Response of Vitis vinifera cell cultures to Phaeomoniella chlamydospora: changes in phenolic production, oxidative state and expression of defence-related genes

Abstract

Cell suspension cultures of Vitis vinifera cv. Vinhão (Vv) were used to study the putative response of V. vinifera to Phaeomoniella chlamydospora (Pc), a fungus frequently associated with esca and grapevine decline. Cells were elicited with a Pc autoclaved biomass extract and methyl jasmonate (MeJ). Phenolic production was evaluated by HPLC-DAD and HPLC-MS/MS. Phenolic production of Vv cells significantly changes after elicitation. Compared to control, Vv cells elicited by Pc extract increase their stilbene production 20-fold and those elicited by MeJ increase stilbenic production 9-fold. In both cases, there is de novo production of viniferin type compounds. We also analyzed the oxidative burst of Vv cells after elicitation with Pc extract and MeJ, using the probe 2′,7′-dichlorodihydrofluorescein diacetate. Adding Pc extract induces an oxidative burst that shows a biphasic pattern in Vv cells. Moreover, the induction of 7 defence-related genes expression in Vv cell cultures upon Pc extract elicitation was investigated employing semi-quantitative RT-PCR. Elicitation increases the expression of class 6 and class 10 pathogenesis-related proteins, β-1,3-glucanase, class III chitinase, lipoxygenase, phenylalanine ammonia lyase and stilbene synthase. Therefore, Vv in vitro cell cultures could be an important tool to study esca disease, since they offer a simple, rapid and selective way to evaluate plant/fungus interactions.

Response of Vitis vinifera cell cultures to Eutypa lata and Trichoderma atroviride culture filtrates: expression of defence-related genes and phenotypes

“…There was a biphasic pattern in the upregulation of PAL and STS transcripts in the cell cultures treated with the pathogen elicitor indicating a coordinated expression of genes of the phenylpropanoid pathway. A biphasic pattern in the expression of PAL and STS was also reported in grapevine cell cultures in response to cell wall extracts of P. chlamydospora (Lima et al 2012). However, the biphasic response could be due to activity of two or more elicitors, all present simultaneously, since in both the current Fig.…”

“…The synthesis of stilbenes is induced through the induced systemic resistance (ISR) pathway involving signalling molecules jasmonic acid and ethylene (Belhadj et al 2008;Lijavetzky et al 2008). Lima et al (2012) showed that grapevine cell cultures respond to methyl jasmonate (methyl ester of jasmonic acid) in a similar pattern as they respond to P. chlamydospora. The application of Trichoderma harzianum to pruning wounds of kiwi fruit (Actinidia deliciosa) has been demonstrated to increase the wood phenolic compounds content which aids wound protection against wood pathogens Phaeacremonium aleophilum and Fomitiporia mediterranea (Neri et al 2008).…”

“…An understanding of these early defence responses would be critical for understanding disease progression and be useful in cultivar comparative studies aimed at the development of pathogen-resistant or pathogen-tolerant cultivars. Cell suspension cultures provide a useful and reproducible model system for the study of early grapevine response to infection, as demonstrated by Lima et al (2012) and Bénard-Gellon et al (2014). These authors showed the usefulness of cell suspension cultures in evaluating the response of grapevine to Phaeomoniella chlamydospora, which is another grapevine trunk pathogen.…”

“…Lima et al (2012) studies, crude extracts (culture filtrate and fungal biomass, respectively) were used for the elicitation of cell cultures. Characterisation of the fungal extracts either by treating the elicitors with proteases, to degrade protein fractions, or cell wall-degrading enzymes, to disrupt the cell wall fragments, could clarify the number and type of elicitors.…”

Morphological and transcript changes in the biosynthesis of lignin in oil palm (Elaeis guineensis) during Ganoderma boninense infections in vitro

“…COMT activities in T3 plantlets were reduced on 4 DPI from (117.24 to 49.93 μg ml −1 ferulic acid equivalents). Comparable biphasic inductions of COMT enzymes were observed in tobacco mosaic virus‐infected tobacco leaves (Toquin et al ) and Phaeomoniella chlamydospora ‐induced grape culture (Lima et al ). The induction pattern of COMT activity in planta upon pathogen infection could be explained as a characteristic of the hypersensitive response (Zurbriggen et al ).…”