this presentation will give useful information about viroids..

1. By,
Devanshu dev
PALB 3256
welcome

2. VIROIDS

3. VIROIDS: The Plant Invaders…
A VIROID is a…
VIR(virus)OID(like) particle.
Viroids are“sub-viruses” composed
exclusively of a single circular strand
of nucleic acid (RNA) that codes for a
single protein.
OR
Small, circular RNA molecules without
a protein coat

4. VIROIDS: DID YOU KNOW VIROIDS…
Very small, covalently closed, circular RNA molecules
capable of autonomous replication and induction of disease.
Range in size from approximately 20 nm
(8 x 10-7 inches) ...
Are infectious particles.
Cause certain plant and human diseases.
Consists only of Nucleic Acid (RNA) .

5. No coding capacity - do not program their own polymerase.
Use host-encoded polymerase for replication.
Mechanically transmitted; often seed transmitted.
More than 40 viroid species and many variants have been characterized.
“Classical” viroids have been found only in plants.
Viroids differ from viruses in that viruses, at their most basic level, consist
of genetic material (DNA or RNA) contained within a protective protein shell.
Viroids differ from prions, another type of subviral infectious agent, in that
prions are made only of protein, lacking nucleic acid.
Contd….

6.  Thesmallestviroididentifiedsofarisa220nucleobase
scRNA(smallcytoplasmicRNA)associatedwiththerice
yellowmottlesobemovirus(RYMV)(Collinsetal.1998).
Incomparison,thegenomeofthesmallestknownviruses
capableofcausinganinfectionbythemselvesarearoundtwo
kilobasesinsize.
Manyviroidsconsistofonly300to400nucleotides.

7. 1974:
Confirmation that
viroids are non-
coding
1973: Electron
micrograph shows
viroid’s hairpin
structure
Discovery
Early 1960s:
Raymer and
O’Brien develop a
bioassay for the
agent causing
potato spindle
tuber disease
1965: Raymer
teams up with
Diener; they show
that agent is not a
typical virion
1971: Diener
demonstrates that
the agent is a free
non-coding RNA,
coins the term
viroid
1968:
Characterization of
chrysanthemum
stunt and citrus
exocortis as non-
typical viruses
1978: PSTVd is
sequenced
1976: EM
shows that
viroids form
closed circular
RNAs
Theodore O. Diener

8. Nomenclature
e
Viroid =
“virus- like”

9. Example Members
Genus Pospiviroids:
PSTVd (potato spindle tuber)
Genus Hostuviroids:
HSVd (hop stunt)
Genus Cocadviroids:
CCCVd (coconut cadang-cadang)
Genus Apscaviroids:
ASSVd (apple scar skin)
Genus Coleviroids:
CbVd 1 (coleus blumei 1)
Genus Avsunviroids:
ASBVd (avocado sunblotch)
Genus Pelamoviroids:
PLMVd (peach latent mosaic)

10. Viroid Diseases
• Potato spindle tuber viroid (PSTVd)
• May be limiting to potato growers
• First viroid characterized
• Many variants described
• Control with detection in mother
stock, clean seed
PSTVd in tomato
PSTVd in potato

11. • Citrus exocortis viroid (CEVd)
• Causes stunting of plants, shelling of bark
• May result in little yield loss
• May be useful to promote dwarfing for
agronomic advantage
• Transmitted through stock, graft
• Control by removal of infected plants,
detection, clean stock
Citrus exocortis viroid
Contd…..

12. 1. Disinfection of cutting tools
2. Cold treatment
E.g storage at 4 °C for 6months or more, followed
by apical shoot-tip-culture, grafting, can be used
to eliminate CSVd and HSVd.
3. Pre-inoculation with protective mild strains of viroid
has proved effective to control PSTVd
Viroid disease control

13. Transmission
 Most mechanically transmitted
 Some seed or pollen transmitted
 Tomato planta macho viroid-aphid
transmitted
(Ricardo Flores et al., 2005)

14. Human pathologies induced by viroids
 The only human disease known to be caused
by a viroid is hepatitis D. This disease was
previously discribed to a defective virus
called the delta agent.
 However, it now is known that the delta
agent is a viroid enclosed in a hepatitis B
virus capsid.
 For hepatitis D to occur there must be
simultaneous infection of a cell with both
the hepatitis B virus and the hepatitis D
viroid.

