Diversification of Vascular Occlusions and Crystal Deposits in the Xylem Sap Flow of Five Tunisian Grapevines

Xylem vessels are essential pivotal organs in bulk hydraulic flow through the whole woody plant. However, environmental constraints generate disagreements in xylem structures, which are characterized by air emboli and occlusions formations, compromising water conductivity in grapevines. The aim of this work was to explore xylem morphology dynamics through the xylem sap flow of five Tunisian grapevine cultivars during the natural bleeding sap periods of 2019, 2021, and 2022. In fact, Sakasly, Khamri, Hencha, Razegui1, and Razegui2 rain-fed grapevine cultivars revealed differential responses towards xylem sap movement. The results demonstrated that the xylem sap flow was significantly more abundant in 2019 than 2021 and 2022 bleeding sap campaigns. A variation was revealed between the cultivars regarding the xylem sap flow. In fact, Sakasly gave the best xylem flow during the three campaigns. Razegui1 and Razegui2 registered approximately similar xylem sap flow, while Hencha and Khamri present the lowest sap fluxes during the three campaigns. Moreover, several vascular occlusions forms were identified from stem cross sections using environmental scanning electron microscopy (ESEM), including tyloses, gels, starch, and gum deposits. The highest occlusion number was observed in Sakasly, Razegui1, and Razegui2 cultivars. Among different biogenic calcium shapes, several were observed for the first time in grapevine, including multi-faceted druse, cubic, crystalline sand, styloids, spherical, or drop-like structures. Considering their lower flow and totally blocked vessels, both Hencha and Khamri confirmed their susceptibility to environmental constraints. However, Sakasly, Razegui1, and Razegui2 cultivars presented higher tolerance according to their sap flow and xylem morphology

Elimination of Grapevine leafroll associated virus-3, Grapevine rupestris stem pitting associated virus and Grapevine virus A from a Tunisian Cultivar by Somatic Embryogenesis and Characterization of the Somaclones Using Ampelographic Descriptors

Prospecting of local grapevine (Vitis vinifera L.) germplasm revealed that Tunisia possesses a rich patrimony which presents diversified organoleptic characteristics. However, viral diseases seriously affect all local grapevine cultivars which risk a complete extinction. Sanitation programs need to be established to preserve and exploit, as a gene pool, the Tunisian vineyards areas. The presence of the Grapevine leafroll associated virus-3 (GLRaV-3), Grapevine stem pitting associated virus (GRSPaV) and Grapevine virus A (GVA), were confirmed in a Tunisian grapevine cultivar using serological and molecular analyses. The association between GRSPaV and GVA viruses induces more rugose wood symptoms and damages. For this reason the cleansing of the infected cultivar is highly advisable. Direct and recurrent somatic embryos of cv. ‘Hencha’ were successfully induced from filament, when cultured on Chée and Pool (1987). based-medium, enriched with 2 mg 1−1 of 2,4-dichlorophenoxyacetic acid and 2.5 mg 1−1 of Thidiazuron, after 36 weeks of culture. After six months of acclimatization, RT-PCR carried on 50 somaplants confirmed the absence of GVA, GRSPa-V as well as GLRaV-3 viruses in all somaplants. Ampelographic analysis, based on eight OIV descriptors, was carried out on two years acclimated somaplants, compared to the mother plant. Results demonstrated that the shape and contours of 46 somaclones leaves are identical to mother plant leaves and four phenotypically off-type plants were observed. The healthy state of 100% ‘Hencha’ somaclones and the high percentage of phenotypically true-to-type plants demonstrate that somatic embryogenesis is a promising technique to adopt for grapevine viruses elimination