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…

Biodiversity and biocontrol ability of Trichoderma natural populations in soil vineyards from Castilla y León region (Spain)

Trichoderma is a microorganism present in many agricultural soils and some of its species could be used as natural biological control agents. In this work, the presence of natural populations of Trichoderma was estimated in soil vineyard and its biocontrol capacity against Phaeoacremonium minimum, one of the main agent causals of grapevine trunk diseases instead of using pesticides. Moreover, physicochemical variables in soil such as pH, organic matter and nutrients were evaluated to determine a possible correlation to natural populations of Trichoderma.

CHEMICAL DRIVERS OF POSITIVE REDUCTION IN NEW ZEALAND CHARDONNAY WINES

According to winemakers, wine experts and sommeliers, aromas of wet stone, mineral, struck match and flint in white wines styles, such as those produced from Vitis vinifera L. cv. Chardonnay, are considered to be hallmarks of positive reduction.1,2 In recent years, the production of Chardonnay styles defined by aroma characteristics related to positive reduction has become more desirable among wine experts and consumers. The chemical basis of positive reduction is thought to originate from the concentration of specific volatile sulfur compounds (VSCs), including methanethiol (MeSH) imparting mineral and chalk notes,3 and benzenemethanethiol (BMT) responsible for struck match and flint.1,4

Analysis of peptide fraction from white wines

Among nitrogen compounds included in white wines, the peptide fraction is certainly the least studied, however this fraction is quantitatively the most important (Feuillat, 1974). Existing studies concern the fraction below 1 kDa and only for white and sparkling wines (Bartolomé et al, 1997, Desportes et al 2000). In this report, we have developed methods to isolate peptides from reference white wines. Then, we have applied this methodology with bitter wine to answer a research question: is there a relation between peptides and the bitterness of white wine as for some cheese for example (Furtado, 1984)?

The use of rootstock as a lever in the face of climate change and dieback of vineyard

As viticulture faces challenges such as climate change or vineyard dieback, the choice of the variety and rootstock becomes more and more crucial. To study rootstock levers in the Bordeaux region, a parcel of Cabernet Sauvignon (CS) was planted with four rootstocks in 2014. Twenty repetitions of each of the following four rootstocks were set up: 101-14 MGt, Nemadex AB, 420A MGt and Gravesac. The number of bunches, yields and pruning weights of the vine shoots were measured individually on 240 vines from 2017 to 2021. Since 2020, nitrogen status assessed by assimilable nitrogen level, hydric status assessed by δ13C and berry maturity were measured on 80 samples taken from 20 repetitions of the four rootstocks. A lower yield was measured for CS grafted onto Nemadex AB due to the lower number of bunches and the lower weight of berries. The differences between the other three rootstocks are small, but CS grafted onto 420A MGt was the most productive. The CS grafted onto Nemadex AB had the lowest pruning weight while 101-14 MGt had the highest. In 2020, δ13C showed a more moderate water stress with 101-14 MGt and 420A MGt than with Nemadex AB. Surprisingly, the Gravesac was under more stress than the 101-14 MGt. The nitrogen status in the berries was better for Nemadex AB but this was perhaps due to the significantly lower weight of the berries.Rootstock 101-14 MGt attained the highest accumulation of sugars in the berries while 420A MGt allows to preserve higher acidity. The parcel is still young which may explain some of the results. These measures must therefore be continued over the next several years to fully assess the effects of these rootstocks on the development of the vines and the quality of the production under new climatic conditions.

Can grapevine tolerance to bunch rot be directly induced by groundcover management?

Botrytis bunch rot occurrence is the most important limitation for the wine industry in humid environments. The effect of grapevine vegetative growth on bunch rot expression results from direct effects (cluster architecture, nitrogen status among others) and indirect ones (via microclimate). Previous studies of our group showed strong differences in bunch rot incidence between floor management treatments: cover crop (CC) vs weed-free strips under the trellis with herbicide (H). We observed that in some circumstances this reduction in bunch rot incidence occurred without major vine growth differences among treatments. The aim of the present study was to test the general hypothesis that other factors unrelated to grapevine vegetative expression could be more relevant to grapevine susceptibility to bunch rot.