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…

Physiological behavior of the Chasselas grape variety under water deficit: 30 years of experiments in Switzerland

In the context of increasingly hot and dry summers, the adoption of innovative irrigation technologies has become essential for maintaining grape production while minimizing water use.

Read More

First report to the fruit fly management in vine on Cafayate-Salta

In the present work it was proposed to evaluate the use of massive trapping for the control of the fruit fly complex that attacks the vine, using two types of food attractants; as well as determine the species and sex captured by them, and the level of crop damage. The adaptive research work was carried out in Tolombón- Cafayate- Salta- Argentina, during the 2015/2016, on a plot of 2 ha of grape variety TorrontésRiojano, conducted in horizontal roof.

Read More

Postharvest ozone treatment in grapevine white cultivars: Effects on grape volatile composition

During postharvest management, the metabolism of fruits remains active and continuous physico-chemical changes occur. Ozone treatment has an elicitor effect on secondary metabolites and the treatment conditions can influence the grape response to the stress (Bellincontro et al., 2017; Botondi et al., 2015). Regarding volatile organic compounds (VOCs), previous studies showed that ozone treatment during postharvest dehydration induces the biosynthesis of terpenes in Moscato bianco grapes (Río Segade et al., 2017). It is well known that grape VOCs greatly influence the organoleptic properties of wines, particularly terpenes in aromatic varieties.

Read More

Influence of nitrogen supply on colorimetric parameters of Lugana wines

Color is one of the main qualitative parameters of a wine. As a matter of fact, immediately after having opened a bottle of wine, color, even before aroma and taste, is the first sensorial parameter to be evaluated by the consumer It can change according to various factors depending on the characteristics of the grapes or on the different production and storage processes.

Read More

Les activités peroxidasiques du raisin de quelques cépages de Roumanie

Les enzymes d’oxydation (polyphénoloxydase, peroxydase) des raisins sont d’origine génétique dépendantes des facteurs climatiques et agrotechniques (Sapis et al, 1983). Dans le processus technologique de l’obtention du moût de raisins, ces enzymes catalysent l’oxydation de certains composés phénoliques naturellement présents dans le raisin, produisant ainsi des modifications indésirables de la couleur et de l’arôme du vin.

Read More