Grapevine Plants Research Articles

Systematic Investigation of the Effects of a Novel Protein Hydrolysate on the Growth, Physiological Parameters, Fruit Development and Yield of Grapevine (Vitis Vinifera L., cv Sauvignon Blanc) under Water Stress Conditions

In the last decade climate change has impacted viticulture and water deficit has become a major concern in fruit production. Many studies have been carried out to determine the grapevine response to environmental changes and to identify key genetic traits to be used in grapevine breeding. However, in order to better manage climate-related risks, novel viticultural practices are urgently needed. A promising solution for a more sustainable model of viticulture involves the use of biostimulants. In this study, the effectiveness of a novel biostimulant (APR®) belonging to the group of protein thermal hydrolysates was tested on grapevine plants subjected to progressive water deficit conditions. Our results showed that this compound applied to roots before imposing water deprivation mitigates the consequences of stress by sustaining the growth of the younger vegetative organs and limiting the extent of cell dehydration; this positive impact on the plant’s physiological state persisted during the recovery phase. Furthermore, at the end of the growing season, plants treated with the biostimulant, both in optimal water conditions and under water stress, exhibited a greater accumulation of biomass in the aerial part (6.8% and 21.3 %, respectively) and a higher berry diameter (3.4 % and 9.5 %, respectively). Additional work through field trials will be necessary to further substantiate these results and to translate this knowledge into specific practices that grape growers can easily adopt.

Update and New Epidemiological Aspects about Grapevine Yellows in Chile.

To date, phytoplasmas belonging to six ribosomal subgroups have been detected to infect grapevines in Chile in 36 percent of the sampled plants. A new survey on the presence of grapevine yellows was carried out from 2016 to 2020, and 330 grapevine plants from the most important wine regions of the country were sampled and analyzed by nested PCR/RFLP analyses. Phytoplasmas enclosed in subgroups 16SrIII-J and 16SrVII-A were identified with infection rates of 17% and 2%, respectively. The vineyards in which the phytoplasma-infected plants were detected were further inspected to identify alternative host plants and insects of potential epidemiological relevance. Five previously unreported plant species resulted positive for 16SrIII-J phytoplasma (Rosa spp., Brassica rapa, Erodium spp., Malva spp. and Rubus ulmifolius) and five insect species were fully or partially identified (Amplicephalus ornatus, A. pallidus, A. curtulus, Bergallia sp., Exitianus obscurinervis) as potential vectors of 16SrIII-J phytoplasmas. The 16SrVII-A phytoplasmas were not detected in non-grape plant species nor in insects. This work establishes updated guidelines for the study, management, and prevention of grapevine yellows in Chile, and in other grapevine growing regions of South America.

Response of Mycorrhizal 'Touriga Nacional' Variety Grapevines to High Temperatures Measured by Calorespirometry and Near-Infrared Spectroscopy.

Heat stress negatively affects several physiological and biochemical processes in grapevine plants. In this work, two new methods, calorespirometry, which has been used to determine temperature adaptation in plants, and near-infrared (NIR) spectroscopy, which has been used to determine several grapevine-related traits and to discriminate among varieties, were tested to evaluate grapevine response to high temperatures. ‘Touriga Nacional’ variety grapevines, inoculated or not with Rhizoglomus irregulare or Funneliformis mosseae, were used in this study. Calorespirometric parameters and NIR spectra, as well as other parameters commonly used to assess heat injury in plants, were measured before and after high temperature exposure. Growth rate and substrate carbon conversion efficiency, calculated from calorespirometric measurements, and stomatal conductance, were the most sensitive parameters for discriminating among high temperature responses of control and inoculated grapevines. The results revealed that, although this vine variety can adapt its physiology to temperatures up to 40 °C, inoculation with R. irregulare could additionally help to sustain its growth, especially after heat shocks. Therefore, the combination of calorespirometry together with gas exchange measurements is a promising strategy for screening grapevine heat tolerance under controlled conditions and has high potential to be implemented in initial phases of plant breeding programs.

Plasma activated water triggers plant defence responses

Nowadays, one of the main challenges is moving towards an eco-sustainable agriculture, able to preserve the food production through a reduced use of pesticides. Current global food sustenance by intensive agriculture is mainly based on economic crop monocultures and drastically reduces the biodiversity, increasing the yield losses due to the presence of biotic and abiotic stresses. A technology based on plasma activated water (PAW), characterized by the presence in liquid of reactive oxygen and nitrogen species, was tested to try to ensure yield stability also enhancing the plant resistance responses and to promote an eco-sustainable management of plant diseases. In PAW-treated micropropagated periwinkle shoots, periwinkle and grapevine plants, qRT-PCR and small RNAs high-throughput sequencing were used to analyse the differential expression of genes involved in the major plant defence pathways. The results indicate that PAW treatment enhances the plant defence responses and provide an encouraging framework for future applications in plant disease management programs.

