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…

Fertility assessment in Vitis vinifera L., cv. Alvarinho

The Portuguese wine production is characterized by wide yield fluctuations, causing considerable implications in the economic performance of this sector. The possibility of predicting the yield in advance is crucial as it enables preliminary planning and management of the available resources. The present work aims to study and evaluate two different techniques for the assessment of vine fertility. vineyards.

Read More

Diversity and internationalization of wine grape varieties: Evidence from a revised global database

Aim: To quantify the extent to which national mixes of wine grape varieties (in terms of vineyard bearing area) have become more or less diversified, and ‘internationalized’, since wine globalization accelerated from the 1990s.

Read More

Les sols du cru de Bonnezeaux, Thouarcé, Anjou, France

Le cru de Bonnezeaux est une des appellations prestigieuses des vins liquoreux et moelleux des Coteaux du Layon et sa réputation est ancienne. L’INAO a effectué sa délimitation en 1953. Le vignoble est situé au nord de la ville de Thouarcé et au sud du village de Bonnezeaux, le long du versant rive droite du Layon, exposé au sud-ouest. La superficie du vignoble est de 156 ha.

Read More

UNCOVERING THE ROLE OF BERRY MATURITY STAGE AND GRAPE GENOTYPE ON WINE CHARACTERISTICS: INSIGHTS FROM CHEMICAL CHARACTERISTICS AND VOLATILE COMPOUNDS ANALYSIS

In a climate change context and aiming for sustainable, high-quality Bordeaux wine production, this project examines the impact of grape maturity levels in various cultivars chosen for their adaptability, genetic diversity, and potential to enhance wine quality. The study explores the effects on wine compo-sition and quality through sensory and molecular methods. We studied eight 14-year-old Vitis vinifera cv. grape varieties from the same area (VITADAPT plots 1 and 5): Cabernet Franc, Cabernet Sauvignon, Carmenère, Castets, Cot, Merlot, Petit Verdot, and Touriga Nacional.

Read More

Ozone treatment: a solution to improve sanitary and physiological quality of vine plant

The vineyard world is faced to a lot of fungal diseases. Grapevine Trunk Diseases (GTD) are some of the major. After exhibiting chronical foliar symptoms, grapevines can die by apoplexy within only few days. A range species of fungi was described to be associated with the apparition of early symptoms of GTD. It is well known that ozone dissolved into water is a powerful disinfectant with no remanence. The main goal of this study was to test the efficiency of this process on different fungal species associated with GTD in vitro and in planta conditions.

Read More