1. Pre-Test
Simple Recall
1. The Father of Plant Virology.
2. The infective part of a viral particle.
3. The most important vector of plant viruses.
4. The first person to purify TMV and was a Nobel Prize awardee in 1946
because of this work.
5. Filipino plant pathologist who conducted numerous studies on abaca
mosaic virus in 1920s.
6. An entity mainly composed of RNA which is capable of causing plant
diseases.
7. The structure used by nematode as it feeds on plant roots.
8. Nematodes that penetrate inside the host as they feed on host plants.
9. A technique in virus detection and identification that involves antigen-
antibody reaction.
10. The most common technique in nematode isolation.
3. What is a Plant Virus?
… a nucleoprotein that has the ability to cause disease in
plants
… all viruses are parasitic in cells and cause a multitude
of disease in all forms of living organisms, from single-
celled microorganisms to large plants and animals
… more than 2000 species of viruses are known and about
1/4 are reported to infect plants
… composed of nucleic acid and protein coat
4. Size
ultramicroscopic
visualization requires an electron microscope
Morphology
filamentous/flexous rods, isometric, geminate, bacilliform
flexous rod – Maize dwarf mosaic virus
rod-shaped – Tobacco mosaic virus
isometric – Cowpea chlorotic mottle virus
Genome
nucleic acid genomes: either RNA or DNA
variations in genome structure (single-stranded or
double-stranded; linear or circular; monopartite or
multipartite)
5. Capsids
have one or more protein coats composed of protein subunits
Obligate Parasite
depend on hosts for existence
have no systems for the accumulation of metabolic materials;
energy generation; protein synthesis; capture of light energy
entirely dependent on hosts for NA and amino acid synthesis
Cell Association
infect cellular structure and control part of the subcellular
systems of the hosts; act as “molecular pirates”; move directly
from cell to cell through plasmodesmata
6. Replication by Assembly
depend on pools of required components from hosts, e.g. the
enzyme systems (polymerases and reverse transcriptase)
Vectors
not capable of causing an entry wound; depend on vectors
to breach the epidermal layer of the plant
include insects, mites, nematodes, fungi, seeds, dodders, human
and animals
9. DNA Viruses (cont.)
ssDNA
Single isometric particles
Banana Bunchy Top Virus
dsDNA
Isometric, circular dsDNA
Genus: Caulimovirus (Cauliflower Mosaic Virus)
Bacilliform (rod-shaped)
Genus: Badnavirus (Rice Tungro Bacilliform Virus)
10. Historical Developments in Plant Virology
1500s - “Tulipomania” in Holland
- unusual case of a virus that increased the value of
infected tulips
- termed as “bizarres”
1886 - Adolf Mayer scientifically confirmed a primary
principle to plant virology when he transmitted
TMV to healthy tobacco plants by rubbing with
sap from infected plants
11. 1892 - Dmitri Ivanoski (Russian)
1898 - M. V. Beijerink (Dutch); they first indicated the
unique nature of viruses; they declared TMV as a
new type of pathogen called “contagium fluidium
vivium” or a contagious living fluid
1935 - W. M. Stanley worked on TMV and was the first
person to purify a virus; awarded the Nobel Prize
in 1946
1936 - Bawden and Pierie found the presence of
phosphorus in virus genome (indicated that the
suspension contains NA)
12. 1956 - Fraenkel-Conrat was able to isolate RNA from
TMV and used it to infect healthy tobacco plants
In the Philippines, Gerardo O. Ocfemia made numerous
studies on abaca bunchy top disease in 1920s to 1940s
21. Latent Symptom – the host is infected by a virus but does
not show any visible symptom (termed as
symptomless carriers)
Masked Symptom – the host develops symptom upon
infection but temporarily becomes
symptomless under certain environmental
conditions (e.g. high or low temperature)
22. Virus Transmission
Mechanical Transmission
allows transmission of viruses without a vector
effective for viruses that infect epidermal cells
use of abrasives e.g. silica carbide, sand, bentonite
and celite
Potato virus X, Tobacco mosaic virus and Cucumber
mosaic virus
23. Insect Transmission
5 Orders of insects
Order Homoptera (aphids, leafhoppers,
planthoppers, whiteflies, mealybugs and
treehoppers)
Order Hemiptera (true bugs)
Order Thysanoptera (thrips)
Order Coleoptera (beetles)
Order Orthoptera (grasshoppers)
24. Modes of Insect Transmission
nonpersistent viruses – persist in the vector for only
a few to several hours (termed as “stylet-borne”)
***APHID is the most important vector and
transmit about 250 known species of plant
viruses
semipersistent viruses – persist in the vector for
about 1 to 4 days
circulative/persistent viruses – accumulate
internally but not multiply
circulative/propagative viruses – multiply in their
vectors
25. Nematode Transmission
transmit about 20 viruses
Longidorus, Paralongidorus and Xiphinema transmit
polyhedral-shaped viruses (nepoviruses) such as
grape fanleaf and tobacco ringspot
Trichodorus and Paratrichodorus transmit rod-
shaped viruses (tobraviruses) such as tobacco rattle
and pea early browning
Mites Transmission
mites species belong Family Eriophyidae
transmit about 6 viruses (wheat streak mosaic)
26. Fungal and Fungal-like Transmission
species belong to Class Chytridiomycetes and
Plasmodiophoromycetes
transmit about 15 species of plant viruses
Root infecting species such as Olpidium, Polymyxa
and Spongospora
Dodder Transmission
Cuscuta sp. (with twining stems forming bridge
between two plants)
