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…

Insight on Lugana flavor with a new LC-MS method for the detection of polyfunctional thiols

The analysis of polyfunctional thiols in wine is challenging due to their low abundance and instability within a complex matrix. However, volatile thiols are highly aroma-active, making their accurate quantification in wine at low concentrations crucial [1].

Functional characterisation of genetic elements regulating bunch morphology in grapevine

Vitis vinifera L., is considered one of the world’s most important cultivated fruit crops. In agriculture, bunch morphology is a grapevine-specific trait, which directly impacts fruit quality and health.
Bunch size, shape, and compactness are major aspects of bunch morphology, with the degree of compactness emerging as an important trait for grapevine genetic enhancement and vineyard management. The importance of this trait stems from its impact on disease susceptibility, berry ripening, and other grape quality properties. However, current knowledge of the genes controlling it remains limited.

Genome editing applications on grapevine cv. Aglianico for the knockout of susceptibility genes related to fungal diseases

Context and purpose of the study. Italy hosts diverse grapevine varieties crucial for viticultural biodiversity. Preserving this biodiversity is essential for maintaining a diversified genetic pool and addressing future challenges such as climate change and emerging plant diseases.

Discrimination of monovarietal Italian red wines using derivative voltammetry

Identification of specific analytical fingerprints associated to grape variety, origin, or vintage is of great interest for wine producers, regulatory agencies, and consumers. However, assessing such varietal fingerprint is complex, time consuming, and requires expensive analytical techniques. Voltammetry is a fast, cheap, and user-friendly analytical tool that has been used to investigate and measure wine phenolics.

Under-vine management effects on grapevine vegetative growth, gas exchange and rhizosphere microbial diversity

The use of cover crops under the vines might be an alternative to the use of herbicides or tillage, improving grapevine quality and soil characteristics. The aim of this research was to study the implications of different management strategies of the soil under the vines (herbicide, cultivation or cover crops) on grapevine growth, water and nutritional status, gas exchange parameters and belowground microbial communities.
The experimental design consisted in 4 treatments applied on 35L-potted Tempranillo vegetative grapevines with 10 replicates each grown in an open-top greenhouse in 2022 and 2023. Treatments included two cover crop species (Trifolium fragiferum and Bromus repens), herbicide (glyphosate al 36%) and an untreated control.