Aggressiveness of Fusarium oxysporum and F. verticillioides isolates on stone and scots pine under greenhouse conditions

Scots pine (Pinus sylvestris) and Stone pine (Pinus pinea) are two important species used in re-forestation that are subject to damage by damping-off fungi in forest nurseries. Twenty-two isolates of Fusarium oxysporum and F. verticillioides from diseased seedlings of eight different hosts were tested for aggressiveness on seeds and seedlings of both pine species, including the effects on seedling emergence and mortality. Scots pine was more susceptible to damping-off than Stone pine, as indicated both by reduced seedling emergence and elevated seedling mortality. The impact of F. oxysporum and F. verticillioides on seeds and seedlings did not differ significantly for either pine species. Our findings support previous studies that found that these are damping-off pathogens on the studied pines. Whereas most isolates proved to be pathogenic, some isolates of both Fusarium species showed to be non-pathogenic.     

Influence of light on the Monilinia laxa–stone fruit interaction

Light represents a signal for the regulation of virulence in many microbial pathogens. Two stone fruits, nectarines and cherries, were used to investigate the influence of light on brown rot caused by Monilinia laxa. Three single-spore isolates were inoculated on nectarines and incubated under different white lights, red light, blue light, green light, and black light with two photoperiods. In addition, to understand the effect of daylight irradiance on brown rot, M. laxa was inoculated on different cherry cultivars and incubated under two simulated solar irradiations. Significantly higher disease severity and sporulation were reported on inoculated nectarines incubated under 58 W white light for 12 hr light/12 hr darkness than on nectarines incubated in continuous darkness. Only red light caused a significant increase in the incidence and severity of the disease in nectarines inoculated with the three M. laxa isolates, compared to fruit incubated under white light. High light intensity (185.45 W/m²), caused greater brown rot severity in all cherry cultivars, both early and late varieties, than low irradiance (145.85 W/m²). Significant up-regulation of the pathogenicity-related MlPNL2 gene was observed as an early response after cherry inoculation under high-intensity light, especially in late cherry cultivars, while MlPG1 expression did not show any changes under different light irradiances. M. laxa was shown to be a light-responsive fungal pathogen, and light seemed to play an active role in brown rot development.     

Parasitism of macroconidia, chlamydospores and hyphae of Fusarium culmorum by mycoparasitic Pythium species

Parasitism of macroconidia and endoconidial chlamydospores of Fusarium culmorum by Pythium oligandrum was studied on water agar (WA), corn-meal agar (CMA) and glass slides. Loss of cytoplasmic content in F. culmorum spores was followed by complete degradation, and P. oligandrum produced an abundance of oogonia on the parasitized macroconidia. A simple method for assessing the relative aggressiveness of isolates is presented, based on the percentage of macroconidial cells devoid of cytoplasm. Parasitism of macroconidia by P. acanthophoron , P. oligandrum and P. periplocum , but not by the plant pathogenic species, P. tracheiphilum , was demonstrated by this method. Interactions between hyphae of P. oligandrum and F. culmorum on WA resulted in an increase in the number of oogonia of P. oligandrum and a decrease in the sporulation of F. culmorum . The ability of isolates of P. oligandrum , P. periplocum , P. acanthophoron and P. mycoparasiticum to suppress disease symptoms caused by F. culmorum on barley seedlings was demonstrated in a greenhouse test.     

Influence of competition and host plant resistance on selection in Phytophthora infestans populations in Michigan, USA and in Northern Ireland

Competition between genotypes of Phytophthora infestans was studied by inoculating potato cultivars with differing susceptibility to late blight in field experiments over three years in Northern Ireland, UK, and Michigan, USA. Multiple isolates of six genotype groups of P. infestans were chosen from the local populations in both N. Ireland and Michigan for inoculation of separate field trials planted in 2003, 2004 and 2005. Four cultivars were used in each trial; two (susceptible cv. Atlantic and the partially resistant cv. Stirling) were common to both locations, whereas the two additional cultivars (with partial resistance to late blight) were cvs Santé and Milagro in N. Ireland and cvs Pike and Jacqueline Lee in Michigan. Single-lesion isolates of P. infestans were obtained from leaves at 1% level of infection, characterized using pre-assigned markers and re-assigned to their respective genotype groups. Extreme selection occurred within the population of genotypes of P. infestans in N. Ireland in each year, with different genotype groups dominating the infection of different cultivars. Selection was observed on all cultivars tested, but was greatest on the more resistant cultivars. Over the 3 years, all of the 114 isolates obtained from cv. Milagro belonged to a single group, whereas among the 118 isolates from cv. Atlantic all six groups were represented. By contrast, in Michigan, the US-8 genotype dominated infection in all cultivars in each year; only 12 of 374 isolates characterized belonged to other genotypes (11 US-14 and a single US-10 isolate).     