Other Modes of Transmission
seed, pollen and vegetative propagation and grafting
31. How are Viruses Controlled?
Host Plant Resistance
Cultural Methods
Control of Vectors
Quarantine and
Eradication
32. Host Plant Resistance
Develop plant cultivars with viral resistance through
traditional breeding.
Cross-protection increases resistance of host plant by
introducing the mild strain of the virus.
Genetic engineering to enhance host plant resistance
using genes coding for coat proteins, replicase,
antisense RNA and ribozymes
33. Vector Control
Apply measures to control vectors.
Quarantine and Eradication
Quarantine – acts as the first line of defense against the
introduction of many foreign viruses
Eradication – done when exclusion of viruses fails
Cultural Control
can be referred to as “wise production practices”
34. Plant Viroids
small, low molecular weight RNA that can infect plant
cells, replicate themselves and cause diseases
differ from viruses in at least 2 main characteristics:
1. size of RNA – viroid has 250 to 370 bases;
virus has 4 to 20 kilobases
2. virus RNA is enclosed with a protein coat while
viroid lacks this structure
Cadang cadang disease of coconut
Potato Spindle Tuber
Citrus Exocortis
Avocado Sunblotch
Apple Scar Skin
35. Cadang cadang Disease (CCCVd)
“cadang cadang” means dying
first recognized in 1930s; similar disease called “tinangaja” caused
by a related viroid occurred in Guam
the pathogen infects coconut and other palm species in the
Philippines (the only known monocot-infecting viroid)
seed transmission is only 0.3%
consists of 246 nucleotides, making this the smallest viroid known
disease develop slowly over 8 to 15 years
first symptom: fruits become rounded and develop
scarifications then leaves show bright yellow spots
3 to 4 years later:inflorescences are killed; only new fronds
develop
5 to 7 years later: severe leafspots will be noted; size of fronds
in the crown continue to reduce
finally: growing buds die, fall off and leaves the palm trunk
standing like telephone posts
36. Control of Plant Viroids
Use of viroid-free
planting materials
Eradication of infected
plants
37. Plant Parasitic Nematodes
derived from the Greek words “nema/nematos” =
thread and “edos” = resembling or likeness
small, 300 to 1000 micrometers with some species up to
4mm long and 15-35 micrometers wide
generally eel-shaped or vermiform and round in cross-
section with smooth unsegmented bodies, without legs
or other appendages
nematodes may either be saprophagous, predaceous or
plant parasitic
10% of the 30,000 described species are known as plant
parasites
plant parasitic species are mostly obligate and
equipped with stylet
38. Morphology
… the cross section of a nematode body looks like a “tube
within a tube”
Outer Tube
Cuticle – semi-permeable outermost covering of the
nematode body; made up of protein (keratin and
collagen); shed off during molting to allow growth
Epidermis – thin layer which functions mainly for
secretion of new cuticle during molting
Muscles – allows movement by alternate contraction
and relaxation of dorsal and ventral muscles
39. Inner Tube
begins from the mouth opening to the anus (for female)
or cloaca (the common opening of digestive and
reproductive systems for male)
possess stylet that they use in puncturing plant cells
and withdrawing their contents
three types of stylets:
Stomatostylet – with distinct cone, shaft and knobs; inside
hallow; possessed by members of the
Order Tylenchida
Odontostylet – a hallow spear; possessed by members of the
Order Dorylaimida
Onchiostylet – a bent solid needle-like stylet possessed by
members of the Order Triplonchida
40. Life Cycle of Nematodes
Adult
Larvae
(4 stages)
Eggs
Note: Nematode cycle is completed within 3 to 4 weeks only.
41. Terms Related to Nematode Reproduction
Amphimictic – nematode species which requires the
presence of both female and male for reproduction to
occur.
Parthenogenic – nematodes reproduce from eggs
without fertilization by sperm cells.
