BIBLIOGRAPHY LAN-EW, JULIET K....

BIBLIOGRAPHY

LAN-EW, JULIET K. May, 2013. Preliminary Documentation of Viral Diseases of
Strawberry (Fragaria ananassa) in Swamp , La Trinidad, Benguet, Benguet State
University, La Trinidad, Benguet.

Adviser: Janet S. Luis, Ph. D.

ABSTRACT

The study was conducted to document, collect and identify virus diseases affecting
strawberry plants in Swamp La Trinidad and to identify this virus diseases through
symptomatology.
Results of the study showed that strawberry grown in Swamp La Trinidad showed
symptoms like crinkling, distortion and yellowing of the plants which is attributed to virus
infection. The viral disease was confirmed through sap inoculation using Gotukola
(Centella asiatica) as indicator plant (Toclo and Ligat, 2012. The Gotokula inoculated with
strawberry mottle virus showed vein clearing and mottling symptoms. On the other hand,
the Gotokula inoculated with strawberry crinkle virus showed distorted crinkling of the
leaves and chlorotic spotting symptoms. This result confirms that the symptoms observed
on the assessed strawberry plants in the Swamp area in La Trinidad was caused by
strawberry mottle virus and strawberry crinkle virus.

RESULT AND DISCUSSION
Disease symptoms of strawberry viruses
The symptoms observed were based and similar to the virus infection of strawberry
mottle virus disease as discussed by Leone et al (1992). The symptoms observed and
identified to be caused by strawberry mottle virus disease were vein clearing, mild mottling
and distortion on the leaves. The leaves showed small chlorotic spots that were scattered,
veins that were cleared, leaves becoming bright in color or mild mottling and distortion of
the leaves as compared to the normal leaves. The other symptoms observedwere the
distorted and crinkled appearance on the strawberry leaves.In addition, the leaves showed
small chlorotic spotting. These symptoms were identified to be caused by strawberry
crinkle virus. Symptoms were similar to the virus disease characteristics as discussed by
Yokishawa et al (1986). Symptoms of collected specimens from the 4 areas are shown in
Figures 1and2.
All commercial strawberry cultivars infected with the virus, although symptomless
were observed to have reduced vigor and yield.

A
B

C
D
Figure 1- Strawberry crinkle virus disease symptom. A- Distorted crinkledsymptom,B-

small chlorotic spotting and distorted crinkled symptom, C&D-distorted crinkled

symptom
C
D
A
B
A
Figure 2-(A, B, C&D) Strawberry mottle virus disease symptom.A-showsleaf mottle

symptom,B-C, & D- shows that there is chlorotic spotting and mottling on the

leaves

Fa

Fa
rm
rm

1

2

Fa

rm
4

Fa

rm

3

Figure 3- BSU ATBI/ IC Farm (2012-2013) Locator map
Farm 3
Farm 4
Farm 1
Farm 2
Farm 2

Incidence of Strawberry Virus and aphid in the Field

Table 1 shows the disease incidence in the four sampling sites. Results revealed
that differences in virus incidence ineach area maybe caused by the way the farmers
manage their farms. The four farmersplanted the same variety of strawberry, which is
Sweet Charlie, and runners were of the second generation from BSU tissue cultured
strawberry runners. Farm 3 had the highest disease and aphid incidence. When
interviewed, the farmercontrols pest and diseases depending on his observation in the field.
He only schedules his spray when he sees that insects like aphids are present and uses green
labeled chemicals like Black hawk. The weeds that are present in the field that may serve
as alternate host for the aphid are removed. As no crop rotation is practiced, the increase
in population of the aphids was observed. Farm 4 had the lowest incidence of virus
diseases. In this farm, the population of aphids was kept under control as spraying done at
15 days interval. The farmer used green labeled chemicals like Ortus. The farm is also kept
clean of weeds that may serve as alternate host for the aphids. Rotation of lettuce before
strawberry was done.

Even though the planting material were free of virus disease infection,
Insects typically aphid are able to transmit virus diseases by acquiring then transmitting
the disease from the infected plant to the un infected plant, (Tarr 1966). Aside from aphid,
mites were also present in the field, colonies of mites have needle-like stylets that pierce
and suck the plant sap, usually from the under surface of the leaves, which properly might
make them vector of plant diseases, (Towsend 1996).

Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013

Table 1. Incidence of strawberry mottle and crinkle viruses in the field
Sampling sites
Strawberry mottle
Strawberry crinkle

Farm 1 (Manangwe, N)
17.33
12.00
Farm 2 (Bartolome, B.)
10.67
16.00
Farm 3 (Pa-at,A)
14.33
18.67
Farm 4 (Puyao, J) 7.33
9.33

Table 2. Aphid incidence in the field

Aphid incidence on
Aphid incidence on
Sampling sites
strawberry mottle
strawberry crinkle
Farm 1 (Manangwe, N)
29.6

30.3
Farm 2 (Bartolome, B.)
44.5

46.3
Farm 3 (Pa-at, A)
46.7

48.3
Farm 4 (Puyao, J)
27.7
27.00

The aphid had the great potential in transferring the disease because of acquiring
the disease from the infected plant then transfer to another healthy plant that makes the
disease to spread. The aphid Chaitosiphon fragaefolii were seen on the leaves of the
strawberry and it is one of the factors to consider why there was virus infection even if the
planting materials were tissue cultured from BSU which are supposed to be free from virus
infection.Aphid as vectors which quickly lose their ability to fly are likely to be less
effectively in spreading viruses than those which retain the ability, Tarr (1966). During
their initial flight, aphids tend to feed on a succession of plants before finally settling, and
Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013

this type of behavior expedites the spread of viruses, particularly those viruses which are
rapidly acquired and transmitted.
Strawberry crinkle virus is transmitted in a persistent propagative manner by the
aphids. The aphid remains infective for day or weeks, often for life. The length of
transmission cycle is in nature dependent on temperature conditions. Low temperature
extends incubation period in strawberry and latent period of the virus in the vector has been
reported to be about 2 weeks. Severe occurrence of strawberry crinkle virus shows the
symptoms of crinkling of leaves, leaflets unequal in size and small irregular shaped,
chlorotic spots. Mild strain result no symptoms on strawberry (Tripathi, 2004).