寄主品种-病原物小种寄生适合度中水平抗性与侵袭力关系的初步研究

 通过8个马铃薯品种与2个晚疫病菌生理小种组合寄生适合度的测定,对品种-小种寄生适合度中水平抗性与侵袭力之间的关系进行了研究。从初步研究结果可基本确定,在没有垂直抗性或垂直抗性被相应毒性基因克服了的组合中,品种水平抗性和小种侵袭力以乘积的方式构成寄生适合度。其关系可表示为:相对寄生适合度=品种水平抗性(以相对感病性表示)×小种侵袭力。     

Cell wall-degrading enzymes produced in vitro by isolates of Phaeosphaeria nodorum differing in aggressiveness

The relationships between in vitro production of cell wall-degrading enzymes and aggressiveness of three Phaeosphaeria nodorum isolates were investigated. When grown in liquid medium containing 1% cell wall from wheat leaves as the carbon source, the isolates secreted xylanase, α-arabinosidase, β-xylosidase, polygalacturonase, β-galactosidase, cellulase, β-1,3-glucanase, β-glucosidase, acetyl esterase and butyrate esterase. Time-course experiments showed different levels of enzyme production and different kinetics between isolates. A highly aggressive isolate produced more xylanase, cellulase, polygalacturonase and butyrate esterase than did the two weakly aggressive isolates. Xylanase was the most active polymer-degrading enzyme produced, suggesting a key role during pathogenesis by P. nodorum .     

Greater aggressiveness in the 2_A1 lineage of Phytophthora infestans may partially explain its rapid displacement of the US-1 lineage in east Africa

The displacement in east Africa of the US-1 clonal lineage of Phytophthora infestans by 2_A1 clonal lineage has been very rapid. This study tested the hypothesis that dominance of 2_A1 could be due, at least in part, to the increased aggressiveness of 2_A1 over US-1, using both a detached leaf assay (DLA) and a tuber slice assay. The assays were conducted in Uganda and Kenya but US-1 was only assayed in Uganda, because isolates could not be moved across borders and no potato US-1 isolates were available in Kenya. All isolates were collected from potato and compared on two potato cultivars (Kachpot-1 and Sarpo Mira), with the 2_A1 isolates also tested on tomato cultivar Rio Grande. Additionally, a tuber slice assay was done to test whether the capacity to infect tubers differed between 2_A1 and US-1. The aggressiveness of the isolates in the DLA varied significantly both within and among isolates classified according to clonal lineage and for type of host. The 2_A1 isolates were significantly more aggressive than US-1 isolates on both potato varieties evaluated. There were no significant effects of clonal lineage or potato cultivar used in the tuber assay. No significant correlation between foliar and tuber pathogenicity was observed. The 2_A1 isolates were significantly more aggressive on potato than on tomato. An effect of location was also observed in the DLA, on both hosts. It can be concluded from this study that greater pathogenicity of 2_A1 is at least partly attributable to its increased aggressiveness on potato.     

Characterization of Sclerotinia minor populations in Texas peanut fields

In the summer of 2004 an epidemic of sclerotinia blight of peanut, a disease caused by Sclerotinia minor, occurred in Texas in fields where the disease was never previously detected. The disease was observed on many plants within one of the fields (>3000 disease foci), although most foci were <1 m. It is hypothesized that these observations were inconsistent with the recent introduction of a monocyclic pathogen, even if disease developed under conducive environmental conditions. The pattern of disease is most suggestive of the presence of foliar (ascospore) infections, although air temperature was above the known limits for apothecia development if the pathogen had arrived in the field in 2004 peanut seed. To further examine this epidemic, 232 isolates were collected, across a variety of spatial scales spanning this field and other Texas peanut fields, and evaluated for aggressiveness, fungicide sensitivity and genotypic diversity. There was wide variation among isolates for the phenotypic characteristics measured, but there was no evidence that a genotypically unique, highly aggressive, and fungicide resistant isolate had been introduced or evolved. The predominant genotype, TX1, which contained 154 isolates, was found in every county and field population.     

Is virulence phenotype evolution driven exclusively by Lr gene deployment in French Puccinia triticina populations?

