Endophytic and rhizosphere actinobacteria isolated from the root system of 1-year-old grafted Vitis vinifera plants were evaluated for their activities against fungi that cause grapevine trunk diseases. A total of 58 endophytic and 94 rhizosphere isolates were tested. Based on an in vitro bioassay, 15.5% of the endophytic isolates and 30.8% of the rhizosphere isolates exhibited antifungal activity against the fungal pathogen Diplodia seriata, whereas 13.8% of the endophytic isolates and 16.0% of the rhizosphere isolates showed antifungal activity against Dactylonectria macrodidyma (formerly Ilyonectria macrodidyma). The strains which showed the greatest in vitro efficacy against both pathogens were further analyzed for their ability to inhibit the growth of Phaeomoniella chlamydospora and Phaeoacremonium minimum (formerly Phaeoacremonium aleophilum). Based on their antifungal activity, three rhizosphere isolates and three endophytic isolates were applied on grafts in an open-root field nursery in a 3-year trial. The field trial led to the identification of one endophytic strain, Streptomyces sp. VV/E1, and two rhizosphere isolates, Streptomyces sp. VV/R1 and Streptomyces sp. VV/R4, which significantly reduced the infection rates produced by the fungal pathogens Dactylonectria sp., Ilyonectria sp., P. chlamydospora, and P. minimum, all of which cause young grapevine decline. The VV/R1 and VV/R4 isolates also significantly reduced the mortality level of grafted plants in the nursery. This study shows that certain actinobacteria could represent a promising new tool for controlling fungal trunk pathogens that infect grapevine plants through the root system in nurseries. IMPORTANCE Grapevine trunk diseases are a major threat to the wine and grape industry worldwide. They cause a significant reduction in yields as well as in grape quality, and they can even cause plant death. Trunk diseases are caused by fungal pathogens that enter through pruning wounds and/or the root system. Although different strategies have recently been developed to protect pruning wounds using antifungal compounds (natural or synthetic) or biocontrol agents, no tools are yet available for controlling soil pathogens that infect plants through their root system. This study shows that different actinobacterial isolates, when applied to grafts in a nursery, can significantly reduce the infection rate caused by fungal pathogens that enter through the root system. This is a new, promising, and green alternative for preventing the decline of young grapevines in nurseries and vineyards.

Grapevine trunk fungal pathogens, such as Diplodia seriata and Phaeomoniella chlamydospora, can infect plants through pruning wounds. They cause grapevine trunk diseases and are involved in grapevine decline. Accordingly, the protection of pruning wounds is crucial for the management of grapevine trunk diseases. The efficacy of different natural antifungals in inhibiting the growth of several fungi causing grapevine trunk diseases was evaluated in vitro. The fungi showing greater in vitro efficacy were tested on autoclaved grape wood assays against D. seriata and P. chlamydospora. Based on results from these assays, chitosan oligosaccharide, vanillin, and garlic extract were selected for further evaluation on pruning wounds inoculated with D. seriata and P. chlamydospora in field trials. A significant decrease in plant mortality was observed after 2 years of growth in the plants treated with the different natural antifungals compared to the mortality rate observed in infected plants that were not treated with antifungals. Also, the infection rate for the inoculated pathogens was significantly reduced in plants treated with the selected natural antifungals. Therefore, natural antifungals represent a promising alternative for disease control and could provide significant economic benefits for the grape-growing industry. Grapevine trunk diseases are a major threat to the wine and grape industry. They have been reported in most grapevineproducing regions worldwide, causing serious economic losses due to a significant reduction of both yields and the quality of the grapes (1-3). The diseases include Botryosphaeria dieback, esca, Eutypa dieback, Petri disease, black foot, and Phomopsis dieback as the most relevant pathologies.Planting material produced in nurseries is frequently infected with fungal pathogens, especially those involved in Petri and black foot diseases (4, 5). In mature vineyards, pathogens such as Ilyonectria sp. mainly penetrate through the root system (6). However, it is widely accepted that for many pathogens, the annually produced pruning wounds are the main route of entry into young and adult plants (7-9). Consequently, numerous studies have been conducted to assess the susceptibility of pruning wounds to fungal pathogens causing trunk diseases. Susceptibility to Botryosphaeriaceae species has been studied by several authors (8-10). Eutypa lata infection has been extensively analyzed by authors like Munkvold and Marois (11) (8) also studied susceptibility to Phomopsis viticola infection. All of these studies support the hypothesis that pruning wounds are the main route of entry of these pathogens in the plant, and, therefore, the development of measures to protect pruning wounds would be an essential tool in the management of vineyards with better sanitation. Accordingly, numerous tests treating pruning wounds with chemical fungicides and/or biological control agents (BCA) have been carried out. Munkvold and Marois (16) checked the efficacy of natural epiphytes and colonizers of pruning wounds fo...

Liming is a common practice used to improve acidic soil properties, as is essential for agricultural quality. A long-term field experiment with one lime rate (6000 kg/ha of carbonate calcium equivalent) and three calcium-based liming amendments (gypsum, limestone and sugar foam) was maintained on a Typic Palexerult for 10 years in order to determine changes in soil acidity and to assess the effects on crop (rye) yields. The soil acidity conditions decreased with all the amendments tested, but the sugar foam and limestone was more effective than gypsum over a long-term period. No significant changes in organic soil matter levels between the treatments tested were found. Interestingly, an increase in the leaching of organic soil matter was observed in limed soils. Lime application significantly increased the total rye biomass compared to the control soils during the whole experiment (2002–2011). Yield trends observed in spike and stem biomass were similar to those reported for total rye biomass. In this respect, at the end of the research, gypsum, limestone and sugar foam increased in relation to the total production of rye biomass by 16%, 32% and 38%, respectively, as compared to the control soils. Additionally, a significant and prolonged difference in calcium concentrations in rye stems between unlimed and limed subplots was observed. However, in spite of the results presented here, further investigations are needed to gain a better understanding of the long-term effects of liming on the chemical properties of soil.

