Relation between cropping frequency of peas and other legumes and foot and root rot in peas

The relation between the frequency of legume crops in a rotation and the root rot severity in pea was examined in a field survey. Additionally, greenhouse experiments were performed with soil samples from legume rotation trials or from farmers' fields. The frequency of pea crops in current rotations proved to be much less than the recommended value of one in six years. The correlation between pea root rot and the number of years that pea or other legumes were not grown on the field under consideration (called crop interval) was weak. Root rot severity correlated better with the frequency of peas or legumes in general over a period of 18 years, but the frequency still explained only a minor fraction of the variation in disease index. Some experimental data pointed to the occurrence of a highly specific pathogen microflora with continuous cropping of only one legume species, but this phenomenon probably does not occur in farmers' fields. In field samples, root disease index for pea correlated well with that for field bean. The survival of resting structures of pathogens such asAphanomyces euteiches probably explains why the frequency of legume cropping has a higher impact than crop interval on root disease incidence. Pea-free periods and legume frequencies have a poor predictive value for crop management purposes.     

Host status and reaction of <Emphasis Type="Italic">Medicago truncatula</Emphasis> accessions to infection by three major pathogens of pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>) and alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis>)

Ditylenchus dipsaci, the stem nematode of alfalfa (Medicago sativa), Mycosphaerella pinodes, cause of Ascochyta blight in pea (Pisum sativum) and Aphanomyces euteiches, cause of pea root rot, result in major yield losses in French alfalfa and pea crops. These diseases are difficult to control and the partial resistances currently available are not effective enough. Medicago truncatula, the barrel medic, is the legume model for genetic studies, which should lead to the identification and characterization of new resistance genes for pathogens. We evaluated a collection of 34 accessions of M. truncatula and nine accessions from three other species (two from M. italica, six from M. littoralis and one from M. polymorpha) for resistance to these three major diseases. We developed screening tests, including standard host references, for each pathogen. Most of the accessions tested were resistant to D. dipsaci, with only three accessions classified as susceptible. A very high level of resistance to M. pinodes was observed among the accessions, none of which was susceptible to this pathogen. Conversely, a high level of variation, from resistant to susceptible accessions, was identified in response to infection by A. euteiches.     

Antagonistic rhizoplane bacteria induce diverse morphological alterations in Peronosporomycete hyphae during <Emphasis Type="Italic">in vitro</Emphasis> interaction

A total of 150 bacteria were isolated from rhizoplanes of the host and non-host plants of a phytopathogenic Peronosporomycete Aphanomyces cochlioides. Upon screening, 5% of the isolates were evaluated as antagonists as they inhibited radial growth of A. cochlioides AC-5 hyphae in a dual culture assay. In addition, those antagonistic bacteria also induced characteristic morphological alterations in the A. cochlioides AC-5 hyphae that grew towards bacterial colonies. Hyphal morphological alterations observed in AC-5 and other tested strains of Peronosporomycetes included excessive branching, curly growth, unusually longer and pointed tip formation and swelling; all of these were comparable to the alterations induced by known antimicrobial compounds. Among the antagonistic bacteria, Pseudomonas sp. strain EC-S101 induced a unique branching pattern (tree-like) in AC-5 hyphae by continuous apical bifurcation of successive hyphae, where increases in number of branches and hyphal area were linearly correlated with time up to 10 h. Our observations suggested that the pathogen might have lost its ability of normal branch production; however maintained the capability of self-branching. Soluble extracts from the culture fluids of Pseudomonas sp. strain EC-S101 and Stenotrophomonas maltophilia EC-S105 induced similar excessive branching and curly growth in A. cochlioides hyphae as the respective bacterium. These results revealed that bacterial metabolites appeared to be responsible for induction of morphological alterations. Interestingly, the antagonistic bacteria that induced hyphal morphological alterations, also efficiently suppressed in vivo damping-off disease caused by AC-5. We suggest that antagonistic rhizoplane bacteria have the capability to induce diverse morphological alterations in Peronosporomycetes hyphae during in vitro interactions. Hyphal morphological alterations associated with growth inhibition and the induction of characteristic morphological changes indicate antagonistic activity against the Peronosporomycete.     

Detection of genomic DNA of the crayfish plague fungus Aphanomyces astaci (Oomycete) in clinical samples by PCR

A diagnostic procedure, based on a polymerase chain reaction method (PCR) was developed to detect infection of crayfish with the Oomycete Aphanomyces astaci. A set of oligonucleotide primers was designed to specifically amplify A. astaci DNA in the ITS region surrounding the 5.8S rDNA gene. The PCR amplifies a 115 bp amplicon. The specificity of the primers was demonstrated by testing on 27 A. astaci strains and against 20 non-A. astaci Oomycetes and 5 fungal species. Most of the non-A. astaci Oomycete or fungal species included in the study are either known parasites of freshwater crayfish cuticle or can be found in their natural environment. Specificity was also tested against crayfish tissue and some known parasites and bacteria infecting crayfish.

A protocol for the extraction of A. astaci DNA from infected crayfish tissue was developed. The optimised method allows the detection of two genome equivalents of purified A. astaci genomic DNA.

The method was tested on noble crayfish (Astacus astacus), artificially infected with A. astaci. Detection of A. astaci was possible at the very first time of sampling, which was 2 days after the beginning of spore exposure.     

Detection of Aphanomyces invadans and epizootic ulcerative syndrome in the Murray‐Darling drainage

Epizootic ulcerative syndrome was diagnosed, and the presence of Aphanomyces invadans confirmed, from an outbreak of clinical disease in wild‐caught bony bream (Nematalosa erebi) from the Darling River near Bourke, in New South Wales, Australia, during 2008. This confirms a significant extension of the agent beyond its historical range.     

