terclim by ICS banner
IVES 9 IVES Conference Series 9 A versatile genome editing platform for grapevine: improving biotic and abiotic stress resilience 

A versatile genome editing platform for grapevine: improving biotic and abiotic stress resilience 

Abstract

New Plant Breeding Techniques (NPBTs) have arisen with the objective of surmounting the constraints inherent in conventional breeding methodologies, thereby enhancing plant resilience against both biotic and abiotic stresses. To date the application of genome editing in grapevine is still limited by the necessity to overcome recalcitrance to produce embryogenic calli and to regenerate plants. In our studies, we developed a smart and versatile genetic transformation system carrying all the most promising features of different genome editing approaches. In specific, we joined the GRF-GIF expression to improve regeneration, the systemic movement of the editing transcripts through tRNA-like sequences (TLS) and the cisgenic-like approach to remove transgenes. In parallel, we were able to assess the efficiencies of several guide RNAs (gRNAs) targeting genes with fall-out on drought stress and pathogen resilience. To the first aim, a gene belonging to glutathione S-transferase (VvGST40) and two gene belonging to pectin-methyl esterase (VvPME1 and VvPME3) have been targeted. Several edited lines were acclimatized and are currently under evaluation. In parallel, two genes belonging to the Mildew Locus-O (VvMLO6 and VvMLO7) and a non-expressor pathogenesis related gene (VvNPR3) were targeted to improve pathogen resilience. Acclimatized plants edited for MLO genes resulted almost resistant to Erysiphe necator (disease incidence reduction up to 80%), whereas NPR3-edited vines showed a significant reduction in disease severity (up to 70%). In conclusion, our approaches allowed to improve stress resilience of several economically-important genotypes such as Pinot noir, Chardonnay, Sangiovese and Glera but also widely used rootstocks (e.g., 110R and K5BB).

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Poster

Authors

Luca Nerva1*, Loredana Moffa1, Anna Narduzzo1, Ivan Bevilacqua1, Irene Perrone2, Chiara Pagliarani2, Giorgio Gambino2, Claudio Lovisolo3, Riccardo Velasco1, Walter Chitarra1

1 Research Centre for Viticulture and Enology, Council for Agricultural Research and Economics (CREA-VE), Via XXVIII Aprile 26, 31015 Conegliano (Italy)
2 Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR), Strada delle Cacce 73, 10135 Torino (Italy)
3 Department of Agricultural, Forest and Food Sciences (DISAFA), University of Torino, Largo Paolo Braccini 2, 10095 Grugliasco (TO), Italy

Contact the author*

Keywords

powdery mildew, climate change, recalcitrance, cisgenic-like approach, New Plant Breeding Techniques

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Towards a European data basis based of advanced multi-isotopic signatures and artificial intelligence: the wine in blue project

Major and trace elements are essential for the development of grapes used for the wine. They are primarily originating from the soil. Some elements are also seldomly added during the wine making process. Therefore, the largest spectrum of major, trace and ultra-trace elements in the final wine product is a good signature of its geographical origin. In the frame of the European tracewindu, we have developed a very original multi-isotopic dilution method using triple quadrupole icp/ms.