Plasmopara viticola infection affects mineral elements allocation and distribution in Vitis vinifera leaves

Plasmopara viticola is one of the most important pathogens infecting Vitis vinifera plants. The interactions among P. viticola and both susceptible and resistant grapevine plants have been extensively characterised, at transcriptomic, proteomic and metabolomic levels. However, the involvement of plants ionome in the response against the pathogen has been completely neglected so far. Therefore, this study was aimed at investigating the possible role of leaf ionomic modulation during compatible and incompatible interactions between P. viticola and grapevine plants. In susceptible cultivars, a dramatic redistribution of mineral elements has been observed, thus uncovering a possible role for mineral nutrients in the response against pathogens. On the contrary, the resistant cultivars did not present substantial rearrangement of mineral elements at leaf level, except for manganese (Mn) and iron (Fe). This might demonstrate that, resistant cultivars, albeit expressing the resistance gene, still exploit a pathogen response mechanism based on the local increase in the concentration of microelements, which are involved in the synthesis of secondary metabolites and reactive oxygen species. Moreover, these data also highlight the link between the mineral nutrition and plants’ response to pathogens, further stressing that appropriate fertilization strategies can be fundamental for the expression of response mechanisms against pathogens.

Growth, heavy metal uptake, and photosynthesis in 'Paulsen 1103' (Vitis berlandieri x rupestris) grapevine rootstocks inoculated with arbuscular mycorrhizal fungi from vineyard soils with high copper contents

Soils in old vineyards in southern Brazil have high copper accumulation due to fungicide applications over the years, which can affect physiology and growth of young grapevine plants. Arbuscular mycorrhizal fungi (AMF) alleviate toxic effects of metals and increase photosynthesis and plant growth. We evaluated whether inoculation with Rhizophagus clarus (Rh) from a mining area or with a trap-culture-enriched AMF community (Tc) isolated from a high-copper vineyard soil, improved growth and photosynthesis in grapevine rootstocks planted in young ( 60 years) vineyards soils of Vale da Uva Goethe, SC, Brazil. Mycorrhizal colonization was higher in grapevines installed in young vineyard soil than those planted in old vineyard soil. Plants grew more in the old vineyard soil than in the soil from a young vineyard, and that was related to plant nutrient concentration in the soil. In both soils, Tc-inoculated grapevines had higher photosynthetic activity, while those inoculated with R. clarus had higher carbon assimilation. In conclusion, grapevines showed a positive response to AMF inoculation in different soil conditions, and the native AMF community from high copper soils are promising for inoculation of grapevines.

The use of artificial habitats increases spider abundance and richness in a vineyard of Argentina

Maintaining biodiversity in agricultural systems is essential, as it plays a key role in ecosystem services provision. However, it is declining at an unprecedented rate. Several methods can contribute to increasing species diversity in agroecosystems, including the use of artificial microhabitats. Studies focusing on biodiversity in vineyards have been carried out recently, mainly in Europe and North America, but are scarce in South America. In this study, we investigate whether introduced environmental variability and habitat complexity, using corrugated cardboard bands, enhances spider abundance and diversity in vineyards. Spiders were collected from grapevine plants, using two collecting methods (foliage beating and direct collection). Plants with and without cardboard bands (N = 30) were sampled in three different phenological stages of the vineyard (veraison, maturity and postharvest). Overall, we collected 904 spiders belonging to 35 species. We found that spider abundance and species richness significantly increased with cardboard bands. In addition, we identified particular species that prefer cardboard bands to build their shelters. Results suggest that cost-effective habitat manipulation can increase spider abundance in vineyards, which implies higher biodiversity conservation value and potential for improving biological control.

