Pathogenicity of Aphanomyces spp. from Different Leguminous Crops in Sweden

Host range and pathogenicity of a range of Aphanomyces spp. isolates obtained from pea roots but also from a range of other field-grown leguminous crops in southern Sweden was investigated. The Aphanomyces euteiches isolates originating from pea and the few obtained isolates originating from alfalfa, green bean and yellow sweet-clover were highly pathogenic only to pea. The A. euteiches isolated from common vetch differed from these isolates by being weakly pathogenic to pea and other legumes, but highly pathogenic to common vetch. Vetch isolates also formed a well-defined separate cluster based on principal component analysis of pathogenicity pattern on tested crops. Oospores of A. euteiches were observed in root tissue of pea as well as common vetch, alfalfa, green bean, broad bean, red clover and yellow sweet-clover in the greenhouse pathogenicity tests. An Aphanomyces sp. that morphologically differed from A. euteiches, was frequently isolated from several leguminous plants, but was non-pathogenic to all tested crops in the pathogenicity tests. In isozyme analysis the banding pattern of these isolates resembled the pattern of A. cladogamus. Another, different and so far unidentified Aphanomyces sp. from roots of green bean and broad bean, was also non-pathogenic to the tested legume species. Based on the isolates tested, the results obtained suggest that the population of Aphanomyces spp. infecting legume roots in Sweden consists of a pea-specific and a vetch-specific group of A. euteiches. Two other groups comprised (i) Aphanomyces sp. isolates that resembled A. cladogamus, and (ii) isolates, which resembled neither A. euteiches nor A. cladogamus. In addition, the host range of Swedish A. euteiches isolates was not as broad as reported for A. euteiches isolates from other countries.     

Population structure,races, and host range of <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis> from alfalfa production fields in the central USA

Aphanomyces euteiches (races 1 and 2) causes root rot of alfalfa; however, its population biology and distribution are poorly understood where alfalfa is a major crop. The objectives of this study were to (1) characterise the distribution and frequency of races of A. euteiches in Illinois alfalfa fields, (2) determine host range of A. euteiches on cultivated and native legumes, and (iii) to describe genetic diversity and population genetic structure of A. euteiches in alfalfa fields. To accomplish this, soil samples (n = 103) were collected from 30 alfalfa fields in 18 Illinois counties. Using the susceptible cv. ‘Saranac’, 148 isolates of A. euteiches were baited from the soil. The virulence phenotype of isolates representing all 18 counties was tested, and 54% were R1 and 46% were R2. Both races were detected in 61% of the counties, whereas only R1 was detected in 22% and R2 in 17%. Thirteen legume hosts for isolates from alfalfa fields were identified based on symptoms and/or production of oospores in roots. In addition to six previously known hosts, seven species were susceptible to infection: kura clover, purple prairie clover, white prairie clover, ladino clover, hairy vetch, Canadian milk vetch, and Illinois tick trefoil. AFLP analysis revealed high levels of genetic diversity among the isolates from different fields and counties and a lack of genetic structuring of populations based on race or geographical origin. The results suggest that populations of A. euteiches in alfalfa fields are diverse, often composed of races 1 and 2, and create risk for alfalfa and to multiple cultivated and native legume species.     

Genetic diversity of the pea root pathogen Aphanomyces euteiches in Europe

The oomycete pathogen Aphanomyces euteiches causes root rot in various legume species. In this study we focused on A. euteiches causing root rot in pea (Pisum sativum), thereby being responsible for severe yield losses in pea production. We aimed to understand the genetic diversity of A. euteiches in Europe, covering a north-to-south gradient spanning from Sweden, Norway and Finland to the UK, France and Italy. A collection of 85 European A. euteiches strains was obtained, all isolated from infected pea roots from commercial vining pea cultivation fields. The strains were genotyped using 22 simple-sequence repeat markers. Multilocus genotypes were compiled and the genetic diversity between individual strains and population structure between countries was analysed. The population comprising strains from Italy was genetically different and did not share ancestry with any other population. Also, strains originating from Finland and the eastern parts of Sweden were found to be significantly different from the other populations, while strains from the rest of Europe were more closely related. A subset of 10 A. euteiches strains from four countries was further phenotyped on two susceptible pea genotypes, as well as on one genotype with partial resistance towards A. euteiches. All strains were pathogenic on all pea genotypes, but with varying levels of disease severity. No correlation between the genetic relatedness of strains and virulence levels was found. In summary, our study identified three genetically distinct groups of A. euteiches in Europe along a north-to-south gradient, indicating local pathogen differentiation.     

