BIBLIOGRAPHY

CATHERINE APALING BAGSAN. NOVEMBER 2006. Management of
Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in Chrysanthemum
(Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and Plant Extracts.
Benguet State University, La Trinidad, Benguet.
Adviser: Luciana M. Villanueva, Ph. D.
ABSTRACT

Selected bacterial antagonists and plant extracts were evaluated as possible
alternatives to synthetic fungicides in managing Fusarium wilt disease in chrysanthemum
caused by Fusarium oxyxporum f. sp. chrysanthemi under the laboratory and greenhouse
conditions. The study was conducted at the Biological Control Laboratory and
greenhouse at Benguet State University, La Trinidad, Benguet from November, 2005 to
October, 2006.

Bioassay test of the two bacterial antagonists Flavobacterium sp. and
Pseudomonas sp. inhibited the growth of the pathogen F. oxysporum f. sp. chrysanthemi
in culture forming inhibition zones. Among the plant extracts tested, garlic extract
(Allium sativum L) significantly suppressed the growth of F. oxysporum f. sp.
chrysanthemi in culture and gave the widest inhibition zone.
Among the fungicides
tested against F. oxysporum f. sp. chrysanthemi, mancozeb (Parafungus) gave
significantly the widest zone of inhibition followed closely by benomyl (Benlate) ,
thiophanate methyl (Fungitox), thiophanate methyl (Topsin-M) and captan (Captan).

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Chlorothalonil (Daconil) and mancozeb (Dithane M-45 ) did not inhibit the growth of the
pathogen.

The test conducted to determine the compatibility of plant extracts and fungicides
with the two bacterial antagonists, garlic extract and mancozeb showed incompatibility
with Flavobacterium sp. and Pseudomonas sp. On the other hand, thiophanate methyl,
gawed extract and table salt (NaCl) did not inhibit the growth of the bacterial antagonists.
Therefore, in an integrated disease management program against Fusarium wilt of
chrysanthemum mancozeb and garlic extract should not be combined with the biocon
agents because these combinations will kill the beneficial bacteria.

Results of the greenhouse experiment conducted to determine the effectiveness of
bacterial antagonists and plant extracts in reducing the soil population of F. oxysporum f.
sp. chrysanthemi showed that the combination of Flavobacterium sp and Pseudomonas
sp. was comparable with the standard fungicide, mancozeb. Although not as effective as
the other treatments, application of garlic extract, Pseudomonas sp and Flavobacterium
sp. alone and table salt significantly reduced the soil population of the pathogen
compared with the untreated - inoculated plants. On the other hand, gawed extract was
inferior with the other treatments in suppressing the pathogen population.

On disease severity, the lowest wilt infection was observed in plants treated with
the fungicides, mancozeb and thiophanate. The result was consistent throughout the
duration of the experiment. Although lower in efficacy, the combination of
Flavobacterium sp. and Pseudomonas sp. and garlic extract significantly reduced wilt
infection compared with the untreated/uninoculated plants. The highest wilt infection
was observed in plants treated with hot water (applied before planting) + garlic extract
ii

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(added after 1 week) + combination of bacterial antagonists (Flavobacterium sp. and
Pseudomonas sp. ) introduced 17 days after transplanting which did not differ
significantly with untreated-inoculated plants.

In terms of yield, plants treated with garlic extract combined with Pseudomonas
sp. and Flavobacterium sp. produced good quality cutflowers. The color of the flowers
was bright orange compared with the untreated inoculated plants which were small and
pale. The treatments produced lower non-marketable comparable to the fungicide treated
plants. The plants applied with the combination of hot water treatment + garlic extract +
combination of bacterial antagonists introduced 17 days after transplanting (DAT)
produced more of class B and non-marketable flowers.



iii

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TABLE OF CONTENTS

Page
Bibliography. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
i
Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
I
Table of Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
iv
INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1
Background of the study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1
Importance of the study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
2
Objectives of the study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3
Time and place of the study . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . .
3
REVIEW OF LITERATURE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4
The Disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . .. . . . . .
4
Symptoms . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5
Sign . . . . . . . . . . . . .. . . .. . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . .
5
Survival . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
6
Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
8
Biological control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
8
Plant Extracts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10
Chloride and Soil pH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
12
MATERIALS AND METHODS . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
13
Collection and isolation of the pathogen . . . . . . . . . . . . . . . . . . . . .
13
Laboratory experiments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
13
Bioassay test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
13
iv

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Preparation of bacterial suspension . . . . . . . . . . . . . . . . . .
14
Preparation of plant extracts . . . . . . . . . . . . . . . . . . . . . . . .
14
Preparation of fungicides . . . . . . . . . . . . . . . . . . . . . . . . . .
15
Compatibility test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
16
Greenhouse experiment . . . . . .. . . . . . . . . . . . . . . . . ………….
17
Effect of bacterial antagonists and plant extracts on the soil
17
population of F. oxysporum f. sp. chrysanthemi . . . . . . . . . .

Efficacy of bacterial antagonists and plant extracts on
19
fusarium wilt infection and chrysanthemum yield . . . . . . . . .

