BIBLIOGRAPHY CODOD, CLARENCE BAGAYAO. ...
BIBLIOGRAPHY
CODOD, CLARENCE BAGAYAO. APRIL 2013. Survey, Assessment, and
Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp. cubense) in Two
Barangays of Kapangan, Benguet. Benguet State University, La Trinidad, Benguet.
Adviser: Asuncion L. Nagpala, Ph.D.
ABSTRACT
Survey and sampling of banana plants infected with Fusarium wilt were done in the
Municipality of Kapangan barangay Sagubo, sitio Timoc, Ampongot, and Dosco, and
barangay Gadang sitio Namun- ao, Gadang Proper, Namat- ikan, Copias, and Lib – libeng,
Benguet from September 2012 to November 2012 to confirm the occurrence of the disease
in the locality; assess the incidence and severity of Fusarium wilt infection in affected
banana plants; isolate and characterize the causal pathogen Fusarium oxysporum f. sp.
cubense from infected samples; and compare the effect of bio control agent and chemicals in
inhibiting the growth of F. oxysporum f. sp. cubense through assay.
Results revealed that Fusarium wilt incidence in barangay Sagubo and Gadang was
16.82 % and 13.57 % respectively. In terms of severity, variety Lakatan was the most
susceptible to Fusarium wilt infection having the most intense discoloration on the
Pseudostem followed by the variety Cavendish (Tomoc) and Bungulan (Kantong) with
mean severity ratings of 5.0, 4.0 and 3.67 respectively. Varieties Saba (Balayang) and
Cardaba (Dippig) did not incur any infection.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Isolates of F. oxysporum f. sp. cubense from Lakatan, Cavendish and Bungulan
designated as isolate 01, 02 and 03 differed in their colony growth in Potato Dextrose Agar
with 02 isolate having the widest mean growth of 65 mm. Isolates 01 and 03 had 62.67
and 59.83 mm growth diameter respectively.
Microscopic measurements of the macro conidia of the three isolates were 30.42 x
3.64 µm, 31.20 x 4.68 µm and 33.80 x 4.81 µm and were found to be within the range of
the published measurement of F. oxysporum which is 27-55 x 3.3-5.5 µm. The micro
conidia of the three isolates measured 8.77 x 2.73 µm, 8.06 x 2.86µm and 7.80 x 3.25 µm
which is also within the range of 5-16 x 2.4-3.5 µm.
Bioassay results showed that Benomyl inhibited the growth of the three isolates of
F.oxysporum f. sp. cubense. It significantly provided the widest zone of inhibition of
32.33mm. Trichoderma KA on the other hand, offers an alternative to chemical control
having inhibited the growth of the fungal isolates at 15.61 mm after seven days of
incubation.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
RESULTS AND DISCUSSION
A. Field Survey
Area surveyed. The survey on the occurrence of Fusarium wilt was done in two
banana growing barangays of Kapangan namely Sagubo and Gadang.
In barangay Sagubo, three sitios that include Ampongot, Dosco, and Timoc were
surveyed for the presence of banana Fusarium wilt. Barangay Gadang on the other hand
included sitiosCopias, Gadang Proper, Lib-libeng, Namon-ao, and Namat-ikan. Fig. 1
shows that the sitios surveyed varied in terms of elevation, and the areas situated in the
lower part of Kapangan (Ampongot and Copias) with higher temperature had higher
Fusarium wilt incidence of 20% and 16.17% (Table 2).
Namon-ao
Timoc
Lib-libeng
Dosco
Gadang Proper
Ampongot
Namat-ikan
Copias
Sagubo
Gadang
Figure 1. Areas surveyed at barangay Sagubo and barangay Gadang
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Area planted with banana. Barangay Sagubo has a total of 13,275 m² area planted
with banana broken down to 4,070 m² in sitio Ampongot (Fig. 3), 2, 285 m² in sitio Timoc
(Fig. 4) including sitio Mocgo, 5,120 m² in sitio Dosco (Fig. 2), and 1,800 m² along the
road. Fig. 10 shows the estimated area planted with bananas in the different sitios.
Figure 2. Banana plants (mixed cultivars), barangay Sagubo, sitio Dosco
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 3. Banana plants (mixed cultivars), barangay Sagubo, sitio Ampongot
Figure 4. Banana plants (mixed cultivars), barangay Sagubo, sitio Timoc
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Barangay Gadang, on the other hand, has a total of 10,920 m² area planted with
banana. As presented in Fig. 10, 2,880 m² is planted with banana in sitio Lib – libeng (Fig.
9), 4,520 m² in sitio Namon – ao (Fig. 5), 2,100 m² in sitio Gadang Proper (Fig. 6), 600 m²
in sitio Namat –ikan (Fig. 7), and 820 m² in sitio Copias (Fig. 8).
Figure 5. Banana plants (mixed cultivars), barangay Gadang, sitio Namon- ao
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 6. Banana plants (mixed cultivars), barangay Gadang, sitio Gadang Proper
Figure 7. Banana plants (mixed cultivars), barangay Gadang, sitio Namat- ikan
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 8. Banana plants (mixed cultivars), barangay Gadang, sitio Copias
Figure 9. Banana plants (mixed cultivars), barangay Gadang, sitio Lib - Libeng
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
6,000
5,120
5,000
4,520
4,070
4,000
2,880
3,000
2,285
2,100
1,800
2,000
820
1,000
600
0
Figure 10. Area (m2) grown with bananas in barangay Sagubo and Gadang
The banana plants in the areas surveyed are mixed with other economic crops such
as “boyboy” or tiger grass (which is processed to soft broom), chayote, corn, ginger, sweet
potato, “pa-o” or bamboo canes, and chili which is either for home consumption or for
market. The farmers say that in the past, when Fusarium wilt and banana bunchy top
diseases were not yet prevalent in their farms, banana was their primary crop; but now it is
only second from tiger grass. Some farmers also planted bananas around their rice fields
as a barrier from strong winds as observed in Fig. 5.
Banana varieties grown. The common banana varieties grown are Lakatan,
Kantong (Bungulan), Lem – en, Cavendish (Tomok), Saba (Balayang), Cardaba (Dippig),
Gloria, Señorita, Malagambang (violet colored), and Latundan. Dippig and Balayang are
less preferred by farmers because they have lower price when sold as compared to Lakatan.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Farmers knowledge about the Fusarium wilt disease. The farmers of barangay
Sagubo claimed that aside from Dippig and Balayang, all the other varieties are susceptible
to Fusarium wilt.
According to Mr. Peter Sangioa (farmer) the disease occurred in the year 2000 with
no known source. Another farmer, Mrs. Remy Anno said that in a sloping farm like the
mountain side, there is higher infection of Fusarium wilt in the lower part as compared to
the upper part. This can be attributed to temperature since it is higher at lower elevation
which could be more favorable to the development of Fusarium wilt thus increasing the
incidence and severity. Mrs. Saenay Fermin also added that the infection is most severe
during the month of May.
During the interview, most of the farmers claimed that Fusarium wilt symptom
appears after eight to twelve months from planting. This is the stage when the banana starts
to flower and develop its fruit. In most case, the fruit will not continue to develop or it may
develop but the fruits will not ripen. Other farmers also claimed that Fusarium wilt
infection will start on the second generation of new planting materials. In this case, the
farmers will have a good harvest during the first year of planting but on the second year,
30% to 40 % of the plants are infected with Fusarium wilt and other diseases such as banana
bunchy top (BBTV) and moko disease. In addition Mr. Doroteo Balawen (farmer) said that
Fusarium wilt infects old plants while BBTV infects young plants.
