terclim by ICS banner
IVES 9 IVES Conference Series 9 Optimizing protocol for a rapid and cost effective DNA isolation for Marker Assisted Selection pipeline

Optimizing protocol for a rapid and cost effective DNA isolation for Marker Assisted Selection pipeline

Abstract

Grapevine is a plant that holds significant socioeconomic importance due to its production of grapes for fresh consumption, wines, and juices. However, climate changes and susceptibility to diseases pose a threat to the quality and yield of these products. The breeding of new genotypes that are resistant/tolerant to biotic and abiotic stresses is essential to overcome the impact of climate changes. In this regard, Marker-assisted selection (MAS), which uses DNA markers, is a crucial tool in breeding programs. The efficiency and economy of this method depend on finding rapid DNA isolation methods. In this study, we compared four different DNA extraction methods to choose the one that quickly isolates DNA from many young vine leaves samples in a single run. The methods used involved Lithium chloride, carboxyl coated magnetic beads, cetyltrimethylammonium bromide (CTAB), and a commercial kit called Red&extract. The results showed that the CTAB method was the best in terms of reliability of the procedure, yield of the extracted DNA, low quantity of inhibitors, and speed of the procedure. Improving the MAS technique will help identify plants containing genes involved in different types of stress and deepen the study of the resistance genes pyramided.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Poster

Authors

Marika Santamaria1,2, Antonella Salerno1,2, Flavia Angela Maria Maggiolini2, Margherita D’Amico2, Carlo Bergamini2, Maria Francesca Cardone2

1 Department of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”, Via Orabona 4, 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, Marker Assisted Selection, DNA isolation, breeding

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Evidence for terroir effect associated with botrytisation relatively to compounds implicated in typical aromas of noble rot sweet wines

Recent studies have demonstrated the role of certain lactones, particularly 2-nonen-4-olide, and volatile thiols (3-sulfanylhexan-1-ol) in the over ripped aromas of noble rot sweet wines (Stamatopoulos et al. 2014ab). These compounds are partly formed during the maturation and under the activity of B. cinerea on grapes. This research was carried out in the vineyard of Sauternes with aim to better understand their genesis depending on the grape over-ripening on two different soil types during 3 vintages. Thus, the study was conducted, with the Sémillon grape, during vintages 2012, 2014 & 2015, at 4 stages of over-maturation of the grapes (healthy, pourri plein, pourri roti, pourri roti + 15 days) considering two vineyard plots with different soil characteristics (calcosol & peyrosol) planted with the 315 Sémillon clone and grafted on 101-14 rootstock respectively in 1981 and 1980 and cultivated with the same vineyard management. Volatile lactones were assayed by liquid-liquid extraction followed by GC/MS analysis and the precursors of 3-sulfanylhexanol by an adaptation of the method by Capone et al. 2010 (SPE-
UPLC/FTMS).

Polyphenols in kombucha: impact of infusion time on extraction and investigation of their behavior during “fermentation”

Kombucha is a non-alcoholic beverage made of sugared tea that is transformed by a symbiotic consortium of yeasts and bacteria. Polyphenols are expected to be responsible of several health benefits attributed to kombucha consumption, among other metabolites. This study investigated the impact of tea infusion time and of kombucha “fermentation”, on total phenolic content,

Above and below: soil moisture and soil temperature interact to alter grapevine water relations

The combined effect of soil moisture and soil temperature on grapevine physiology is gaining interest in the context of global warming.

Is it relevant to consider remote sensing information for targeted plant monitoring?

An experiment was carried out to test the relevance of using satellite images (NDVI) to define locations of plant monitoring systems.

Topographic modeling with GIS at Serra Gaúcha, Brazil: elements to study viticultural terroir

Brazil is historically known at the international wine market as an importer, eventhough in the last decades there was an increase in quantity and quality of the internal production. Nowadays, about 40% of fine wines comsuption of the country are national ones. The main production region is called Serra Gaúcha, where the natural conditions are heterogeneous and viticulture is develloped in small properties, mainly done by the owners family.