Identification,pathogenicity and community dynamics of fungi and oomycetes associated with pea root rot in northern France

The pea root rot complex is a major concern for green pea production worldwide. This study aimed at characterizing its composition and dynamics throughout a cropping season in northern France. To this end, fungi and oomycetes were isolated from green pea plant roots with symptoms sampled at the flowering stage in 22 fields in 2017, and at the pea emergence, elongation and flowering stages in two fields in 2018. Out of 646 isolates collected, 317 were identified using molecular markers. Fusarium oxysporum, F. solani and F. redolens were highly predominant. Pathogenicity tests separated the isolates into four aggressiveness groups. F. solani isolates were the most aggressive. Phylogenetic analysis of their TEF1 sequences showed that they mainly belonged to the F. pisi lineage, and that F. oxysporum isolates were genetically close to isolates from the UK that did not belong to the forma specialis pisi. In addition, several Clonostachys rhizophaga isolates are reported for the first time to cause pea root rot. The oomycetes were rarely found and were represented by a few Pythium spp. isolates. Lastly, this study shows that the fungal and oomycete communities associated with pea root rot change during the cropping season. The level of dissimilarity of the root-rot-associated communities decreased throughout the cropping season towards a more similar composition at the flowering stage, dominated by F. solani, F. oxysporum and F. redolens. The proportion of nonpathogenic to weakly pathogenic isolates decreased progressively during the growing season in favour of moderately to highly pathogenic isolates.     

Risk assessment of Aphanomyces euteiches root rot disease: quantification of low inoculum densities in field soils using droplet digital PCR

European Journal of Plant Pathology - Fusarium verticillioides is a soil-borne plant pathogen of maize plants (Zea mays L.) responsible for major yield losses by causing root, stalk, and ear rot...     

Diversity of microorganisms associated with atypical superficial blemishes of potato tubers and pathogenicity assessment

Skin blemishes of potato (Solanum tuberosum L.) tubers can cause severe economical losses to production. Some blemishes are due to known pathogens and others whose causes are unknown are called atypical blemishes. The present work aims at determining the origin of superficial atypical blemishes on a set of 204 tubers coming from 12 different French regions producing potato. The diversity of fungi and Streptomyces bacteria associated with blemishes was investigated by systematic isolation followed by identification by sequencing the internal transcribed spacer of the ribosomal DNA for fungi and by sequencing the 16S ribosomal DNA for bacteria. We found a high microbial diversity represented by 349 fungal isolates belonging to at least 47 different species and 21 bacterial strains of Streptomyces sp. The most represented fungi belonged to the genera Fusarium, Rhizoctonia, Alternaria, Penicillium, and Clonostachys. The pathogenicity of representative isolates was assessed in three bioassays; two bioassays based on single inoculations in previously sterilized potting mixture, and one bioassay based on both single and double inoculations under hydroponic conditions. We fulfilled the Koch’s postulates for Rhizoctonia solani AG 3 producing sclerotia. For other fungal and bacterial strains, our results did not show any causality or relationship between a single isolate or a complex and the occurrence of the blemishes. Moreover, the observation of irregular polygonal sunken corky lesions (polygonal lesions)—the most frequent atypical blemish—on non-inoculated tubers, suggested that the atypical blemishes could as well be a reaction of the plant to stressful environmental conditions.     

Cause and duration of mustard incorporation effects on soil-borne plant pathogenic fungi

Two fungal plant pathogens, Rhizoctonia solani AG 2-2 and Fusarium oxysporum f.sp. lini, were studied in relation to general responses of soil fungi and bacteria following incorporation of Brassica juncea. Our aim was to understand to what extent the changes in the biological and physicochemical characteristics of the soil could explain the effects on the studied pathogens and diseases, and to determine the temporal nature of the responses. Short-term effects of mustard incorporation (up to 4 months) were investigated in a microcosm experiment, and compared with a treatment where composted plant material was incorporated. In a field experiment, the responses were followed up to 11 months after removal or incorporation of a mustard crop. In general, responses in the variables measured changed more after incorporation of fresh mustard material than after addition of similar amounts of composted plant material (microcosms) or after removal of the mustard crop (field). The soil inoculum potential of R. solani AG 2-2 decreased directly after incorporation of mustard, but increased later to disease levels above those in the untreated soil. Neither of these effects could be explained by changes in the population density of R. solani AG 2-2. Fusarium spp. were less influenced, although an increase in the suppressiveness to Fusarium wilt was observed after mustard incorporation as compared with the treatment where mustard was removed. The microbial responses to mustard incorporation were more pronounced for bacteria than for fungi. After an initial substantial increase, the bacterial density decreased but remained above the levels in the control treatment throughout the experimental periods. The bacterial community structure was modified up to 8 months after mustard incorporation. We conclude that incorporation of fresh mustard influences soil microbial communities, especially the bacteria, and has a potential to control the pathogenic activity of R. solani 2-2 on a short-term perspective. The time dependency in microbial responses is important and should be taken into consideration for the evaluation of the potential of Brassicas to control plant disease on a field scale.     

