terclim by ICS banner
IVES 9 IVES Conference Series 9 Optimized protocol for high-quality RNA extraction from grape tissues using sorbitol pre-wash

Optimized protocol for high-quality RNA extraction from grape tissues using sorbitol pre-wash

Abstract

Obtaining high-quality RNA from grape tissues, including berry pulp, berry skins, stems, rachis, or roots, is challenging due to their composition, which includes polysaccharides, phenolic compounds, sugars, and organic acids that can negatively affect RNA extraction. For instance, polyphenols and other secondary metabolites can bind to RNA, making it difficult to extract a pure sample. Additionally, RNA can co-precipitate with polysaccharides, leading to lower extraction yield. Also, sugars and organic acids can interfere with the pH and ionic properties of the extraction buffer. To address these challenges, we optimized a protocol for RNA isolation from grape tissues. Although commercial kits can provide a rapid extraction, they were inefficient for these plant materials. Similarly, protocols that work well for other vegetal tissues were also inefficient and time-consuming on grape tissues. To overcome these limitations, we added a sorbitol pre-wash step to both a three-day long protocol based on LiCl precipitation and a commercial kit. Our results showed that the addition of a sorbitol pre-wash improved multiple parameters: the A260/280 absorbance ratio, integrity and quality (IQ), and RNA integrity number (RIN). Sorbitol played a crucial role in ensuring high-quality RNA extraction from grape tissues. It inhibits RNase, thereby preserving RNA integrity and stability. It also helps in disrupting cellular membranes, facilitating the release of RNA, and maintains the osmotic pressure through hypertonicity, which is beneficial to RNA extraction. By using sorbitol, commercial kits can be used to extract RNA from challenging grape tissues, leading to an efficient and time-saving procedure.

Download the abstract

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Poster

Authors

Annalisa Prencipe1, Antonella Salerno1,2, Marco Vendemia2, Carlo Bergamini2, Margherita D’Amico2, Lucia Rosaria Forleo2, Teodora Basile2, Maria Francesca Cardone2, Antonio Domenico Marsico2, Riccardo Velasco2, Mario Ventura1, Flavia Angela Maria Maggiolini2*

1 Department of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”, 70125 Bari, Italy
2 Council for Agricultural Research and Economics – Research Center Viticulture and Enology (CREA-VE), Via Casamassima 148-70010 Turi (Ba), Italy

Contact the author*

Keywords

Vitis vinifera, RNA, sorbitol, extraction protocol

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

“Q & A” of the european commission for labeling and desalcoholization for wines: european wine “soft-law”?

Recently, the European Commission seems to have inaugurated a new mechanism for regulating the wine sector. Through two communications, articulated in the form of “Questions & Answers”, concerning the new rules for labeling (24.11.2023) and dealcoholization of wine (15.01.2024), the Commission is not simply “explaining” the new rules but, in an approach close to the theory of “Circulaire Normative” established in comparative law, chooses among different interpretations and even adds Praeter Legem constraints.

Read More

Impact of dried stems in winemaking on Veneto Passito wines

The use of stems during fermentation is generally avoided due to the herbaceous off-odors they can impart to the wine. [1].

Read More

Contribution of grape seeds to evolution of acetaldehyde, pigments and tannins reactive towards salivary proteins of red wine over time

This study investigated the impact of the gsk/gse ratio on the evolution of acetaldehyde and of major phenolic compounds of aglianico wine in wine like solution and real wine. Four model solutions and the correspondant control wines were prepared. The natural weight ratio between grape skins and seeds was determined on the real grapes, and a control wine was obtained from those.

Read More

Climate effect on ripening process in Vitis vinifera, L. cv. Cencibel

A seven years survey (2003 to 2009) has been carried out over old traditional vineyards cv. Cencibel in La Mancha region (Spain). Seven plots with more than 35 years old were sampled from veraison to harvest, measuring soluble solids (ºBaumé) and acid concentration (g/l in tartaric acid).

Read More

SENSORY CHARACTERIZATION OF COGNAC EAUX-DE-VIE AGED IN BARRELS REPRESENTING DIFFERENT TOASTING PROCESS

Cognac is an outstanding french wine spirit appreciated around the world and produced exclusively in the Nouvelle-Aquitaine region, and more precisely in the Cognac area. According to AOC regulations (Appellation D’origine Controlée), the spirit required at least 2 years of continuous ageing in oak barrels to be granted the title of Cognac. The oak wood will import color, structure and organoleptic complexity. The different steps during barrel-making process, such as seasoning and toasting, influence the above quality attributes in both wines and spirits.

Read More