Data to be gathered . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
22
RESULTS AND DISCUSSION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
24
Laboratory experiment . . . . .. . . . . . . . . . . . . . . . . . . . . .
24
Effect of bacterial antagonists and plant extracts and
24
fungicides on the growth of the pathogen . . . . . . . . . . . . . . .

Compatibility test . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . .
28
Greenhouse experiments . . . . . . .. . . . . . . . . . . . . . . . . . . . .
30
Effects of bacterial antagonists and plant extracts on the soil
30
population of Fusarium oxysporum f. sp. chrysanthemi . .

Effects of bacterial antagonists and plant extracts on the
33
fusarium wilt infection and yield of chrysanthemum . . . . . .

Fusarium wilt infection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
33
Vascular discoloration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
35
Fresh weight . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
38
Cutflower yield . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
39
v

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SUMMARY, CONCLUSION AND RECOMMENDATION . . . . . . .
42
Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
42
Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
43
Recommendation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
44
LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
45
APPENDICES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
48





vi

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INTRODUCTION

Background of the Study

Fusarium wilt represents a continuing challenge for worldwide production
of chrysanthemum. It causes significant losses in chrysanthemum crops. These
losses may occur year after year because of the carry over of the organisms in
infected propagation stocks, the persistence of the fungus in the soil and the
difficulty in controlling the fungus once it becomes established in the soil
(Engelhard and Woltz, 1973). This is caused by the fungus, Fusarium oxysporum
f. sp. chrysanthemi. The disease is frequently complicated by the variability of
symptoms on various cultivars and because the symptoms may resemble those of
nutrient deficiencies, Pythium root rot or excess water ( Engelhard and Woltz,
1971). Infection results in plugging of the xylem vessel elements with gum and
pectinaceous materials, hypertrophy and hyperplasia of xylem parenchyma cells,
abnormal activity of the vascular cambium derivatives, the formation of cavities
within xylem tissues and eventual colonization of phloem and cortex parenchyma
cells that results in their collapse ( Bowers and Locke, 2000).
Most of the growers are aware of this dreaded disease , however, very
few know how to manage the disease properly. When the disease attacks, it
usually appears at reproductive stage thus normally affects the quality and
quantity of the produce. Farmers apply control measures however, they do it
improperly and at the wrong time. Agri-chemicals play an important role in
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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improving the quality of cutflowers harvested. However, use and misuse can
give detrimental effect to the environment and to public health. Chemicals
pollute the environment, enter the food chain, harm beneficial or non-target
organisms and contaminate the water supply. Moreover, chemical pesticides are
ineffective against most soil-borne pathogens and the control action of the
effective ones is often short-lived (Madamba et al., 1999; Agrios, 1997).

Importance of the Study
Biologically-based and environmentally-safe alternatives such as the use
of biological control agents, natural plant products, and cultural methods are
being investigated for possible use in integrated management program (Bowers
and Locke, 2000). Biological control along with accurate disease detection,
diagnosis, and sound cultural management techniques offer the best alternative
measure to reduce pesticide use in chrysanthemum production.
In the highlands of Northern Philippines, the first report on the use of
bacterial antagonists and plant extracts for the control of Fusarium wilt in garden
pea was done by Villanueva and Lirio (2000). Flavobacterium sp. isolated from
healthy beans at Sagada, Mt. Province provided the best control of the disease
and the highest yield. Among the plant extracts, Psidium guajava provided the
highest percentage control followed by Piper betle. The efficacy of garlic extract
and bacterial antagonists against F. oxysporum f. sp. chrysanthemi has been
shown in the previous trials (Villanueva et al , 2004).
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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Objectives of the Study


This study was therefore conducted with the following objectives:
1. to determine the effect of selected bacterial antagonists,
plant extracts, and fungicides against F. oxysporum f. sp. chrysanthemi
under laboratory conditions;
2. to determine the compatibility of selected plant extracts
and fungicides with the bacterial antagonists;
3. to determine the effectiveness of the bacterial antagonists and

plant extracts in reducing the soil population of F.oxysporum f. sp

chrysanthemi; and
4. to determine the effect of bacterial antagonists and plant extracts
on Fusarium wilt infection and cutflower yield.

Time and Place of the Study

The experiments were conducted at the Biocon Laboratory and
greenhouse of the Horticulture Research and Training Institute (HORTI),
Benguet State University, La Trinidad, Benguet from November, 2005 to
October, 2006.



Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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REVIEW OF LITERATURE

The Disease

In chrysanthemum, the fungus that caused Fusarium wilt is composed of
two races of Fusarium oxysporum f. sp. chrysanthemi (Forsberg, 1976), each of
which attacks specific chrysanthemum varieties. In addition, disease
development depends on the crop or variety, the race of Fusarium present and
environmental factors such as temperature, nitrogen and soil reaction (Nyvall,
1979; Singh, 1978; Agrios, 1988).

