terclim by ICS banner
IVES 9 IVES Conference Series 9 Open-GPB 9 Open-GPB-2024 9 Flash - New biotechnological tools 9 Deciphering the function and regulation of VviEPFL9 paralogs to modulate stomatal density in grapevine through New Genomic Techniques

Deciphering the function and regulation of VviEPFL9 paralogs to modulate stomatal density in grapevine through New Genomic Techniques

Abstract

Stomata are microscopic pores mainly located in leaf epidermis, allowing gas exchanges between plants and atmosphere. Stomatal initiation relies on the transcription factor SPEECHLESS which is mainly regulated by the MAP kinase cascade, in turn controlled by small signaling peptides, the Epidermal Patterning Factors (EPF and EPF-Like), namely EPF1, EPF2 and EPFL9. While EPF1 and EPF2 induce the inhibition of SPEECHLESS, their antagonist, EPFL9, stabilizes it, leading to stomatal formation. In grapevine, there are two paralogs for EPFL9, VviEPFL9-1 and VviEPFL9-2. Despite their structural similarity, it remains unclear whether they are differentially regulated and have distinct roles. In our study we showed that while VviEPFL9-1 is expressed only in the apex, VviEPFL9-2 is expressed both in the apex and in mature leaves along the plant axis and is significantly repressed by ABA. To support experimental data, an in-silico transcriptomic analysis has been carried out using publicly available datasets. In addition, both genes were functionally characterized using knock-out (KO) mutants generated via CRISPR/Cas9. Leaf stomatal density and gas exchange parameters were significantly different between ‘Sugraone’ WT plants and VviEPFL9-2 KO lines, whereas the differences were negligible between WT and VviEPFL9-1 KO lines. Moreover, a water stress experiment was carried out to deeply compare the physiology of edited lines and WT plants under drought conditions. Our results suggest that VviEPFL9 paralogs have distinct roles in determining stomatal plasticity during leaf growth, and that VviEPFL9-2 may be considered a key target to increase grapevine resilience to water deficiency.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Article

Authors

Umar Shahbaz1,2, Pierre Videau3, Katerina Labonova3, David Navarro-Payá4, Alvaro Vidal1,2, José Tomás Matus4, Mickael Malnoy1, Olivier Zekri3, Fabio Fiorani5, Michele Faralli2, Lorenza Dalla Costa1*

1 Research and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, Italy
2 Center Agriculture Food Environment (C3A), University of Trento, 38098, San Michele all’Adige, Italy
3 Novatech, Mercier Groupe, Le Champ des Noels, France
4 Institute for Integrative Systems Biology, Universitat de València-CSIC,46980 Paterna, Valencia, Spain
5 Institute of Bio- and Geo-Sciences, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany

Contact the author*

Keywords

Vitis vinifera, Epidermal Patterning Factors, CRISPR/Cas9, gas exchange, water stress

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Quelles cibles moléculaires pourraient expliquer l’effet du terroir sur la composition des baies en sucres et acides?

Le manque de connaissances concernant la physiologie de la maturation du raisin a longtemps interdit d’interpréter l’effet du terroir ou du millésime sur la qualité des vendanges en termes moléculaires. L’hypothèse selon laquelle c’est la perméabilité membranaire qui contrôlerait le sens comme l’intensité du stockage des acides est pourtant déjà ancienne (1). L’étude du transport des acides organiques et de son coût énergétique permet d’avancer certaines hypothèses concemant les sites potentiels de la régulation du contenu en sucres et acides du raisin sous l’effet de paramètres environnementaux.

Vine selection in France: An assessment after more than 60 years of work

It was at the end of the second world war that professor Branas laid the foundations of french vine selection. He was also behind the creation of domaine de vassal (1949) and antav (1962), which were to become the bridgeheads of the french strategy for the conservation, selection and multiplication of viticultural diversity. Initially based on visually virus-symptom-free massal selections, with the main aim of providing healthy, clearly-identified plant material, the process evolved as knowledge gained towards clonal selection.

First insights on the intra-species diversity in V. berlandieri: environmental adaptation and agronomic performances when used as rootstock

In grafted plants, such as grapevine, increasing the diversity of rootstocks available to growers is an ideal strategy to get adaptation to climate change. The rootstocks used for grapevine are hybrids of various American Vitis, including V. berlandieri. The rootstocks currently used in vineyards are derived from breeding programs involving very small numbers of parental individuals.

Composition and molar mass distribution of different must and wine colloids

A major problem for winemakers is the formation of proteinaceous haze after bottling. Although the exact mechanisms remain unclear, this haze is formed by unfolding and agglomeration of grape proteins, being additionally influenced by numerous further factors.

Differences in the chemical composition and “fruity” aromas of Auxerrois sparkling wines from the use of cane and beet sugar during wine production.

The main objective of this study was to establish if beet sugar produces a different concentration of “fruity” volatile aroma compounds (VOCs), compared to cane sugar when used for second alcoholic fermentation of Auxerrois sparkling wines. Auxerrois base wine from the 2020 vintage was separated into two lots; half was fermented with cane sugar and half with beet sugar (both sucrose products and tested for sugar purity). These sugars were used in yeast acclimation (IOC 2007), and base wines for the second fermentation (12 bottles each). Base wines were manually bottled at the Cool Climate Oenology and Viticulture Institute (CCOVI) research winery. The standard chemical analysis took place at intervals of 0, 4 weeks, and 8 weeks post-bottling. Acidity and pH measurements were carried out by an auto-titrator. Residual Sugar (g/L) (glucose (g/L), fructose (g/L)), YAN (mg N/L), malic acid, and acetic acid (g/L) were analyzed by Megazyme assay kits. parameters were analyzed by Megazyme assay kits. Alcohol (% v/v) was assessed by GC-FID. VOC analysis of base wines, finished sparkling wines, as well as the two sugars in model sparkling wine solutions, was carried out by GC-MS. VOCs included ethyl octanoate, ethyl hexanoate, ethyl butanoate, ethyl decanoate, ethyl-2-methylbutyrate, ethyl-3-methylbutyrate, ethyl 2-methyl propanoate, ethyl 2- hydroxy propanoate, 1-hexanol, 2-phenylethan-1-ol, ethyl acetate, hexyl acetate, isoamyl acetate and 2-phenylethyl acetate.