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…

Effects of water and nitrogen uptake, and soil temperature, on vine development, berry ripening and wine quality of Cabernet-Sauvignon, Cabernet franc and Merlot (Saint-Emilion, 1997)

Wine quality depends largely on berry ripening conditions in relation to soil and climat. The influence of the soil has been studied in Bordeaux since the early Seventies (SEGUIN, 1970; DUTEAU et al., 1981; VAN LEEUWEN, 1991; VAN LEEUWEN et SEGUIN, 1994) and, more recently, in the Val de Loire (MORLAT, 1989), the Alsace (LEBON, 1993) and the Costières de Nîmes regions (MARTIN, 1995).

Read More

Qualitative and productive characterization of a minority variety: ‘Branco lexítimo’ in DO Ribeira Sacra (Spain)

The actual climate changes, together with the strong regulation of the European Union and Spanish government, in search of sustainable viticulture, have forced the recovery of minority varieties, expanding the range of grape varieties, as well as the possible development of wines with unique profiles. In the Ribeira Sacra DO (Spain), a comparative study of the agronomic and qualitative behavior of the ‘Branco lexítimo’ variety has been carried out, compared to the majority white variety in the DO: ‘Godello’, located in the same study plot, with identic soil and climatic conditions. The study contemplated the analysis of phenology and leaf water potential, as well as the productive results and the analysis of the must quality, during four seasons: 2018 – 2021.

Read More

Texas terroir: gis characterization of the texas high plains ava

The Texas High Plains AVA is one of eight officially recognized wine regions in Texas, established in 1993. Six local wineries, including the second-largest in Texas, are supported by approximately 50 vineyards, which are also major suppliers of grapes to Texas wineries outside the region.

Read More

Influence of social interaction levels on panel effectiveness in developing wine sensory profiles using consensus method

The development of sensory profiles is crucial for quality control and innovation in the wine industry. If quantitative descriptive analysis is the most commonly used method for establishing sensory profiles due to its robustness, it presents significant limitations.

Read More

Using multifactorial analysis to evaluate the contribution of terroir components to the oenological potential of grapes at harvest

The oenological potential of grapes at harvest depends on a combination of the major components of Terroir: the climate, the soil, the plant material, the training system and the crop management.

Read More