Reaction of genotypes from several species of grain and forage legumes to infection with a French pea isolate of the oomycete <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis>

The susceptibility/resistance to Aphanomyces euteiches of various genotypes (cultivars and breeding lines) of several grain legume species was assessed in controlled conditions. A total of 279 genotypes from the major grain legumes grown in temperate climates (faba bean, chickpea, lentil, lupin and common vetch) and three other legumes frequently cultivated in France (French bean, clover and alfalfa) were screened with one pea-infecting isolate from France. Four different categories of susceptibility/resistance were identified among the legume species/cultivars tested with the pea A. euteiches isolate: (1) susceptible legume species (lentil, alfalfa, French bean) among which low levels of partial resistance was observed; (2) legume species including susceptible genotypes and genotypes with high levels of resistance (common vetch, faba bean and clover), (3) species with a very high level of resistance (chickpea) and (4) species displaying no symptoms (lupin). It is therefore important to consider pathogen-species and pathogen-genotype interactions when defining the host specificity of A. euteiches and considering the possible role of different legume species in increasing or decreasing the soil inoculum potential.     

Effects of hydrostatic pressure,agitation and CO2 stress on Phytophthora nicotianae zoospore survival

BACKGROUND: Phytophthora nicotianae Breda de Haan is a common pathogen of ornamental plants in recycled irrigation systems. In a previous study, annual vinca (Catharanthus roseus Don) inoculated with zoospore suspensions using a CO₂‐pressurized sprayer had less foliage blight than plants inoculated using a hand sprayer. Here, the impact of hydrostatic pressure, agitation and aeration with CO₂ on the survival of P. nicotianae zoospores was examined. RESULTS: Exposure of zoospores to 840 kPa hydrostatic pressure for 8 min or agitation at a mixing intensity (G) of 6483 s^?1 for 4 min at 22–23 °C did not kill zoospores, but resulted in viable cysts. Motile and forcefully encysted zoospores of P. nicotianae were equally infectious on vinca or lupine (Lupinus polyphylus Lindl.). Bubbling CO₂ into zoospore‐infested water at 110.4 mL (0.2 g) min^?1 for 5 min caused 81% reduction in the number of germinated zoospores. Pressure at 630 kPa (16.3 g CO₂) or 70 kPa (3.85 g CO₂) facilitated CO₂ injection and shortened the zoospore inactivation time to 30 s. When air was bubbled through the suspension, germination was similar to the control. CONCLUSIONS: Exposure to CO₂ killed P. nicotianae zoospores in water. Neither pressure nor agitation had an effect on zoospore viability or infectivity. Based on results of this study, the authors designed a recycling CO₂ water treatment system that is currently under evaluation. Copyright © 2010 Society of Chemical Industry     

Testing of an integrated regime for effective and sustainable control of invasive Crofton weed (Ageratina adenophora) comprising the use of natural inhibitor species,activated charcoal,and fungicide

Crofton weed is a major invasive species in China. It exhibits superior growth characteristics and can outcompete with native species via allolepathic effects and modulation of the soil fungal microbiome. The simple removal of invading plants will not ensure restoration of the habitat due to the persistence of allelochemicals and viable seeds in the surrounding soil. An orthogonal experimental design was employed to evaluate the effects of three control factors (A, powdered natural inhibitor species to retard growth; B, activated charcoal to absorb allelochemicals; and C, fungicide to reduce fungal modulation effects), applied at three levels, on the growth and competitive ability of Crofton weed against two native species, in a pot‐culture experiment. All treatments reduced all measured growth parameters (P < 0.05) except for a specific leaf area, when compared with control plants. Furthermore, the competitive capacity of Crofton weed was decreased in the treatments while that of the native species was improved. Application to soil of the powdered natural inhibitor species and of activated charcoal significantly inhibited plant growth and competitive ability of Crofton weed (P < 0.05). Application of fungicide was less effective, but significantly reduced the specific leaf area of Crofton weed plants (P < 0.05). The specific combination of factors producing the greatest decrease in plant growth and competitive ability (compared with the control) included the addition of Delavaya toxocarpa powder (37.5 g per kg soil), addition of activated charcoal to soil at a ratio of 1:3 (v/v) (62.5 g per kg soil), and application of fungicide (Thiophanate‐Methyl) (0.28 g per kg soil).     