First record of epizootic ulcerative syndrome from the Upper Congo catchment: An outbreak in the Bangweulu swamps,Zambia

We report on the first outbreak of epizootic ulcerative syndrome (EUS) amongst wild fish populations in the Bangweulu swamps, an inland delta, in the north of Zambia during 2014. The area supports a large and diverse fish fauna related to, but distinct from, that of the Zambezi River system where EUS outbreaks have occurred since 2006. A sizeable artisanal fishery, based on extensive fish weirs, is sustained by the annual flooding of the swamps, and observations of the disease outbreak by fishermen were recorded. Signs typical of infection with Aphanomyces invadans were observed in a number of species. Clinical observations, histology and molecular diagnostic methods were used to confirm infection with A. invadans in two of the most commonly and severely affected species. Several features of the wetland may have contributed to the outbreak and the annual recurrence of the disease. Modes by which the disease may have been introduced into the swamps are discussed. The outbreak is of great significance as the Bangweulu swamps drain into the Congo River in neighbouring Democratic Republic of Congo, Africa's largest drainage system with an extensive and diverse fish fauna previously unaffected by EUS.     

Critical factors driving aphanomyces damping‐off and root disease in clover revealed and explained using linear and generalized linear models and boosted regression trees

Subterranean clover (Trifolium subterraneum) is an important forage legume in Mediterranean regions worldwide. Aphanomyces damping‐off and root disease (Aphanomyces trifolii) poses significant threat to its persistence and productivity. Studies were conducted to define how environmental explanatory variables (temperature, soil type, moisture, nutrition) and variety influence disease severity and consequent forage productivity and persistence. Relationships were modelled using linear and generalized linear models and boosted regression trees. Linear modelling highlighted complex relationships between environmental variables and each dependent variable (emergence, tap and lateral root disease, dry shoot and root weight). All environmental variables produced significant interaction and/or main effects within each dependent variable. Boosted regression trees supported the complex nature of relationships in linear models, with temperature and either soil or variety most, and nutrition least, influential. Heat maps showed more disease for low temperatures. Least tap root disease was under high temperatures, while least lateral root disease was under medium or high temperatures, low moisture, and in sand‐based soil. These are the first studies using modelling approaches to reveal the complexities of how fluctuating soil temperature, moisture and nutrition conditions, and soil type and variety, determine aphanomyces damping‐off and root disease severity and resultant adverse impacts on forage legume productivity and persistence. Outcomes are widely applicable across soilborne oomycete pathogens of forage legumes. Studies highlighted how warming temperatures and drying climate associated with climate change should reduce future impact and importance of this and other soilborne oomycete diseases of forage legumes favoured by cold temperatures and wet and waterlogged conditions.     

Variation in the hyphal growth rate and the virulence of two genotypes of the crayfish plague organism Aphanomyces astaci

Crayfish plague, a devastating disease of freshwater crayfish, is caused by an oomycete organism, Aphanomyces astaci. Currently five genotypes of A. astaci are known, but variable features between the strains or genotypes have not been studied extensively. This study analysed 28 isolates of the As genotype and 25 isolates of the Ps1 genotype and reveals that the radial growth rate is significantly (P < 0.001) different between these two genotypes, although highly variable inside the genotype As. Two Ps1 genotype isolates and two As genotype isolates with different radial growth rates were tested in an infection trial. Clear differences were detected in the development of mortality in the test groups. The representatives of the Ps1 genotype caused total mortality within a short time span. The As genotype isolates were much less virulent. The slow‐growing As isolate showed higher virulence than the As isolate with a high growth capacity. Although slow growth could be one survival strategy of the pathogen, several other mechanisms are involved in the pathogenicity and warrant further studies.     

Host resistances to Aphanomyces trifolii root rot of subterranean clover: first opportunity to successfully manage this severe pasture disease

Subterranean clover (Trifolium subterraneum) is an important pasture legume in Australia (29 million ha) and elsewhere. However, severe pasture decline occurs in association with several root pathogens, including Aphanomyces trifolii, that has been misidentified for decades as A. euteiches until recently confirmed as A. trifolii. A series of controlled environment experiments was undertaken to identify host resistance to A. trifolii in subterranean clover and to compare virulence and phylogeny of isolates. In experiment 1, Dalkeith, Bacchus Marsh, Riverina and Yarloop were the most resistant of 38 cultivars with a percentage disease index (PDI) ≤10 for both tap and lateral roots. Experiment 2 confirmed resistance of Yarloop, but a change in some relative varietal resistances suggested physiological specialization among A. trifolii isolates. Experiment 3 confirmed extensive variation in virulence and physiological specialization across 23 isolates of A. trifolii, with three distinct clades, two of which were distinct from isolates collected previously. Experiment 4 identified host resistance(s) effective against a mixture of 20 A. trifolii isolates, but the most resistant cultivars (Antas, Uniwager, Leura) still showed significant disease. This is the first study to show physiological specialization in A. trifolii and to identify host resistance. This study defines A. trifolii as a significant but largely unknown contributor to severe root disease of subterranean clover in southern Australia. Finally, development and calibration of a new soil commercial DNA test not only enables field quantification of the disease, but development of appropriate breeding, selection and farm management strategies to reduce its impact.     

Plant secondary metabolites regulating behaviour of zoospores of the phytopathogenic fungus Aphanomyces cochlioides

A number of compounds isolated from various plant species were tested for their ability to affect the mobility of zoospores of the fungus Aphanomyces cochlioides which causes root rot in spinach (Spinacia oleracea). Compounds may act as attractants, repellents or stimulants of zoosphore movement or they may halt movement by causing the spore to clump and settle. Bioassay revealed compounds with these methods of action, as well as some which acted directly on the fungus. ©1999 Society of Chemical Industry