Plant viruses Plant viruses • Nucleic acid in protein capsid (no membrane envelop) • Protein capsid – protection and transfer of NA • Nucleic acid – infectious (in some viruses together with polymerases) • Encode just few genes (x bacteriophages up to 70) • Other necessary processes (enzymatic activities) by host cell Viral genome - compact - various arrangement and strategies of expression – formation of polyproteins – segmented genome (alt. more virions - e.g. Tobacco rattle virus) – alt. read-through stop codon (translational readthrough) – alternative frameshift during translation – overlapping reading frames: alt. translation starts (transcription from both strands) – IRES (cap independent initiation of translation) Proteins encoded by plant viruses • Polymerases of NA (helicases) • Movement proteins - transport through plasmodesmata • Capsid proteins • Proteases - cleavage of polyproteins • Suppressors of silencing Different representation of these proteins in different viruses Suppressors of RNA silencing - independently in the majority of viruses – various mechanisms Burgyán, Havelda 2011 - participate in symptoms of infection through repression of RNAi regulated developmental steps! Example: suppressor P19 (tombusvirus) – dual function - homodimers P19 bind siRNA - induce expression of miR168 – block of AGO1 translation Burgyán, Havelda 2011 Viral infection symptoms: depletion of metabolites, defence reactions, suppressor side effects, … - chronic degenerative desease decreasing fitness chlorotic lesions intervein chloroses mozaic necroses growth reduction leaf curling Spreading of viral infection Within a plant - plasmodesmata (movement proteiny) - vascular tissue (phloem) Movement proteins: - interaction with virion - interaction with plasmodesmata (increase of size exclussion limit) Spreading of viral infection Between plants – natural barriers of entrance: cuticle, cell wall - mechanical injury, direct contact (wind) - vectors – sucking insects, other insects, nematods, fungi - grafting, root coalescence, - parasitic plants (Cuscuta) - vegetative propagation - some viruses also via seeds and polen! Protection – elimination of infected plants and vector insects! Transmission via sucking insects Non-persistant • adsorbtion on styletes (specific binding sites on acrostyle) • infectiousness: immediate, persists only minutes to hours Circulative • circulation of virus in insect body – salivary glands • infectiousness: latent period (hours to days), gradually decreasing many days Propagative • virus replication in transmittor • infectiousness: latent period (hours to days), life-long (also transmission to progeny) Viral capsids Capsomers – structural subunits (one or more capsid proteins) Basic shapes: A. Helical – capsomers in helical arrangement (e.g. Tobacco mosaic virus) TMV EM of helical capsids Viral capsids B. Polyhedral – capsomers form usually triangles arranged to polyhedron (usually icosahedr – twenty sides) - various number of proteins in a capsomer 12 pentagons 20 hexagons Classification of plant viruses - genom/replication • ssRNA, also dsRNA, ssDNA, dsDNA • ssRNA - coding ssRNA(+) - non-coding ssRNA(-) - replication via RT (also dsDNA viruses) DNA viruses - transcription by RNA polymerase II from dsDNA dsDNA viruses – replication through RNA intermediate (reverse transcription) ssDNA viruses – replication through dsDNA intermediate (by host DNA polymerase) Caulimoviridae - derived from LTR retroTE (order of ORF, replication, tRNA primer) - rarely integrated = „endogenous pararetroviruses“ – integrase? - 35S transcript > full genomic - circularization Replication cycle of ssDNA viruses (Geminiviridae) – ability to activate cell cycle! Why? RNA viruses dsRNA viruses e.g. Phytoreoviridae - 12 dsRNA segments, - viral polymerase - transcription in cytoplasma (viroplasma) - minus strands synthetized after encapsidation ssRNA viruses - RT – Pseudoviridae – again derived from retrotransposons Classical RNA viruses – enkapsidation of + or –RNA - RNA- : Rhabdo- a Bunyaviridae - all propagate also in insect vectors - RNA dep. RNA-polymerase in capsid – why? - RNA+ : most frequent (Tombusviridae, Bromoviridae, Potyviridae) Replication of RNA(+) viruses ssRNA(+) = mRNA and replication template Replication cycle of RNA+ viruses • e.g. tobacco mosaic virus (TMV) – Release of RNA – Translation of polymerase – RNA replication – Translation of viral proteins (polymerase, capsid, ….) – new virions spontaneously through „polymeration“ of capsid proteins on NA VIROIDs – circular ssRNA, no protein envelop (capsid) – genom size insufficient to encode proteins (359 b = 1/10 of smallest RNA viruses) VIROIDs - symptoms of infection – likely results from induced RNAi non-specifically affecting expression of plant genes - common features (origine?) with HDV (hepatitis D virus) Replication with host DNA dep.(!) RNA Pol II - probably rolling circle - concatemers of some viroids autocatalytically cleaved by hammer-head ribozyme e.g. Potato Spindle Tuber (the first sequenced eucaryotic patogen) Hammerhead ribozyme yellow NTs + 3 short dsRNA regions necessary for cleavage (but also sufficient = possible induction of cleavage in trans) N N C G A A U A H G N N N N N G N C N’ N’ N’ N’ C N’ C N’ G A U G G N A cleavage site H = A,C,T cleavage site minimal requirements of cleaved RNA: N N C G A A U A H cleaved RNA G N N N N N G N C N’ N’ N’ N’ C N’ C N’ G A U introduced inducing RNA G G N A H = A,C,T