ABSTRACT
Previous works on grapevine-trunk diseases indicate that minimal or non-pruning of the grapevine under certain circumstances can significantly reduce the risk of symptom expression. Nevertheless, knowledge of the mechanisms behind these observations are limited. Therefore, it was the aim of this study to investigate in more detail the effect of pruning intensity on the grapevine trunk by means of trunk integrity and the fungal community in the wood tissue. Two German vineyards partially trained in vertical-shoot position and semi-minimally pruned hedges were chosen for this survey due to the accessibility of multi-annual esca-monitoring data. The results revealed that only in one of the two vineyards was the incidence of external esca symptoms significantly reduced over a period of five years (2017-2021) by minimal pruning, which was up to 73.7% compared to intensive pruning. In both vineyards, the trunks of intensively pruned vines not only had more pruning wounds on the trunk (by 86.0% and 72.9%, respectively) than minimally pruned vines, but also exhibited a larger (by 19.3% and 14.7%, respectively) circumference of the trunk head. In addition, the percentage of white rot and necrosis in the trunks of esca-positive and esca-negative vines was analyzed and compared between the two pruning intensities; hereby, significant differences were only found for esca-negative 'Dornfelder' vines, in which the proportion of necrosis was higher for intensively pruned vines (23.0%) than for minimally pruned vines (11.5%). The fungal communities of the differently pruned vine trunks were mainly dominated by four genera, which are also associated with GTDs: Diplodia, Eutypa, Fomitiporia and Phaeomoniella. All in all, the fungal diversity and community composition did not differ between minimally and intensively pruned, esca-positive vines.
ABSTRACT
RNA interference (RNAi) was established in Nicotiana benthamiana plants by introducing constructs containing a defective interfering (DI) sequence from Tomato bushy stunt virus (TBSV) in combination with a conserved sense-sequence from the target Grapevine fanleaf virus (GFLV). Silencing in plants was confirmed by Agrobacterium-mediated infiltration of a GFP-sensor containing the GFLV-derived target sequence. The transgene-induced RNAi led to silencing of the GFP-sensor and GFP fluorescence was absent post-infiltration. In plants without GFP fluorescence after infiltration with the GFP-sensor, siRNA specific to GFP and the target virus sequence could not be detected. In contrast, infiltrated leaves of wild type and transgenic plants showing GFP fluorescence after infiltration revealed accumulation of siRNA specific to the sequence of the sensor. Silencing could be inhibited by co-infiltration using a p19 silencing suppressor construct together with the GFP-sensor, which always resulted in bright GFP fluorescence. In parallel, virus resistance of transgenic Nicotiana benthamiana was investigated via challenge inoculation with GFLV. Our results indicate that efficient RNAi in transgenic plants does not necessarily lead to a detectable accumulation of siRNA.