Fusarium wilt is one of the most widespread and destructive diseases of
major ornamental and horticultural crops. The soil-borne fungus causes vascular
wilts by infecting plants through the roots and growing internally through the
cortex to the stele. The vascular tissues of the root and then the stems, are
colonized by growth of hyphae and movement of conidia in the transpiration
stream. Initial symptoms appear as chlorosis, and distortion of the lower leaves,
often on one side of the plant. Foliar chlorosis, necrosis and plant stunting become
more pronounced as the disease progresses. Wilting occurs on the affected side of
the plant, followed by vascular discoloration and stem necrosis. The entire plant
wilts and dies as the pathogen moves into the stem (Bowers and Locke, 2000).

According to Agrios (1997) Fusarium wilt damages plant by causing
stunting until it soon wilts and finally die. These pathogens do not reach the
apical meristem until the very late stages of the disease. This is characterized by
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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5
drying of the entire plant due to the toxic substances secreted by the pathogen
attacking the plant (Forsberg, 1976).

Symptoms

According to Nyvall (1979) the typical symptoms appear as yellowing
(chlorosis) and distortion of leaves on one side of the plant. Chlorosis begins on
the lower leaves followed by wilting that progresses up the plant. In the early
stages of the disease, the roots are not rotted. In many plants such as carnation
and gladiolus, the infection may be one sided at first. Infected plants wilt, the
lower leaves turn yellow and dry and the xylem tissues turn brown followed by
death of the plant (Agrios, 1997 ; Bowers and Locke, 2000).

The occurrence of specific symptoms and their severity depends
primarily on the interaction of the cultivar and temperature of the soil and air.
The pattern of symptom development in cultivars also varies wherein symptoms
appear first at the plant apex and move down in some cultivars, rather than
starting at the base of the plant as in most vascular wilts. Temperature above
28ºC favor disease development and play a major role in symptom development
(Stuehling et. al. 1981).

Sign
The fungus F. oxysporum f. sp. chrysanthemi has two types of conidia:
macroconidia ( large, multi-celled spores) and microconidia (small, one-celled
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
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Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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spores) (Bowers and Locke, 2000). According to Agrios (1997) under the
microscope, macroconidia appear as crescent-shaped, hyaline, slimy, banana-
shaped and septated while microconidia appear as hyaline and single-celled.

Survival
F. oxysporum f. sp. chrysanthemi like other species survived in the soil
saprophytically almost forever ( Agrios, 1988 and 1997; Roberts and Boothroyd,
1972and Bowers and Lock, 2000). It is generally spread through water splash,
irrigation, contaminated tools and contaminated planting materials. Once within
the plant, the fungus grows and multiplies in the vascular system (water and food
conducting tissues) of the roots. It then moves upward in the plant by spores
(macrocinidia and micro conidia) that are transported in the sap stream where they
become lodged, germinate and affect new plant parts; or the fungus extends its
colonization as it grows in the vascular tissues of the host. The normal flow of
liquids and nutrients from the roots to the foliage is greatly reduced or stopped
because the conducting tissue becomes partially plugged or killed by fungal
mycelium and spores, or by the overgrowth of the neighboring cells. Toxic
substances are believed to be secreted by interaction of the fungus and the host
plant. These materials apparently cause the wilting and eventual death of the
plant. Wilt symptoms typically are not observed until the fungus has colonized
the underground parts of the plant.
Resting structures (chlamydospores) are formed within infected plant
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
6
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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7
parts. After the host plant dies or the growing season ends, this Fusarium fungi
survive as mycelia and chlamydospores, overwintering in dead plant parts; or may
live in the soil indefinitely in the absence of the host plant, especially if the soil is
warm, as in the greenhouse. Chlamydospores are stimulated to germinate by
exudates from the roots of a host plant which they then infect (Forsberg, 1976;
Bowers and Locke, 2000). Fusarium wilts are much more common and
destructive in the warmer temperate regions and in the tropics and subtropics
becoming lesser damaging or rare in colder climates except for greenhouse crops
in these areas ( Agrios, 1997). According to Forseberg (1976) severe symptoms
develop at constant temperatures of 80ºF and 90ºF. They are mild at 70ºF and
absent at 60ºF or below. Soil temperatures of about 25ºC (77ºF) and higher and
low soil moisture generally favor wilt development (Singh , 1978). On the other
hand, (Nyvall , 1979) also reported that disease severity is greater during high
temperatures (35ºC day and 29 ºC night), moist weather and in plants grown in
acidic and light textured soils that are high in nitrogen. Constant high
temperatures are known to favor development of Fusarium wilt of
chrysanthemum caused by Fusarium oxysporum f. sp. chrysanthemi. In a
previous study, a constant temperature of 35ºC was most favorable for
development of this disease ( Gardiner et. al. 1989).

In the presence of roots, chlamydospores or conidia germinate and
penetrate susceptible plants. The fungus enters the xylem and grows upward
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
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Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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plugging the tissues and reducing the movement of water. Toxins are produced
that cause the foliage to turn yellow but an infected plant may remain
symptomless at lower temperatures.

Management
Control
of
Fusarium wilt of chrysanthemum involves the use of culture
indexed cuttings, treatment of the growing medium and sanitation. Culture-
indexing is widely used in the industry to assure that cuttings are free of the
pathogens that cause Fusarium and Verticillium wilts. Sanitation is used to keep
the medium free of the pathogen during the growing period (Toop, 1963).