The farmers of barangay Gadang observed that the banana varieties Lakatan and
Kantong or Bungulan are the most susceptible to Fusarium wilt while variety Gloria and
Malagambang are slightly resistant, with Cardaba and Saba varieties as highly Resistant.
During the interview, Mr. Thomas Atoway, a farmer stated that Fusarium wilt of banana
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
was first observed in 1998. According to Virginia Dizon (farmer) they acquired planting
materials of variety Señorita from Aurora Province, Ilocos Sur in 1995 and they suspected
these planting materials to have been the source of inoculum of Fusarium wilt infection.
The farmers also confirmed that at lower elevation of Kapangan, a higher Fusarium
wilt infection was observed. According to Mr. Nobel Catalio, a farmer, the disease is most
severe during the months of April to May when the temperature is high. According to a
publication by Wikipedia.org, Fusarium wilt progresses rapidly in soil that is 70 ºF or
warmer. The months of April to May is already summer season, which means the
temperature is higher than normal thereby enabling fast development of Fusarium wilt
infection.
Management practices done by the farmers. As soon as the symptom of Fusarium
wilt is observed, the farmers eradicate the banana tree by cutting and use it as supplement
feeds for cattle and carabao. They also noted that when the mother plant is Fusarium wilt
infected, the suckers when planted will later show Fusarium wilt infection.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Incidence and Severity of Fusarium Infected Bananas.
Incidence. Four (4) randomly selected survey profiles, in each sitio, were
statistically analyzed to determine if there is a significant difference of Fusarium wilt
incidence. Data presented in Table 2 shows that barangay Sagubo had 16.82 % while
barangay Gadang had 13.57 % Fusarium wilt infected plants.
On the other hand, sitio Ampongot had the highest Fusarium wilt incidence of 20.21
% among the sitios surveyed while 16.17 % was noted in sitio Copias (Table 2). Recorded
incidence of Fusarium wilt between Sagubo and Gadang, and the different sitios surveyed
did not vary significantly from each other.
Table 2. Banana Fusarium wilt incidence (%) in the different areas surveyed
AREAS SURVEYED
PERCENT (%)
INCIDENCE
BARANGAY
Barangay Sagubo
16.82
Barangay Gadang
13.57
SITIO
Barangay Sagubo, Sitio Ampongot
20.21
Barangay Sagubo, Sitio Timoc and Mocgo
12.56
Barangay Sagubo, Sitio Dosco
14.98
Barangay Gadang, Sitio Lib-libeng
13.44
Barangay Gadang, Sitio Namon-ao
14.17
Barangay Gadang, Sitio Copias
16.17
Barangay Gadang, Sitio Gadang Proper
10.72
Barangay Gadang, Sitio Namat-ikan
15.88
C.V. = 17.40 %
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Fusarium wilt severity. As shown in Table 3, the severity of Fusarium wilt in the
different banana variety is significantly different. Fusarium wilt is most severe in Lakatan
(Fig. 12) with an infection rate of 5 which describe a Fusarium wilt infected banana having
all leaves brown/ dried and reddish – brown discoloration on the pseudo stem. Severity of
infection on variety Lakatan of 5.00 is significantly different to infection on variety
Kantong (Fig. 11) with a rate of 3.67.
Meanwhile, Fusarium wilt infection was not observed on Saba and Cardaba
(Dippig) throughout the survey period in all areas. It appears that the existing race of
Fusarium wilt in does not infect Saba and Dippig. Based from the result, it appeared that
the race responsible in affecting Dippig and Saba are not present in Kapangan. Dippig and
Saba were identified by PCARRD (2004) to be susceptible only to F. oxysporum f. sp.
cubense Race 2 and Race 4.
Figure 11. (A) Kantong or Bungulan variety showing infection of F. oxysporum f. sp.
cubense; (B) Pseudo stem showing vascular discoloration
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 12. (A) Lakatan variety showing infection of F. oxysporum f. sp. cubense;
(B) Pseudo stem showing vascular discoloration
Figure 13. (A) Tomok or Cavendish variety showing infection of F. oxysporum f. sp.
cubense;(B) Pseudo stem showing vascular discoloration
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 3. Fusarium wilt severity as observed on the infected pseudo stems
Banana Variety
Severity of Infection
Lakatan
5.00a
Bungulan(Kantong)
3.67b
Cavendish (Tomok)
4.00ab
Saba (Balayang)
0.00c
Cardaba (Dippig)
0.00c
C.V. = 26.97 %
B. LABORATORY ACTIVITIES
Colony diameter. F. oxysporum f. sp. cubense that was isolated from banana
varieties Cavendish (Tomok), or Bungulan (Kantong), and Lakatan were designated as
isolate 01, 02, and 03.
Table 4 presents the mean colony diameter of the three isolates after three, five, and
seven days. Based on the result isolate 02 had the highest colony diameter of 65.00 mm,
followed by isolate 01 with 62.67 mm. Isolate 03 gave the smallest colony diameter of
59.83 mm. In terms of pigment in Potato Dextrose Agar (PDA) isolate 01 had a light purple
pigment, dark purple on isolate 02, and light orange on isolate 03.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 4. Colony diameter of F. oxysporum f. sp. cubense in fresh Potato Dextrose Agar
(PDA) after three, five, and seven days
COLONY DIAMETER (mm)
Isolates
3 day
5 day
7 day
MEAN
Pigment in PDA
Isolate 01
55.50
63.50
69.00
62.67
light purple
Isolate 02
51.50
59.83
83.67
65.00
dark purple
Isolate 03
47.50
55.83
76.17
59.83
light orange
Figure 14. Three day old cultures of F. oxysporum f. sp. cubense on fresh Potato Dextrose
Agar (PDA). (A) Isolate 01, (B) Isolate 02, (C) Isolate 03
Table 5 below shows the sizes of conidia of the three isolates. Based on microscopic
observation, there are more macro conidia in isolate 03 compared to the other isolates,
while more micro conidia was noted in isolate 02. Figures 15, 16, and 17 show the macro
and micro conidia of each isolate. According to Ploetz (2000) F. oxysporum f.
sp. cubense has a micro conidia measuring 5 - 16 x 2.4 - 3.5 µm while the macro conidia
measures 27 - 55 x 3.3 - 5.5 µm.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Microscopic measurement of the macro conidia of the three isolates were 30.42 x
3.64 µm, 31.20 x 4.68 µm and 33.80 x 4.81 µm, while micro conidia of the three isolates
measured 8.77 x 2.73 µm, 8.06 x 2.86 µm and 7.80 x 3.25 µm. The result was found to
be within the range of the measurement reported by Ploetz in 2000.
Table 5. Morphological characteristics of F. oxysporum f. sp. cubense.