2,4-D em videiras: estudo de caso no Município de Jaguari-RS, estado do Rio Grande do Sul, Brasil

Atualmente plantas de videira no município de Jaguari, localizado na região centro-oeste do estado do Rio Grande do Sul - Brasil, apresentam sintomas de fitotoxidez, possivelmente oriundos da aplicação de agrotóxicos em áreas de produção agrícola. A uva é um produto de grande valor econômico, social e cultural para a região. Neste sentido a Lei Municipal N° 3.163 restringe o uso de herbicidas derivados da composição química 2,4-D (diclorofenoxiacético) em todo o território do município de Jaguari no período de 15 de setembro a 15 de março de cada ano agrícola, independente da modalidade de sua aplicação. Portanto, o objetivo deste trabalho foi identificar e quantificar resíduos de 2,4-D nas folhas de videira pelo método QuEChERS e análise por HPLC-MS/MS. A coleta das amostras foi realizada em seis propriedades localizadas dentro do perímetro delimitado pela Lei Municipal N° 3.163. O período de amostragens foi semanal de 29/09/2017 a 06/12/2017, perfazendo um total de 62 amostras. Foram coletadas folhas de videiras e armazenadas em sacos plásticos devidamente identificados e encaminhados para o Laboratório de Análises de Resíduos e Pesticidas (LARP), na Universidade Federal de Santa Maria (UFSM). Durante o período de amostragem observou-se problemas de fitotoxidez das plantas. Comprovou-se a presença de resíduos do herbicida em 69% das amostras, concluindo-se que a deriva do produto pode ser a causa dos danos nas videiras apontados pelos produtores de Jaguari.

Overview of Kaolin Outcomes from Vine to Wine: Cerceal White Variety Case Study

Kaolin protective effect was assessed in a white grapevine cultivar ‘Cerceal’ in ‘Alentejo’ Region (southeast Portugal) where plants face extreme conditions during the summer season. We addressed the hypothesis that kaolin effects lead to several changes in leaves, fruits, and wine characteristics on the primary and secondary metabolism. Results showed that kaolin reduces leaf temperature which provokes an improvement in physiological parameters such as net photosynthesis and water use efficiency. This protection interferes with berry color, leaving them more yellowish, and an increase in phenolic compounds were observed in all fruit tissues (skin, seed, and pulp). Additionally, both berry and wine characteristics were strongly affected, with an increase of tartaric and malic acid and consequently high total acidity, while the sugar concentration decreased 8.9% in berries provoking a low wine alcohol level. Results also showed that kaolin induces high potassium, magnesium, and iron, and low copper and aluminum concentrations. Moreover, the control wine showed higher content of esters related with hostile notes whereas wine from kaolin treated vines presented higher content of esters associated with fruity notes. Overall, the results strengthen the promising nature of kaolin application as a summer stress mitigation strategy protecting grapevine plants and improving fruit quality and creating more balanced wines.

A supplement of ultraviolet-B radiation under field conditions increases phenolic and volatile compounds of Tempranillo grape skins and the resulting wines

Ultraviolet-B (UV-B) supplements have been applied to grapevine in controlled environments but not under field conditions, which would be the only way to manipulate UV-B to provide tangible agronomic benefits. Further, the effects of UV-B supplements should be investigated not only in grapes but also resulting wines, to check whether possible beneficial effects incurred by the raw material are conserved in the elaborated product. In this study, a UV-B supplement was applied to Tempranillo grapes under commercial field conditions, to test its effects on (1) the phenolic composition of grape skins and the resulting wines in two different seasons and (2) the volatile organic compounds (VOCs) of the wines. The UV-B supplement was applied mechanically, using a manufactured lamp mounted on a tractor, to make the application method more realistically viable at a crop scale. Flavonols and antioxidant capacity, and to a lesser extent phenolic acids and flavanols, increased in UV-B-supplemented grape skins and their resulting wines, thus enhancing the quality of both components in wine production. In particular, the augmented flavonols could improve the health properties of grapes and wines, while also contributing to wine color stabilization. In addition, six VOCs were found elevated in the wines elaborated with UV-B-supplemented grapes. Following from these results, we discuss whether these changes were more likely attributable to the irradiance peaks of the supplemental UV-B than to the additional UV-B dose applied. In conclusion, we propose the mechanical application of UV-B supplements on grapevine plants at a crop scale as a new agricultural practice with promising utility, such as counteracting deficits of natural UV-B in specific locations or years.

