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…

Metabolomic profiling of botrytized grape berries: unravelling the dynamic chemical transformations during noble rot

Botrytis cinerea, a fungal pathogen commonly known as grey mold, which under specific climatic conditions can develop into a desirable form known as noble rot. In this process the fungus penetrates the grape skin, allowing water evaporation and concentration of sugars and flavors, while profoundly affects the metabolite composition of grapes, leading to the production of unique and desirable compounds in the resulting wines. The result is a unique and complex wine with a luscious sweetness, heightened aromatics, and a distinct character.

Evaluation of Valdadige DOC “Terra dei Forti” vineyards by zoning approach

La conoscenza dell’interazione genotipo x ambiente e pertanto della caratterizzazione territoriale è di prioritaria importanza nella valutazione dei siti. Grazie alla combinazione di dati GIS

Comparing different vineyard sampling densities and patterns for spatial interpolation of intrinsic water use efficiency

The need to rationalize agricultural inputs has recently increased interest in assessing vineyard variability in order to implement variable rate input applications, so-called ‘precision viticulture’. In many viticultural areas globally, precision viticulture is already widely used such as for selective harvesting and variable rate application (VRA) of inputs such as irrigation and/or fertilizer. Robust VRA relies on having a geostatistically accurate map (of one or more vineyard attributes) requiring high sampling densities, which can be cost- and time-prohibitive to obtain. Previous work on spatial interpolation using kriging have upscaled ground-based measurements, but such upscaling strategies are applicable only when vineyard conditions are spatially continuous and satisfies the assumption of second-order stationary processes. Alternatively, mixed models that combine kriging and auxiliary information, such as the regression kriging (RK) method, are more instructive for spatial predictions. In order to improve prediction accuracies, it is therefore necessary to incorporate additional information to achieve accurate spatial patterns with low error.

Protection of grapevines from red blotch by understanding mechanistic basis of its infection

Currently, grapevine is host to a large number of pathogenic agents, including 65 viruses, five viroids and eight phytoplasmas. Needless to say, these pathogens, especially viruses responsible for several ‘infectious degeneration’ or ‘decline’ cause great distress to wine makers and grape growers, let alone the large economic losses incurred by the wine industry. A recent addition to this wide repertoire of grapevine viruses is a new viral disease known as Red Blotch in viticulture parlance. Its causal organism, Grapevine red blotch associated virus (GRBaV), discovered in 2008 is a newly identified virus of grapevines and a putative member of a new genus within the family Geminiviridae.

Effect of malolactic fermentation in barrels or stainless steel tanks on wine composition. Influence of the barrel toasting

Ellagitannin, anthocyanin and woody volatile composition of Cabernet Sauvignon wines aged in oak barrels for 12 months was evaluated. Depending on the container where malolactic fermentation (MLF) was carried out, two wine modalities were investigated: wines with MLF carried out in stainless steel tanks and barrel-fermented wines. Three toasting methods (medium toast, MT; medium toast with watering, MTAA; noisette) were considered for ageing of each wine modality. Sensory analyses (triangle and rating tests) were also performed. Two-way ANOVA of the raw experimental data revealed that the toasting method and the container where MLF took place, as well as the interaction between both factors, have a significant influence (p < 0.05) on ellagitannin, anthocyanin and woody volatile profiles of Cabernet Sauvignon wines.