Variations of soil attributes in vineyards influence their reflectance spectra

Knowledge on the reflectance spectrum of soil is potentially useful since it carries information on soil chemical composition that can be used to the planning of agricultural practices. If compared with analytical methods such as conventional chemical analysis, reflectance measurement provides non-destructive, economic, near real-time data. This paper reports results from reflectance measurements performed by spectroradiometry on soils from two vineyards in south Brazil. The vineyards are close to each other, are on different geological formations, but were subjected to the same management. The objective was to detect spectral differences between the two areas, correlating these differences to variations in their chemical composition, to assess the technique’s potential to predict soil attributes from reflectance data.To that end, soil samples were collected from ten selected vine parcels. Chemical analysis yield data on concentration of twenty-one soil attributes, and spectroradiometry was performed on samples. Chemical differences significant to a 95% confidence level between the two studied areas were found for six soil attributes, and the average reflectance spectra were separated by this same level along most of the observed spectral domain. Correlations between soil reflectance and concentrations of soil attributes were looked for, and for ten soil traits it was possible to define wavelength domains were reflectance and concentrations are correlated to confidence levels from 95% to 99%. Partial Least Squares Regression (PLSR) analyses were performed comparing measured and predicted concentrations, and for fifteen out of 21 soil traits we found Pearson correlation coefficients r > 0.8. These preliminary results, which have to be validated, suggest that variations of concentration in the investigated soil attributes induce differences in reflectance that can be detected by spectroradiometry. Applications of these observations include the assessment of the chemical content of soils by spectroradiometry as a fast, low-cost alternative to chemical analytical methods.

Effect of the winemaking technology on the phenolic compounds, foam parameters in sparklig wines

Contribution Sparkling wines elaborated following the traditional method undergo a second fermentation in closed bottles of base wines, followed by aging of wines with lees for at least 9 months. Most of the sparkling wines elaborated are white and rosé ones, although the production of red ones is highly increasing. One of the initial problems in red sparkling wine processing is to obtain suitable base wines that should have moderate alcohol content and astringency and adequate color intensity; which is difficult to obtain when grapes must be harvested at low phenolic and industrial maturity stage. The low phenolic maturity degree in the red grapes makes essential to choose an adequate winemaking methodology to obtain the base wines because the extracted polyphenols will vary according the winemaking technique: carbonic maceration or destemmed-crushed grapes.

Successful technology transfer of the early defoliation technique in cv. Mandó, an autochthon variety from south-east Spain

In the old-world viticulture autochthonous varieties are an important inheritance because they can provide wines with authenticity and distinction. Cultivar Mandó is an almost extinguished variety from the south-east

Influence of cork density upon cork stopper resiliency after opening a sparkling wine bottle

After Champagne popping, the first consumer’s observation is the shape of the cork stopper. Consumers expect a “mushroom shape”. Nevertheless, we sometimes observe a “barrel” shape due to inappropriate cork’s elastic properties. The aim of this study was to follow the loss of cork stopper resiliency during 26 months according to the density (d) of the cork in contact with the wine. 1680 disks were weighed + measured and divided in 6 density classes: High (H1 d= 0,19 g/cm3 – H2 d= 0,21 g/cm3), Medium (M, not studied) and Low (L1 d= 0,13 g/cm3 – L2 d= 0,14 g/cm3). Then, 138 technical cork stoppers were produced for each of the 4 studied groups. These corks consisted of an agglomerated natural cork granule body to which two natural cork disks were glued. A total of 552 bottles of sparkling wine were closed with these corks and open after 13, 19 and 26 months to follow cork resiliencies. Wine bottles were stored horizontally; thus, the external natural cork disks were in contact to the wine. During the 26 months of the study, highly significant differences (ANOVA) were observed between the resiliencies of H-corks and those of L-corks, whatever the time studied. The diameters of the L-corks were statistically higher than those of the H-corks. No significant differences were observed between L1 and L2 corks. At the opposite, differences were noted between H1 and H2 at 19 and 26 months. This could be explained by the heterogeneity of the resiliency that was higher for H-corks than for L-corks. Finally, the corks were visually (12 judges) divided in 3 classes corresponding to high (expected mushroom shape, i.e high resiliency), medium (irregular shape of the disk in contact with the wine and/or low premature deterioration of the expected resiliency) and low qualities (barrel shape = premature deterioration of the resiliency). The corks were also divided in 3 categories corresponding to 0-33%, 34-66% and 67-100% resiliency. A strong correlation was noted between the visual and the instrumental categorizations. This study strongly evidenced 1) the importance of the cork density on the cork stopper behaviour when opening the bottle and 2) the interest of an instrumental approach reflecting the consumer’s perception.