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1.
Biological role of Grapevine fanleaf virus (GFLV) in winegrowing region of Northern Primorska
Anastazija Jež Krebelj, 2015, doktorska disertacija

Opis: Grapevines (Vitis vinifera L.) represent one of the most important crops in the world in terms of both production and economic importance. Grapevines are exposed to many types of abiotic stresses (e.g., drought, flooding, low and high temperature, salinity) and biotic stresses (e.g., viruses, bacteria, phytoplasma, fungal disease) during their life-cycle. Therefore, grapevines elicit the appropriate defence mechanisms. In the first part of this study, we monitored the occurrence of Grapevine fanleaf virus (GFLV) infection, which causes progressive decline of infected grapevines and lowers their yield. Grapevines were also tested for the presence of other viruses important for grapevines: Arabis mosaic virus (ArMV), Grapevine leafroll associated virus (GLRaV)-1, -2, -3, -4, -9, Grapevine virus A (GVA), Grapevine fleck virus (GFkV) in this study; and by Cigoj (2015): Grapevine virus B (GVB), Tomato black ring virus (TBRV), Grapevine chrome mosaic virus (GCMV), Tomato ringspot virus (ToRSV), Raspberry ringspot virus (RpRSV), Strawberry latent ringspot virus (SLRSV), and Tobacco ringspot virus (TRSV). Using ELISA, the presence of the following grapevine viruses were detected: GFLV, (GFkV), (GVA), and Grapevine leafroll associated viruses- 1, -2, -3,. A wide range of GFLV symptoms caused by grapevine fanleaf disease in naturally infected vineyards were observed, including leaf, shoot and cluster malformations and leaf yellowing. GFLV is disseminated by its biological vector X. index, and through vegetative propagation of virus-infected material. The spread of GFLV in the vineyards was investigated here. We constructed a spatio-temporal study of the GFLV titres during the seasons and throughout the grapevine, for its distribution in different grapevine organs through the season. This study shows that young leaves have high virus titres through the whole vegetative period, while mature leaves, tendrils and flower/ berry clusters only have high titres at the beginning of the vegetative period. The seeds retain high virus titres after berry colouring. Phloem scrapings were shown to contain lower virus titres during the vegetative period, with an increase outside and at the beginning of the vegetative period. In flower/ berry clusters, mature leaves and tendrils, the GFLV titres decrease significantly over the vegetative period. Additionally, different GFLV titres were shown in five different cultivars, and different combinations of mixed infections with other grapevine viruses influenced the GFLV titre differently. Finally, correlation between the magnitude of symptom appearance and GFLV titres was analysed. Grapevines adapt to abiotic stresses and biotic stresses by the expression of a wide range of stress-responsive genes, which are thought to have key roles in stress tolerance and survival. SWP of the infected grapevines through the season was lower than SWP measured for healthy grapevines. For both seasons, there were significant differences in SWP measurements between healthy and GFLV-infected grapevines of ‘Schioppettino’ trained using the single Guyot training system. SWP and RHC of the GFLV-infected grapevines were reduced compared to the healthy controls. The water deficit triggered the production of ABA, which induced the expression of the stress-related gene RD22. Additionally, this study shows that the WRKY gene that is involved in the ABA signalling network is regulated by water deficit. Plant defence responses to water stress also included up-regulation of the F3H2 and LDOX genes, which are involved in anthocyanins synthesis. GFLV infection significant impacted upon the expression of genes involves in ABA biosynthesis, as NCED1 and NCED2, and upon two genes involved in the early stages of anthocyanins synthesis, as CHS2 and F3H1. We also showed that the combination of grapevine cultivar, training system, and environmental conditions impacts on gene expression
Najdeno v: ključnih besedah
Ključne besede: Vitis vinifera L., grapevine, Grapevine fanleaf virus, GFLV, grapevine disease, virus titre, distribution, fluctuation, ELISA, qPCR, ABA, drought, water status, water deficit, SWP, RHC, anthocyanins, gene expression
Objavljeno: 27.07.2015; Ogledov: 2938; Prenosov: 122
.pdf Polno besedilo (4,19 MB)

2.
DISTRIBUTION OF ENTERIC VIRUSES IN THE GULF OF TRIESTE AND THEIR INTERACTIONS WITH ENVIRONMENTAL AND BIOLOGICAL PARAMETERS
José Manuel Carita Gonçalves, 2018, doktorska disertacija

Opis: The available classical diagnostic methods, due to many disadvantages, do not allow effective detection of pathogenic enteric viruses in environmental samples. Due to low concentrations of pathogenic viruses in the sea, it is important to develop an effective concentration procedure for their successful detection. In the first part of the doctoral thesis, we focused primarily on the development of a protocol for an effective concentration of pathogenic enteric viruses in coastal water samples. Monolithic chromatographic columns (BIAseparations) were used for the concentration of rotaviruses and noroviruses, prior to the detection with reverse transcription quantitative PCR in real time (RT-qPCR). We tested the efficiency of concentration using columns of various chemical properties and selected pathogenic enteric viruses (rotavirus and norovirus). Among them, hydrophobic interaction monolithic column (CIM® C4) was the most effective. CIM C4 was used to optimize the concentration step and tested in waters with different salinities. The presence of concentrated viruses was confirmed by RT-qPCR and transmission electron microscope. We have developed a protocol that enables rapid concentration of viruses in coastal waters of various salinities and can be used on-site. The presence of RoV and NoV was surveyed, using the developed concentration protocol, prior to one-step RT-qPCR molecular detection, in the inner part of the Bay of Koper, in mussel farming areas and a swimming area. Rotaviruses, noroviruses and fecal indicator bacteria were frequently detected in the inner part of the Bay of Koper. Rotaviruses and noroviruses were detected in the studied area, with higher rates close to the outfall of the wastewater treatment plant in the estuary of river Rižana and were also detected in the middle of the Bay of Koper and in areas used for recreation and mussel farming. The results show that water bodies, which are otherwise defined as suitable for bathing or mussel farming, based on the results of fecal indicator bacteria, still contain low concentrations of pathogenic enteric viruses. In addition to human pathogenic enteric viruses and faecal coliforms, changes in abundance of bacteria and virus particles were studied in relation to temperature, salinity, inorganic and organic nutrient concentrations in the organically polluted Rižana estuary. Preliminary results showed spatially and seasonally changes in bacterial and viral particles abundance, and bacterial composition spatially and seasonally. However, seasonality plays a greater role in bacterial dynamics.
Najdeno v: ključnih besedah
Povzetek najdenega: ...reverse transcription quantitative PCR in real time ( RT-qPCR). We tested the efficiency of concentration using...
Ključne besede: Concentration of viruses, Enteric viruses, Rotavirus, Norovirus, Feacal coliforms, Feacal contamination, qPCR, RT-qPCR, Monolithic columns, Sewage, Seasonal dynamics, Concentration, Coastal environment, Gulf of Trieste
Objavljeno: 02.07.2018; Ogledov: 128; Prenosov: 28
.pdf Polno besedilo (1,95 MB)

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