Sensitivity of Plasmopara viticola to selected fungicide groups and the occurrence of the G143A mutant in Australian grapevine isolates.

BACKGROUND: Grapevine downy mildew, caused by Plasmopara viticola, is an economically important disease in Australia and worldwide. The application of fungicides is the main tool to control this disease. Frequent fungicide applications can lead to the selection of resistant P. viticola populations, which has negative impacts on the management of the disease. Identification of resistance and its prevalence is necessary to inform resistance management strategies. RESULTS: A total of 86 P. viticola isolates were collected between 2017 and 2022 from vineyards in 15 growing regions across Australia for four fungicide groups; phenylamide (PA, group 4), carboxylic acid amide (CAA, group 40), quinone outside inhibitor (QoI, group 11) and quinone outside inhibitor stigmatellin binding type (QoSI, group 45). Decreased phenotypic sensitivity was detected for all four groups, and resistance to metalaxyl-M (PA) and pyraclostrobin (QoI), was detected. Genetic analysis to detect the G143A (QoI) and G1105S (CAA) mutations using amplicon-based sequencing was performed for 239 and 65 isolates collected in 2014-2017 and 2017-2022, respectively. G143A was detected in 8% and 52% of isolates, respectively, with strong association to phenotypic resistance. However, G1105S was not detected in any isolates. CONCLUSION: Plasmopara viticola isolates in Australia with resistance to at least two fungicide groups have been detected, therefore it is necessary to adopt resistance management strategies where resistance has been detected. Vineyards should continue to be monitored to improve management strategies for downy mildew. © 2024 Society of Chemical Industry.

Molecular detection and identification of Diatrypaceous airborne spores in Australian vineyards revealed high species diversity between regions.

The grapevine trunk disease, Eutypa dieback (ED), causes significant vine decline and yield reduction. For many years, the fungus Eutypa lata was considered the main pathogen causing ED of grapevines in Australia. Recent studies showed other Diatrypaceous fungi were also associated with vines exhibiting dieback symptoms but there is limited information on how these fungal pathogens spread in vineyards. Thus, information on the spore dispersal patterns of Diatrypaceous fungi in different wine regions will assist in identifying high-risk infection periods in vineyards. Using more than 6800 DNA samples from airborne spores collected from eight wine regions in south-eastern Australia over 8 years using a Burkard spore trap, this study investigated the diversity and abundance of Diatrypaceous species, using multi-faceted molecular tools. A multi-target quantitative PCR (qPCR) assay successfully detected and quantified Diatrypaceous spores from 30% of the total samples with spore numbers and frequency of detection varying between regions and years. The high-resolution melting analysis (HRMA) coupled with DNA sequencing identified seven species, with E. lata being present in seven regions and the most prevalent species in the Adelaide Hills, Barossa Valley and McLaren Vale. Cryptovalsa ampelina and Diatrype stigma were the predominant species in the Clare Valley and Coonawarra, respectively while Eutypella citricola and Eu. microtheca dominated in the Hunter Valley and the Riverina regions. This study represents the first report of D. stigma and Cryptosphaeria multicontinentalis in Australian vineyards. This study further showed rainfall as a primary factor that triggers spore release, however, other weather factors that may influence the spore release in different climatic regions of Australia still requires further investigation.

Population genomics of the grapevine pathogen Eutypa lata reveals evidence for population expansion and intraspecific differences in secondary metabolite gene clusters.

Eutypa dieback of grapevine is an important disease caused by the generalist Ascomycete fungus Eutypa lata. Despite the relevance of this species to the global wine industry, its genomic diversity remains unknown, with only a single publicly available genome assembly. Whole-genome sequencing and comparative genomics was performed on forty Australian E. lata isolates to understand the genome evolution, adaptation, population size and structure of these isolates. Phylogenetic and linkage disequilibrium decay analyses provided evidence of extensive gene flow through sexual recombination between isolates obtained from different geographic locations and hosts. Investigation of the genetic diversity of these isolates suggested rapid population expansion, likely as a consequence of the recent growth of the Australian wine industry. Genomic regions affected by selective sweeps were shown to be enriched for genes associated with secondary metabolite clusters and included genes encoding proteins with a role in nutrient acquisition, degradation of host cell wall and metal and drug resistance, suggesting recent adaptation to both abiotic factors and potentially host genotypes. Genome synteny analysis using long-read genome assemblies showed significant intraspecific genomic plasticity with extensive chromosomal rearrangements impacting the secondary metabolite production potential of this species. Finally, k-mer based GWAS analysis identified a potential locus associated with mycelia recovery in canes of Vitis vinifera that will require further investigations.