Successive legumes tested in a greenhouse crop rotation experiment modify the inoculum potential of soils naturally infested by Aphanomyces euteiches

The consequence of 10 successive monocultural cycles involving different legume species/cultivars on the inoculum potential (IP) of soils naturally infested by Aphanomyces euteiches was investigated under greenhouse conditions. The results showed that the IP of a soil naturally infested by A. euteiches can be significantly modified not only by the non‐host or host status of crop species but also by the level of resistance of the cultivar. Susceptible species/cultivars (pea, lentil and susceptible cultivars of vetch and faba bean) are very favourable to pathogen multiplication, and continuous cultivation of each of these increased the IP values of a soil with a moderate initial IP (from 1·9 to 3·5 after 10 cycles). Conversely, non‐host species and resistant cultivars of vetch or faba bean contributed to reducing the IP values of soils irrespective of the initial IP (from 1·9 to 0·5 and from 4 to 2, respectively, after 10 cycles). Aphanomyces root rot severity values on the resistant legume species/cultivars were not affected by the successive cultural cycles. This study, which showed that the IP of A. euteiches in soil can be reduced by planting appropriate legume species and cultivars in greenhouse conditions, will be useful for defining better crop successions for legumes.     

Activity of the New Fungicide Benthiavalicarb Against Plasmopara viticola and its Efficacy in Controlling Downy Mildew in Grapevines

Benthiavalicarb is a new fungicide active against Oomycetes fungal plant pathogens. The present study shows that benthiavalicarb is effective for controlling the Oomycete fungal pathogen Plasmopara viticola, which causes downy mildew in grapevines. The fungicide did not affect zoospore discharge from sporangia of P. viticola, but strongly inhibited zoospore encystment, cystospore germination in vitro and mycelial growth, together with sporangial production in vivo. Benthiavalicarb showed strong prophylactic and local activity in intact plants or detached leaves and low translaminar activity. The compound was not translocated from leaf to leaf in either a acropetal or basipetal direction. Benthiavalicarb applied at 1, 3 and 6 days post-inoculation protected grapevine plants against downy mildew and inhibited sporulation of the pathogen. Similar results were obtained on leaf disks if benthiavalicarb was applied up to 96 h post-inoculation. Benthiavalicarb diminished the sporulation of P. viticola when applied to established disease in the tissue. Benthiavalicarb remained active on leaves for a period up to 28 days. Two foliar applications of benthiavalicarb, 2 weeks apart, to field-grown grapevines inhibited downy mildew development and were as effective as the standard metalaxyl-Cu treatment in controlling the disease. A formulated mixture of benthiavalicarb + Folpet was similar or superior in performance to metalaxyl-Cu and the new strobilurin trifloxystrobin in controlling downy mildew. The effectiveness of benthiavalicarb makes it well suited for integration into a control programme against downy mildew disease in vineyards, and as a component to delay resistance buildup.     

Selection for decreased sensitivity to phosphite in Phytophthora cinnamomi with prolonged use of fungicide

To test the hypothesis that resistance in Phytophthora cinnamomi to control by the fungicide phosphite (phosphonate) would arise in sites with prolonged use of phosphite, 30 P. cinnamomi isolates were collected from a range of sites with different phosphite‐use histories, including phosphite‐treated and untreated avocado orchards, and phosphite‐treated and untreated native vegetation sites. The colonizing ability of these isolates was tested by different inoculation methods against a range of host tissues, treated and untreated with phosphite, including mycelial stem inoculation on clonally propagated Leucadendron sp., mycelial root inoculation of lupin seedlings and zoospore inoculation of Eucalyptus sieberi cotyledons. Isolates from avocado orchards with a long history of phosphite use were, on average, more extensive colonizers of the phosphite‐treated Leucadendron sp., lupin seedling roots and Eucalyptus sieberi cotyledons. These isolates did not colonize untreated plant tissue (Leucadendron sp.) more extensively than isolates from sites with no history of phosphite use and no isolates were resistant to control by phosphite. Analysis of all isolates with microsatellite markers revealed the majority were from a single clonal lineage. Selection for decreased sensitivity to phosphite in planta has taken place within asexual clonal lineages of P. cinnamomi in sites with prolonged use of phosphite.     

