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Introduction to Plant Virology • History • Definitions • Classification • Structure Additional program (student‘s presentation): Mimivirus (Raoult et al.

, 2004, SCIENCE 306, p. 1344 ff, Xiao et al., J. Mol. Biol. (2005) 353, 493–496)

K. Richert-Pöggeler, WS 05/06

Roger Hull: Matthews‘ Plant Virology (2nd edition, 2004)

http://www.apsnet.org/ http://www.virology.net

First plant virus description 752 AD

Eupatorium yellow-vein (gemini) virus (EpYVV)

Saunders et al., 2003

Dr Robert G. Milne, CNR, Instituto di Fitovirologica Applicata, Torino, Italy

Tulip breaking (Poty) virus

Ambrosio Bosschaert, Dutch painter, 1573-1621

Brunt and Walsh, 2005

1892: Dmitri Iwanowski. Russian scientist. nor grow on microbial media -concluded he had found infectious agent smaller than bacterium . works with tobacco plants with Tobacco Mosaic Disease -discovers filtration does not remove infectious agent for TM disease -could not visualize agent with microscope.

Beijerinck (1851-1931) Über ein contagium vivum fluidum als Ursache der Fleckenkrankheit der Tabaksblätter. slime Martinus W. Amsterdam 65:3-21 (1898) John Shaw. Wetensch.“…infection is not caused by microbes but by a contagium vivum fluidum” “…reproduces itself in the diseased plants” “…other diseases of unknown cause may be ascribed to a contagium fluidum” VIRUS (lat. 2002 . Verhandel. Acad.): venom.

The causal agent must be reisolated. The causal agent must be associated in every case with the disease as it occurs naturally. Nobel Prize 1905 1. The causal agent must be isolated in pure culture 3. 2. . 4.Koch’s Postulates 1843-1910. When the host is inoculated the characteristic symptoms of the disease must develop.

Ontario . Toronto.Visualization of viruses made possible by electron microscope Magnification up to 106 fold !!!! nm=10-9 meter The original electron microscope as developed in 1938 in McLennan Laboratories of the University of Toronto is now on permanent exhibition at the Ontario Science Centre.

it became obvious that a sharp line dividing living from non-living things could not be drawn and this fact served to add fuel for discussion of the age-old question of 'What is life?'" . 1904-1971 By courtesy of the Molecular Biology & Virus Laboratory. 1935 Wendell Stanley. Berkeley Stanley achieved the first crystallization of a virus (1935). He later remarked on the unique position of viruses at the junction of life and non-life: "The fact that. University of California. with respect to size.Isolation of a crystalline protein possessing the properties of tobacco mosaic virus. Science 81:644-645. Then too. the viruses overlapped with the organisms of the biologist at one extreme and with the molecules of the chemist at the other extreme only served to heighten the mystery regarding the nature of viruses. the basis for his Nobel Prize of 1946 (Chemistry).

1982: TMV (ssRNA) • 1984-Polymerase chain reaction (PCR) (Mullis) • 1995-Entire genome sequenced (Haemophilus influenzae) .Molecular Biology and Biotechnology • 1969-Restriction endonuclease cloned (Arber & Smith) • 1970-Reverse transcriptase (Temin & Baltimore) • 1973-Recombinant plasmid (Cohen & Boyer) • 1977-DNA sequencing (Gilbert & Sanger) 1977: bacteriophage (ssDNA). 1980: CaMV (dsDNA).

Size The Mimivirus has a size larger than the smallest bacteria and.Distinguishing viruses from other organisms What fails to distinguish viruses from cellular organisms? 1. a genetic complexity greater than that of the most reduced bacteria http://news.bbc.co.uk/1/hi/health/2895165.stm . with about 900 genes.

E. R. F. Closteroviridae: 1. Academic Press. Plant Virology.Matthews. (1981).9x104 Nanoviridae: 1x103 Mimiviridae (amoeba):1x106 .

Stable. reticulate bodies (bilayer membrane. ATP production. DNA no cell wall. Rickettsiae: nonmotile bacteria (typhus fever). plasmamembrane.Distinguishing viruses from other organisms What fails to distinguish viruses from cellular organisms? 2. elementary-. Chlamydiae: psittacosis. inert phase in life cycle Many bacterial spores are more stable than some virus particles . Rickettsiae and Chlamydiae Mycoplasma: 150-300 nm diamenter. binary fission. Obligate intracellular parasite Fails to distinguish viruses from many bacteria. DNA. ribosomes. binary fission) 3. Mycoplasma. Replication by binary fission. bilayer membrane. CW. ribosomes.

Acellular. Viruses do not grow.envelope= stolen host cell membrane) B. . Genome = RNA or DNA C. Ability of replicated viruses to be released from host cell H. Can’t generate own energy therefore are “metabolic parasites” F. No ribosomes. Viruses direct synthesis of viral nucleic acid and viral proteins by host cell. Protein coat = capsid D. Host cell specificity: all cellular organisms may be attacked 1. Lack ability to synthesize organic molecules E. Viral adhesins must bind specific host cell surface receptors 2.General characteristics of viruses A. Obligate intracellular parasites-can only replicate inside another host cell G. Viruses are “assembled”. nor divide. No metabolism. Appropriate host enzymes for viral replication 3. don’t synthesize a cell membrane (+/.

Intact non-replicating virus particles.Distinguishing “virion” from “virus” Virion the particle that is the extracellular phase of the infection cycle. including replication intermediates . no signs of life Virus virion plus intracellular aspects. typically composed of the genomic nucleic acid and coat protein but may have a lipid membrane and other components.

Alive? viruses reproduce. property of life occur as populations have variation that is inherited Why viruses are non-living Lack a complete protein synthesis system Lack a complete energy generation system .