Isolates
Size of Macro conidia
Size of Micro conidia
No. of Septation
(µm)
(µm)
of Macro conidia
Isolate 01
30.42 um x 3.64 um
8.77 um x 2.73 um
3-4
Isolate 02
31.20 um x 4.68 um
8.06 um x 2.86 um
3-4
Isolate 03
33.80 um x 4.81 um
7.80 um x 3.25 um
3-4
Figures 15, 16, and 17 show the macro and micro conidia of the different isolates
under 400X magnification, it is clearly shown that isolate 3 had more macro conidia than
isolate 01 and 02. The macro and micro conidia have green color. Further, the macro
conidia of the three isolates have three to four septations.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 15. Macro and micro conidia of isolate 01 (400X)
Figure 16. Macro and micro conidia of isolate 02 (400X)
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 17. Macro and micro conidia of isolate 03 (400X)
Bioassay
Table 6 shows the result of bioassay of isolate 01 after three, five, and seven days
of incubation. On day three, Benomyl had the highest inhibition zone with 38.67 mm
diameter which is significantly different to Lysol with an inhibition zone of 26.17 mm
diameter. The inhibition zone produced by Trichoderma, Mancozeb (Dithane M45),
Mancozeb (Parafungus), and Captan which measured 16.17 mm, 20.00 mm, 18.50mm, and
19.17 mm did not vary significantly. Sterilized distilled water did not produce inhibition
zone.
On day five, Benomyl still had the highest inhibition zone of 31.33 mm, which is
significantly different to the rest of the treatments. The inhibition zone produced by
Trichoderma, Mancozeb, and Captan still did not vary significantly.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 6. Inhibition zones (mm) formed on Potato Dextrose Agar (PDA) by isolate 01
after three, five, and seven days
TREATMENTS
DAY 3
DAY 5
DAY 7
MEAN
Sterile Distilled Water (SDW)
0.00d
0.00e
0.00d
0.00
Trichoderma (biocon)
16.17c
15.67d
15.00c
15.61
Mancozeb (Dithane M45)
20.00c
18.67c
16.17c
18.28
Lysol
26.17b
25.50b
21.67b
24.45
Mancozeb (Parafungus)
18.50c
17.67cd
17.33c
17.83
Benomyl (Benlight)
38.67a
31.33a
27.00a
32.33
Captan
19.17c
17.33cd
16.67c
17.72
On day seven, the inhibition zones produced by Benomyl and Lysol remained
significantly different with the inhibition zones of Trichoderma, Mancozeb, and Captan.
Table 7 shows the result of the bioassay on Isolate 02, after 3, 5, and 7 days
incubation using biological control agent, disinfectant, and fungicides. On day three, the
inhibition zones of Benomyl and Lysol having a diameter of 24.00 mm and 25.83 mm are
the widest and were significant compared to the inhibition zone of 20.67 mm and 21.17
mm formed by Dithane M45 and Parafungus. Trichoderma and Captan produced inhibition
zones that are almost equal (15.67 mm and 18.00 mm).
The same trend was observed after five days, the inhibition zones of Benomyl and
Lysol slightly decreased but were still the widest, having a diameter of 23.00 mm and 24.50
mm. The inhibition zones of Dithane M45, Parafungus, Trichoderma and Captan were
comparable to each other.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 7. Inhibition zones (mm) formed on Potato Dextrose Agar (PDA) by isolate 02
after three, five, and seven days
TREATMENTS
DAY 3
DAY 5
DAY 7
MEAN
Sterile Distilled Water (SDW)
0.00e
0.00e
0.00d
0.00
Trichoderma (biocon)
15.67d
15.17d
15.17c
15.61
Mancozeb (Dithane M45)
20.67bc
18.67bc
16.83c
18.28
Lysol
25.83a
24.50a
20.00b
24.45
Mancozeb (Parafungus)
21.17bc
19.50b
17.17c
17.83
Benomyl (Benlight)
24.00a
23.00a
22.17a
32.33
Captan
18.00cd
16.50cd
15.67c
17.72
On day seven, the inhibition zone of Benomyl 22.17 mm, continued to decrease
slightly but still the widest and is significantly higher than the inhibition zone of Lysol
which is 20.00 mm. The inhibition zones of Trichoderma, Dithane M45, Parafungus, and
Captan remained comparable.
Table 8 shows the result of bioassay on Isolate 03, after 3, 5, and 7 days of
incubation. On day three, the inhibition zones of Benomyl and Lysol of 26.83 mm and
27.33 mm were significantly different over the other treatments. The inhibition zone of
Captan (21.33 mm) was next to Benomyl and Lysol, while the rest were comparable in
inhibiting F. oxysporum f. sp. cubense.
The same observation was noted after five days. Benomyl and Lysol still provided
the best inhibition zones measuring 25.17 mm and 24.67 mm followed by the inhibition
zone produced by Captan which is 19.83 mm, the rest of the treatments remained
comparable.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 8. Inhibition zones (mm) formed on Potato Dextrose Agar (PDA) by isolate 03 after
three, five, and seven days
TREATMENTS
DAY 3
DAY 5
DAY 7
MEAN
Sterile Distilled Water (SDW)
0.00e
0.00d
0.00d
0.00
Trichoderma (biocon)
15.17d
15.00c
14.33c
14.83
Mancozeb (Dithane M45)
17.00d
15.50c
14.33c
15.61
Lysol
27.33a
24.67a
22.33a
24.78
Mancozeb (Parafungus)
17.50d
15.83c
14.35c
15.89
Benomyl (Benlight)
26.83a
25.17a
23.00a
25.00
Captan
21.33b
19.83b
18.00b
19.72
On day seven, Benomyl and Lysol still had the highest inhibition zones measuring
23.00 mm and 22.33 mm. Captan had an inhibition zone of 18.00 mm which is significantly
different with the inhibition zones of Trichoderma, Dithane M45, and Parafungus.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Other observations
Laboratory. Table 9 shows the colony growth of F. oxysporum f. sp. cubense grown
in a commercially prepared Potato Dextrose Agar (PDA). Results revealed that isolates 01,
02, and 03 had lower colony growth of 32.75 mm, 32.78 mm, and 39.53 mm compared to
the colony growth of 62.67 mm, 65.00 mm, and 59.83 mm obtained from the fresh PDA
(Table 4). The pigment on PDA also varied from light purple to light pink, dark purple to
pink, and light orange to dark pink.
Table 9. Colony diameter of F. oxysporum f. sp. cubense in commercially prepared Potato
Dextrose Agar (PDA) after three, five, and seven days
COLONY DIAMETER (mm)
Isolates
3 day
5 day
7 day
MEAN
Pigment on PDA
Isolate 01
28.67
32.50
37.08
32.75
light pink
Isolate 02
28.92
34.25
35.17
32.78
pink
Isolate 03
34.83
40.33
43.42
39.53
dark pink
Figure 18. Three day old cultures of F. oxysporum f. sp. cubense on the commercially
prepared Potato Dextrose Agar(PDA); (A) Isolate 01, (B) Isolate 02,
(C) Isolate 03
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Incidence of Banana Bunchy Top Virus (BBTV). As presented in Table 10, bunchy
top disease is also present in the areas surveyed. The incidence recorded in the barangays
and the different sitios showed that while the virus disease exists, the distribution does not
differ significantly.