Grapevine VlbZIP30 improves drought resistance by directly activating VvNAC17 and promoting lignin biosynthesis through the regulation of three peroxidase genes

Drought stress severely affects grapevine quality and yield, and recent reports have revealed that lignin plays an important role in protection from drought stress. Since little is known about lignin-mediated drought resistance in grapevine, we investigated its significance. Herein, we show that VlbZIP30 mediates drought resistance by activating the expression of lignin biosynthetic genes and increasing lignin deposition. Transgenic grapevine plants overexpressing VlbZIP30 exhibited lignin deposition (mainly G and S monomers) in the stem secondary xylem under control conditions, which resulted from the upregulated expression of VvPRX4 and VvPRX72. Overexpression of VlbZIP30 improves drought tolerance, characterized by a reduction in the water loss rate, maintenance of an effective photosynthesis rate, and increased lignin content (mainly G monomer) in leaves under drought conditions. Electrophoretic mobility shift assay, luciferase reporter assays, and chromatin immunoprecipitation-qPCR assays indicated that VlbZIP30 directly binds to the G-box cis-element in the promoters of lignin biosynthetic (VvPRX N1) and drought-responsive (VvNAC17) genes to regulate their expression. In summary, we report a novel VlbZIP30-mediated mechanism linking lignification and drought tolerance in grapevine. The results of this study may be of value for the development of molecular breeding strategies to produce drought-resistant fruit crops.

Community succession of the grapevine fungal microbiome in the annual growth cycle.

Microbial ecology and activity in wine production influences grapevine health and productivity, conversion of sugar to ethanol during fermentation, wine aroma, wine quality and distinctiveness. Fungi in the vineyard ecosystem are not well described. Here, we characterized the spatial and temporal dynamics of fungal communities associated with the grapevine (grapes, flowers, leaves, and roots) and soils over an annual growth cycle in two vineyards to investigate the influences of grape habitat, plant developmental stage (flowering, fruit set, veraison, and harvest), vineyards, and climatic conditions. Fungi were influenced by both the grapevine habitat and plant development stage. The core microbiome was prioritized over space and time, and the identified core members drove seasonal community succession. The developmental stage of veraison, where the grapes undergo a dramatic change in metabolism and start accumulating sugar, coincided with a distinct shift in fungal communities. Co-occurrence networks showed strong correlations between the plant microbiome, the soil microbiome, and weather indices. Our study describes the complex ecological dynamics that occur in microbial assemblages over a growing season and highlight succession of the core community in vineyards.

(Z)-3-Hexenyl Butyrate Induces Stomata Closure and Ripening in Vitis vinifera

Agronomy solutions for modifying pre-harvest grape ripening are needed for a more sustainable viticulture. Field experiments were performed in Vitis vinifera L. vines to study the effect of the previously described stomata-closing compound (Z)-3-hexenyl butyrate (HB). Exogenous treatments at different doses were periodically carried out using a randomized block design. Firstly, we observed that HB was able to induce stomatal closure in grapevine plants. Under field conditions, the application of HB around veraison induced a higher color intensity in berries, and vines treated at higher doses reached this stage earlier than the un-treated controls. There was also a clear increase in both grape anthocyanin concentration and total soluble solids without having a negative impact on total yield. We therefore, confirm the role of HB as a universal natural stomatal closure compound and propose a new use for HB in viticulture as a ripening inducer, by accelerating anthocyanin accumulation.

The arbuscular mycorrhizal fungus Funneliformis mosseae induces changes and increases the concentration of volatile organic compounds in Vitis vinifera cv. Sangiovese leaf tissue

Arbuscular mycorrhizal fungi (AMF) are beneficial obligate symbionts of plant roots. Volatile organic compounds (VOCs) participate in plant communication and defence. The aim of this study was to analyse the effects of the arbuscular mycorrhizal fungus Funneliformis mosseae IMA1 on VOCs in Vitis vinifera cv. Sangiovese leaf tissue. Grapevine plants inoculated with F. mosseae IMA1 were incubated for 23 weeks. VOCs were extracted from leaves and identified using headspace solid-phase microextraction (HS–SPME) coupled to GC-MS. VOCs in leaf tissue were strongly enhanced (85%) by F. mosseae IMA1. The mycorrhizal fungus IMA1 modified the levels of specific VOCs synthesised in different anabolic pathways. An increase in volatiles that have been related to plant defences under pathogen/herbivore attack or linked to water stress, such as (E)–2–hexenal, 3–hexenal, geraniol, benzaldehyde and methyl salicylate, was observed in mycorrhizal plants. In contrast, some C13–norisoprenoids decreased strongly in mycorrhizal plants. The study of the effects of AMF on VOCs in grapevine plants may provide useful information to establish sustainable viticultural practices.

Flashes of UV-C light: An innovative method for stimulating plant defences.

