BIBLIOGRAPHY SABAWAY FREDERICK A. APRIL...
BIBLIOGRAPHY
SABAWAY FREDERICK A. APRIL 2012. Pathogenecity Test of Colletotrichum
Isolates on Arabica Coffee Seedlings and Berries. Benguet State University, La Trinidad,
Benguet.
Adviser: Andres A. Basalong, MSc.
ABSTRACT
The study was conducted from October 2011 to February 2012 at the Department
of Plant Pathology Laboratory, Benguet State University, Plant Pathology Laboratory.
Colletotrichum spp. isolated from leaf(
Colletotrichum coffeanum), stem/twig
(
Colletotrichum gleosporioides) and berry (
Colletotrichum kahawae) of Arabica coffee
were inoculated on leaves and stem of Arabica coffee seedlings and berries, to determine
their infectivity.
All the three
Colletotrichum species infected the leaf, stem/twig and berry of
Arabica coffee but at varying degrees of severity.
Colletotrichum coffeanum which was
isolated from the leaves showed the highest infection on leaves and severe infection in
berries.
Colletotrichum gleosporioides isolated from stem/twig had the highest infection
when inoculated in stem/twig and very severe infection in berries. Likewise,
Colletotrichum kahawae isolated from berries had very severe infection to berries, but were
likewise infectious to leaves and stem/twig.
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
RESULTS AND DISCUSSIONS
Symptoms on naturally infected plant parts
The symptoms are most visible on leaves and ripe fruits. At first, anthracnose
generally appears on leaves as small and irregular yellow, brown, dark-brown or black
spots. The spots can expand and merge to form larger affected area. The color of the
infected part darkens as it ages.
The disease can also produce brown to black color lesions on the stem and
blackening of the whole stem.
Infected fruit has small, water-soaked, sunken, circular spots that may increase in
size up to 1 cm in diameter. As it ages, the center of an older spot becomes blackish and
emits gelatinous pink spore masses (Figure 5).
Figure 5.
Colletotrichum spp. on leaf, stems, and fruit
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
Cultural and Morphological Characterization of the Pathogen
Table 1 summarizes the cultural and morphological characteristics of the
Colletotrichum isolates.
Colony size. Isolates differed in the diameter of their colony growth in culture
media within one week.
Colletotrichum coffeanum attained the largest colony diameter of
40.3 mm, followed by
Colletotrichum gloesporiodes at 35.5 mm and the least was isolate
Colletotrichum kahawae at 30.5 mm. Based on the size of colony,
Colletotrichum
coffeanum obtained from the infected leaves was fast growing compared to other isolates.
Colony color. The colony of
Colletotrichum coffeanum and
Colletotrichum
kahawae was white to black, (Figure 11. A and C), while
Colletotrichum gleosporioides
was white colony (Figure 11. B).
Rate of growth.
Colletotrichum coffeanum was observed to be fast growing as
compared to
C. gleosporioides and
C. kahawae which were observed to be moderately fast.
These observed growth characteristics conform to their differences in colony size in one
week.
Mycelia. The mycelia of
Colletotrichum coffeanum were hyaline branched,
septated and elongated. Mycelia of
Colletotrichum gleosporioides were observed to be
hyaline, septated, and large walled, while mycelia of
Colletotrichum kahawae were
slender, septated and hyaline.
Conidia. Shape of conidia varied between isolates. Conidia of
Colletotrichum
coffeanum is oblong,
Colletotrichum gloesporiodes is elliptic, and cylindrical for
Colletotrichum kahawae. Conidia colors were observed similar to all the isolates as
hyaline. In terms of the conidia size, length and width of Colletotrichum coffeanum has a
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
size of 5.31 x 2.06 mm (Figure 12. A), Colletotrichum gleosporioides 4.25 mm x 1.40 mm,
and Colletotrichum kahawae 5.25 mm x 0.50 mm (Figure 11. B and C). This
characterization confirmed the study of Coyasan (2011).
Table 1
. Cultural and morphological characteristics
of
Colletotrichum isolates at
temperature range of 24-26 oC on
PDA.
COLLETOTRICHUM ISOLATES
CRITERIA
C. coffeanum C. gloesporiodes C. kahawae
Colony diameter 40.3 mm
35.5 mm
30.5 mm
(mm)
Colony color
White to black
White
White to black
Rate growth in one
Fast growing
Moderately fast
Moderately fast
week
Presence of septa on
Present
Present
Present
mycelia
Shape of conidia
Oblong
Elliptic
Cylindrical
Color of conidia
Hyaline
Hyaline
Hyaline
Size of conidia
5.31 mm x 2.06 mm 4.25 mm x 1.4
5.25 mm x 0.50mm
mm
A
B
C
Figure 6.
Colletotrichum spp. Isolate on A. leaf, B. twig and C. berry
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
A
B
C
Figure 7.
Colletotrichum spp. conidia of isolate A. leaf, B. twig and C. berry (400x)
Pathogenecity Test and Symptom Development
Symptoms on inoculated seedlings. Symptom appearance varied between each
plant parts. Irregular yellowish spots appeared on the leaves eight days after inoculation
(Figure 8). The spot was observed mostly on the older leaves. The infection expands as it
age (Figure 9)
.
Figure 8. Slight infection
Figure 9. Severe infection on leaf
on leaf surface surface
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
Symptoms on inoculated Stem/twig. Sunken lesion with slightly raised rim was
observed on the stem of the seedling twelve days after inoculation. As it ages, this causes
stem and twig dieback (Figure 10).