Table 10. Incidence (%) of Banana Bunchy Top Virus (BBTV) in the areas surveyed
AREAS SURVEYED
PERCENT (%) INCIDENCE
BARANGAY
Barangay Sagubo
14.76
Barangay Gadang
14.36
SITIO
Barangay Sagubo, Sitio Ampongot
19.79
Barangay Sagubo, Sitio Timoc and Mocgo
7.94
Barangay Sagubo, Sitio Dosco
15.69
Barangay Gadang, Sitio Lib-libeng
19.25
Barangay Gadang, Sitio Namon-ao
18.46
Barangay Gadang, Sitio Copias
16.33
Barangay Gadang, Sitio Gadang Proper
7.61
Barangay Gadang, Sitio Namat-ikan
10.63
CV= 1.5064 %
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Banana Bugtok or Blood disease. This disease was noted in barangay Gadang, sitio
Lib-libeng. The disease was observed to infect the banana variety Dippig or Cardaba.
Infected fruits do not ripen and are not fit for consumption.
According to Soguilon et.al (1995) bugtok is an endemic and a widely distributed
bacterial disease of cooking banana cultivars in the Philippines. Bugtok is a local term in
the southern Philippines used to describe the infected fruit which are discolored and hard
even when ripe. It was noted as a minor disorder more than 40 years ago, but was reported
by Roperos in 1965 as a developing problem of importance. It is caused by Pseudomonas
solanacearum E. F. Smith.
Bugtok can be controlled by bagging the inflorescence at the bending stage just
after emergence. The bagging material can be a polyethylene bag, muslin cloth, or a fine
nylon mesh bag. Bags can be removed after all the fruits have set followed by removal of
the male inflorescence. This practice should also include mat and field sanitation, and
removal of old, dead leaves.
Figure 17. Banana bugtok or blood disease on Cardaba (Dippig)
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
SUMMARY, CONCLUSIONS AND RECOMMENDATIONS
Summary
The study was conducted to survey the occurrence of Fusarium wilt in two banana
producing barangays of Kapangan, Benguet, namely Gadang, and Sagubo; asses the
incidence and severity of infection of the affected banana plants; isolate and characterize
the causal pathogen Fusarium oxysporum f. sp. cubense from infected samples, and
compare the effect of bio control agent and chemicals in inhibiting the growth of F.
oxysporum f. sp. cubense through assay.
Results show that Fusarium wilt of banana is prevalent in the areas surveyed.
Fusarium wilt incidence of 16.82% and 13.57 % in barangays Sagubo and Gadang
including the incidence noted on the different sitios does not vary significantly. Among the
different sitios surveyed, sitio Ampongot has the highest incidence of 20.21 % while sitio
Gadang Proper has the lowest incidence of 10.72 %.
Fusarium wilt severity based on a five point rating scale showed that Lakatan was
severely affected followed by Tomok or Cavendish and then Kantong or Bungulan. Banana
cultivars Dippig and Balayang were not infected by Fusarium wilt in the area surveyed.
Based on the result of the cultural and morphological characterization of F.
oxysporum assigned as isolate 01, 02, and 03, isolate 02 had the highest colony diameter
of 65.00 mm, followed by isolate 01 with 62.67 mm. Isolate 03 gave the smallest colony
diameter of 59.83 mm. In terms of pigment on fresh Potato Dextrose Agar (PDA) isolate
01 has a light purple pigment, dark purple on isolate 02, and light orange on isolate 03.
Macro conidia measurements of isolate 01, 02, and 03 of 30.42 x 3.64 µm, 31.20 x
4.68 µm, and 33.80 x 4.81 µm are statistically comparable. On the other hand, the micro
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
conidia of the three isolates that measured 8.77 x 2.73 µm, 8.06 x 2.86 µm, and 7.80 x 3.25
µm was found to be within the range of the reported measurement size of F. oxysporum.
The results of the bioassay on isolate 01 revealed that Benomyl and Lysol
significantly produced the highest inhibition zones of 32.33 mm and 24.45 mm. The
inhibition zones of Trichoderma, Dithane M45 (Mancozeb), Parafungus (Mancozeb), and
Captan were comparable.
On the other hand, the result of bioassay on isolate 02 showed that the widest
average inhibition zones were observed on Benomyl and Lysol having 23.06 mm and 23.44
mm, respectively. The inhibition zones produced by Trichoderma, Dithane M45
(Mancozeb), Parafungus (Mancozeb), and Captan were also comparable.
The same trend was observed on isolate 03 where in Benomyl and Lysol still
provided the widest inhibition zones measuring 25.00 mm and 24.78 mm. Captan had
higher inhibition zone as compared to the inhibition zones produced by Trichoderma,
Dithane M45, and Parafungus after seven days.
Conclusions
Banana Fusarium wilt incidence in barangay Sagubo and Gadang is low ranging
from 10 to 20 %. The varieties grown in the area are Lakatan, Kantong, Lemen, Tomok,
Balayang, Dippig, Latundan, Gloria, Señorita, and Malagambang. Lakatan, Kantong, and
Tomok are susceptible to the existing race of F. oxysporum f. sp. cubense existing in
Kapangan.
Banana is a secondary crop in barangay Sagubo and barangay Gadang with a total
area planted of 24,195 m².
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Isolates 01, 02 and 03 of F. oxysporum f. sp. cubense differed in their colony growth
in Potato Dextrose Agar including the size of macro and micro conidia.
Benomyl is effective in inhibiting the growth of F. oxysporum. Trichoderma is not
as effective as benomyl but can be used as alternative for sustainable Fusarium wilt
management.
Recommendations
Bioassay should be tested in pot experiments to confirm if the same results will be
obtained. Planting materials should not be sourced out from areas, such as Mindanao,
where Race 4 exists.
Similar study should be done to other banana producing areas of Benguet and
Mountain Province to determine the presence and incidence of F. oxyporum f. sp. cubense.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
LITERATURE CITED
BANCROFT, J. 1876. Report in the Board Appointed to Enquire into the Cause of Disease
Affecting Livestock and Plants. Retrieved October 18, 2012 from
http://www.plantdiseases.org.
Bureau of Agricultural Statistics (BAS). 2012. Baseline Commodity Production Data.
Retrieved December 11, 2012 from www.bas.gov.ph.
DALY, A. 2006. Fusarium Wilt of Bananas (Panama Diseases). CABI Publishing.
Wallingford, UK. Pp. 1-18.
DHILLON, B.S., R.K. TIYAGI, S. SAXENA, and G. J. RANDHAWA. 2005. Plant
Genetic Resources: Horticultural Crops. NAROSA Publishing House PVT. LTD.
Pp. 127.
LACORTE, G. and J. QUIROS. Panama Disease. Retrieved December 10, 2012 from
newsinfo.inquirer.net
MOORE, N. Y., P. A. HARGREAVES, K. G. PEGG, AND J.A.G. IRWIN. 1991.
Characterization of strains of Fusarium oxysporum f. sp. cubense by production of
volatiles. Australian Journal of Botany. P. 39.
PADILLA, L. D. E. 2012. Optimizing the production of chemical free banana. Bureau of
Agricultural Research Chronicle. 2(8):14-15.
Philippine Council for Agriculture, Forestry and Natural Resources Research and
Development (PCARRD). 1986. Banana and Plantain Research and Development.
PCARRD Los Baños, Laguna, Philippines. Pp. 18 – 21.
Philippine Council for Agriculture, Forestry and Natural Resources Research and
Development (PCARRD). 2004. Banana Production Manual. PCARRD Los Baños,
Laguna, Philippines. P. 25.
