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…

How small amounts of oxygen introduced during bottling and storage can influence the metabolic fingerprint and SO2 content of white wines

The impact of minute amounts of headspace oxygen on the post-bottling development of wine is generally considered to be very important, since oxygen, packaging and storage conditions can either damage or improve wine quality. This is reflected in the generalised use of inert bottling lines, where the headspace between the white wine and the stopper is filled with an inert gas. This experiment aimed to address some open questions about the chemistry of the interaction between wine and oxygen, crucial for decisions regarding optimal closure. While it is known that similar amounts of oxygen affect different wines to a variable extent, our knowledge of chemistry is not sufficient to construct a predictive method.

Green berries on Gewürztraminer (Vitis vinifera L.) in South Tyrol (Italy)

The grape variety Gewürztraminer is known to be affected by two physiological disorders namely berry shrivel and bunch stem necrosis. During the season 2014 we noticed a new symptomatology type of ripening disorder on the variety. The new symptom showed not all berries fallowing the normal maturation stages, but single berries remaining at a soft but green stage till harvest. The broad distribution of these so called “green berries” symptoms in different production sites of our region, caused huge damage due to the difficulty of eliminating single berries per bunch before harvesting. Therefore, the Research Centre Laimburg began to investigate the reasons and origins of this new symptom. This work shows the results of first attempts to find causes for the symptom as well as the resulting approach to mitigate symptoms. Applications of magnesium leaf fertilizer showed first promising results against this putative disorder. To study the causal effect of the green berries 30 symptomatic vineyards in 2014 have been selected for a monitoring during the season 2016. To evaluate the foliar nutrient treatment two vineyards have been selected for application of magnesium sulfate and magnesium chloride. Leaf and berry nutrient analysis, as well as the main quality parameters during ripening have been performed. As soon as “green berries” symptoms appeared, incidence and severity have been evaluated. Most of the symptomatic vineyards of the 2016 monitoring showed light to clear magnesium deficit symptoms on their foliage. Only during the seasons 2020 and 2021 “green berries” symptoms could be found in the leaf fertilizer treatment vineyards. Both seasons showed a significant effect of the magnesium treatments to reduce the incidence and severity of the symptom. It seems that the appearance of the “green berries” symptom on Gewürztraminer is correlated to a disturbed uptake of magnesium of the vines.

Sugar accumulation disorder Berry Shrivel – from current knowledge towards novel hypothesis

In contrast to fruit and grape berry ripening, the biological processes causing ripening disorders are often much less understood, although shriveling disorders of fruits are manifold and contribute to yield losses and reduced fruit quality worldwide. Shrinking berries are a common feature for all shriveling disorders in grapevine although their timing of appearance during the berry ripening process and their underlying induction processes distinct them from each other. The sugar accumulation disorder Berry Shrivel (BS) is characterized by a suppression of sugar accumulation short after veraison resulting in berries low in sugar content and anthocyanins in berry skins, while the organic acid content is similar. Recent studies analyzed the biochemical, morphological and molecular processes affected in BS berries and linked early changes to the period of ripening onset [1,2].

Influence of trellis system and shoot density in yield and grape composition of a vineyard of Cabernet Sauvignon, in warm climate

In vineyards grown in warm areas, it is usual that the stage of maturity of the grapes is fast and easily reach a high concentration of sugar and low acidity, but not a adequate phenolic maturation. The geometry of the trellis system and the shoot density can modify the microclimate of the cluster and, therefore, the maturation process.

An automated cooling system to mitigate thermal and radiative stresses in Pignoletto white grapes

In the context of increasingly hot and dry summers, the adoption of innovative irrigation technologies has become essential for maintaining grape production while minimizing water use.