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Pawan Kumar
M.Sc. Bioinformatics
1st semester
PK
1
Learning Objectives
 Introduction to Plant viruses
 Infections
 Virus life cycle
 Transmission
 Structure
 Classification
 Replication
 Symptom
 Identification
 Control.
PK
2
Plant viruses
 Non-cellular form of life.
 Nucleic acid in protein capsid (no membrane envelop).
 Virus is a nucleo-protein having RNA or DNA as a genetic
material.
 RNA or DNA may be ss or ds, ssRNA may be +ve or –ve sense.
 The fundamental characteristic is their absolute dependence
on a living host organism for their reproduction.
 Other necessary processes (enzymatic activities) by host cell.
 Encode just few genes (x bacteriophages up to 70)
 Most of plant virus infect a limited number of different plant
species and a few have a wide host range.
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3
chronic degenerative desease decreasing fitness
growth reductionchlorotic lesions mozaic
intervein chloroses necroses leaf curling
Viral infection
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Plant response to inoculation
with a virus
 Plant is immune: virus does not replicate in protoplasts nor
in cells of the intact plant
 Infection is limited to initially infected cells: replication, but
no cell-to-cell movement (could be due to an ineffectual
virus movement protein)
 Infection is limited to initial leaf: replication, cell-to-cell
movement, but no long distance movement (inability to
enter phloem or due to a host resistance gene-mediated
response, i.e. hypersensitive response when the plant
resistance gene product recognizes some viral product,
local lesions formation)
PK
6
MODE OF ENTRY
 Mechanically TMV, BCMV, PVX, PYDV.
 Vectors PLRV,TLCV, WTV.
 Pollen Grain necrotic ring spot, ring spot of soybean.
 Seeds or vegetative:
Propagation BCMV, CMV, Pea stripe virus.
 Pinocyte CPMV, Pea enation mosaic
 Fungal parasite TNV, Barley yellow mosaic virus,
Tobacco stunt virus.
 Ectodesmata TMV.
 Epidermal hair TMV.
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7
Virus life cycle
Invasion
Genome
uncoating,
expression
and
replicationPK
8
Systemic transport through phloem
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9
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)
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10
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!
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11
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)
Transmission via sucking insects
PK
12
Proteins encoded by plant viruses
 Polymerases of NA (helicases)
 Movement proteins
- transport through plasmodesmata
 Capsid proteins
 Proteases
- cleavage of polyproteins.silencing
Different representation of these proteins in
different viruses
PK
13
TMV
Viral capsids
Capsomers – structural subunits (one or more capsid
proteins)
Basic shapes:
A. Helical – capsomers in helical arrangement
(e.g. Tobacco mosaic virus)
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14
capsomers form usually triangles arranged to polyhedron
(usually icosahedra – twenty sides)
- various number of proteins in a capsomer.
12 pentagons
20 hexagons
Viral capsids
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15
Classification of plant viruses
- genom/replication
 ssRNA, also dsRNA, ssDNA, dsDNA
 ssRNA - coding ssRNA(+)
- non-coding ssRNA(-)
- replication via RT (also dsDNA viruses)
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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)
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Replication cycle of ssDNA viruses
(Geminiviridae)
PK
18
RNA viruses
PK
19
dsRNA viruses
e.g. Phytoreoviridae - 12 dsRNA segments,
- viral polymerase
- transcription in cytoplasma (viroplasma)
- minus strands synthetized after encapsidation
PK
20
ssRNA viruses
 Ss RNA(RT):Pseudoviridae
again derived from retrotransposons.
 Classical RNA viruses:
enkapsidation of + or –RNA
 RNA+ : most frequent (Tombusviridae,
Bromoviridae, Potyviridae)
 RNA- : Rhabdoviridae
all propagate also in insect vectors
RNA dep. RNA-polymerase in capsid.
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21
Ss RNA (RT) viruses:
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22
Reverse transcription is applied to first make ds DNA
and then transcription and translation.
Ss 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 pack with coating protein.