PLOETZ, R. C. 2000. Panama Disease: A Classic and Destructive Disease of Banana.
Online Plant Health Progress. Retrieved October 18, 2012 from
http://www.plantmanagementnetwork.org/pub/php/management/bananapanama/.
PURSS, G.S. 1953. A Disease in Williams Hybrid Produced by Fusarium spp. Queensland
Journal of Agricultural Science. P. 126.
SOGUILON, C. E., L. V. MAGNAYE, and M. P. NATURAL. 1995. Bugtok Disease of
Banana.
Retrieved
March
18,
2013
from
http://www.bioversityinternational.org/fileadmin/bioversity/publications/pdfs/701_
Bugtok_disease of banana_.pdf
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
STOVER, R.H.l. and S.E. MALO. 1972. The Occurrence of Fusarial Wilt in Normally
Resistant ‘Dwarf Cavendish’ Banana. Longmans Scientific and Technical, London.
P. 23.
SUN, E.J. and H.J. SU. 1978. Rapid Method for Determining Differential Pathogenicity of
Fusarium oxysporum f. sp. cubense using Banana Plantlets. Trop. Agric. (Trinidad).
Pp. 7-8.
VALMAYOR, R. V. 1990. Proceedings of INIBAP Banana Plantain R & D in Asia and
the Pacific. Philippines, November 20 – 24, 1989. Pp. 46 – 52.
VALMAYOR, R. V., R. G. DAVIDE, J. M. STANTON, N. L. TREVERROW, and V.
N. ROA. 1994. INIBAP Banana Nematodes and Weevil Borers in Asia and the
Pacific. UPLB Philippines. Pp. 32 – 37.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
CODOD, CLARENCE BAGAYAO. APRIL 2013. Survey, Assessment, and
Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp. cubense) in Two
Barangays of Kapangan, Benguet. Benguet State University, La Trinidad, Benguet.
Adviser: Asuncion L. Nagpala, Ph.D.
ABSTRACT
Survey and sampling of banana plants infected with Fusarium wilt were done in the
Municipality of Kapangan barangay Sagubo, sitio Timoc, Ampongot, and Dosco, and
barangay Gadang sitio Namun- ao, Gadang Proper, Namat- ikan, Copias, and Lib – libeng,
Benguet from September 2012 to November 2012 to confirm the occurrence of the disease
in the locality; assess the incidence and severity of Fusarium wilt infection in affected
banana plants; isolate and characterize the causal pathogen Fusarium oxysporum f. sp.
cubense from infected samples; and compare the effect of bio control agent and chemicals in
inhibiting the growth of F. oxysporum f. sp. cubense through assay.
Results revealed that Fusarium wilt incidence in barangay Sagubo and Gadang was
16.82 % and 13.57 % respectively. In terms of severity, variety Lakatan was the most
susceptible to Fusarium wilt infection having the most intense discoloration on the
Pseudostem followed by the variety Cavendish (Tomoc) and Bungulan (Kantong) with
mean severity ratings of 5.0, 4.0 and 3.67 respectively. Varieties Saba (Balayang) and
Cardaba (Dippig) did not incur any infection.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Isolates of F. oxysporum f. sp. cubense from Lakatan, Cavendish and Bungulan
designated as isolate 01, 02 and 03 differed in their colony growth in Potato Dextrose Agar
with 02 isolate having the widest mean growth of 65 mm. Isolates 01 and 03 had 62.67
and 59.83 mm growth diameter respectively.
Microscopic measurements of the macro conidia of the three isolates were 30.42 x
3.64 µm, 31.20 x 4.68 µm and 33.80 x 4.81 µm and were found to be within the range of
the published measurement of F. oxysporum which is 27-55 x 3.3-5.5 µm. The micro
conidia of the three isolates measured 8.77 x 2.73 µm, 8.06 x 2.86µm and 7.80 x 3.25 µm
which is also within the range of 5-16 x 2.4-3.5 µm.
Bioassay results showed that Benomyl inhibited the growth of the three isolates of
F.oxysporum f. sp. cubense. It significantly provided the widest zone of inhibition of
32.33mm. Trichoderma KA on the other hand, offers an alternative to chemical control
having inhibited the growth of the fungal isolates at 15.61 mm after seven days of
incubation.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
RESULTS AND DISCUSSION
A. Field Survey
Area surveyed. The survey on the occurrence of Fusarium wilt was done in two
banana growing barangays of Kapangan namely Sagubo and Gadang.
In barangay Sagubo, three sitios that include Ampongot, Dosco, and Timoc were
surveyed for the presence of banana Fusarium wilt. Barangay Gadang on the other hand
included sitiosCopias, Gadang Proper, Lib-libeng, Namon-ao, and Namat-ikan. Fig. 1
shows that the sitios surveyed varied in terms of elevation, and the areas situated in the
lower part of Kapangan (Ampongot and Copias) with higher temperature had higher
Fusarium wilt incidence of 20% and 16.17% (Table 2).
Namon-ao
Timoc
Lib-libeng
Dosco
Gadang Proper
Ampongot
Namat-ikan
Copias
Sagubo
Gadang
Figure 1. Areas surveyed at barangay Sagubo and barangay Gadang
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Area planted with banana. Barangay Sagubo has a total of 13,275 m² area planted
with banana broken down to 4,070 m² in sitio Ampongot (Fig. 3), 2, 285 m² in sitio Timoc
(Fig. 4) including sitio Mocgo, 5,120 m² in sitio Dosco (Fig. 2), and 1,800 m² along the
road. Fig. 10 shows the estimated area planted with bananas in the different sitios.
Figure 2. Banana plants (mixed cultivars), barangay Sagubo, sitio Dosco
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 3. Banana plants (mixed cultivars), barangay Sagubo, sitio Ampongot
Figure 4. Banana plants (mixed cultivars), barangay Sagubo, sitio Timoc
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Barangay Gadang, on the other hand, has a total of 10,920 m² area planted with
banana. As presented in Fig. 10, 2,880 m² is planted with banana in sitio Lib – libeng (Fig.
9), 4,520 m² in sitio Namon – ao (Fig. 5), 2,100 m² in sitio Gadang Proper (Fig. 6), 600 m²
in sitio Namat –ikan (Fig. 7), and 820 m² in sitio Copias (Fig. 8).
Figure 5. Banana plants (mixed cultivars), barangay Gadang, sitio Namon- ao
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 6. Banana plants (mixed cultivars), barangay Gadang, sitio Gadang Proper
Figure 7. Banana plants (mixed cultivars), barangay Gadang, sitio Namat- ikan
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 8. Banana plants (mixed cultivars), barangay Gadang, sitio Copias
Figure 9. Banana plants (mixed cultivars), barangay Gadang, sitio Lib - Libeng
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
6,000
5,120
5,000
4,520
4,070
4,000
2,880
3,000
2,285
2,100
1,800
2,000
820
1,000
600
0
Figure 10. Area (m2) grown with bananas in barangay Sagubo and Gadang
The banana plants in the areas surveyed are mixed with other economic crops such
as “boyboy” or tiger grass (which is processed to soft broom), chayote, corn, ginger, sweet
potato, “pa-o” or bamboo canes, and chili which is either for home consumption or for
market. The farmers say that in the past, when Fusarium wilt and banana bunchy top
diseases were not yet prevalent in their farms, banana was their primary crop; but now it is
only second from tiger grass. Some farmers also planted bananas around their rice fields
as a barrier from strong winds as observed in Fig. 5.