Fungicide drenches and growing potted chrysanthemums in a high lime,
all nitrate nitrogen cultural regime have been shown to provide complete
management of Fusarium wilt. However, plants may still be colonized by the
fungus and should not be used as a source of cuttings for propagation Engelhard
and Woltz (1971).

Biological Control

Biological control is becoming an urgently needed component in
agriculture. Chemical pesticides have been the objects of substantial criticism in
recent years due to the adverse environmental effects causing health hazards to
human and other non-target organisms including beneficial natural enemies. It is
now safer and environmentally feasible control alternatives the use of existing
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
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Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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9
living organisms to provide protection against a large range of plant pathogenic
fungi without damage to the ecological system ( Madamba et. al., 1999).

Biological control is the use of any organisms to control a pathogen. It is
the reduction in attack of a crop species by a pathogen achieved using another
living organisms. This includes both direct and indirect effects due either to
introduced antagonist or manipulation of existing populations to reduce disease
(Baker and Cook, 1974).
Becker et. al (1993) obtained fluorescent pseudomonads strain from
wheat roots grown in take-all decline soils and tested for a possible control of the
disease in the field and found out several strains were effective in reducing
severity and increasing yield of wheat. On the other hand, Arie et al (1987) also
obtained strain M-2196 of Pseudomonas gladioli that was effective in reducing
the severity of Fusarium wilt of bottle gourd when the roots of Allium spp.
seedlings were dipped in the bacterial suspension before being transplanted along
side bottle gourds plants.
In addition, strains of fluorescent Pseudomonads PGPR that promoted
potato yield induced reduction in root populations of the potato soft-rot pathogen,
Erwinia carotovora of 95-100% and a reduction in the percentage of daughter
tubers infected with the pathogen ranging from 28-92% compared to non-treated
controls ( Bolayo, 1996). PGPR has been reported as potential biological control
agents for many root and crown rot pathogens including Aphanomyces, Fusarium
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
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Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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solani, Gauanomyces graminis var.tritici, Phytophthora megasperma f. sp.,
glycinia, Sclerotium rolfsii and Thielaviopsis bassicola. These provided
protection by diverse mechanisms including production of antibiotics, production
of extra - cellular lytic enzymes, and production of hydrogen cyanide (Bowers
and Locke, 2000).
Villanueva and Lirio (2000) reported that out of 160 bacterial isolates
collected from vegetable growing areas in Benguet and Mt. Province, two isolates
showed strong potential against Fusarium wilt pathogen. Flavobacterium sp.
obtained from healthy beans at Sagada, Mt. Province provided the best control
(53.76%) followed by the standard fungicide, mancozeb and Bacillus sp. with
53.3 and 48.10 % control, respectively. Likewise, the highest pod yield was
obtained from plants applied with Flavobacterium sp followed closely with plants
treated with Bacillus sp.

Plant Extracts

Antimicrobial activity of plant extract is attributed to the presence of
bioactive compound (s) as reported by some researchers (Favaron et. al., 1993:
Onu, 1995; Rao and Singh, (1992) and Villanueva and Lirio, (2000). Aqueous
extract of Piper betle was reported to demonstrate antifungal activity against
several test fungi. The active volatile principle was identified as eugenol. There
was in fact, the first report of the antimicrobial activity of P. betle ethanol extract
at concentration of 1.0 g per liter and 2.5 g per liter when tested both in - vitro and
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
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Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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11
in - vivo respectively against three fungal pathogens namely, Pyricularia oryzae,
Cochliobolus miyabeanus Deschler and Rhizoctonia solani Kuhn which incite
blast, brown spot and sheath blight diseases of rice, respectively (Villanueva and
Lirio, 2000).
The high levels of polyphenols from P. betle leaves ( chavicol, chavibetol,
allylpyro catechol, chabetol acetate and allylpyrocatecol diacetate) were
considered responsible for this fungicidal and nematicidal activities (Villanueva
and Lirio ( 2000).

According to Saxena ( 1983) plants are virtually “nature’s chemical
factories” providing practically unlimited natural resources of botanical
pesticides. Plant themselves produced highly sophisticated defense chemicals
which contain abundance of natural defense systems. Different societies in the
world have continually employed plants to kill or repel pests since civilization
began. However, due to appearance of synthetic chemicals, this kind of
indigenous technology was ignored (Wagang, 1999).
According
to
Villanueva and Lirio (2000) Psidium guajava provided the
highest percentage control (44.81%) of Fusarium wilt in garden pea. This was
not, however, significantly different from the degree of control exhibited by P.
betle (21.68%). Greenhouse experiment conducted by Villanueva and Masangcay
(2004) showed that 10% garlic extract significantly reduced the soil population
density of F. oxysporum f. sp. chrysanthemi.
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
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Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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Chloride and Soil pH.
Chloride nutrition has proven useful in the suppression of Fusarium
diseases in many crops that have tolerance to Cl. A single application of 0.25 –
0.5g of Na CL/L soil applied to cyclamen plugs grown in soil infested with
Fusarium oxysporum decreased mortality and increased fresh weight and leaf
area. The most noticeable effect was its ability to postpone the onset of wilt
symptoms and delay disease severity. Plant tissue analysis revealed elevated
levels of Na, Cl, and Mn. Since Mn is associated with the defense mechanism in
plant tissue, this may be one mechanism by which NaCl suppresses Fusarium
wilt. Adding lime to a potting mix to raise to pH from 6.5 to 7.5 did not result in
any significant effect on the disease, plant weight or flower number. Also, no
significant differences in growth or disease were noted when the pH of medium
was lowered to 5.5 with sulfuric acid. When NaCl was combined with the
different pH treatments, NaCl improved growth, but the greatest benefit was seen
at a pH of 7.5 pH. This may be due to the NaCl increasing Mn and other trace
elements at the higher pH (Elmer, 2000).

Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
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Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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MATERIALS AND METHODS

Collection and Isolation of the Pathogen

Plants infected with Fusarium wilt were collected from greenhouses where
chrysanthemum plants are grown. The diseased specimens were brought to the
biocontrol laboratory for isolation and washed thoroughly. From the advancing
lesion, two sections were cut and disinfected with 1% chlorox for three minutes
then rinsed three times in sterile distilled water. The lesions were blotted dry in
sterilized tissue paper. After which, four sections of the cut specimen were
transferred equidistantly in previously plated potato dextrose agar (PDA) and then
placed in the incubation chamber with an average temperature of 27ºC for 5 days.

Laboratory Experiment

Bioassay Test

The effect of bacterial antagonists, plant extracts and selected fungicides
on the growth of F. oxysporum f. sp. chrysanthemi was determined.

In preparing inoculum suspension, the plates containing pure culture of F.
oxysporum f. sp. chrysanthemi were dispensed with 10 ml sterile distilled water
(SDW). Using sterilized wire loop, the fungal growth was scraped and the
inoculum suspension was pre-standardized by counting the number of spores per
ml using a haemacytometer. A fungal suspension with spore concentration of
1.2 x 106 was used in the bioassay test. On sterilized petri plate, 0.1 ml of the
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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14
fungal suspension was placed and evenly distributed into each sterilized plate.
Later, PDA medium was then poured and spread uniformly in each plate and
incubated for 5 days.


Preparation of bacterial suspension

Pure cultures of the bacterial antagonists maintained in the Biocon
Laboratory were sub-cultured by re-streaking into solidified agar slants containing
nutrient agar medium. After incubation at 28ºC for 24 hours, the bacterial cells
were suspended in SDW and standardized to contain ca. 1 x 107 cfu/ml. Nine
sterile paper discs were dipped into the suspension of the antagonistic
microorganisms. Three discs were equidistantly placed at the center of the plate.
The treatments were replicated three times and arranged randomly in the
improvised incubation chamber following the completely randomized design
(CRD). The diameter of inhibition zone was measured for each replication after
24 hours of incubation.

Preparation of Plant Extracts

Leaves of gawed (Piper betle) and kutsai (Allium schoenoprasum L.) ,
bulbs of garlic (Allium sativum L) and red onions (Allium cepa) and hot pepper
fruits (Capsicum frutescens L.) were brought to the laboratory and washed
thoroughly. Twenty grams of the materials were added to 20 ml of sterilized
distilled water. Using mortar and pestle, the materials were macerated and the
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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15
sap was extracted using sterilized cheese cloth (Plate 1).












Plate1. Extraction of plant extracts for bioassay tests.



Preparation of Fungicides

To prepare the fungicide solution to be used in the bioassay test, the
computation was based on recommended rates. The desired amount was weighed
using an electronic balance. Sterile filter paper discs were soaked in each
chemical solution and placed at the center of the solidified PDA medium prepared
earlier. The treatments were replicated three times and arranged randomly in the
improvised incubation chamber. The diameter of inhibition zone was measured
after three days.





Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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16

The different treatments were:

TREATMENTS
ISOLATES/
SCIENTIFIC NAME/
CONCENTRATIONS/
RECOMMENDED

LOCAL NAME
CHEMICAL NAME
RATE




T1
Control


T2
Isolate 31
Bacillus sp.
107 cfu/ml
T3
Isolate 131
Bacillus pumilus
107 cfu/ml
T4
Isolate 158
Pseudomonas sp.
107 cfu/ml
T5
Isolate 94
Flavobacterium sp.
107 cfu/ml
T6
Isolate 73
Bacillus pumilus
107 cfu/ml
T7

Verticillium sp.
107 spores/ml
T8
Garlic
Allium sativum L.
50%
T9
Gawed
Piper betle
50%
T10
Kutsai
Allium schoenoprasum L. 50%
T11
Hot pepper
Capsicum frutescens L.
50%
T12
Red onion
Allium cepa
50%
T13
Fungitox
Thiophanate methyl
0.03g/20ml
T14
Topsin-M
Thiophanate methyl
0.03g/20ml
T15
Parafungus
Mancozeb
0.09 g/20ml
T16
BLB stopper
Thiodazole copper
0.06ml/20ml
T17
Captan
Captan
0.1g/20ml
T18
Benlate
Benomyl
0.03g/20ml
T19
Daconil
Chlorothalonil
0.06g/20ml
T20
Dithane M- 45
Mancozeb
0.08g/20ml
T21 Salt
Sodium chloride
.5g/li H20






Compatibility Test


The compatibility of the plant extracts and selected fungicides with the
bacterial antagonists was tested. This was necessary to determine if plant extracts
and fungicides could be integrated to effectively control fusarium wilt of
chrysanthemum.

Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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17
The different treatments are:
T1
Flavobacterium sp. + SDW
T2
Flavobacterium sp. + Mancozeb (Parafungus)
T3
Flavobacterium sp. + Thiophanate Methyl (Fungitox)
T4
Flavobacterium sp. + Garlic extract (Allium sativum L.)
T5
Flavobacterium sp. + Gawed extract (Piper betle)
T6
Flavobacterium sp. + Table salt(NaCl = .5g/L H20)
T7
Pseudomonas sp. + SDW
T8
Pseudomonas sp. + Mancozeb (Parafungus)
T9
Pseudomonas sp. + Thiophanate Methyl (Fungitox)
T10
Pseudomonas sp. + Garlic extract (Allium sativum L.)
T11
Pseudomonas sp. + Gawed extract (Piper betle)
T12
Pseudomonas sp + Table salt (NaCl = .5g /L H20)
T13
Pseudomonas sp. + Flavobacterium sp.

Greenhouse Experiments

Effect of bacterial antagonists and plant extracts
on the soil population of Fusarium oxysporum
f.sp. chrysanthemi

Two hundred g heat-sterilized soil was placed in each cup and
inoculated with F. oxysporum f. sp. chrysamthemi. Later, the different treatments
were applied (Plate 2). Soil samples were taken at 0 ( before soil treatment), 1,
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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3, 7, 14, 21 and 28 days after soil treatment (Plate 3). The population densities of
the pathogen were determined using dilution plate techniques (Plate 4).







Plate 2. Application of different treatments. Plate 3. Collection of soil samples


(10g/

replicate)









Plate 4. Determination of pathogen population using soil
dilution technique






Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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The treatments were:
T1 Uninoculated-Untreated
T2
Uninoculated – Treated
T3
F.o.c. + Hot water treatment
T4
F.o.c. + Hot water treatment + garlic extract after a week +
Flavobacterium sp. + Pseudomonas sp. after 17 DAT

T5 F.o.c.
+ Flavobacterium sp.
T6
F.o.c. + Pseudomonas sp.
T7
F.o.c. + Pseudomonas sp. + Flavobacterium sp.
T8
F.o.c. + Mancozeb (Parafungus)
T9
F.o.c. + Garlic extract (Allium sativum L)
T10
F.o.c + Gawed extract (Piper betle)
T11
F.o.c + Table salt (NaCl = .5g/Li)



Effect of bacterial antagonists and plant extracts on
Fusarium wilt infection and chrysanthemum yield

The soil used in the experiment was obtained from an area not previously
planted with chrysanthemum. This was brought to the greenhouse for sterilization
using a pressure cooker at 15 psi for 2 hours to ensure that all microorganisms in
the soil were killed. About two kg soil was placed in 10 cm-diameter plastic pots.
The procedure used for preparing the fungal inoculum, bacterial
antagonists and plant extracts was followed. The different treatments were as
follows:
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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T1 Uninoculated/Untreated
T2 Uninoculated/Treated
T3
F.o.c. + Hot water treatment + garlic extract after a week +
Pseudomonas sp. + Flavobacterium sp. after 17 days

T4
F.o.c. + Pseudomonas sp. + Flavobacterium sp.
T5
F.o.c + Mancozeb (Parafungus)
T6
F. o.c. + Thiophanate Methyl (Fungitox)
T7
F. o.c. + Garlic extract (Allium sativum L)
T8
F. o. C. + Gawed extract (Piper betle)
T9
F. o.c. + Table salt (1 g/L H20)


Seedlings of chrysanthemum cv. Toledo were transplanted and arranged
randomly in the greenhouse using the randomized complete block design. The
treatments were replicated four times with three sample plants per replicate. All
cultural management practices for chrysanthemum production were employed
uniformly in each treatment such as lighting three hours at night, fertigation,
pinching, disbudding (Plate 5), weeding and control of insect pests and foliar
diseases. The disease severity was assessed weekly using the modified rating
scale by Gardiner (1987) as follows:






Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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21
SCALE DESCRIPTION


1
Normal or no infection


2
5-10% leaf foliage showed symptoms of chlorosis or infection


3
11-25% leaf foliage showed symptoms of chlorosis or infection


4
26-50% leaf foliage showed symptoms of chlorosis and show
slight symptom of wilting



5
51-75% leaf foliage showed symptoms of chlorosis and show
moderate symptom of wilting and a slight vascular rotting


6
76-100% most of the leaf foliage showed symptom of chlorosis
and plants are wilting and visual vascular stem rotting






















Plate 5. Disbudding of plants



Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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22
Data Gathered

