COURSE OBJECTIVES:
Students will be introduced to the types and features of subviral infectious molecules, which are mostly RNA and to a lesser extent proteins. The molecular basis of the interactions of subviral molecules with hosts will be studied, thus educating students about the mechanisms of causing diseases of subviral aetiology, and enabling consideration of the functioning of subviral and non-coding RNAs within the framework of basic molecular biological knowledge. Students will be able to understand the significance of non-coding RNA molecules as regulators of cellular processes, defence against pathogens through RNA silencing, and their importance in the development of evolutionary theories and the origin of life on Earth.
COURSE CONTENTS:
LECTURES
1) Introducing the content and structure of the course, teachers and arrangements for conducting exercises;
2) Definition of subviral infectious molecules according to the types of molecules and the relationship with the host cell and the helper virus within the concept of satellitism;
3) Satellites of different types;
4) Lethal necrosis disease of tomato ? a case study of the influence of viral RNA satellites on the expression of helper virus symptoms in the plant host;
5) Viroids as causative agents of plant diseases and non-coding genomes;
6) Molecular mechanisms of viroid diseases - RNA silencing;
7) Viroids with ribozyme activity and mechanisms of their action in plants;
8) The concept of quasispecies and the evolution of RNA, viroids as relics of the RNA world;
9) Growing group of viroid-like RNAs with ribozyme activity and problems of their taxonomy;
10) Molecular biology of hepatitis D virus (HDV);
11) Molecular basis of hepatitis D pathogenesis;
12) Growing group of HDV-like viruses and their phylogeography;
13) Discovery and molecular features of prions, prion hypothesis;
14) Prion diseases of humans and animals - biological basis of transmissible spongiform encephalopathies;
15) Prevention and methods for detecting prion diseases.
EXCERSISES
1) Teacher's proposal for several variants of student projects for viroid or satellite RNA research, discussion, student agreement and project selection;
2) Literature research and project development by students, satellite and/or viroid genome searches in databases, preparation of experimental plants and inoculation (if necessary);
3) Presentation of the project plan for purification, detection, identification of viroids and/or satellite RNAs, and research on their characteristics;
4) Ordering the necessary chemicals, design of primers, getting to know the software for the planned analyses;
5) Preparation of solutions, buffers, primers, samples of host plants infected with satellites or viroids;
6) Isolation of the subviral pathogen from plant tissue (Part 1);
7) Isolation of the subviral pathogen from plant tissue (Part 2);
8) Quality and quantity measurements of isolated RNA, preparation for detection;
9) Detection of subviral pathogen by RT-PCR method;
10) Electrophoretic analysis of viroid/satellite amplicons;
11) Preparation and submission of amplicon samples for Sanger sequencing;
12) Alternative methods for detecting subviral pathogens (e.g. RT-LAMP);
13) Analysis of project results, agreement on the preparation of a project report in the form of an original research article;
14) Independent student work on writing a report, sending the report to the teacher for assessment;
15) Discussion with the teacher after feedback, revision of the report and saving the final version with corrections.
Exercises are held in small groups of up to 4 students in a condensed manner due to the lab space restrictions and contamination issues. The topic of the student project is related to current research problems in the laboratory involving viruses with satellites or viroids.
|
-
Fundamentals of Viroid Biology, odabrana poglavlja, Adkar Purushotama et al., Elsevier, Academic Press, 2024.
-
Viroids and Satellites, odabrana poglavlja, Hadidi et al., Elsevier, Academic Press, 2017.
-
Interna skripta, Prezentacije s predavanja i odabrani znanstveni članci., Škorić Dijana, 2025.
|
Pristupačnost
