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IVES 9 IVES Conference Series 9 OIV 9 OIV 2024 9 Orals - Viticulture, table grapes, dried grapes and unfermented grape products 9 An efficient protocol for long-term maintenance of embryogenic calluses of Vitis vinifera

An efficient protocol for long-term maintenance of embryogenic calluses of Vitis vinifera

Abstract

New breeding techniques (NBTS) could play a significant role in the genetic improvement of grapevine by producing new grape varieties with improved quantitative and qualitative characteristics. However, the application of these new techniques faces some technical challenges. One of the challenges is the generation of embryogenic calluses, which are not only difficult to obtain but it is also difficult to maintain their competence during in vitro cultivation, and thus regenerate plants without defects. We have developed a protocol for improving the production of embryogenic calluses by testing three substances: citric acid (CA), silver thiosulphate (STS), and salicylic acid (SA). Ca is an antioxidant, STS is an ethylene inhibitor, while SA is an elicitor with different effects depending on the concentration of use beyond the ethylene inhibitor activity. These three substances were tested on callus cultures obtained from different grapevine varieties, with or without embryogenic competence, showing different responses to the treatments. The STS and ac treatments generally enhanced the embryogenic competence of the calluses, while at the same time having a growth-depressing effect on the non-embryogenic calluses. The effect of SA was more dose-dependent and produced different effects. We found that among the tested varieties, in some cases the highest tested concentration of SA showed a decrease in the growth of both embryogenic and non-embryogenic calluses, while for other varieties, it only decreased the growth of embryogenic calluses. Our results show how CA, STS, and SA treatments showed promising results in enhancing embryogenic competence, even if they showed a variety-dependent outcome. The regeneration of embryos from callus after these treatments is currently under evaluation in our laboratories. Our results highlight the importance of testing the efficacy of different treatments on multiple grapevine varieties to identify the most effective strategies for increasing the production of embryogenic calluses, which is a fundamental aspect for NBTS applications.

Un protocollo efficiente per il mantenimento a lungo termine dei calli embriogenici di Vitis vinifera

Le new plant techniques (NBTS) potrebbero svolgere un ruolo significativo nel miglioramento genetico della vite producendo nuove varietà di uva con caratteristiche quantitative e qualitative migliorate. Tuttavia, l’applicazione di queste nuove tecniche comporta il superamento di alcune problematiche tecniche. Una prima difficoltà è la produzione di calli embriogenici, che non solo sono difficili da ottenere ma di cui è anche difficile mantenere la competenza durante la coltura in vitro per poter poi rigenerare piante senza difetti. Abbiamo sviluppato un protocollo per migliorare la produzione di calli embriogenici testando tre sostanze, l’acido citrico (AC) il tiosolfato d’argento (STS) e l’acido salicilico (AS). Ac è un antiossidante, STS è un inibitore dell’etilene, mentre as è un elicitore con effetti diversi a seconda della concentrazione impiegata oltre ad avere attività di inibizione dell’etilene. Queste tre sostanze sono state testate su colture di callo ottenute da diverse varietà di vite, con o senza competenza embriogenica, mostrando diverse risposte ai trattamenti. Il trattamento con AC e STS generalmente ha migliorato la competenza embriogenica dei calli, mentre allo stesso tempo ha un avuto un effetto di depressione della crescita sui calli non embriogenici. L’effetto dell’as è stato più dose-dipendente ed ha prodotto effetti diversi per le differenti varietà. Abbiamo scoperto che tra le varietà testate, in alcuni casi la più alta concentrazione testata di sa ha mostrato una diminuzione nella crescita sia dei calli embriogenici che non embriogenici, mentre per altre varietà ha solo diminuito la crescita dei calli embriogenici. I nostri risultati mostrano come i trattamenti ac, sts che quelli as abbiano mostrato risultati promettenti nel migliorare la competenza embriogenica, anche se hanno evidenziato un risultato dipendente dalla varietà. La rigenerazione degli embrioni dal callo dopo questi trattamenti è attualmente in fase di valutazione nei nostri laboratori. I nostri risultati evidenziano l’importanza di testare l’efficacia di diversi trattamenti su più varietà di vite per identificare le strategie più efficaci per aumentare la produzione di calli embriogenici, che è un aspetto fondamentale per le NBTS.

Un protocolo eficiente para el mantenimiento a largo plazo de callos embriogénicos de Vitis vinifera

Las new breeding techniques (NBTS) podrían desempeñar un papel importante en la mejora genética de la vid al producir nuevas variedades de uva con características cuantitativas y cualitativas mejoradas. Sin embargo, la aplicación de estas nuevas técnicas enfrenta algunos desafíos técnicos. Uno de los desafíos es la generación de callos embriogénicos, que no sólo son difíciles de obtener sino que también mantienen su competencia durante el cultivo in vitro, y así regenerar plantas sin defectos. Hemos desarrollado un protocolo para mejorar la producción de callos embriogénicos probando tres sustancias, acido citico (AC), tiosulfato de plata (STS) y ácido salicílico (SA). AC es un antioxidante, sts es un inhibidor de etileno, mientras que sa es un inductor con diferentes efectos dependiendo de la concentración de uso más allá de la actividad inhibidora de etileno. Estas tres sustancias fueron probadas en cultivos de callos obtenidos de diferentes variedades de vid, con o sin competencia embriogénica, mostrando diferentes respuestas a los tratamientos. Lss tratamientos AC y STS generalmente mejoró la capacidad embriogénica de los callos, al mismo tiempo que tuvo un efecto depresor del crecimiento en los callos no embriogénicos. El efecto de sa dependió más de la dosis y produjo efectos diferentes. Descubrimos que entre las variedades probadas, en algunos casos la concentración más alta de sa mostró una disminución en el crecimiento de callos embriogénicos y no embriogénicos, mientras que para otras variedades, solo disminuyó el crecimiento de callos embriogénicos. Nuestros resultados muestran cómo tanto los tratamientos ac, sts como sa mostraron resultados prometedores en la mejora de la competencia embriogénica, incluso si mostraron un resultado dependiente de la variedad. Actualmente se encuentra en evaluación en nuestros laboratorios la regeneración de embriones a partir de callos tras estos tratamientos. Nuestros resultados resaltan la importancia de probar la eficacia de diferentes tratamientos en múltiples variedades de vid para identificar las estrategias más efectivas para aumentar la producción de callos embriogénicos, que es un aspecto fundamental para las aplicaciones de NBTS.

Publication date: November 18, 2024

Issue: OIV 2024

Type: Article

Authors

Lucia Rosaria Forleo¹, Bruna Suriano¹, Flavia Angela Maria Maggiolini¹, Margherita D’Amico¹, Annalisa Prencipe², Teodora Basile¹, Riccardo Velasco¹, Maria Francesca Cardone¹, Carlo Bergamini¹

¹ CREA Viticoltura ed Enologia, Via Casamassima, 148, Turi, Italy
² Università degli Studi di Bari, Italy

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Tags

IVES Conference Series | OIV | OIV 2024

Citation

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