15. HEPATITIS D

16. Host factors RNA silencing
Viroid RNA structural elements
Factors responsible – viroid pathogenesis

17. Viroids and RNA silencing
The evidence indicates that when viroids replicate
via a double stranded intermediate RNA, they are
targeted by a dicer enzyme and cleaved into siRNAs
that are then loaded onto the RNA-induced
silencing complex.
The viroid siRNAs actually contain sequences
capable of complementary base pairing with the
plant's own messenger RNAs and induction of
degradation or inhibition of translation is what
causes the classic viroid symptoms.

18. • Some viroids are ribozymes, having RNA enzyme
properties that allow self-cleavage and ligation of unit-
size genomes from larger replication intermediates. It
has been proposed that viroids are "escaped introns."
Not all viroids are known to be pathogenic, but some
are serious pathogens of plants. Viroids are usually
transmitted by seed or pollen, but may be transported by
farm implements as well.
Infected plants can show distorted growth and
sometimes are killed by the viroid.

19. Viroid structures
Two main groups of viroids: self-cleaving and non-self-
cleaving
Non-self cleaving viroids replicate in nucleus and fold into
“dog bone” or rod-like structure
All are covalently closed circular RNAs fold to tightly
base-paired structures
Five domains identifiable in non-self-cleaving
Terminal left (TL)
Terminal right (TR)
Pathogenicity (P)
Central (C)
Variable (V)

20. The closed single-stranded RNA circle has
extensive intrastrand base pairing and
interspersed unpaired loops.
Viroids have five domains. Most changes in
viroid pathogenicity seem to arise from
variations in the P and TL domains.

21. Contd…..

22. Schematic models of viroid structures
i. (A) Rod-like secondary structure proposed for PSTVd, the
type member of family Pospiviroidae.
ii. (B) Quasi rod-like secondary structure proposed for ASBVd,
the type member of family Avsunviroidae.
iii. (C) Complex branched conformation proposed for PLMVd.

23. Pospiviroidae:
 Intra cellular movement
 cell-to-cell through plasmodesmata
 long distance through phloem
Viroid movement

24. Host factors responsible for viroid movement
 Cellular proteins
 Phloem proteins- Phloem lectin PP2(CsPP2)
 Viroid binding protiens-Virp1
 specific sequence or Structural motifs.

25. Viroid replication
 Viroid RNA does not code for any known protein;
some even lack the AUG initiation codon.
 Nonetheless, they replicate autonomously in host
cells.
 The replication mechanism involves interaction
with RNA polymerase II, an enzyme normally
associated with synthesis of messenger RNA, and
"rolling circle" synthesis of new RNA.

26. Asymmetric rolling circle replication
Viroid replication
(Biao Ding et al., 2005)
Member of Pospiviroidae replicate via an asymmetric
 Incoming (+)-circular RNA initially is transcribed into
concatemeric linear (-)-strand RNA
 Which then serves as the replication intermediate for
the synthesis of concatemeric, linear (+)- strand RNA
 This (+)- strand RNA subsequently is cleaved into unit
length monomers that are ligated into circles
viroids replicate autonomously by using host-encoded RNA
polymerase
Concatemer: Multiple copies of a DNA sequence arranged
end to end in tandem.

27. Enzymes involved in
replication
RNA polymerase
RNA cleavage
RNA ligase

28. Asymmetric Symmetric

29. Symmetric rolling circle replication
 Member of Avsunviroidae replicate via an symmetric
 The circular (+)-RNA is transcribed into linear, concatemeric (-)-
strand RNA
 Instead of serving as the direct template for the synthesis of
linear concatemeric (+)- strand RNA
 The concatemeric (-)- strand RNA is cleaved into unit length
molecules followed by circularization
 The circular (-)- RNA then serves as the template for the
synthesis of linear, concatemeric (+)- strand RNA
 When subsequently is cleaved into unit-length monomers and
circularized