Further Contributions to the Development of a Differential Set of Pea Cultivars (Pisum sativum) to Investigate the Virulence of Isolates of Aphanomyces euteiches

Common root rot (Aphanomyces euteiches Drechs.) has become a very destructive disease in the French pea crops since 1993. For an accurate investigation of the virulence variability among French A. euteiches populations and between French and foreign populations, a new set of differential pea genotypes was developed. Thirty-three American and European pea lines, displaying different levels of resistance, were screened in a growth chamber against two French isolates. Symptoms (disease severity from 0 to 5, evaluating symptom surface on roots and epicotyl) and percentage of top fresh weight (inoculated/uninoculated top fresh weight ratio) were measured. From this screening 12 relatively resistant lines, from various genetic backgrounds, were identified along with a highly susceptible control. This set of 13 genotypes was inoculated under controlled conditions with 14 isolates from France, Sweden, USA, Canada and New Zealand, to investigate genotype–isolate interactions. Root symptoms were rated (disease severity), and a susceptibility/resistance threshold was established at disease severity = 1. Significant quantitative interactions were observed, and five 'resistance patterns' were identified, leading to a set of six pea genotypes: Baccara (susceptible), Capella, MN313, 902131, 552 and PI180693. Fields trials of this set in 1999 and 2000 gave the same resistance rankings than in growth chamber conditions. This set will allow more accurate assessments of the variability in virulence/aggressiveness of A. euteiches isolates from France and foreign countries, and further investigations of the epidemiological and genetic basis of pea–A. euteiches interactions.     

Plant extracts containing caffeic acid and rosmarinic acid inhibit zoospore germination of Phytophthora spp. pathogenic to Theobroma cacao

The three most important species of Phytophthora worldwide causing black pod disease of cacao are P. palmivora, P. megakarya, and P. capsici. Chemicals are effective in controlling this disease but more natural methods would be preferred. One alternative is to use natural plant extracts. Rosemary and lavender leaf extracts were found to be effective in reducing germination of P. capsici, P. megakarya, and P. palmivora zoospores when supplemented to agar plates at different dilutions. The extracts displayed the biggest impact on P. megakarya zoospores where it completely inhibited germination at a 25% dilution of the prepared extract. When applied to cacao leaf disks, rosemary extract reduced necrosis caused by P. megakarya zoospores. In a bioassay, pears first treated with lavender extract showed no symptoms of P. megakarya infection compared with the non-treated controls. Based upon HPLC analyses, the active compound in these extracts was determined to be caffeic acid, rosmarinic acid or some simple derivative thereof. When added to agar plates, synthetic caffeic acid and rosmarinic acid completely inhibited germination of P. capsici, P. megakarya, and P. palmivora zoospores at concentrations of 3 and 6 g l⁻¹, respectively. In addition, sage and rice bran extracts, which both contain caffeic acid, were also effective in reducing zoospore germination. In contrast, inhibition of Botrytis cinerea or Trichoderma asperellum conidia germination did not occur, displaying some level of specificity. These extracts could provide an economically safe method for reducing damage caused by black pod disease on cacao until resistant varieties are developed and released.     

Variability in Virulence Among Asexual Progenies of Phytophthora infestans

ABSTRACT One hundred two single zoospore isolates of Phytophthora infestans, derived asexually from four parental isolates of US-8 genotype and one isolate of US-1 genotype, were characterized for their virulence phenotypes to determine changes in virulence during asexual reproduction. Potato differentials, each containing a major gene for resistance to P. infestans (R1 to R11), were used to characterize the virulence patterns. Isolates were also characterized for mating type, glucose-6-phosphate isomerase (Gpi) banding pattern, and DNA fingerprints using probe RG57 to determine any genotypic changes in the single zoospore isolates. A subset of these single zoospore isolates was tested for response to mefenoxam to determine any shifts in sensitivity. Results showed that single zoospore isolates derived from parent PI-1 (US-8, 11 isolates) were identical to their parental virulence. Isolates derived from parent PI-191 (US-8, 29 isolates) showed some differences in virulence, mainly toward R8 and R9. Isolates derived from parent PI-126 (US-8, 14 isolates) demonstrated a higher level of virulence diversity. Isolates derived from parents PI-52 (US-1, 28 isolates) and PI-105 (US-8, 20 isolates) showed the highest level of virulence variability among the single zoospore isolates. Mating type, Gpi banding pattern, and DNA fingerprints for the single zoospore isolates were, in most cases, identical to the parental isolates. Single zoospore isolates showed different levels of sensitivity to mefenoxam. Virulence and other genetic changes during asexual reproduction are likely to play a major role in changing the race structure of P. infestans populations. This continuous change in the race structure is a serious problem and now poses a new challenge for utilization of race-specific resistance to manage late blight of potato.     