1. Diameter of inhibition zone for bioassay and compatibility tests. This is
the area where pathogen growth is inhibited and was measured in mm using a foot
rule to indicate the degree of fungitoxicity of the selected bacterial antagonists,
plant extracts and fungicides.
2. Colony forming units (CFU/ml). This was obtained by counting the
total number of fungal colonies per plate and computed using the following
formula:
Cfu = Average plate count x dilution factor (104)





Volume plated (0.1 ml)
3. Disease severity rating. This was be assessed using the modified rating
scale of Gardiner (1989).
4. Marketable and non-marketable yield. This was determined by
counting the cutflowers produced without damage and non-marketable using the
classification of King Louis Flowers and Plants, Inc. for spray type cultivars as
follows:





Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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23

STEM

CLASSIFICATION
LENGTH
FLOWER DESCRIPTION

Class AA
> 71
Large flower(s), clean leaves with straight stem
Long
65 – 71
Large medium flower(s), clean leaves with
Straight stem

Medium
55 – 61
Medium – small flower(s), clean leaves
with straight stem

Short
51
Small flower, clean leaves with straight stem
Class B
41
Small flower(s) with leaf disease
Non marketable

Damaged plants (wilted and diseased)

Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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RESULTS AND DISCUSSION


Laboratory Experiment


Effect of bacterial antagonists, plant extracts
and fungicides on the growth of the pathogen

Among the bacterial isolates tested, only Flavobacterium sp and
Pseudomonas sp. showed potential in controlling Fusarium oxysporum f.sp.
chrysanthemi with 1.0 and 0.66 mm inhibition zone, respectively. However, this
was comparable with sterile distilled water. Bacillus sp., Verticillium sp and the
two strains of Bacillus pumilus did not affect the growth of the pathogen (Table 1
and Plate 6).
Among the plant extracts tested, the widest zone of inhibition was
obtained from garlic extract with 9.65 mm. This was significantly different from
gawed (Piper betle), kutsai (Allium schoenoprasum) and hot pepper (Capsicum
frutescens L.) which were comparable with the control. Similarly, red onion
extract (Allium cepa) did not inhibit the growth of the pathogen. Antimicrobial
activity of plant extracts is attributed to the presence of bioactive compound(s) as
reported by most researchers (Favaron et al., 1993). The efficacy of the garlic
extract may be attributed to the constituents of the garlic which has a broad
spectrum anti-bacterial and anti-fungal activity. This antibiotic property is due to
the presence of allicin, a S- containing compound in the bulb (Rimando and
de Guzman, 1986).
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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25
Table 1. Effect of bacterial antagonists, plant extracts and selected fungicides
on the growth of Fusarium oxysporum f. sp. chrysanthemi a.

A.
BIOCONTROL
AGENTS
INHIBITION ZONE (MM




T1 SDW
0e

T2
Bacillus pumilus (73)
0e

T3
Bacillus pumilus (131)
0e
T4
Bacillus sp. (31)
0e

T5
Flavobacterium sp. (94)
0.66e

T6
Pseudomonas sp. (158)
1.0e

T7
Verticillium sp.
0e






B. PLANT EXTRACTS






T8 Garlic
(Alliumsativum L)
9.65ab

T9 Gawed
(Piper betle) 2.11e

T10 Kutsai
(Allium schoenoprasumL.)
1.25e
T11
Hot Pepper (Capsicum frutescens) 1.66e
T12
Red Onion (Allium cepa) 0e





C.
FUNGICIDES






T13 Thiophanate
Methyl
(Fungitox)
7.55c

T14
Thiophanate Methyl (Topsin – M)
5.01d

T15 Mancozeb
(Parafungus)
11.01a
T16
Thiodazole copper (BLB Stopper)
0e

T17 Captan
(Captan)
4.91d

T18
Benomyl (Benlate )
8.78bc

T19 Clorothalonil
(Daconil)
0e

T20 Mancozeb
(Dithane
M-45)
0e


D. OTHERS


T21
0.5 g Table salt/Liter
0e

CV = 42.48%
a Data are means of four replications. Means followed by a common letter are
not significantly different at 5% level using DMRT.





Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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26



bb
c
a





d
e
f




g
h
j




j
k




Plate 6. Effect of bacterial antagonists, plant extracts and fungicides on the
growth of F. oxysporum f. sp. chrysanthemi. a= Bacillus pumilus;
b = Bacillus pumilus ; c = Bacillus sp.; d = Verticillium sp.; e =
Pseudomonas sp.; f = Flavobacterium sp.; g = Garlic extract (
Allium sativum L.) ; h = Gawed extract (P. betle); I = Mancozeb
(Parafungus); j = Benomyl (Benlate); and k = Thiophanate Methyl
(Fungitox)

Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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27