Strawberry mottle virus is known to transmit the virus in semi-persistent manner.
The virus is lost by the vector when it molts and it does not multiply in vectors gut. The
advantages of virus persistence in the vector are partly nullified by the short flight period
and short life of migrating aphids, especially if there is a prolonged latent period during
which the aphid is not infective, Frazier et al, (1988).
Viruses which infect strawberry plants include strawberry mottle virus and crinkle
virus are transmitted from plant to plant by insects, typically aphid.

Figure 4- The aphid from the strawberry leaf showing symptom of virus infection
Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013

Aside from the aphid, mites were also present in all plants in the area, Towsend
(1996) stated that colonies of mites occur mainly on the underside of leaves, sheltered by
strands of fine webbing. They have needle-like stylets that pierce and suck the plant sap,
usually from the under surface of the leaves, which properly might make them vector of
plant diseases, Agrios (2005) stated that mites carry plant viruses on their stylets and can
acquire and inoculate the virus after feeding from the virus infected plants
.

Figure 5- Mites present in the underside of the strawberry leaves.
Confirmation of Virus Disease Using Indicator Plant
The indicator plant Gotukola exhibited similar symptoms of the strawberry
showing the virus infection. The infection showed finely cleared veins, mild mottling and
distorted and crinkled leaves.
The Gotukola as indicator plant was inoculated with the extracted sap of strawberry
leaves that showed the symptoms of strawberry mottle virus disease infection. The
indicator plant also showed the symptoms of mottle virus disease infection, the same as the
symptoms were observed in the strawberry plant. The symptoms did not appear from the
first day of inoculation but it started to appear 2 days after inoculation at 1.80 and 1.60
Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013

respectively which is equivalent to 1-10% infection then the severity continued to increase,
(Table 3).
Table 3. Disease severity of virus infection in Gotukola
Observation period in days after inoculation
Virus
1
2
3
4
Strawberry crinkle virus disease
0.00
1.80
4.65
6.65
Strawberry mottle virus disease
0.00
1.60
3.90
5.90

Figure 6. Gotukola as indicator plant

A
B

Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013

D
C
C
D
A
B
Figure 7-.A-not inoculated test plant. B, C&D- inoculated test plant showing mild

mottling and minor leaf distortion
Figure 8-A- shows uneven leaf blade and distorted crinkled leaves, B, C&D- showed

chlorotic spotting, distorted crinkled leaf and un even leaf blades.

C

C
D
Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013

SUMMARY, CONCLUSION AND RECOMMENDATION
Summary

Fourstrawberry farms measuring 500 square meters served as collection sites.
Strawberry plants having the symptoms of virus disease infection like mottling and
crinkling on the leaves werecollected then brought in the laboratory for confirmatory
studies using Gotukola as indicator plant.
The infection of crinkle and mottle viruses in strawberry was confirmed using
Gotukola (Centella asiatica) as indicator plant (Toclo and Ligat, 2012). Result showed that
the inoculated Gotukola showed more distinct symptoms three days after
inoculation.Symptoms includemild mottling and minor leaf distortion, chlorotic spotting,
distorted crinkled leaves and uneven leaf blades.

Data on aphid incidence gathered in every 6 plants surrounding the virus infected
strawberry plant revealed the presence of aphids which may have caused the infection of
other strawberry plants. The differences in virus incidence in each area maybe caused by
the way the farmers control the aphids which is viruses’ vector.

Conclusion

Confirmation test showed that symptoms of strawberry mottle virus and the
strawberry crinkle virus infection were present in the four strawberry farms. Confirmation
of infection using indicator plant showed symptoms like mottling and crinkling of
leaves.Virus infection of crinkle virus disease and mottle virus disease appeared on the
indicator plant 3 days after inoculation. In addition, the Gotukola plant that was used as
indicator plant was effective as indicator plant for virus symptom specifically strawberry
Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013

mottle virus and strawberry crinkle virus. The presence aphids must be controlled tolessen
if not to prevent the spread of virus disease infection in the field.
Recommendation
It is recommended that plants showingsymptoms of viral infection in the field should
be removed and burned to avoid the spread of the disease. The use of virus free planting
materials for establishment of the new plantation is needed to decrease potential sources of
infection. Weeds must be removed in the field because it may serve as alternate host for
the viruses like strawberry mottle virus and strawberry crinkle virus that will allow for
these viruses to survive even in the absence of the host. Vector control can help reduce the
chances of disease spread to some extent because they transmit the disease through
transferring from one place to another. Gotukola can be used for indexing crinkle and
mottle virus of strawberry in the production of clean planting materials of strawberry.

Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013

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Preliminary Documentation of Viral Diseases of Strawberry (Fragaria ananassa) in Swamp
La Trinidad, Benguet | LAN-EW, JULIET K. May, 2013