Preliminary steps of a protocol to isolate transcription factors bound to a specific DNA locus in grapevine using CRISPR-dCas9 system

UV-Visible spectroscopy in conjunction with chemometrics, was successfully applied to objectively differentiate sparkling wine press juice fractions of Pinot noir. Two measurements methods were applied: reflectance using a fibre optic probe in-line and transmission using a benchtop spectrophotometer.

The chemical composition of grape berries at harvest is one of the most important factors that should be considered to produce high quality wines. Among the different chemical classes which characterize the grape juice, the polyphenolic compound, such as flavonoids, contribute to the final taste and color of wines. Recently, an innovative non-destructive method, based on chlorophyll fluorescence, was developed to estimate the phenolic maturity of red grape varieties through the evaluation of anthocyanins accumulated in the berry skin. To date, only few data are available about the application of this method on white grape varieties.

Esca complex is a disease affecting grapevine trunks, characterized by the colonization of the wood by xylem-residing fungi (Phaeomoniella chlamydospora, Phaeoacremonium minimum and Fomitiporia mediterranea), and posing significant risks to vineyard longevity since no efficient treatment is available. Despite its prevalence, the mechanisms beyond symptomatic manifestations like interveinal chlorosis and leaf necrosis remain unclear. Preliminary findings indicated a more pronounced metabolic reprogramming in fruits compared to vegetative organs and a putative impact on wine quality by using fruits from symptomatic grapevines.

In this study macro and micro nutrients of plants (N = NH4 + NO3 , P, K, Ca, Na, Zn, Mn, Fe and Cu) were determined both in soil solution and bleeding sap and compared each other. Bleeding sap was collected from the nine varieties of grapevine Cvs. grafted on 5BB rootstock and grown in different soil conditions. For all varieties, plant nutrients content in bleeding sap as higher than in soil solution except for Ca and Na. While in soil solution Ca content was found at 10209 ppm, this value in bleeding sap was 49.20 ppm (Kozak Beyazy), 55.38 ppm (Trakya Ylkeren), 50.37 (Cardinal) and 74.27 ppm (Tekirdaô Çekirdeksizi) respectively. For the same varieties the Na values were as follows : 7.16 ppm (in soil solution) : 4.8, 3.23, 4.21,4.58 ppm (in bleeding sap) respectively. K content in bleeding sap was higher than in soil solution for a few varieties, and lower in some varieties. Traces of Fe and Cu were found in both media.

In table grapes seedlessness is a crucial breeding target, mainly results from stenospermocarpy, linked to the Thompson Seedless variety. Several studies investigated the genetic control of seedlessness identifying AGL11, a MADS-box transcription factor, as a crucial gene.
We performed a deep investigation of the whole AGL11 gene sequence in a collection of grapevine varieties revealing three different promoter-CDS combinations. By investigating the expression of the three AGL11 alleles and evaluating their ability to activate the promoter region, we show that AGL11 regulates its transcription in a specific promoter-CDS manner. By a multi-AGL11 co-expression analysis we identified a methyl jasmonate esterase, an indole-3-acetate beta-glucosyltransferase, and an isoflavone reductase as top AGL11 candidate targets. In vivo experiments further confirmed AGL11 role in regulating these genes, demonstrating its significant influence in seed development and thus in seedlessness trait.