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23
Infection cycle of TMV.
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24
Ss RNA- viruses:
 -ve ss RNA require RDRP(RNA dependent RNA polymerase).
 Which make +ve ss RNA from its original stand.
 These newly formed +ve ss RNA will make –ve ss RNA and
also translate to form protein capsid.
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Some example
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Symptoms:
PK
28
Systemic Symptoms:
 General:
Abnormal growth and developmental
malformations (reduced growth, dwarfing,
stunting of plants, etc.)
Reduced life span of plants, death.
 Foliage symptoms:
Mosaics
Yellows Ring spots.
 Other:
Symptoms on stems, fruit, roots.
 Local symptoms:
Necrotic lesions.
Detection and identification
PK
29
 Symptoms (possible causes by nutrient deficiencies,
toxicities, insects, etc.
 dsRNA isolation.
 PCR, RT-PCR (for known viruses).
 Virus genome sequencing.
 Microarray.
Control of plant viruses
 Keep out of an area through quarantine, inspection, and
certification programs that would prevent or limit local or
international virus movement and warranty the use of virus-free
seed, tubers, budwood .
 Eradication of diseased plants
 Controlling the insect vectors (oil sprays, repellents, insecticides,
predators…)
 Breeding plants for resistance
 Transgenic resistance (natural resistance genes, viral sequences,
genes from other sources).
PK
30
Reference:
 Prescott Harley Klein's Microbiology - Willey, Sherwood & Woolvertion
(7th ed.).
 Molecular Biology of the Gene - James D. Watson, Tania A. Baker,
Stephen P. Bell (5th ed.).
 Wikipedia source.
 YouTube source.
 Characteristic of Plant viral pathogen by Svetlana Folimonova (Ass.
Prof. -Dept. of Plant Pathology, University of Florida).
 Introduction to Plant Viruses, the Invisible Foe (www.apsnet.org).
 Viron | Biology Field Guide | Biowars (www.biowars.com).
 Plant Virus replication by P.N. Sharma Department of Plant Pathology,
CSK HPKV, Palampur (H.P.).
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32

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Plant virus

  • 2. Learning Objectives  Introduction to Plant viruses  Infections  Virus life cycle  Transmission  Structure  Classification  Replication  Symptom  Identification  Control. PK 2
  • 3. Plant viruses  Non-cellular form of life.  Nucleic acid in protein capsid (no membrane envelop).  Virus is a nucleo-protein having RNA or DNA as a genetic material.  RNA or DNA may be ss or ds, ssRNA may be +ve or –ve sense.  The fundamental characteristic is their absolute dependence on a living host organism for their reproduction.  Other necessary processes (enzymatic activities) by host cell.  Encode just few genes (x bacteriophages up to 70)  Most of plant virus infect a limited number of different plant species and a few have a wide host range. PK 3
  • 4. chronic degenerative desease decreasing fitness growth reductionchlorotic lesions mozaic intervein chloroses necroses leaf curling Viral infection PK 4
  • 6. Plant response to inoculation with a virus  Plant is immune: virus does not replicate in protoplasts nor in cells of the intact plant  Infection is limited to initially infected cells: replication, but no cell-to-cell movement (could be due to an ineffectual virus movement protein)  Infection is limited to initial leaf: replication, cell-to-cell movement, but no long distance movement (inability to enter phloem or due to a host resistance gene-mediated response, i.e. hypersensitive response when the plant resistance gene product recognizes some viral product, local lesions formation) PK 6
  • 7. MODE OF ENTRY  Mechanically TMV, BCMV, PVX, PYDV.  Vectors PLRV,TLCV, WTV.  Pollen Grain necrotic ring spot, ring spot of soybean.  Seeds or vegetative: Propagation BCMV, CMV, Pea stripe virus.  Pinocyte CPMV, Pea enation mosaic  Fungal parasite TNV, Barley yellow mosaic virus, Tobacco stunt virus.  Ectodesmata TMV.  Epidermal hair TMV. PK 7