Banana varieties grown. The common banana varieties grown are Lakatan,
Kantong (Bungulan), Lem – en, Cavendish (Tomok), Saba (Balayang), Cardaba (Dippig),
Gloria, Señorita, Malagambang (violet colored), and Latundan. Dippig and Balayang are
less preferred by farmers because they have lower price when sold as compared to Lakatan.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Farmers knowledge about the Fusarium wilt disease. The farmers of barangay
Sagubo claimed that aside from Dippig and Balayang, all the other varieties are susceptible
to Fusarium wilt.
According to Mr. Peter Sangioa (farmer) the disease occurred in the year 2000 with
no known source. Another farmer, Mrs. Remy Anno said that in a sloping farm like the
mountain side, there is higher infection of Fusarium wilt in the lower part as compared to
the upper part. This can be attributed to temperature since it is higher at lower elevation
which could be more favorable to the development of Fusarium wilt thus increasing the
incidence and severity. Mrs. Saenay Fermin also added that the infection is most severe
during the month of May.
During the interview, most of the farmers claimed that Fusarium wilt symptom
appears after eight to twelve months from planting. This is the stage when the banana starts
to flower and develop its fruit. In most case, the fruit will not continue to develop or it may
develop but the fruits will not ripen. Other farmers also claimed that Fusarium wilt
infection will start on the second generation of new planting materials. In this case, the
farmers will have a good harvest during the first year of planting but on the second year,
30% to 40 % of the plants are infected with Fusarium wilt and other diseases such as banana
bunchy top (BBTV) and moko disease. In addition Mr. Doroteo Balawen (farmer) said that
Fusarium wilt infects old plants while BBTV infects young plants.
The farmers of barangay Gadang observed that the banana varieties Lakatan and
Kantong or Bungulan are the most susceptible to Fusarium wilt while variety Gloria and
Malagambang are slightly resistant, with Cardaba and Saba varieties as highly Resistant.
During the interview, Mr. Thomas Atoway, a farmer stated that Fusarium wilt of banana
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
was first observed in 1998. According to Virginia Dizon (farmer) they acquired planting
materials of variety Señorita from Aurora Province, Ilocos Sur in 1995 and they suspected
these planting materials to have been the source of inoculum of Fusarium wilt infection.
The farmers also confirmed that at lower elevation of Kapangan, a higher Fusarium
wilt infection was observed. According to Mr. Nobel Catalio, a farmer, the disease is most
severe during the months of April to May when the temperature is high. According to a
publication by Wikipedia.org, Fusarium wilt progresses rapidly in soil that is 70 ºF or
warmer. The months of April to May is already summer season, which means the
temperature is higher than normal thereby enabling fast development of Fusarium wilt
infection.
Management practices done by the farmers. As soon as the symptom of Fusarium
wilt is observed, the farmers eradicate the banana tree by cutting and use it as supplement
feeds for cattle and carabao. They also noted that when the mother plant is Fusarium wilt
infected, the suckers when planted will later show Fusarium wilt infection.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Incidence and Severity of Fusarium Infected Bananas.
Incidence. Four (4) randomly selected survey profiles, in each sitio, were
statistically analyzed to determine if there is a significant difference of Fusarium wilt
incidence. Data presented in Table 2 shows that barangay Sagubo had 16.82 % while
barangay Gadang had 13.57 % Fusarium wilt infected plants.
On the other hand, sitio Ampongot had the highest Fusarium wilt incidence of 20.21
% among the sitios surveyed while 16.17 % was noted in sitio Copias (Table 2). Recorded
incidence of Fusarium wilt between Sagubo and Gadang, and the different sitios surveyed
did not vary significantly from each other.
Table 2. Banana Fusarium wilt incidence (%) in the different areas surveyed
AREAS SURVEYED
PERCENT (%)
INCIDENCE
BARANGAY
Barangay Sagubo
16.82
Barangay Gadang
13.57
SITIO
Barangay Sagubo, Sitio Ampongot
20.21
Barangay Sagubo, Sitio Timoc and Mocgo
12.56
Barangay Sagubo, Sitio Dosco
14.98
Barangay Gadang, Sitio Lib-libeng
13.44
Barangay Gadang, Sitio Namon-ao
14.17
Barangay Gadang, Sitio Copias
16.17
Barangay Gadang, Sitio Gadang Proper
10.72
Barangay Gadang, Sitio Namat-ikan
15.88
C.V. = 17.40 %
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Fusarium wilt severity. As shown in Table 3, the severity of Fusarium wilt in the
different banana variety is significantly different. Fusarium wilt is most severe in Lakatan
(Fig. 12) with an infection rate of 5 which describe a Fusarium wilt infected banana having
all leaves brown/ dried and reddish – brown discoloration on the pseudo stem. Severity of
infection on variety Lakatan of 5.00 is significantly different to infection on variety
Kantong (Fig. 11) with a rate of 3.67.
Meanwhile, Fusarium wilt infection was not observed on Saba and Cardaba
(Dippig) throughout the survey period in all areas. It appears that the existing race of
Fusarium wilt in does not infect Saba and Dippig. Based from the result, it appeared that
the race responsible in affecting Dippig and Saba are not present in Kapangan. Dippig and
Saba were identified by PCARRD (2004) to be susceptible only to F. oxysporum f. sp.
cubense Race 2 and Race 4.
Figure 11. (A) Kantong or Bungulan variety showing infection of F. oxysporum f. sp.
cubense; (B) Pseudo stem showing vascular discoloration
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 12. (A) Lakatan variety showing infection of F. oxysporum f. sp. cubense;
(B) Pseudo stem showing vascular discoloration
Figure 13. (A) Tomok or Cavendish variety showing infection of F. oxysporum f. sp.
cubense;(B) Pseudo stem showing vascular discoloration
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 3. Fusarium wilt severity as observed on the infected pseudo stems
Banana Variety
Severity of Infection
Lakatan
5.00a
Bungulan(Kantong)
3.67b
Cavendish (Tomok)
4.00ab
Saba (Balayang)
0.00c
Cardaba (Dippig)
0.00c
C.V. = 26.97 %
B. LABORATORY ACTIVITIES
Colony diameter. F. oxysporum f. sp. cubense that was isolated from banana
varieties Cavendish (Tomok), or Bungulan (Kantong), and Lakatan were designated as
isolate 01, 02, and 03.