β-Tubulins in Gibberella zeae: their characterization and contribution to carbendazim resistance

BACKGROUND: Fusarium head blight caused by Gibberella zeae is an important disease of wheat and barley because it reduces grain yield and quality and results in the contamination of grain with mycotoxins. Recent studies have shown that carbendazim resistance in field strains of G. zeae is not caused by mutation of the β‐tubulin gene (β₁tub), which is the case with other filamentous fungi, but that fungicide resistance is greatly increased by deletion of β₁tub. The aim of the present study was to clarify the function of β₁tub and its role in carbendazim resistance in G. zeae by artificial gene operation. RESULTS: Deletion of β₁tub reduced vegetative growth and pathogenicity but increased asexual reproduction in G. zeae. All the mutants were more resistant to carbendazim than parent strains. A three‐dimensional model of β₁tub was constructed, and the possible carbendazim binding site was analysed. CONCLUSION: β₁tub is not an essential gene in G. zeae, but it affects the sensitivity of the fungus to carbendazim. Copyright © 2012 Society of Chemical Industry     

Aphanomyces euteiches as a component of the complex of foot and root pathogens of peas in Dutch soils

The occurrence ofAphanomyces euteiches Drechs. in Dutch soils is reported for the first time. Isolates of the pathogen were obtained from peas (Pisum sativum L.). A bioassay was used that baited the pathogen from soil into the cortex of stem and root of seedlings of a highly susceptible pea cultivar. The pathogen could subsequently be isolated on a semi-selective medium. Screening of soil samples from 13 fields known to be infested with fungi causing foot and root rot demonstrated the presence ofA. euteiches in 10 cases. In a second screening on soil samples from 43 fields, the pathogen was present in 16 cases. A positive correlation was found between the disease severity caused byA. euteiches in the seedling bioassay and the disease severity caused by the complex of foot and root pathogens in the same soils as evidenced by a mature plant bioassay. It is considered probable thatA. euteiches has since long been a common component of the foot and root rot complex in Dutch soils but has not been detected previously due to inadequate sampling and isolation techniques.Samenvatting De aanwezigheid vanAphanomyces euteiches Drechs. in Nederlandse gronden is voor het eerst aangetoond. Isolaten van het pathogeen werden verkregen van erwten (Pisum sativum L.). De pathogene schimmel werd in petrischalen uit grond in het schorsweefsel van wortel en stengel van een zeer vatbaar erwteras gelokt. Met behulp van een semiselectief medium konden vervolgens isolaten van de schimmel worden verkregen. Toetsing van grondmonsters afkomstig van 13 percelen, waarvan bekend was dat ze besmet waren met schimmels die voetziekten in erwten veroorzaken, toonde de aanwezigheid vanA. euteiches aan in 10 gevallen. In een tweede biotoets op grondmonsters van 43 percelen bleken 16 monsters het pathogeen te herbergen. Er werd een positieve correlatie gevonden tussen de ernst van de aantasting doorA. euteiches van kiemplanten en de aantasting van volwassen planten in een biotoets in de kas. Het is waarschijnlijk dat de schimmel reeds lang in Nederlandse akkers voorkomt, maar door inadequate bemonsterings- en isolatietechnieken over het hoofd is gezien.     