  • 10. 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) PK 10
  • 11. 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! PK 11
  • 12. 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) Transmission via sucking insects PK 12
  • 13. Proteins encoded by plant viruses  Polymerases of NA (helicases)  Movement proteins - transport through plasmodesmata  Capsid proteins  Proteases - cleavage of polyproteins.silencing Different representation of these proteins in different viruses PK 13
  • 14. TMV Viral capsids Capsomers – structural subunits (one or more capsid proteins) Basic shapes: A. Helical – capsomers in helical arrangement (e.g. Tobacco mosaic virus) PK 14
  • 15. capsomers form usually triangles arranged to polyhedron (usually icosahedra – twenty sides) - various number of proteins in a capsomer. 12 pentagons 20 hexagons Viral capsids PK 15
  • 16. Classification of plant viruses - genom/replication  ssRNA, also dsRNA, ssDNA, dsDNA  ssRNA - coding ssRNA(+) - non-coding ssRNA(-) - replication via RT (also dsDNA viruses) PK 16
  • 17. 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) PK 17
  • 18. Replication cycle of ssDNA viruses (Geminiviridae) PK 18
  • 20. dsRNA viruses e.g. Phytoreoviridae - 12 dsRNA segments, - viral polymerase - transcription in cytoplasma (viroplasma) - minus strands synthetized after encapsidation PK 20
  • 21. ssRNA viruses  Ss RNA(RT):Pseudoviridae again derived from retrotransposons.  Classical RNA viruses: enkapsidation of + or –RNA  RNA+ : most frequent (Tombusviridae, Bromoviridae, Potyviridae)  RNA- : Rhabdoviridae all propagate also in insect vectors RNA dep. RNA-polymerase in capsid. PK 21
  • 22. Ss RNA (RT) viruses: PK 22 Reverse transcription is applied to first make ds DNA and then transcription and translation.
  • 23. Ss 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 pack with coating protein. PK 23
  • 24. Infection cycle of TMV. PK 24
  • 25. Ss RNA- viruses:  -ve ss RNA require RDRP(RNA dependent RNA polymerase).  Which make +ve ss RNA from its original stand.  These newly formed +ve ss RNA will make –ve ss RNA and also translate to form protein capsid. PK 25
  • 27. PK 27
  • 28. Symptoms: PK 28 Systemic Symptoms:  General: Abnormal growth and developmental malformations (reduced growth, dwarfing, stunting of plants, etc.) Reduced life span of plants, death.  Foliage symptoms: Mosaics Yellows Ring spots.  Other: Symptoms on stems, fruit, roots.  Local symptoms: Necrotic lesions.
  • 29. Detection and identification PK 29  Symptoms (possible causes by nutrient deficiencies, toxicities, insects, etc.  dsRNA isolation.  PCR, RT-PCR (for known viruses).  Virus genome sequencing.  Microarray.
  • 30. Control of plant viruses  Keep out of an area through quarantine, inspection, and certification programs that would prevent or limit local or international virus movement and warranty the use of virus-free seed, tubers, budwood .  Eradication of diseased plants  Controlling the insect vectors (oil sprays, repellents, insecticides, predators…)  Breeding plants for resistance  Transgenic resistance (natural resistance genes, viral sequences, genes from other sources). PK 30
  • 31. Reference:  Prescott Harley Klein's Microbiology - Willey, Sherwood & Woolvertion (7th ed.).  Molecular Biology of the Gene - James D. Watson, Tania A. Baker, Stephen P. Bell (5th ed.).  Wikipedia source.  YouTube source.  Characteristic of Plant viral pathogen by Svetlana Folimonova (Ass. Prof. -Dept. of Plant Pathology, University of Florida).  Introduction to Plant Viruses, the Invisible Foe (www.apsnet.org).  Viron | Biology Field Guide | Biowars (www.biowars.com).  Plant Virus replication by P.N. Sharma Department of Plant Pathology, CSK HPKV, Palampur (H.P.). PK 31
  • 32. PK 32