Table 4 presents the mean colony diameter of the three isolates after three, five, and
seven days. Based on the result isolate 02 had the highest colony diameter of 65.00 mm,
followed by isolate 01 with 62.67 mm. Isolate 03 gave the smallest colony diameter of
59.83 mm. In terms of pigment in Potato Dextrose Agar (PDA) isolate 01 had a light purple
pigment, dark purple on isolate 02, and light orange on isolate 03.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 4. Colony diameter of F. oxysporum f. sp. cubense in fresh Potato Dextrose Agar
(PDA) after three, five, and seven days
COLONY DIAMETER (mm)
Isolates
3 day
5 day
7 day
MEAN
Pigment in PDA
Isolate 01
55.50
63.50
69.00
62.67
light purple
Isolate 02
51.50
59.83
83.67
65.00
dark purple
Isolate 03
47.50
55.83
76.17
59.83
light orange
Figure 14. Three day old cultures of F. oxysporum f. sp. cubense on fresh Potato Dextrose
Agar (PDA). (A) Isolate 01, (B) Isolate 02, (C) Isolate 03
Table 5 below shows the sizes of conidia of the three isolates. Based on microscopic
observation, there are more macro conidia in isolate 03 compared to the other isolates,
while more micro conidia was noted in isolate 02. Figures 15, 16, and 17 show the macro
and micro conidia of each isolate. According to Ploetz (2000) F. oxysporum f.
sp. cubense has a micro conidia measuring 5 - 16 x 2.4 - 3.5 µm while the macro conidia
measures 27 - 55 x 3.3 - 5.5 µm.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Microscopic measurement of the macro conidia of the three isolates were 30.42 x
3.64 µm, 31.20 x 4.68 µm and 33.80 x 4.81 µm, while micro conidia of the three isolates
measured 8.77 x 2.73 µm, 8.06 x 2.86 µm and 7.80 x 3.25 µm. The result was found to
be within the range of the measurement reported by Ploetz in 2000.
Table 5. Morphological characteristics of F. oxysporum f. sp. cubense.
Isolates
Size of Macro conidia
Size of Micro conidia
No. of Septation
(µm)
(µm)
of Macro conidia
Isolate 01
30.42 um x 3.64 um
8.77 um x 2.73 um
3-4
Isolate 02
31.20 um x 4.68 um
8.06 um x 2.86 um
3-4
Isolate 03
33.80 um x 4.81 um
7.80 um x 3.25 um
3-4
Figures 15, 16, and 17 show the macro and micro conidia of the different isolates
under 400X magnification, it is clearly shown that isolate 3 had more macro conidia than
isolate 01 and 02. The macro and micro conidia have green color. Further, the macro
conidia of the three isolates have three to four septations.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 15. Macro and micro conidia of isolate 01 (400X)
Figure 16. Macro and micro conidia of isolate 02 (400X)
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Figure 17. Macro and micro conidia of isolate 03 (400X)
Bioassay
Table 6 shows the result of bioassay of isolate 01 after three, five, and seven days
of incubation. On day three, Benomyl had the highest inhibition zone with 38.67 mm
diameter which is significantly different to Lysol with an inhibition zone of 26.17 mm
diameter. The inhibition zone produced by Trichoderma, Mancozeb (Dithane M45),
Mancozeb (Parafungus), and Captan which measured 16.17 mm, 20.00 mm, 18.50mm, and
19.17 mm did not vary significantly. Sterilized distilled water did not produce inhibition
zone.
On day five, Benomyl still had the highest inhibition zone of 31.33 mm, which is
significantly different to the rest of the treatments. The inhibition zone produced by
Trichoderma, Mancozeb, and Captan still did not vary significantly.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 6. Inhibition zones (mm) formed on Potato Dextrose Agar (PDA) by isolate 01
after three, five, and seven days
TREATMENTS
DAY 3
DAY 5
DAY 7
MEAN
Sterile Distilled Water (SDW)
0.00d
0.00e
0.00d
0.00
Trichoderma (biocon)
16.17c
15.67d
15.00c
15.61
Mancozeb (Dithane M45)
20.00c
18.67c
16.17c
18.28
Lysol
26.17b
25.50b
21.67b
24.45
Mancozeb (Parafungus)
18.50c
17.67cd
17.33c
17.83
Benomyl (Benlight)
38.67a
31.33a
27.00a
32.33
Captan
19.17c
17.33cd
16.67c
17.72
On day seven, the inhibition zones produced by Benomyl and Lysol remained
significantly different with the inhibition zones of Trichoderma, Mancozeb, and Captan.
Table 7 shows the result of the bioassay on Isolate 02, after 3, 5, and 7 days
incubation using biological control agent, disinfectant, and fungicides. On day three, the
inhibition zones of Benomyl and Lysol having a diameter of 24.00 mm and 25.83 mm are
the widest and were significant compared to the inhibition zone of 20.67 mm and 21.17
mm formed by Dithane M45 and Parafungus. Trichoderma and Captan produced inhibition
zones that are almost equal (15.67 mm and 18.00 mm).
The same trend was observed after five days, the inhibition zones of Benomyl and
Lysol slightly decreased but were still the widest, having a diameter of 23.00 mm and 24.50
mm. The inhibition zones of Dithane M45, Parafungus, Trichoderma and Captan were
comparable to each other.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 7. Inhibition zones (mm) formed on Potato Dextrose Agar (PDA) by isolate 02
after three, five, and seven days
TREATMENTS
DAY 3
DAY 5
DAY 7
MEAN
Sterile Distilled Water (SDW)
0.00e
0.00e
0.00d
0.00
Trichoderma (biocon)
15.67d
15.17d
15.17c
15.61
Mancozeb (Dithane M45)
20.67bc
18.67bc
16.83c
18.28
Lysol
25.83a
24.50a
20.00b
24.45
Mancozeb (Parafungus)
21.17bc
19.50b
17.17c
17.83
Benomyl (Benlight)
24.00a
23.00a
22.17a
32.33
Captan
18.00cd
16.50cd
15.67c
17.72
On day seven, the inhibition zone of Benomyl 22.17 mm, continued to decrease
slightly but still the widest and is significantly higher than the inhibition zone of Lysol
which is 20.00 mm. The inhibition zones of Trichoderma, Dithane M45, Parafungus, and
Captan remained comparable.
Table 8 shows the result of bioassay on Isolate 03, after 3, 5, and 7 days of
incubation. On day three, the inhibition zones of Benomyl and Lysol of 26.83 mm and
27.33 mm were significantly different over the other treatments. The inhibition zone of
Captan (21.33 mm) was next to Benomyl and Lysol, while the rest were comparable in
inhibiting F. oxysporum f. sp. cubense.
The same observation was noted after five days. Benomyl and Lysol still provided
the best inhibition zones measuring 25.17 mm and 24.67 mm followed by the inhibition
zone produced by Captan which is 19.83 mm, the rest of the treatments remained
comparable.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Table 8. Inhibition zones (mm) formed on Potato Dextrose Agar (PDA) by isolate 03 after
three, five, and seven days
TREATMENTS
DAY 3
DAY 5
DAY 7
MEAN
Sterile Distilled Water (SDW)
0.00e
0.00d
0.00d
0.00
Trichoderma (biocon)
15.17d
15.00c
14.33c
14.83
Mancozeb (Dithane M45)
17.00d
15.50c
14.33c
15.61
Lysol
27.33a
24.67a
22.33a
24.78
Mancozeb (Parafungus)
17.50d
15.83c
14.35c
15.89
Benomyl (Benlight)
26.83a
25.17a
23.00a
25.00
Captan
21.33b
19.83b
18.00b
19.72
On day seven, Benomyl and Lysol still had the highest inhibition zones measuring
23.00 mm and 22.33 mm. Captan had an inhibition zone of 18.00 mm which is significantly
different with the inhibition zones of Trichoderma, Dithane M45, and Parafungus.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Other observations
Laboratory. Table 9 shows the colony growth of F. oxysporum f. sp. cubense grown
in a commercially prepared Potato Dextrose Agar (PDA). Results revealed that isolates 01,
02, and 03 had lower colony growth of 32.75 mm, 32.78 mm, and 39.53 mm compared to
the colony growth of 62.67 mm, 65.00 mm, and 59.83 mm obtained from the fresh PDA
(Table 4). The pigment on PDA also varied from light purple to light pink, dark purple to
pink, and light orange to dark pink.