Effect of fungicides on the complex of <Emphasis Type="Italic">Fusarium</Emphasis> species and saprophytic fungi colonizing wheat kernels

Fusarium head blight of wheat, often associated with mycotoxin contamination of food and feed is caused by various Fusarium species. The efficacy of fungicide sprays for the control of the disease and mycotoxins varies from being highly effective to even increasing mycotoxin levels. The potential role of antagonistic fungi in this variability was investigated assessing sensitivity of Fusarium species and saprophytic fungi colonizing wheat kernels to fungicides. Saprophytes were tested for their antagonistic activity to the prevalent Fusarium species Fusarium avenaceum, Fusarium culmorum, Fusarium graminearum, and Fusarium poae. Fungal isolates from mature winter wheat kernels were Alternaria alternata, Arthrinium sp., Aspergillus niger, Epicoccum sp., Microdochium spp., Rhizopus oryzae and Trichoderma sp. In dual culture A. niger, R. oryzae and Trichoderma hamatum were more effective in reducing mycelial growth of Fusarium species than Microdochium majus; A. alternata and Epicoccum sp. were ineffective because of slow growth rates. Saprophytic fungi were sensitive to triazoles; however, prothioconazole and tebuconazole had stronger effects on mycelial growth of Fusarium species. ED₅₀ values also indicated significant differences in the sensitivity of Fusarium species to triazoles (range 0.1–1.7 mg l⁻¹). Azoxystrobin and fluoxastrobin were largely ineffective in inhibiting in vitro growth of Fusarium spp.; sensitivity of the other fungi was generally lower, except for M. majus which was highly sensitive. Due to differences in fungicide sensitivity among Fusarium spp. and ear-colonizing fungi antagonistic to Fusarium spp. fungicides are likely to modify the balance within the mycoflora of wheat ears which may also affect the mycotoxin contamination of grain.     

Effects of fungicides on in vitro spore germination and mycelial growth of the phytopathogens Leptosphaeria maculans and L. biglobosa (phoma stem canker of oilseed rape)

BACKGROUND: Phoma stem canker, caused by the coexisting related fungal pathogens Leptosphaeria maculans (Des.) Ces. & de Not and L. biglobosa Shoemaker & H Brun, is a major disease of winter oilseed rape in the UK. Annually, over 90% of UK crops receive at least one foliar application of fungicide, but little is known about the sensitivity of the more damaging L. maculans and the less damaging L. biglobosa to these fungicides. The effects of flusilazole, tebuconazole and Methyl Benzimidazole Carbamate (MBC) fungicides (benomyl and carbendazim) on the germination of ascospores, conidia and germ tube growth of both species were examined. Isolates collected from different oilseed rape crops in England and Wales were assessed for their mycelial growth on fungicide‐amended medium, and ED₅₀ values were calculated. RESULTS: Leptosphaeria maculans and L. biglobosa differed in their sensitivity to fungicides. Conidial germination of L. maculans was more sensitive to these fungicides than that of L. biglobosa. Isolates of L. maculans had smaller ED₅₀ values for mycelial growth for all fungicides tested than isolates of L. biglobosa. CONCLUSION: These results suggest that fungicide applications might affect the structure of L. maculans/L. biglobosa populations in UK winter oilseed rape crops. Copyright © 2009 Society of Chemical Industry     

Extracellular self-DNA plays a role as a damage-associated molecular pattern (DAMP) delaying zoospore germination rate and inducing stress-related responses in Phytophthora capsici

Phytophthora capsici is a highly destructive pathogen of crops. Although chemical pesticides are the most widely used strategy to counter phytopathogens, they have been inefficient to combat P. capsici and have produced significant environmental and health problems. Therefore, sustainable alternatives to control soilborne pathogens, such as the inhibitory effect of self-extracellular DNA (eDNA), have been proposed. This inhibition phenomenon has been attributed to the action of self-eDNA as a damage-associated molecular pattern (DAMP). Here, we describe the effect of self-eDNA on P. capsici zoospore germination rate, antioxidant enzymes activity and MAPK gene expression. Also, the effect of P. capsici eDNA on the protection of chilli pepper (Capsicum annuum) plants against P. capsici was investigated. The results highlight that P. capsici can sense 2–500 µg/ml self-eDNA and induce stress-related responses like SAK1 gene expression, and superoxide dismutase and catalase activities. Moreover, in vitro zoospore germination rate was suppressed with self-eDNA concentrations ranging from 50 to 500 µg/ml. Interestingly, drench applications of P. capsici eDNA at 60 and 100 µg/ml on chilli pepper plants did not show any protective effect against the phytopathogen, whereas 2 µg/ml of P. capsici eDNA drench application showed a lower percentage of plants with symptoms and lower disease severity. Moreover, phenols and total flavonoids were increased in chilli pepper plants, therefore inducing plant immunity. This study showed that self-eDNA acts as a DAMP in P. capsici and provides insight into the use of eDNA for the protection of crops of agronomic interest.     

