Functional characterization of grapevine MLO genes to define their roles in Powdery mildew susceptibility by CRISPR/Cas9 genome editing

Volatile organic compounds (VOCs) constitute a diverse group of secondary metabolites key for the communication of plants with other organisms and for their adaptation to environmental and biotic stresses. The emission of these compounds through leaves is also affected by the interaction of plants with symbiotic microorganisms, arbuscular mycorrhizal fungi (AMF) among them [1]. Our objective was to know the concentration and profile of VOCs emitted by the leaves of two grapevine varieties (Tempranillo, T, and Cabernet Sauvignon, CS, grafted onto R110 rootstocks), inoculated or not with a consortium of five AMF (Rhizophagus irregularis, Funneliformis mosseae, Septoglomus deserticola, Claroideoglomus claroideum and C. etunicatum).

Carbaryl is an acetylcholine esterase inhibitor-type insecticide used for pest control on grapevine. We repeatedly observed the occurrence of interveinal leaf necrosis following carbaryl spray application in a Vitis rupestris x Vitis riparia F1 hybrid progeny vineyard. Spray applications induced necrosis in this progeny under both Missouri and New York field conditions an approximate one-to-one sensitive-to-insensitive segregation ratio and with 42% concordance. Results of subsequent in vitro experiments established causality between carbaryl treatment and leaf necrosis and confirmed the pattern of segregation observed in the field. We consistently map this phenotype to a major QTL on chromosome 16 of the female parent V. rupestris ‘B38’ regardless of whether we used field or in vitro-generated phenotype data.

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The presence of grape-derived heat unstable proteins can lead to haze formation in white wines [1], an instability prevented by removing these proteins by adding bentonite, a hydrated aluminum silicate that interacts electrostatically with wine proteins leading to their flocculation. Despite effective, using bentonite has several drawbacks as the costs associated with its use, the potential negative effects on wine quality, and its environmental impact, so that alternative solutions are needed.

Most of the effects of ultrasound (US) result from the collapse of bubbles due to cavitation. The shockwave produced is associated with shear forces, along with high localised temperatures and pressures. However, the high-speed stream, radical species formation, and heat generated during sonication may also affect the stability of some enzymes and proteins, depending on their chemical structure. Recently, Ce-lotti et al. (2021) reported the effects of US on protein stability in wines. To investigate this further, the effect of temperature (40°C and 70°C; 60s), sonication (20 kHz and 100 % amplitude, for 20s and 60s, leading to the same temperatures as above, respectively), in combination with Aspergillopepsins I (AP-I) supplementation (100 μg/L), was studied on unstable protein concentration (TLPs and chitinases) using HPLC with an UV–Vis detector in a TLPs-supplemented model system and in an unstable white wine.