Campylocarpon fasciculare (Nectriaceae, Sordariomycetes); Novel Emergence of Black-Foot Causing Pathogen on Young Grapevines in China

The black-foot disease of grapevine is one of the most destructive diseases in viticulture and it is caused by a complex of soil-borne fungi. This study aimed to identify the species associated with black-foot disease in young grapevines in vineyards of China. Fungal isolates were identified as Campylocarpon fasciculare, based on both morphological and multi-locus phylogenetic analysis of ITS, tef1–α and ß-tubulin sequence data. For the first time in China, we report Campylocarpon fasciculare associated with symptomatic young grapevines. Koch’s postulates were performed on Vitis vinifera cv. Summer Black (SB) in a greenhouse and to confirm the pathogenicity on grapevines. This work improves the knowledge of black-foot disease in Chinese vineyards and will be helpful to growers in their decisions regarding vinicultural practices, planting and disease management.

The relative susceptibility of grapevine rootstocks to black foot disease is dependent on inoculum pressure

Black foot disease of grapevines is a major economic issue for the viticulture industry worldwide. The disease is mainly associated with a complex of pathogen species within the genera Dactylonectria and Ilyonectria. The susceptibility of six grapevine rootstock cultivars to black foot disease under field conditions was assessed. Callused rootstocks of 101-14, 5C, 420A, Riparia Gloire, Schwarzmann and 3309C were planted into soil containing low natural pathogen populations or inoculated with isolates representing the species diversity in New Zealand. Disease incidence, disease severity and dry weight accumulation were assessed after 8 months of growth. Root and shoot dry weights were not significantly affected by inoculation treatment, but differed among rootstock cultivars, with cultivar 420A having the lowest root and shoot dry weight, cultivar 3309C having the largest shoot dry weight and cultivar 5C the largest root dry weight. The relative susceptibility of rootstocks differed significantly depending on whether they were grown under low natural inoculum pressure or a higher pressure in artificially inoculated soil. Schwarzmann and Riparia Gloire rootstock cultivars were the least susceptible under natural low inoculum pressure, but were the most susceptible in inoculated soil. In contrast, 5C was one of the most susceptible under low inoculum levels but was the least susceptible under high pathogen pressure. The result of the study indicate that black foot pathogen inoculum levels in soil affect the relative susceptibility of grapevine rootstocks to infection, and may have implications for the selection of rootstocks for planting.

Droplet Digital PCR Technology for Detection of Ilyonectria liriodendri from Grapevine Environmental Samples

Black-foot disease is one of the most important soilborne diseases affecting planting material in grapevine nurseries and young vineyards. Accurate, early, and specific detection and quantification of black-foot disease causing fungi are essential to alert growers and nurseries to the presence of the pathogens in soil, and to prevent the spread of these pathogens through grapevines using certified pathogen-free planting material and development of resistance. We comparatively assessed the accuracy, efficiency, and specificity of droplet digital PCR (ddPCR) and real-time PCR (qPCR) techniques for the detection and quantification of Ilyonectria liriodendri in bulk and rhizosphere soils, as well as grapevine endorhizosphere. Fungal abundance was not affected by soil-plant fractions. Both techniques showed a high degree of correlation across the samples assessed (R2 = 0.95) with ddPCR being more sensitive to lower target concentrations. Roots of asymptomatic vines were found to be a microbial niche that is inhabited by black-foot disease fungi.

Occurrence and Diversity of Black-Foot Disease Fungi in Symptomless Grapevine Nursery Stock in Spain

In this study, 3,426 grafted grapevines ready to be planted from 15 grapevine nursery fields in Northern Spain were inspected from 2016 to 2018 for black-foot causing pathogens. In all, 1,427 isolates of black-foot pathogens were collected from the asymptomatic inner tissues of surface sterilized secondary roots and characterized based on morphological features and DNA sequence data of the nuclear ribosomal DNA-internal transcribed spacer region, histone H3, translation elongation factor 1-alpha and β-tubulin genes. Eleven species belonging to the genera Dactylonectria, Ilyonectria, Neonectria, and Thelonectria were identified, including Dactylonectria alcacerensis, D. macrodidyma, D. novozelandica, D. pauciseptata, D. torresensis, Ilyonectria liriodendri, I. pseudodestructans, I. robusta, Neonectria quercicola, Neonectria sp. 1, and Thelonectria olida. In addition, two species are newly described, namely D. riojana and I. vivaria. Twenty-four isolates representing 13 black-foot species were inoculated onto grapevine seedlings cultivar ‘Tempranillo’. The pathogenicity tests detected diversity in virulence among fungal species and between isolates within each species. The most virulent species was D. novozelandica isolate BV-0760, followed by D. alcacerensis isolate BV-1240 and I. vivaria sp. nov. isolate BV-2305. This study improves our knowledge on the etiology and virulence of black-foot disease pathogens, and opens up new perspectives in the study of the endophytic phase of these pathogens in grapevines.

Dactylonectria and Ilyonectria Species Causing Black Foot Disease of Andean Blackberry (Rubus Glaucus Benth) in Ecuador

Andean blackberry (Rubus glaucus Benth) plants from the provinces of Tungurahua and Bolivar (Ecuador) started showing symptoms of black foot disease since 2010. Wilted plants were sampled in both provinces from 2014 to 2017, and fungal isolates were obtained from tissues surrounding necrotic lesions in the cortex of the roots and crown. Based on morphological characteristics and DNA sequencing of histone 3 and the translation elongation factor 1α gene, isolates were identified as one of seven species, Ilyonectria vredehoekensis, Ilyonectria robusta, Ilyonectria venezuelensis, Ilyonectria europaea, Dactylonectria torresensis, or Dactylonectria novozelandica. Pathogenicity tests with isolates from each species, excluding I. europaea and D. novozelandica whose isolates were lost due to contamination, confirmed that the four species tested can produce black foot disease symptoms in Andean blackberry. This is the first report of Dactylonectria and Ilyonectria species causing black foot disease of Andean blackberry.

Does Inoculation with Arbuscular Mycorrhizal Fungi Reduce Trunk Disease in Grapevine Rootstocks?

Ilyonectria is a weak pathogen known for causing black foot disease in young vines, infecting roots and vascular tissues at the basal end of the rootstock and restricting the movement of water and nutrients. This negatively impacts vine establishment during transplant into the vineyard. Arbuscular mycorrhizal (AM) fungi are symbiotic fungi that associate with most plants and have been shown to mitigate the infection and effect of pathogens. This greenhouse study was designed to determine if the mycorrhizal fungi could mitigate Ilyonectria infection and whether this was dependent on inoculation timing. ‘Riparia gloire’ grapevine rootstocks (Vitis riparia) were infected with Ilyonectria either after AM fungi, at the same time as AM fungi, or to roots that were not inoculated by AM fungi. We measured the abundance using specific markers for both the pathogen and AM fungi. Colonization by AM fungi did not suppress Ilyonectria, but instead increased the abundance of Ilyonectria. Further, mycorrhizal rootstocks did not have enhanced growth effects on physiological parameters when compared to non-mycorrhizal rootstocks. These findings stand in contrast to the general perception that AM fungi provide protection against root pathogens.