Leaves of lettuce, pepper, tomato and grapevine plants grown in greenhouse conditions were exposed to UV-C light for either 60 s or 1 s, using a specific LEDs-based device, and wavelengths and energy were the same among different light treatments. Doses of UV-C light that both effectively stimulated plant defences and were innocuous were determined beforehand. Tomato plants and lettuce plants were inoculated with Botrytis cinerea, pepper plants with Phytophthora capsici, and grapevine with Plasmopara viticola. In some experiments we investigated the effect of a repetition of treatments over periods of several days. All plants were inoculated 48 h after exposure to the last UV-C treatment. Lesions on surfaces were measured up to 12 days after inoculation, depending on the experiment and the pathogen. The results confirmed that UV-C light stimulates plant resistance; they show that irradiation for one second is more effective than irradiation for 60 s, and that repetition of treatments is more effective than single light treatments. Moreover a systemic effect was observed in unexposed leaves that were close to exposed leaves. The mechanisms of perception and of the signalling and metabolic pathways triggered by flashes of UV-C light vs. 60 s irradiation exposures are briefly discussed, as well as the prospects for field use of UV-C flashes in viticulture and horticulture.

Quantifying ecological and economic value of pest control services provided by bats in a vineyard landscape of central Chile

The top-down suppression of pest insects by their natural enemies, such as insectivorous bats, represents an important ecosystem service in agricultural systems. Recognizing the importance of bats to suppress pest populations and further conserve their populations near agricultural systems could reduce damage to crops thereby potentially increasing the monetary gain of farmers. Viticulture represents one of the most extensive and economically important agricultural crops in the Mediterranean biome. While pest consumption by bats has been recently revealed in vineyards, direct evidence of the reduction of grapevine pests by bats in vineyards is lacking, and their benefits to winegrape production remains to be quantified. Using large nocturnal exclosures in vineyards, we examined the top-down effects of aerial insectivorous bats in suppressing insect populations by assessing leaf and grape cluster damage; we also examined the benefit of these natural pest predators by quantifying resulting increases in potential vineyard yield. Grapevine plants excluded from bats had significantly higher leaf herbivory and greater grape cluster damage than control plots. Grape cluster damage was 7% lower on control plots, yielding an average economic benefit of US$188-$248/ha/year due to bat predation. These results provide the first experimental evidence that bats reduce grapevine pest insect infections and thus increase vineyard yield and winegrowers’ income. Therefore, bats should be included in future biodiversity conservation plans in vineyards and be considered within agricultural management strategies based on natural pest suppression.

Actinobacteria Associated with Vineyard Soils of Algeria: Classification, Antifungal Potential Against Grapevine Trunk Pathogens and Plant Growth-Promoting Features

Grapevine trunk diseases(GTDs) are among the most destructive diseases of vineyards worldwide, including Algeria. In the fungal complex involved in GTD symptoms, referred as grapevine trunk-pathogens, Paeomoniella chlamydospora and Phaeoacremonium minimum have a determining infecting role as pioneer fungi. Due to the lack of efficiency of conventional disease management practices, a search for alternative strategies, such as biocontrol, is needed. Taking the approach of looking for biocontrol candidates in the environment surrounding the plant, the present study explored actinobacteria diversity within vineyard soils of six grape-producing regions in Algeria. Based on their 16S rRNA gene sequence, identification and phylogenic analysis were performed on the 40 isolates of actinobacteria obtained. Forty percent of strains were attached to Streptomyces, including two evidenced new species, and 32.5% were affiliated to Saccharothrix. The other less represented genera were Actinoplanes, Nocardia, Nocardiopsis, Lentzea, Nonomuraea, Promicromonospora, Saccharopolyspora and Streptosporangium. Screening based on antagonistic and plant growth promotion (PGP) abilities of the strains showed that 47.5% of the isolates exhibited appreciable antagonistic activities against both Pa. chlamydospora and Pm. minimum, with the two best strains being Streptomyces sp. Ms18 and Streptomyces sp. Sb11. Screening for plant growth promoting properties demonstrated that majority of the strains were able to produce indole acetic acid, siderophores, ammonia, ACC deaminase, cellulase and amylase, and fix N2. Through a PGP-traits-based cluster analysis, the most interesting strains were highlighted. Taking into account both antagonistic and PGP properties, Streptomyces sp Sb11 was selected as the most promising candidate for further evaluations of its efficiency in a GTDs context.

