The most commonly used methods for the detection of viroids, such as molecular hybridization and RT-PCR, are based on nucleic acids. In the case of aggressive hop stunt disease present in Slovenia, the presence of three different viroids on the diseased plants leads to higher costs of diagnosis and indicates the need for methods that allow simultaneous detection of all viroids. For this purpose, we have developed the diagnostic method mRT-PCR for detection of all four hop viroids. Use of mRT-PCR enables shorter analysis, reduces the use of reagents and, consequently, the cost of analysis. The method can be used for routine testing of symptomatic plants from a field, for testing in the context of a certification scheme for planting material, as well as for epidemiological studies.
B.03 Paper at an international scientific conference
COBISS.SI-ID: 8259705Artificial plant inoculation methods are an important tool in plant virology and can be used in various studies, such as resistance screening, biological indexing of host plants, epidemiological studies and analysis of plant-pathogen interactions. With the aim of developing a reliable bioassay for CBCVd for hop genotype resistance screening and analysis of the CBCVd host range, we compared four different types of inoculum based on RNA and sap extract obtained from CBCVd infected donor plants. The inoculum was mechanically introduced using stem injections or rubbing the inoculum onto leaves treated with carborundum. The assay was performed on six indicator plants: two varieties of cucumber (Cucumis sativus), two varieties of tomato (Solanum lycopersicum), eggplant (Solanum melongena) and purple passion plant (Gynura aurantiaca), with 10 repetitive plants per treatment. The results of our preliminary study show that the best bioassay for CBCVd is rubbing the sap extract inoculum onto leaves treated with carborundum, using Heinz 1370 tomato variety as an indicator plant. Using inoculum based on sap extract, we were able to infect both varieties of cucumber and tomato, as well as eggplant.
B.03 Paper at an international scientific conference
COBISS.SI-ID: 8301433In the frame of scientific conference we present viroid management studies in Slovenia. Until now 4 viroids are known on hop from which Hop latent viroid (HLVd), Hop stunt viroid (HSVd) and Citrus bark cracking viroid (CBCVd) are present in Slovenia. The most harmful is CBCVd which causes severe hop stunt disease. We present epidemiological studies which were focused into study of critical viroids spreading points. These studies significantly contribute in improvement of management strategy and in eradication process. The results were also base for changes in legislation which cover hop viroids in Slovenia and prepositions for EU measures.
B.03 Paper at an international scientific conference
COBISS.SI-ID: 8301689