The Influence of Water Deficit Stress on the Grapevine Trunk Disease Pathogens Eutypa lata and Diplodia seriata.

The increasing prevalence of the grapevine trunk diseases Eutypa and Botryosphaeria dieback has been attributed, in part, to abiotic stresses imposed on vineyards as production intensifies worldwide. The aim of this study was to evaluate the influence of water deficit irrigation practices on the infection of pruning wounds by Eutypa lata and Diplodia seriata and the subsequent rate of colonization. Two vineyard trials were conducted over two consecutive seasons in South Australia, one in the Riverland with 'Cabernet Sauvignon' with four irrigation treatments (100, 50, 25, and 12.5% of the standard irrigation program) and another in the Barossa Valley with 'Shiraz' on six rootstocks and own roots, either irrigated or not irrigated. According to leaf water potential assessments, vines with reduced irrigation were generally in water deficit and therefore subjected to stress. On the whole, incidence of wound infection and distance of colonization were similar between irrigation treatments for both pathogens, except in the Riverland, where E. lata colonized canes to a greater extent in well-watered vines than those in water deficit. Only vines on rootstock 'Ramsey' in the Barossa Valley had greater extent of colonization by E. lata in the nonirrigated vines. There was no correlation between internal staining and colonization, with both pathogens recovered to nearly 20 cm ahead of the staining. Water deficit did not increase the susceptibility of grapevine pruning wounds to infection or colonization of the subtending tissue by E. lata and D. seriata. In fact, there was evidence of lower susceptibility to colonization by E. lata in vines subjected to severe water deficit.

Pruning Wound Protection Strategies for Simultaneous Control of Eutypa and Botryosphaeria Dieback in New Zealand.

The grapevine trunk diseases Eutypa and Botryosphaeria dieback threaten the sustainability of vineyards worldwide. This study aimed to develop practical and efficient wound protection strategies, which will lead to widespread adoption of preventative control for these diseases and increased longevity of vineyards. Five fungicides (tebuconazole, carbendazim, fluazinam, mancozeb, and flusilazole) were evaluated for efficacy against Eutypa lata and Neofusicoccum luteum infection on pruning wounds of 'Sauvignon blanc' in a New Zealand vineyard. All five fungicides controlled infection by both pathogens to varying degrees, with mean percent disease control of 70 to 90% for E. lata and 33 to 93% for N. luteum. This has led to the first known registration of a fungicide, fluazinam (Gem), for simultaneous control of both Eutypa and Botryosphaeria dieback. Furthermore, application of carbendazim with tractor-driven sprayers provided similar control of E. lata and N. luteum to that when applied by hand with a paintbrush. This constitutes the first report of Botryosphaeria dieback control with fungicides applied with tractor-driven sprayers.

Managing Grapevine Trunk Diseases With Respect to Etiology and Epidemiology: Current Strategies and Future Prospects.

Fungal trunk diseases are some of the most destructive diseases of grapevine in all grape growing areas of the world. Management of GTDs has been intensively studied for decades with some great advances made in our understanding of the causal pathogens, their epidemiology, impact, and control. However, due to the breadth and complexity of the problem, no single effective control measure has been developed. Management of GTD must be holistic and integrated, with an interdisciplinary approach conducted in both nurseries and vineyards that integrates plant pathology, agronomy, viticulture, microbiology, epidemiology, biochemistry, physiology, and genetics. In this review, we identify a number of areas of future prospect for effective management of GTDs worldwide, which, if addressed, will provide a positive outlook on the longevity of vineyards in the future.

