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…

From vineyard to bottle. Rationalizing grape compositional drivers of the expression of valpolicella aroma ‘terroir’

AIM: Valpolicella is a renowned Italian wine-producing region (Paronetto, 1981). Wines produced in its different sub-regions are anecdotally believed to be aromatically different, although there is no systematic study addressing the chemical bases of such diversity

Read More

Quality assessment of partially dealcoholized and dealcoholized red, rosé, and white wines: physicochemical, color, volatile, and sensory insights

The global non-alcoholic wine market is projected to grow from USD 2.7 billion in 2024 to USD 6.97 billion by 2034, driven by health awareness, lifestyle shifts, and religious factors [1-3]. Consequently, the removal of alcohol can significantly alter the key quality parameters of wine.

Read More

INTENSE PULSED LIGHT FOR VINEYARD WASTEWATER: A PROMISING NEW PROCESS OF DEGRADATION FOR PESTICIDES

The use of pesticides for vine growing is responsible for generating an important volume of wastewater. In 2009, 13 processes were authorized for wastewater treatment but they are expensive and the toxicological impact of the secondary metabolites that are formed is not clearly established. Recently photodecomposition processes have been studied and proved an effectiveness to degrade pesticides and to modify their structures (Maheswari et al., 2010, Lassale et al., 2014). In this field, Pulsed Light (PL) seems to be an interesting and efficient process (Baranda et al., 2017). Therefore, the aim of this work was to investigate the PL technology as a new process for the degradation of pesticides.

Read More

Organic mulches improve vine vigour, yield and physiological response in a semi-arid region

Recycled organic mulch within the row in vineyard floor management has become an interesting ecological strategy to adapt the crop to climate change consequences in semi-arid regions.
This study aimed to assess the impact of three recycled organic mulches [straw (STR), grape pruning debris (GPD), and spent mushroom compost (SMC)] and two conventional soil management practices [herbicide (HERB) and under-row tillage (TILL)] on vegetative vigour (NDVI), production (kg/plant), and physiological parameters (δ13C in grapes and leaf gas exchange during four grapevine phenology stages). Additionally, temperature and water soil parameters were collected at three soil depths. Data was collected during the 2021 and 2022 grapevine growing seasons in La Rioja, Spain.

Read More

Impact of red blotch disease on Cabernet Sauvignon and Merlot grape and wine composition and wine sensory attributes

Grapevine Red Blotch disease (RB) is a recently discovered disease that has become a major concern for the viticulture and winemaking industry in California, USA. The causal
agent, Grapevine Red Blotch Virus (GRBV) was identified in 2011 and its presence was confirmed in several states in the US, in Canada, and in Switzerland. It has been demonstrated that RB compromised the regulation of ripening by suppressing specific ripening events, altering the expression patterns of transcription factors and causing hormonal imbalances in Zinfandel.

Read More