Hermaphroditic – possessing both functional male and
female reproductive organs.
Sexual dimorphism – phenomenon wherein the male
and female nematodes of the same species have entirely
different morphology.
Sex reversal – phenomenon observed in juveniles of
parthenogenic species of Meloidogyne during
unfavorable condition. Depending on the onset of the
unfavorable condition, the gonad of “destined to be”
female juveniles develops into non-functional testis.
42. Groups of Nematodes
Based on Feeding Position
Ectoparasites – feed from the outside and only the
stylet enters the plant cells
Belonolaimus – sting nematode
Paratylenchus – pin nematode
Trichodorus – stubby-root nematode
Xiphinema – dagger nematode
Longidorus – needle nematode
Semi-endoparasites – feed by burying the front part
of the body into the host cells while the posterior
portion is outside the host
Pratylenchus – lesion nematode
Radopholus – burrowing nematode
43. Hoplolaimus – lance nematode
Helicotylenchus – spiral nematode
Ditylenchus – stem and bulb nematode
Aphelenchoides – foliar nematode
Endoparasites – the entire nematode body enters the
plant cells while it feeds
Meloidogyne – root-knot nematode
Heterodera – cyst nematode
44. Based on Movement while Feeding
Migratory – move from one part of the plant to
another portion of the host or move from plant to
the soil and back
Radopholus
Pratylenchus
Ditylenchus
Aphelenchoides
Sedentary – attach themselves to the roots or
burrow into the root
Meloidogyne
Rotylenchulus – reniform nematode
Tylenchulus – citrus nematode
47. Important Nematode Diseases in the Philippines
1. Slow decline of citrus – Tylenchulus semipenetrans
2. Spreading decline of citrus – Radopholus similis
3. Toppling of banana - Radopholus similis
4. Root knot of various crops – Meloidogyne spp.
5. False root knot – Naccobus spp.
6. Ufra disease of rice – Ditylenchus angustus
7. White tip disease of rice – Aphelenchoides besseyi
8. Rice root knot (mentek) – Hirschmaniella oryzae
9. Yellow dwarf disease of black pepper
– Radopholus similis
10. Red ring disease of coconut
– Radinaphelenchus cocophilus
48. Sampling Techniques
Reasons for Sampling:
nematodes are too small to be seen with bare eyes
symptoms caused are not typical
need for quantitative estimation of density
Sampling Theories:
1. Spatial Distribution – nematode distribution within
the field is generally patchy owing to their small size
and slow rate of active movement.
2. Vertical Distribution – refers to the dispersal of
nematodes in a soil profile; mainly influenced by
density of feeder roots which is higher in the upper 30
cm of the soil.
49. 3. Temporal/Seasonal Distribution – influenced by
climatic pattern (wet and dry) and host availability;
nematode density is highest during near harvest stage
of crop
Extraction Techniques
1. Motility-dependent/active methods – Baermann
funnel and its modifications; H2O2 incubation
method
2. Motility-independent/passive methods – wet sieving
method; maceration; elutriation method; centrifugal
flotation
50. Disease Complex with Nematode
Nematode and Viruses – Trichodorus/Paratrichodorus
and Tobacco Rattle Virus
Nematode and Bacteria – Meloidogyne and
Ralstonia solanacearum
Nematode and Fungi - Meloidogyne and
Fusarium oxysporum
51. Methods of Nematode Management
1. Prevention – quarantine at farm, country and
international level
2. Crop rotation – alternating susceptible host with non-
host crop
3. Trap crops – susceptible crops intentionally planted
but are uprooted before the nematode complete its life
cycle.
e.g. Crotolaria
4. Antagonistic crops – exudes chemicals that are either
repellant or nematicidal
e.g Tagetes patula – produces a-thertienyl
Brassicas – produces isothiocyanates
52. 5. Land management and cultural practices
Early planting
Plowing
Flooding
Organic soil amendment
6. Physical methods
Steaming
Solarization
Field burning
Heat treatment of planting materials
7. Resistant/Tolerant varieties
8. Biological control e.g. BIOACT
9. Chemical control
Fumigants e.g. methyl bromide, ethylene dibromide,
chloropicrin
Non-fumigants e.g. fenamiphos, thionazin, aldicarb, oxamyl,
methomyl
54. Post-Test
Give the specific causal agent of the following diseases:
1. Tomato root-knot infection
2. Tungro
3. Pepper Mosaic
4. Bunchy Top
5. Cadang cadang of coconut
True or False
1. All plant parasitic nematodes are obligate.
2. The completion of the life cycle of nematode is only 3-4 weeks.
3. Viruses can be detected and identified based on their protein
components.
4. ELISA is one of the serological techniques for virus identification.
5. Viroids are apparently naked entities.