Virus Disease Symptoms Local lesions Stunting Barley yellow dwarf virus Yellowing Beet mild yellowing virus Ringspot PV-Y Necrosis Mosaic Abutilon mosaic virus Tomato spotted wilt virus Developmental abnormalities Tobacco mosaic virus Alfalfa mosaic virus Zucchini yellow mosaic virus .

potyviruses) .Microsymptoms Chloroplast Degeneration (tymoviruses) Enlarged Nuclei (rhabdoviruses) Disorganized Mitochondria (aggregation (potyvirus). modification (tombusvirus) Inclusion Bodies (caulimoviruses.

1973. Phytopathology 63 Protein enoded by gene 6 of CaMV Host range Symptom expression Translation (transactivator) DNA replication .Cytoplasmic inclusion bodies Pinwheel Inclusions (Potyvirus) Inclusion bodies (caulimovirus) Originate and develop in association with the plasma membrane CI protein of potyviruses RNA replication Lesemann and Casper.

1992): PVCV. PV-Y. CMV. TMV . but different viruses: Tobacco mosaic virus (ssRNA) Cauliflower mosaic virus (dsDNA) Abutilon mosaic virus (ssDNA) mixed infected petunia (Richert.Symptoms are not sufficient to classify virus: • mixed infections • distinct strains that cause different symptoms in same host • same symptoms.

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Virus classification Nucleic acid Morphology Genome organisation Transmission vector .

no DNA stage Satellites ssDNA viruses unclassified bacteriophages Comments and suggestions to: [genomes@ncbi.gov/genomes/VIRUSES/viruses.nlm. 2005 .ncbi. Deltavirus dsDNA viruses.nlm.gov] Revised: October 18.html Entrez Genomes currently contains 2139 Reference Sequences for 1486 viral genomes and 36 Reference Sequences for viroids.http://www.nih. no RNA stage ssRNA negativestrand viruses unclassified viruses Retro-transcribing viruses dsRNA viruses ssRNA positive-strand viruses.nih.

Comparitive abundance of different viruses +ssRNA 600 dsDNA -ssRNA dsRNA 300 ssDNA rtDNA H. Scholthof .

Relative abundance of different plant viruses +ssRNA dsDNA -ssRNA dsRNA ssDNA rtDNA H. Scholthof .

Classification of plant viruses .

Genome (DNA or RNA) dsDNA-RT Caulimoviridae (pararetroviruses. Tobravirus. fungi) ssRNA(+) Bromoviridae*. Furovirus Pecluvirus * Alphavirus. vertebrates) ssDNA Geminiviridae Nanoviridae dsRNA Partitiviridae (fungi) Reoviridae (invertebrates. Comoviridae. vertebrates) Tenuivirus. vertebrates. Tymoviridae Flexivirdae. Varicosavirus ssRNA-RT Pseudoviridae (invertebrates. Closteroviridae Ourmiavirus Tobamovirus. Hordeivirus. Pomovirus. Potyviridae#. fungi) ssRNA(-) Rhabdoviridae (invertebrates. vertebrates) Bunyaviridae (invertebrates. Luteoviridae. fungi) Metaviridae (invertebrates. Benyvirus. Sesquiviridae. # Picornaviridae . Tombusviridae.

Virus Particle Structure Bacilliform Flexuous rod Geminate Spherical Rigid rod .

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Morphology .

500 * The virion RNA has a 5' cap structure at the 5' nucleotide residue .COMPOSITION OF TMV VIRIONS TMV virions are rod shaped.3 x 106 17. 300nm long and about 18nm in diameter. cylindrical core. Component Number of molecules in virion 1 about 2140 Molecular weight RNA [6395 residues]* Capsid protein 2. The virions have helical symmetry and a hollow.

Hull. p. 132 .

3 fold symmetry axis midpoint of each edge .The regular icosahedron: Symmetry 12 vertices.2 fold symmetry axis . 20 identical triangular faces 5 fold rotational symmetry center of face .

Variations on a Theme Objective: Make particles larger and more spherical Strategy: Divide the original 20 faces in smaller faces. which each again can be filled with subunits Triangulation: T=Px(f)2 Many viruses: P=3. f=1. (pentamers and hexamers of subunits) Page 136 . T=3.

. where h & k are any distinct. is defined by:T = f 2 x P where f is the number of subdivisions of each side of the triangular face. nonnegative integers. • The triangulation number. number T. the symmetry of the particle is defined by the triangulation number of the icosahedron. f 2 is the number of subtriangles on each face & P = h2 + hk + k2.Icosahedral Symmetry • In higher order icosahedra.

page 137 Each original face is divided up in 6x1/2 new faces: P=3 Each new sub-triangle again can handle 3 protein units T=Pxf2: 3x1=3===>60x3=180 protein subunits . 5.17.Fooling around with P Fig.

Academic Press .Triangulation Numbers Molecular Virology. 3rd edition.

icosahedral Cowpea mosaic (como) virus virion Three copies of L coat protein Five copies of S coat protein (12 vertices) x (5 S protein per vertex) = 60 copies of S protein (20 faces) x (3 L protein per face) = 60 copies of L protein Capsid is composed of equal molar amounts of two coat proteins. L and S .

M.Virus Structure (Reconstruction based on X-Ray crystal structures) Tomato Bushy Stunt Virus Cowpea Chlorotic Mottle Virus D. Canada . Rochon.

Genome organisation .

bbc. with about 900 genes.uk/1/hi/health/2895165.co.The Mimivirus has a size larger than the smallest bacteria and.stm . a genetic complexity greater than that of the most reduced bacteria http://news.