Aluminium toxicity has been recognized as one of the most common causes of reduced grape yields in vineyard acid soils. The main aim of this study was to evaluate the effect of two liming materials, i.e. dolomitic lime and sugar foam, on a vineyard cultivated in an acid soil. The effects were studied in two soil layers (0-30 and 30-60 cm), as well as on leaf nutrient contents, must quality properties and grape yield, in an agricultural soil dedicated to Vitis vinifera L. cv. ‘Mencía’ cultivation. Data management and analysis were performed using analysis of variance (ANOVA). As liming material, sugar foam was more efficient than dolomitic limestone because sugar foam promoted the highest decrease in soil acidity properties at the same calcium carbonate equivalent dose. However, potassium contents in vines organs, including leaves and berries, seemed to decrease as a consequence of liming, with a concomitant increase in must total acidity. Soil available phosphorus also decreased as a consequence of liming, especially with sugar foam, though no effects were observed in plants. For these reasons fertilization of this soil with K and P is recommended along with liming. Grape yields in limed soils increased, although non-significantly, by 30%. This research has therefore provided an important opportunity to advance in our understanding of the effects of liming on grape quality and production in acid soils.

Aluminium phytotoxicity is considered the main limiting factor for crop productivity in agricultural acid soils. Liming is a common practice used to improve acidic soil properties, but an appropriate liming material is essential for both agricultural productivity and environmental sustainability. A long-term field experiment with two liming amendments (dolomitic limestone and limestone) was developed during 10 years to determine the changes in soil acidity and assess the effects on crop (rye) yields. Although the adverse effects of the soil acidity conditions were alleviated with both amendments tested, dolomitic limestone was the most effective in the short- and long-term period. In terms of the saturation of exchange complex, dolomitic limestone had a better efficiency, likely based on its rate of dissolution. No significant changes in soil organic matter and exchangeable potassium levels between the treatments tested were found. Both liming materials significantly increased the rye total biomass, but interestingly, significant correlations were showed between tissue levels of magnesium and biomass production, but not between the latter and calcium. The increases in rye biomass production compared with control soils at the end of the research were the following: dolomitic limestone, 47%, and limestone, 32%. A link between an increase in magnesium bioavailability and biomass production was found, as well as between magnesium rye content and total, spike and stem biomass. Hence, it could conceivably be hypothesized that since magnesium is crucial for the transport of assimilates from source leaves to sink organs, alleviating its deficiency leads to avoiding the reducing growth rate of sink organs. Although further investigations are needed to gain a better understanding of liming on the biological, chemical and physical soil properties in the long term, our research provides support for the conceptual premise that an appropriate selection of liming material is crucial for the productivity of acid soils.

Aims: Soil acidity decreases soil fertility and grapevine growth. Aluminum toxicity has been recognized as one of the most common causes of reduced grape yields in acid vineyard soils. The main aim of this study was to evaluate the effect of two liming materials, i.e. dolomitic lime and sugar foam, on a vineyard cultivated in an acid soil. Methods and results: The effects were studied in two soil layers (0-30 and 30-60 cm), as well as on leaf nutrient contents, grape yield, and must and wine quality properties, in a vineyard dedicated to Vitis vinifera L. cv. Mencía cultivation. The data management and analysis were carried out using ANOVA. Conclusion: Sugar foam was more efficient than dolomitic limestone as liming material since it induced the highest decrease in soil acidity properties at the same calcium carbonate equivalent dose. Effects of liming on leaf nutrient contents, grape yield, and must and wine quality properties were barely observed. Significance and impact of the study: Until recently, little was known about the effects of liming on both vine nutritional status and must/wine quality properties. Thus, this research fills an important knowledge gap. Abstract manuscript

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Vineyard calcareous soils are usually low in organic matter, which makes them prone to physical, chemical, and biological degradation. Besides, these soils are also usually poor in various nutrients in plant-available form, e.g., iron. To make up for this lack of soil fertility, on the one hand, manures, and on the other, iron chelates are usually used. However, the soil application of these materials is not free from problems, and other amendments based on leonardites could be advantageously used as an alternative. Therefore, two organic amendments, one leonardite alone (1 Mg/ha), and the other leonardite (1 Mg/ha) plus ferrous sulphate heptahydrate (0.5 Mg/ha), were tested for three years in a commercial vineyard calcareous plot under Mediterranean climate. The effects of these amendments on soil fertility, plant nutrient contents, and berry quality were studied against a control of bare soil by means of a fully randomized trial with three repetitions per treatment. Soil organic matter (SOM) increased as a consequence of both leonardite treatments, but much more than expected on the basis of a simple mass transfer from the amendments. With the ferrous-sulphate-heptahydrate-supplemented leonardite, the increase in SOM was noticeably higher. This is explained on the basis of nutrient quantity and intensity-pH-related effects, which increased soil nutrient plant-availability and presumably enhanced vine root growth. In response to the higher plant availability of nutrients, the petiole nutrient concentrations were observed to increase under the leonardite treatments. However, only a trend to increase potassium in petioles and in grape must, linked to a decrease of grape must pH, was observed in harvest quality under the leonardite treatments. Leonardite and adequately supplemented leonardite seem to have potential for increasing SOM contents and nutrient plant-availability, thus improving the soil fertility of vineyard calcareous soils.