Biotic changes in relation to local decrease in soil conduciveness to disease caused by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

The relationships between biotic changes and local decrease in soil conduciveness in disease patches towards the disease incited by Rhizoctonia solani AG 2-2 in a sugar beet field in France were investigated. Soil samples from healthy and diseased areas were analysed for bacterial and fungal densities, molecular and physiological microbial community structures, and antagonistic abilities of Trichoderma isolates collected from diseased and healthy areas. Although the inoculum density was higher inside the disease patches, the respective soil was less conducive towards disease incited by R. solani AG 2-2. It was concluded that the pathogen was present in healthy areas but did not incite disease in field conditions. Conversely, the response of the microflora to previous development of R. solani in diseased areas prevented further pathogenic activity. Indeed, genetic and physiological structures of the fungal communities and physiological structures of the bacterial communities were modified in disease patches compared to healthy areas. The terminal restriction fragment length polymorphism (T-RFLP) analysis revealed that the peaks corresponding to R. solani and Trichoderma spp. were higher inside the patches than in the healthy areas. Trichoderma isolates from the disease patches were more antagonistic than those from the healthy areas. These results suggest that disease caused by R. solani AG 2-2 induced changes in genetic and physiological structure of microbial populations and development of antagonists. The decreased conduciveness inside the patches may help explain patch mobility in the following season.     

Genetic Diversity of Pathogenic and Nonpathogenic Populations of Fusarium oxysporum Isolated from Carnation Fields in Argentina

ABSTRACT In order to elucidate the origin of Fusarium oxysporum f. sp. dianthi in Argentina, the genetic diversity among pathogenic isolates together with co-occurring nonpathogenic isolates on carnation was investigated. In all, 151 isolates of F. oxysporum were obtained from soils and carnation plants from several horticultural farms in Argentina. The isolates were characterized using vegetative compatibility group (VCG), intergenic spacer (IGS) typing, and pathogenicity tests on carnation. Seven reference strains of F. oxysporum f. sp. dianthi also were analyzed and assigned to six different IGS types and six VCGs. Twenty-two Argentinean isolates were pathogenic on carnation, had the same IGS type (50), and belonged to a single VCG (0021). The 129 remaining isolates were nonpathogenic on carnation and sorted into 23 IGS types and 97 VCGs. The same VCG never occurred in different IGS types. Our results suggest that the pathogen did not originate in the local populations of F. oxysporum but, rather, that it was introduced into Argentina. Given the genetic homogeneity within Argentinean isolates of F. oxysporum f. sp. dianthi, either IGS type or VCG can be used for the identification of the forma specialis dianthi currently in Argentina.     

Fingerprinting methods to approach multitrophic interactions among microflora and microfauna communities in soil

The soil functioning and the response of the biota to external perturbations such as organic input are based on multitrophic interactions among a wide range of organisms. However, the various components of the soil microflora and microfauna are rarely addressed in ecological studies. We have developed a molecular approach based on terminal restriction fragment length polymorphism (T-RFLP) analysis to assess the community structures of protozoa and nematodes, together with bacteria and fungi that share the same soil environment. Two soils were characterized by a specific fingerprint for each of the four groups of organisms, showing the potential of all T-RFLP procedures to differentiate the community structures. The response of the soil biota to organic inputs was addressed using T-RFLP fingerprints together with physiological profiles of bacteria communities and global microbial activities and densities. Although the impact of compost or manure on the soil biota was only slightly noticeable from the global parameters measured, it was obvious from the community level analyses. However, the different components of the soil biota were altered to various extents, depending on the group of organisms and the soil–organic matter combination. The potential of the T-RFLP strategy to analyze simultaneously different biotic groups from the same soil DNA extract will facilitate the more systematic integration of eukaryotic organisms in ecological studies to investigate multitrophic interactions among the microflora and microfauna in relation with soil processes.     

High Genetic Diversity Among Strains of Fusarium oxysporum f. sp. vasinfectum from Cotton in Ivory Coast

ABSTRACT Seventeen isolates of Fusarium oxysporum f. sp. vasinfectum from the Ivory Coast were characterized using vegetative compatibility group (VCG), restriction fragment length polymorphism of the ribosomal inter-genic spacer region (IGS), and mating type (MAT) idiomorph, and compared with a worldwide collection of the pathogen containing all available reference strains. Some of the isolates were identical to known reference strains for all three traits, whereas others had previously unknown varieties of IGS and (possibly) VCG. One or the other MAT idiomorph was present in each of the new isolates and the reference strains. The new isolates and reference strains were grouped based upon the three traits. Strains from the Ivory Coast were found in 7 of 11 groups detected, suggesting multiple sources for Fusarium wilt in the country. Despite the presence of both MAT idiomorphs among isolates, no evidence for recombination was found.