Table 9. Colony diameter of F. oxysporum f. sp. cubense in commercially prepared Potato
Dextrose Agar (PDA) after three, five, and seven days
COLONY DIAMETER (mm)
Isolates
3 day
5 day
7 day
MEAN
Pigment on PDA
Isolate 01
28.67
32.50
37.08
32.75
light pink
Isolate 02
28.92
34.25
35.17
32.78
pink
Isolate 03
34.83
40.33
43.42
39.53
dark pink
Figure 18. Three day old cultures of F. oxysporum f. sp. cubense on the commercially
prepared Potato Dextrose Agar(PDA); (A) Isolate 01, (B) Isolate 02,
(C) Isolate 03
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Incidence of Banana Bunchy Top Virus (BBTV). As presented in Table 10, bunchy
top disease is also present in the areas surveyed. The incidence recorded in the barangays
and the different sitios showed that while the virus disease exists, the distribution does not
differ significantly.
Table 10. Incidence (%) of Banana Bunchy Top Virus (BBTV) in the areas surveyed
AREAS SURVEYED
PERCENT (%) INCIDENCE
BARANGAY
Barangay Sagubo
14.76
Barangay Gadang
14.36
SITIO
Barangay Sagubo, Sitio Ampongot
19.79
Barangay Sagubo, Sitio Timoc and Mocgo
7.94
Barangay Sagubo, Sitio Dosco
15.69
Barangay Gadang, Sitio Lib-libeng
19.25
Barangay Gadang, Sitio Namon-ao
18.46
Barangay Gadang, Sitio Copias
16.33
Barangay Gadang, Sitio Gadang Proper
7.61
Barangay Gadang, Sitio Namat-ikan
10.63
CV= 1.5064 %
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Banana Bugtok or Blood disease. This disease was noted in barangay Gadang, sitio
Lib-libeng. The disease was observed to infect the banana variety Dippig or Cardaba.
Infected fruits do not ripen and are not fit for consumption.
According to Soguilon et.al (1995) bugtok is an endemic and a widely distributed
bacterial disease of cooking banana cultivars in the Philippines. Bugtok is a local term in
the southern Philippines used to describe the infected fruit which are discolored and hard
even when ripe. It was noted as a minor disorder more than 40 years ago, but was reported
by Roperos in 1965 as a developing problem of importance. It is caused by Pseudomonas
solanacearum E. F. Smith.
Bugtok can be controlled by bagging the inflorescence at the bending stage just
after emergence. The bagging material can be a polyethylene bag, muslin cloth, or a fine
nylon mesh bag. Bags can be removed after all the fruits have set followed by removal of
the male inflorescence. This practice should also include mat and field sanitation, and
removal of old, dead leaves.
Figure 17. Banana bugtok or blood disease on Cardaba (Dippig)
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
SUMMARY, CONCLUSIONS AND RECOMMENDATIONS
Summary
The study was conducted to survey the occurrence of Fusarium wilt in two banana
producing barangays of Kapangan, Benguet, namely Gadang, and Sagubo; asses the
incidence and severity of infection of the affected banana plants; isolate and characterize
the causal pathogen Fusarium oxysporum f. sp. cubense from infected samples, and
compare the effect of bio control agent and chemicals in inhibiting the growth of F.
oxysporum f. sp. cubense through assay.
Results show that Fusarium wilt of banana is prevalent in the areas surveyed.
Fusarium wilt incidence of 16.82% and 13.57 % in barangays Sagubo and Gadang
including the incidence noted on the different sitios does not vary significantly. Among the
different sitios surveyed, sitio Ampongot has the highest incidence of 20.21 % while sitio
Gadang Proper has the lowest incidence of 10.72 %.
Fusarium wilt severity based on a five point rating scale showed that Lakatan was
severely affected followed by Tomok or Cavendish and then Kantong or Bungulan. Banana
cultivars Dippig and Balayang were not infected by Fusarium wilt in the area surveyed.
Based on the result of the cultural and morphological characterization of F.
oxysporum assigned as isolate 01, 02, and 03, isolate 02 had the highest colony diameter
of 65.00 mm, followed by isolate 01 with 62.67 mm. Isolate 03 gave the smallest colony
diameter of 59.83 mm. In terms of pigment on fresh Potato Dextrose Agar (PDA) isolate
01 has a light purple pigment, dark purple on isolate 02, and light orange on isolate 03.
Macro conidia measurements of isolate 01, 02, and 03 of 30.42 x 3.64 µm, 31.20 x
4.68 µm, and 33.80 x 4.81 µm are statistically comparable. On the other hand, the micro
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
conidia of the three isolates that measured 8.77 x 2.73 µm, 8.06 x 2.86 µm, and 7.80 x 3.25
µm was found to be within the range of the reported measurement size of F. oxysporum.
The results of the bioassay on isolate 01 revealed that Benomyl and Lysol
significantly produced the highest inhibition zones of 32.33 mm and 24.45 mm. The
inhibition zones of Trichoderma, Dithane M45 (Mancozeb), Parafungus (Mancozeb), and
Captan were comparable.
On the other hand, the result of bioassay on isolate 02 showed that the widest
average inhibition zones were observed on Benomyl and Lysol having 23.06 mm and 23.44
mm, respectively. The inhibition zones produced by Trichoderma, Dithane M45
(Mancozeb), Parafungus (Mancozeb), and Captan were also comparable.
The same trend was observed on isolate 03 where in Benomyl and Lysol still
provided the widest inhibition zones measuring 25.00 mm and 24.78 mm. Captan had
higher inhibition zone as compared to the inhibition zones produced by Trichoderma,
Dithane M45, and Parafungus after seven days.
Conclusions
Banana Fusarium wilt incidence in barangay Sagubo and Gadang is low ranging
from 10 to 20 %. The varieties grown in the area are Lakatan, Kantong, Lemen, Tomok,
Balayang, Dippig, Latundan, Gloria, Señorita, and Malagambang. Lakatan, Kantong, and
Tomok are susceptible to the existing race of F. oxysporum f. sp. cubense existing in
Kapangan.
Banana is a secondary crop in barangay Sagubo and barangay Gadang with a total
area planted of 24,195 m².
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
Isolates 01, 02 and 03 of F. oxysporum f. sp. cubense differed in their colony growth
in Potato Dextrose Agar including the size of macro and micro conidia.
Benomyl is effective in inhibiting the growth of F. oxysporum. Trichoderma is not
as effective as benomyl but can be used as alternative for sustainable Fusarium wilt
management.
Recommendations
Bioassay should be tested in pot experiments to confirm if the same results will be
obtained. Planting materials should not be sourced out from areas, such as Mindanao,
where Race 4 exists.
Similar study should be done to other banana producing areas of Benguet and
Mountain Province to determine the presence and incidence of F. oxyporum f. sp. cubense.
Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013
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Survey, Assessment, and Characterization of Banana Fusarium Wilt (Fusarium oxysporum f. sp.
cubense) in Two Barangays of Kapangan, Benguet | CODOD, CLARENCE BAGAYAO. APRIL 2013