Pathogenicity of Diatrypaceous Fungi on Grapevines in Australia.

In addition to Eutypa lata, which causes Eutypa dieback, numerous other fungi in the Diatrypaceae family have been isolated from diseased grapevines (Vitis vinifera) and other woody hosts. Pathogenicity trials comprising 70 strains of diatrypaceous fungi representing nine species in six genera were conducted to determine whether these fungi, collected in Australia, were pathogenic to grapevines. When inoculated into wounded trunks of 'Cabernet Sauvignon', eight species, including E. lata, E. leptoplaca, Cryptovalsa ampelina, C. rabenhorstii, Eutypella citricola, E. microtheca, Diatrypella vulgaris, and a Diatrype sp. produced necrotic lesions significantly longer than on controls. In addition, all nine species (including a Cryptosphaeria sp.) were reisolated from the margins of developing lesions and at varying distances above and below the point of inoculation. Diatrypaceous fungi were frequently isolated from asymptomatic or otherwise healthy tissue several centimeters ahead of the disease margin. Methods to control diseases associated with diatrypaceous fungi must take into account their propensity to colonize woody tissues ahead of or in the absence of visible symptoms. Current recommendations for the management of Eutypa dieback using remedial surgery and pruning wound protection appear sufficient for the control of the other diatrypaceous fungi included in this study.

Evaluating Treatments and Spray Application for the Protection of Grapevine Pruning Wounds from Infection by Eutypa lata.

Eutypa dieback threatens the sustainability of vineyards worldwide and limited treatments are available for control of the disease in grapevine. Following the loss of the effective benzimidazole fungicides, benomyl and carbendazim, there is a need to identify alternatives for the protection of pruning wounds against infection by ascospores of Eutypa lata. In this study, 24 fungicide and natural treatments were evaluated in the laboratory and field. Tebuconazole and carbendazim were the most effective fungicides for reducing colonization of pruning wounds inoculated with E. lata. Pyrimethanil and fluazinam also provided some control but were less effective than tebuconazole at the rates tested. Other treatments, such as cyprodinil + fludioxionil, pyraclostrobin, a garlic extract, and lactoferrin, reduced colonization of wounds by E. lata but require further evaluation at higher concentrations. Carbendazim applied to pruning wounds using tractor-driven sprayers reduced the incidence of pruning wound infection by E. lata to levels similar to that achieved by application with a paint brush.

Evaluation of Fungicides for the Management of Botryosphaeria Canker of Grapevines.

The family Botryosphaeriaceae comprises a number of species that are associated with the dieback disease of grapevine (Vitis vinifera), referred to as Botryosphaeria canker. To date, there are few effective agents available for the management of this disease. In this study, fungicides were evaluated for controlling the disease using a combination of in vitro tests and field trials. Twenty fungicides registered for use on other diseases in Australian viticulture were tested in vitro for their effect on mycelial growth of four species within the Botryosphaeriaceae. The concentrations of fungicide at which 50% of mycelial growth is inhibited (EC50 values) were significantly affected both by fungicide and isolate (P < 0.001). Differences in sensitivities of the four species to the fungicides were negligible (0.41 to 0.59 mg/liter). The most effective fungicides were fludioxonil, carbendazim, fluazinam, tebuconazole, flusilazole, penconazole, procymidone, iprodione, myclobutanil, and pyraclostrobin, for which EC50 values were <1.0 mg/liter. These fungicides were evaluated under field conditions, in addition to the pruning wound protectants Bacseal Super, Garrison, and ATCS tree wound dressing, as well as the biological control agent Vinevax. In field trials, carbendazim (Bavistin), fluazinam (Shirlan), tebuconazole (Folicur), Garrison, and ATCS tree wound dressing applied to freshly cut pruning wounds were the most effective and reduced infection by Diplodia seriata and D. mutila by 41 to 65%. These results suggest that the occurrence of Botryosphaeria canker on grapevines may be reduced via treatment of pruning wounds with selected fungicides as soon as possible after pruning.