10³ cfu g⁻¹ soil to 1.4 × 10⁴ cfu g⁻¹ soil and were highest at 0–5 cm soil depths compared to 10–15 cm. Soil population levels were negatively correlated with S content in soil and positively correlated with Zn levels. Barley or wheat straw added to soil significantly increased population levels under laboratory conditions. Mycelial growth in culture was highest at 26–30°C and at pH 7.2–7.8. Samples of flowers and berries, and harvested seed, contained DNA of F. equiseti detected using a Fusarium-specific DNA array and the fungus was isolated from these tissues on agar medium. A high degree of genetic variation in the EF-1 alpha gene sequence was present among 52 isolates of F. equiseti which originated from ginseng fields. At least seven clades were identified. Inoculum dispersal from straw mulch used in ginseng gardens can result in seed contamination by the fungus. In addition, fungal growth near the soil surface under warm summer conditions can result in infection and crown discolouration of ginseng roots.     

Discovery of an isolate of Fusarium oxysporum with potential to control Striga hermonthica in Africa

Native fungi of West Africa were evaluated as a means to control Striga hermonthica (Del.) Benth., a troublesome parasitic weed of several gramineous crops. An isolate of Fusarium oxysporum, grown on sorghum straw and incorporated into pots, successfully prevented all emergence of S. hermonthica, and resulted in a 400% increase of sorghum dry weight. The fungus inhibited germination and attachment of S. hermonthica to sorghum roots in Plexiglas root chambers. Several crop species, including sorghum, inoculated with the fungus showed neither any disease symptoms nor any reduction in biomass.