Management strategies and distribution of Aphanomyces root rot of alfalfa (Medicago sativa), a continuing threat to forage production in the United States

Alfalfa (Medicago sativa) is one of several legumes that is affected by Aphanomyces root rot (ARR) caused by Aphanomyces euteiches. Symptoms of ARR on alfalfa seedlings include a yellow-grey discolouration of roots, rotting and loss of lateral roots, stunted growth, chlorotic foliage and reduction of nitrogen-producing nodules on roots. Infection can also occur on adult plants leading to loss of lateral roots and nodules. At the seedling stage, ARR decreases alfalfa stand establishment, and field longevity is reduced when adult plants are infected. A. euteiches is an oomycete pathogen that has motile zoospores and thick-walled oospores that can survive for many years in soil. Two races are currently recognized by pathogenicity on differential alfalfa check cultivars. Most alfalfa cultivars contain race 1 resistance, but there is an increasing development of cultivars with resistance to race 2. Management strategies include planting resistant cultivars, avoiding planting in fields with poor drainage and rotating crops with nonhost plants.     

Cross-Resistance Patterns Among Sterol Biosynthesis Inhibiting Fungicides (SBIs) in Cercospora beticola

Thirty single-spore isolates of Cercospora beticola, collected from several fields in northern Greece, representing a broad spectrum sensitivity to the sterol demethylation-inhibiting (DMIs) fungicide flutriafol, were tested for sensitivity to eleven other sterol biosynthesis-inhibiting (SBI) fungicides and to the guanidine fungicide dodine. Sensitivity was measured as EC₅₀ values for each fungicide and log-transformed EC₅₀ values to each fungicide were pairwise correlated and the correlation coefficient estimated. These pairwise comparisons showed high correlation coefficients between the DMIs suggesting a cross-resistance relationship between these fungicides. However, the degree of cross-resistance between DMIs varied greatly. Conversely, low correlation coefficients were obtained for the pair-wise comparisons with the morpholine fungicide fenpropimorph suggesting a lack of cross-resistance between morpholines and DMIs in C. beticola. Similarly, there was no correlation between the sensitivity (EC₅₀ values) to dodine and all the other fungicides tested, indicating that there was no negative cross-resistance relationship between dodine and SBIs in C. beticola. Based on these results, combinations or alternations of fungicides which show no cross-resistance relationship should be used to control the disease in areas where reduced sensitivity to DMIs has been already observed.     

Hydrogen peroxide scavenging mechanisms are components of Medicago truncatula partial resistance to Aphanomyces euteiches

The biochemical processes underlying the expression of resistance in the roots of Medicago truncatula against Aphanomyces euteiches infection was investigated, with emphasis on oxidative stress. The levels of H₂O₂, superoxide dismutase, peroxidase, ascorbate peroxidase, catalase, soluble phenolics and lignin were measured in the roots of two lines, A17 partially resistant and F83005.5 susceptible to A. euteiches at three infection stages; penetration of the epidermis (1 dpi), colonization of the cortex (3 dpi) and invasion of the root stele (6 dpi). A rapid and large decrease of the H₂O₂ levels in A17 roots occurred. However, in F83005.5 roots, the decrease in H₂O₂ levels was delayed until 3 dpi. In A17 roots, the activities of ascorbate peroxidase, peroxidase and catalase were induced as early as 1 dpi, whereas a general decrease in the activity of the four antioxidant enzymes was observed in F83005.5 roots. The levels of soluble phenolics and lignin were increased in A17 roots at 3 and 6 dpi, respectively. The H₂O₂ levels were negatively correlated to ascorbate peroxidase, catalase and lignin production at 1, 3 and 6 dpi, respectively in A17 roots. Physiological concentrations of H₂O₂ found in M. truncatula infected roots had no detrimental effect on the in vitro growth of this oomycete. Our data suggest that H₂O₂ does not have a direct antimicrobial effect on M. truncatula resistance to A. euteiches, but is involved in cell wall strengthening around the root stele, preventing pathogen invasion of the vascular tissues.