.
.
.
Figure 10. Brown lesions on the stem
Symptoms on inoculated berries. Four days after inoculation, the appearance of
symptom was observed on the berry. Symptom on berry isolate occurs as a small pit at the
middle part of the berry that usually starts as a dark lesion on the surface of the berry
(Figure 11). As it ages, the infection occupies the whole berry and produces pink spore
masses and mycelia (Figure 12).
Figure 11. Dark lesion on Figure 12. Severe infection
Arabica coffee berries on Arabica coffee berries
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
Disease Severity Ratings on Arabica coffee Seedlings and Berries
Table 2. Initial and final mean disease severity rating on leaves samples of inoculated
seedlings
TREATMENT
Initial rating
Final rating
T1= UNTREATED
1
1c
T2=
Colletotrichum coffeanaum
2.5
6a
T3=
Colletotrichum gleosporioides
1.5
3.5b
T4=
Colletotrichum kahawae
2
3b
Cv= Coefficient variance
40.75%
50.61%
*means with the same letters are not significantly different at 0.05% level of significance
Initial disease severity rating on Leaves. Table 2 shows the initial severity rating on
leaf samples of inoculated seedlings. The highest rating was obtained from
Colletotrichum
coffeanum of 2.5 but not significantly different from the rating obtained from the other
species. The lowest severity rating was observed from
Colletotrichum gleosporioides
isolate which is not significantly different from
Colletotrichum coffeanum and
Colletotrichum kahawae isolates. The control had not infected with a rating of 1.0.
Final disease severity rating on leaves. The highest final disease severity rating on
leaves was obtained from
Colletotrichum coffeanum which was significantly different from
the other species (Figure 13). The result indicates that the
Colletotrichum species either
from leaves, stem/twig, and berries are pathogenic to leaves of Arabica coffee (Table 2).
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
Table 3. Initial and final disease severity rating on stem/ twig of Arabica coffee seedling
TREATMENT
Initial rating
Final rating*
T1= UNTREATED
1
1b
T2=
Colletotrichum coffeanum
1
2b
T3=
Colletotrichum gleosporioides
2
5.5a
T4=
Colletotrichum kahawae
2
2.5b
Cv= Coefficient variance
50.77%
52.61%
*means with the same letters are not significantly different at 0.05% level of significance
Initial disease severity rating on Stem/Twig. Initial Infection ratings on stem/twig
showed no infection to very slight infection with ratings of 1 and 2, respectively.
Colleotrichum gleosporioides and
colletotrichum kahawae have the same infection ratings
were not significantly different from
colletotrichum coffeanum (Table 3).
Final disease severity rating on stem/twig. The highest final disease rating on
stem/twig was obtained from
Colletotrichum gleosporioides (Figure 13), which was
significantly different from the other species (Table 3).
Table 4. Initial and final disease severity rating on berries
TREATMENT
Initial rating
Final rating
T1= UNTREATED
1
1b
T2= Colletotrichum coffeanum
1
8.33a
T3= Colletotrichum gleosporioides
1.67
9a
T4= Colletotrichum kahawae
2.33
9a
Cv= Coefficient variance
13.64%
8.41%
*means with the same letters are not significantly different at 0.05% level of significance
Initial disease severity rating on Berries. Table 4 shows the initial infection ratings
on berries. The highest infection rating was obtained from Colletotrichum kahawae of 2.33
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
but not significantly different from the ratings obtained from the other species.
Colletotrichum coffeanum shows no infection which is not significantly different from the
other species.
Final disease severity rating on berries. The highest final severity disease rating was
obtained from
Colletotrichum kahawae and
Colletotrichum gleosporioides (Figure 13), but
not significantly different from rating of
Colletotrichum coffeanum (Table 4)
Untreated
Colletotrichum coffeanaum
Colletotrichum gleosporioides
Colletotrichum kahawae
Figure 13. Graph showing the Summary of Final Ratings on leaves, stem/twig and
berries
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
SUMMARY, CONCLUSION AND RECOMMENDATION
Summary
The study was conducted from October 2011 to February 2012 at the Department
of Plant Pathology Laboratory, Benguet State University.
Isolates of
Colletotrichum spp. isolated from leaf (
Colletotrichum coffeanum),
stem/twig (
Colletotrichum gleosporioides) and berry (
Colletotrichum kahawae) of
Arabica coffee are inoculated on leaves and stem of Arabica coffee seedlings and berries
to determine their pathogenecity on either leaves and stem of Arabica coffee seedling and
detached berries.
The three isolates of
Colletotrichum spp. were found to be pathogenic to Arabica
coffee leaves, stem/twig and berries. Spots, and canker were the symptom developed on
the Arabica coffee seedling and berries.
Conclusion
Colletotrichum coffeanum was identified as the main causal pathogen in the leaves,
Colletotrichum glesporioides in stem/twigs, and
Colletotrichum kahawae in the berries.
However all Colletotrichum spp. are able to infect the leaves, stem/twig, and berries.
Recommendation
It is recommended that a verification study will be done in a greater number of test
samples and in different varieties of Arabica coffee. It should also be done in the field, not
on seedlings only. Likewise, trials on berries should also be done on intact berries to the
mother plants.
Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012
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Pathogenecity Test of Colletotrichum Isolates on Arabica Coffee Seedlings and Berries |
SABAWAY FREDERICK A. APRIL 2012