Puccinia triticina is a highly damaging wheat pathogen. The efficacy of leaf rust control by genetic resistance is mitigated by the adaptive capacity of the pathogen, expressed as changes in its virulence combinations (pathotypes). An extensive P. triticina population survey has been carried out in France over the last 30 years, describing the evolutionary dynamics of this pathogen in response to cultivar deployment. We analysed the data set for the 2006–2016 period to determine the relationship between the Lr genes in the cultivars and virulence in the pathotypes. Rust populations were dominated by a small number of pathotypes, with variations in most of the virulence frequencies related to the corresponding Lr gene frequencies in the cultivated landscape. Furthermore, the emergence and spread of a new virulence matched the introduction and use of the corresponding Lr gene (Lr28), confirming that the deployment of qualitative resistance genes is an essential driver of evolution in P. triticina populations. However, principal component analysis (PCA) revealed that certain pathotype–cultivar associations cannot be explained solely by the distribution of Lr genes in the landscape. This conclusion is supported by the predominance of a few pathotypes on some cultivars, with the persistence of several other compatible pathotypes at low frequencies. Specific interactions are not, therefore, sufficient to explain the distribution of virulence in rust populations. The hypothesis that quantitative interactions between P. triticina populations and bread wheat cultivars—based on differences in aggressiveness—is also a driver of changes in pathotype frequencies deserves further investigation.     

Testing the resistance of potato varieties to common scab

Summary Resistance to common scab was tested by growing potatoes in dry sand or soil in 3 l pots placed on irrigated subsoil. Inoculated coarse sand (0.5–0.005 Petri dish culture ofStreptomyces scabies per pot) proved more reliable than naturally infested soil. The incidence of scab on the seed tubers did not significantly affect the results. Tests in the greenhouse were most reliable, although those in open air also often gave satisfactory results. For screening purposes satisfactory results can be obtained by using only two or three pots per variety. Thirty-six tyrosinase positive isolates ofStreptomyces from potato and carrot from different parts of Norway were tested on four potato varieties and some of the isolates were tested on carrot. Large differences in aggressiveness were found, but no difference in virulence could be demonstrated, i.e. the same varieties were the most resistant to all isolates.

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Zusammenfassung Es wurden Versuche angestellt, um das Verfahren von Wiersema (1970) für den lokalen Gebrauch anzupassen oder zu verbessern. Er prüfte die Resistenz gegen den gew?hnlichen Schorf durch Anbau von Kartoffeln in natürlich verseuchtem, trockenem Boden in T?pfen auf bew?ssertem Untergrund. In unseren Versuchen brachte die Verwendung von natürlich verseuchtem Boden unterschiedliche Ergebnisse (Tabelle 1), haupts?chlich wegen der kapillaren Aufw?rtsbewegung des Wassers aus dem Untergrund. Die Verwendung von inokuliertem, grobem Sand erwies sich als zuverl?ssiger (Tabellen 2 und 4). Drei Wochen alte Kulturen vonStreptomyces scabies auf PDA in 9 cm-Petrischalen wurden in Wasser zerkleinert (100 ml pro Topf) und gründlich mit grobem Sand vermischt. Für grosse Quantit?ten bei Routinetests wurde ein Zementmischer gebraucht. Der Sand wurde dann in Plastikt?pfe (3 1 Inhalt) abgefüllt. W?hrend des Knollenwachstums konnten relativ trockene Bedingungen in den T?pfen aufrechterhalten werden. Vor der Bonitierung auf Schorfbefall konnte der Sand leicht von den Knollen gewaschen werden. Wenn agressive Isolate gebraucht wurden, war die Schorfentwicklung über die ganze Skala der getesteten Inokulumkonzentrationen, von 0,5 bis zu 0,005 Petrischalenkulturen pro Topf, stark (Tabelle 2). Inokulierter Sand konnte in den folgenden Jahren in Mischungen mit grossen Mengen von nicht inokuliertem Sand wiederholt verwendet werden (Tabelle 3). Sowohl die Schwere der L?son (l = Oberfl?chenschorf; 2 = mittlerer Tief- oder Buckelschorf; 3 = Tiefschorf) als auch die relative Bedeckung der Oberfl?che (0–9) wurden bestimmt; auf Basis dieser Ergebnisse wurde ein Schorfindex errechnet (0–100) (Tabelle 5). Jede Knolle, die gr?sser als 2,5 cm Durchmesser war, wurde einzeln bonitiert. Der Genauigkeitsverlust war jedoch klein, wenn für alle Kartoffeln in einem Topf nur eine Bewertung gegeben wurde (Tabelle 4). Unterschiedliche Schorfgrade auf den Saatknollen beeinflussten die. Ergebnisse nicht signifikant (Tabelle 6). Die Untersuchungen im Glashaus waren meistens zuverl?ssig, aber es konnten auch im freien Feld befriedigende Ergebnisse erzielt werden (Tabelle 8), wo das geeignetste Verfahren darin bestand, die T?pfe fast bis zu ihrem Rand in D?mme einzugraben (Tabelle 7). Die erhaltenen Ergenissee waren über Jahre übereinstimmend, und für gut bekannte Sorten waren sie in gutem Einklang mit früheren Ergebnissen aus Feldversuchen und praktischer Erfahrung. Befriedigende Ergebnisse für Auslesezwecke dürften durch Verwendung von nur zwei oder drei T?pfen pro Klon erzielt werden. Mit der beschriebenen Methode wurden 36 tyrosinasepositiveStreptomyces-Isolate von Kartoffeln und Karotten aus verschiedenen Teilen Norwegens an vier Kartoffelsorten getestet, neun davon auch an Karotten. Bei Anwendung der Terminologie von Van der Plank (1968) wurden grosse Unterschiede in der Agressivit?t festgestellt, aber es konnte kein Unterschied in der Virulenz bewiesen werden, d.h. gegenüber allen Isolaten bestand gleichbleibende, sortenbedingte Resistenz (Tabellen 9 und 10). Zwei der Isolate Karotten.verursachten die st?rkste Infektion auf Karotten. Sie geh?rten auch gegenüber Kartoffel zu den am meisten pathogenen Isolaten. Die Ergebnisse weisen darauf hin, dass die Isolate vor Anwendung im Ausleseverfahren auf Schorfresistenz auf ihre Agressivit?t untersucht werden sollten. Als Alternative k?nnte eine Mischung von verschiedenen Isolaten benützt werden.