Microbial Functional Diversity in Vineyard Soils: Sulfur Metabolism and Links With Grapevine Plants and Wine Quality

The quality of the vineyard soils has a direct impact on grapes and wine quality and represents a key component of the “Terroir concept”. However, information on the impact of soil microbiota on grapevine plants and wine quality are generally lacking. In fact, over the last few years most of the attempts made to correlate soil microbial communities and wine quality were limited by overlooking both the functional traits of soil microbiota and the spatial variability of vineyards soils. In this work, we used a functional gene microarray approach (GeoChip) and soil enzymatic analyses to assess the soil microbial community functional potential related to the different wine quality. In order to minimize the soil variability, this work was conducted at a “within-vineyard” scale, comparing two similar soils (BRO11 and BRO12) previously identified with respect to pedological and hydrological properties within a single vineyard in Central Tuscany and that yielded highly contrasting wine quality upon cultivation of the same Sangiovese cultivar (BRO12 exhibited the higher quality). Our results showed an enrichment of Actinobacteria in BRO12, whereas Alfa- and Gamma-Proteobacteria were more abundant in BRO11, where an enrichment of bacteria involved in N fixation and denitrification occurred. Overall, the GeoChip output revealed a greater biological activity in BRO11 but a significant enrichment of sulfur-oxidation genes in BRO12 compared to BRO11 soil, where a higher level of arylsulfatase activity was also detected. Moreover, the low content of sulfates and available nitrogen found in BRO12 suggested that the reduced availability of sulfates for vine plants might limit the reduced glutathione (GSH) synthesis, which plays an important role in aroma protection in musts and wines. In conclusion, in addition to nitrogen availability, we propose that soil microbial sulfur metabolism may also play a key role in shaping plant physiology, grapes and wine quality. Overall, these results support the existence of a “microbial functional terroir” effect as a determining factor in vineyard-scale variation among wine grapes.

Detection of Grapevine Leafroll-Associated Virus 1 and 3 in White and Red Grapevine Cultivars Using Hyperspectral Imaging

Grapevine leafroll disease (GLD) is considered one of the most widespread grapevine virus diseases, causing severe economic losses worldwide. To date, six grapevine leafroll-associated viruses (GLRaVs) are known as causal agents of the disease, of which GLRaV-1 and -3 induce the strongest symptoms. Due to the lack of efficient curative treatments in the vineyard, identification of infected plants and subsequent uprooting is crucial to reduce the spread of this disease. Ground-based hyperspectral imaging (400–2500 nm) was used in this study in order to identify white and red grapevine plants infected with GLRaV-1 or -3. Disease detection models have been successfully developed for greenhouse plants discriminating symptomatic, asymptomatic, and healthy plants. Furthermore, field tests conducted over three consecutive years showed high detection rates for symptomatic white and red cultivars, respectively. The most important detection wavelengths were used to simulate a multispectral system that achieved classification accuracies comparable to the hyperspectral approach. Although differentiation of asymptomatic and healthy field-grown grapevines showed promising results further investigations are needed to improve classification accuracy. Symptoms caused by GLRaV-1 and -3 could be differentiated.

Efficiency of chemotherapy combined with thermotherapy for eliminating grapevine leafroll-associated virus 3 (GLRaV-3)

Grapevine leafroll-associated virus 3 (GLRaV-3) is distributed in all grapevine-growing regions worldwide, resulting in significant economic losses. In this study, ribavirin (15 and 25 μg/mL, R15 and R25), high temperature (37 °C, T), and a combined treatment (R15 + T and R25 + T) were used to eliminate the virus from in vitro GLRaV-3-infected Vitis champini plantlets. The phytotoxicity of ribavirin on plantlets was found in R15 and R25 lasted 30 days, but the damage of high temperature on grapevine plantlets was found in T, R15 + T and R25 + T at the beginning of treatment. Moreover, high temperature could enhance the growth of grapevine plantlets. The survival rate was lowest for the regenerated T plants (23.3%). In contrast, the survival rates for the regenerated plants that underwent the chemotherapy (R15 and R25) and the combined treatment (R15 + T and R25 + T) were 47.0% (31/66) and 41.0% (50/122), respectively. Reverse transcription polymerase chain reaction and quantitative real-time PCR assays were used to evaluate the efficiency of the GLRaV-3 elimination in the regenerated grapevine plants. The combined treatment was most effective for eliminating GLRaV-3. Specifically, GLRaV-3 was detected in all regenerated plants after the 40-day R15 and R25 treatments, whereas the virus elimination rates for the 20-day R15 + T and R25 + T treatments exceeded 80%. Additionally, the virus elimination rate for the regenerated T plants was 42.9% for the same treatment periods.