The grapevine plants from the collection vineyard in which we proved the presence of singe GLRaV-3 virus infection were rooted and transplanted in pots for the purpose of biological indexing. Since we did not find all genetic variants of GLRaV-3 as a single infection, we have obtained individual GLRaV-3 genetic variants which we will use to infect indicator plants. The cuttings containing monophyletic variants of GLRaV-3 were gained by the courtesy of dr. Rachelle Bester from the Stellenbosch University (Department of Genetics), South Africa. These cuttings are properly stored until spring next year when they will be used as a source of virus inoculum for grafting of indicator plants.
In parallel with laboratory analyzes of GLRaV-3 genetic diversity that we had detected in autochthonous varieties, since spring we have been monitoring the occurrence and intensity of symptoms of all viral diseases on grapevine plants in collection vineyard. The first symptoms we noticed were the yellowing and leaf malformations associated with viruses from the infectious degeneration group. With the beginning of the summer, symptoms of leaf redness between leaf veins began to develop in grapevines infected with viruses from the leafroll associated virus group. Over time, these leaves became redder and twisted. Because of this symptom of leaf downrolling, leafroll viruses justifiably bear the name given.
Mate Čarija was in the short scientific training from 17.06.-30.06. 2019 at the Institute for Sustainable Plant Protection in Bari, Italy. The host was prof. dr. sc. Pasquale Saldarelli, also a collaborator on the project.
While we in Croatia were celebrating two holidays during that period, Mate was in the Laboratory of prof. Saldarelli learning and adopting the Real-Time PCR method, advanced analytical detection and quantification method for grapevine viruses. In the second week of his stay in Bari, Mate was hosted at the Agritest Laboratory to perform ELISA on the Croatian autochthonous grapevine samples.
Bravo Mate for great learning and results and we owe you two days off the work :-)!
We attended an internal methodological workshop at the Faculty of Science in Zagreb. At the workshop, we were theoretically and practically introduced with the method of SSCP (Single-Strand Conformation Polymorphism) used to determine genetic variants present in the sample. The method was used to determine the genetic structure of GLRaV.-3 virus in grapevine plants. The workshop has lasted for 5 days and was led by PhD Silvija Černi, associate on the project.
The program of the workshop was adapted on an everyday basis in accordance with the protocol for the implementation of the SSCP method and results obtained.
Set of virus-free cuttings of indicator plants were put to rooting for the purpose of GLRaV-3 biological detection and characterization. Indexing plants includes following grapevine cultivars: Pinot noir, Merlot and Cabernet franc. Cuttings of autochthonous cultivar Tribidrag were also put to rooting so the symptoms occurrence as well as the morphological, physiological, biochemical changes after the virus infection could be observed and described. Special attention will be paid on the interaction between virus and host plants by exploring the dynamic of virus replication throughout the infection processes.
Some rooted seedlings were already transplanted into pots and placed in a greenhouse chamber, under the proper irrigation, nutrition and protection regime.
In the Laboratory for Molecular Characterization at the Institute for Adriatic Crops and Karst Reclamation we have introduced a protocol for the detection of grapevine viruses so we have successfully detected all 5 viruses included in the EU directive. European Commission directive 2005/43/EC considers infectious degeneration [Grapevine fanleaf virus(GFLV), Arabis mosaic virus (ArMV)], leafroll [Grapevine leafroll associated virus 1 (GLRaV-1), Grapevine leafroll associated virus 3 (GLRaV-3)], fleck [Grapevine fleck virus (GFkV) only for rootstocks] and their causal viruses, as harmful diseases whose presence must be monitored and if found, excluded from nursery stocks upon implementation of national certification schemes. Plenty of viruses in our autochtonous Croatian grapevine cultivars!
Mate Čarija, MSc in hort. held a lecture “All colors of Grapevine Viruses” at the Split Science Festival. Mate spoke in a scientifically-popular way about the consequences of different viruses on grapevine plants and the ways of preventing the infection. The lecture was held on April 10, 2019 in the Library of the Institute for Adriatic Crops and Karst Reclamation in Split. The lecture was also the first presentation by Mate as a collaborator on the GRAPE-PATH3 project. It was a very educational, interesting and colorful lecture!
For more information, you can download the program at http://novosti.krs.hr/festival-znanosti-2019/
Mate Čarija, MSc in horticulture, has started working at the Institute for Adriatic Crops and Karst Reclamation at the Department of Plant Sciences within the project : “Young researchers’ career development project –training ofdoctoral students” of the Croatian Science Foundation. Graduated from the Faculty of Agriculture, University of Zagreb, in 2019, he was studiing viticulture and enology. Already during the Study, Mate worked actively in the field of plant virology, spending a month at the Institute National De La Recherche Agronomique, Montpellier, France. His research work will be focused on the pathogenic potential of GLRaV – 3 virus and its interaction with plant host.
Kick off meeting was held on March 11 and 12, 2019 at the Institute’s lecture hall. Project objectives, research methods and expected results were presented. Research plan were discussed as well as activities per each project associate. The key stages for the successful implementation of the project were defined.
Monday (start at 9:00)
1. Project presentation
2. Associates activity review
Tuesday (start at 9:00)
1.Lecture of prof. Sandre Radić Brkanac entitled “Mechanisms of Plant Response to High Salinity”
2. Work in the working group