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Résumé L’expérimentation a été faite dans le but d’améliorer ou d’adapter aux conditions locales la méthode de Wiersema (1970) qui consiste à tester la résistance à la gale commune par une culture de pommes de terre en sol sec contaminé naturellement, dans des pots placés sur un sous-sol irrigué. Dans nos essais, l’emploi de sol naturellement contaminé a donné des résultats irréguliers (tableau 1), d?s principalement à une remontée de l’eau du sous-sol par capillarité. L’utilisation de sable grossier contaminé artificiellement s’est a vérée plus viable (tableaux 2 et 4). Des cultures agées de 3 semaines deStreptomyces scabies sur milieu PDA en boites de pétri de 9 cm étaient broyées dans de l’eau et mélangées soigneusement au sable grossier (à raison de 100 ml par pot) en utilisant une bétonnière quand des quantités importantes étaient nécessaires pour les tests de routine. Le sable était ensuite mis en pots plastiques de 3 1. Durant la croissance du tubercule on pouvait maintenir dans les pots un milieu relativement sec. Avant d’effectuer les notations concernant la gale, on pouvait aisément laver les tubercules du sable qui y adhérait. Quand des souches agressives étaient utilisées, le développement de la gale était très important sur toute la gamme de concentration d’inoculum testée, de 0,5 à 0,005 contenu de boite de pétri par pot (tableau 2). Le sable contaminé pouvait être réutilisé plusieurs fois les années suivantes en mélange avec de grandes quantités de sable sain (tableau 3). Pour calculer un indice de gale (0–100), on a utilisé le produit de l’estimation de l’importance des lésions (1 = gale superficielle; 2=gale moyennement profonde ou en relief; 3=gale profonde) par l’estimation de la surface relative atteinte (0–9) (tableau 5). Tous les tubercules de plus de 25 mm ont été notés individuellement. Cependant, une notation globale de toutes les pommes de terre d’un pot n’entra?ne pas une grande perte de précisions (tableau 4). Différentes intensités de gale sur les tubercules de semence n’affectent pas significativement les résultats (tableau 6). Les essais en serre se sont avérés plus fiables mais on a pu obtenir des résultats satisfaisants en plein air (tableau 8), la meilleure méthode consistant alors à enterrer les pots dans des sillons presque jusqu’à leur bord supérieur (tableau 7). Les résultats obtenus sont logiques d’une année à l’autre et pour les variétés bien connues concordent avec les résultats antérieurs des essais en plein champ et de la pratique. Pour des essais de Screening, on peut obtenir des résultats satisfaisants avec seulement deux ou trois pots par variété. Par la méthode décrite, on a testé sur quatre variétés de pommes de terre 36 souches deStreptomyces de pommes de terre et de carotte ayant une réaction positive à la tyrosinase et provenant de différentes régions de Norvège. 9 d’entre elles ont été également estées sur carotte. En utilisant la terminologie de Van der Plank (1968), on a trouvé de grandes différences d’agressivité, mais on n’a pu mettre en évidence aucune différence de virulence, ce qui montre qu’il y a une résistance variétale uniforme à toutes les souches (tableau 9–10). Deux des souches de la carotte ont provoqué l’attaque la plus grave sur carotte et étaient également parmi les plus pathogènes pour la pomme de terre (tableau 10). Les résultats montrent que l’on doit d’abord déterminer l’agressivité des souches avant de les utiliser pour des Screenings de résistance à la gale. D’autre part, il faudrait utiliser un mélange de plusieurs souches.