On the other hand, aqueous extract of Piper betle was reported to
demonstrate antifungal activity against several test fungi. The active volatile
principle was identified as eugenol (Dubey and Tripathi, 1987). There was in fact
a first report of the antimicrobial activity of P. betle ethanol extract at
concentrations of 1.0 g per liter and 2.5 g per liter when tested both in-vitro and
in-vivo, respectively against the three fungal pathogens namely, Pyricularia
oryzae Cav., Cochliobolus miyabeanus Dreschler and Rhizoctonia solani Kuhn
which incite blast, brown spot and sheath blight of rice, respectively (Tewari and
Nayak, 1991).
Among the fungicides tested, the most effective was mancozeb
(Parafungus) with inhibition zone of 11.01mm. This was significantly more
effective than thiophanate methyl (Fungitox), benomyl (Benlate), thiophanate
methyl (Topsin –M) and captan (Captan) with inhibition zones of 7.55, 8.98 5.01
and 4.91 mm, respectively. On the other hand, thiodazole copper (BLB Stopper),
chlorothalonil (Daconil) and mancozeb (Dithane M-45) did not affect the growth
of the pathogen. This is quite surprising because the active ingredient of dithane
M-45 and parafungus are both mancozeb. Table salt applied at 0.5g/liter did not
inhibit the growth of F. oyxsporum f. sp. chrysanthemi.





Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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28
Compatibility Test
The effect of plant extracts and selected fungicides using their
recommended rate on the growth of the bacterial antagonists is shown in
Fig. 1 and Plate 7. Garlic extract and mancozeb significantly inhibited the
growth of Flavobacterium sp. and Pseudomonas sp. On the other
hand, thiophanate methyl, gawed extract and table salt did not affect the
growth of the above bacterial antagonists. This implies that plant extracts and
fungicides are compatible with the biocontrol agents and could therefore be used
in a sustainable management of Fusarium wilt of chrysanthemum.

Flavobacterium sp.
10
a
8
Pseudomonas sp.
6
b
ne (mm)
b
b
4
2
t
i
on Zo
c c
c c
c c
bi
0
Inhi
eb
a
te
e
d
/L)
a
r
lic
w
t
h
yl
G
Ga
.
5g
Mancoz
h
iophan
me
T
S
a
lt (
Treatment

Fig 1. Effect of plant extracts and selected fungicides on the growth of
bacterial antagonists. Bars with a common letter are not
significantly different at 5% level using DMRT.


Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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29





Flavobacterium sp. + +
Flavobacterium sp. ssp.p. +

Garlic extract
+ Mancozeb (Parafungus)








Pseudomonas sp. +
Pseudomonas sp.p.

Mancozeb (Parafungus)
+ + Garlic extract


Plate No.7 . Effect of garlic extract (A. sativum L.) and mancozeb
(Parafungus) on the growth of bacterial antagonists:
Pseudomonas sp. and Flavobacterium sp.











Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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30
Greenhouse Experiment

Effect of Bacterial Antagonists and Plant Extracts
on the Soil Population of F.oxysporum
f. sp. chrysanthemi

Fig. 2 shows the effect of the different treatments on the soil population
of F. oxysporum f.sp. chrysanthemi. Apparently, soil treated with hot water,
Flavobacterium sp., combination of Flavobacterium sp. and Pseudomonas sp. and
the standard fungicide, mancozeb significantly reduced the population density of
the pathogen a day after application. Three days after soil treatment, a decrease in
population was noted in all the treatments except those applied with gawed extract
and table salt. However, aside from mancozeb and the combination of the two
bacterial antagonists, Flavobacterium sp. and Pseudomonas sp., all the treatments
reduced their efficacy after a week. This implies that the efficacy of the treatments
was only good for three to seven days. One possible explanation is that most of
the pathogens died after treatment but the surviving populations multiplied rapidly
resulting to the increase of the population on the seventh day after treatment. At
this time the pathogen had reached its maximum growth so competition for food
and space occurred leading to the death of the organisms. On the 14th day, the
population started to decline due to lack of food. Application of garlic extract a
week after hot water treatment resulted in significant reduction in soil
population of F. oxysporum f. sp. chrysanthemi. This is reflected in the data
14 days after treatment.
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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31


T1
T2
T3
T4
T5
T6
T7
T8
T9
T10
T11

1.40E+06
a

1.20E+06
a
t 1.00E+06

coun 8.00E+05
on
ab
a
l
a
ti 6.00E+05
ab
a
a

ab
a
opu
a
bc
4.00E+05
P
abc
bc
a
ab
a
bc
bc
abc
bc
a
bc

a
bc
abc
ab
2.00E+05
bcd
ab
a
bcd
bc
a
bc
c
cd
bc
ab
c
c
cd
a
cd
bc
a
d
d
ab
bcd
a
d
d
a
ab
b
d
e
e
e
e
b
ab
0.00E+00

0
1
3
7
14
21
28

Days after inoculation

Fig. 2. Effect of bacterial antagonists and plant extracts on the soil population of
F. oxysporum f. sp. chrysanthemi. Lines with a common letter are not
significantly different at 5 % level using DMRT. Note: 1-
Uninoculated/untreated, 2-untreated/inoculated, 3-F.o.c. + Hot H2O
Treatment . 4- F.o.c. + Hot H2O + after 1week add garlic extract + Comb.
Of Pseudomonas sp. + Flavobacterium sp 17DAT., 5- F.o.c. +
Flavobacterium sp., 6- F.o.c. + Pseudomonas sp., 7- - F.o.c. +
Pseudomonas sp. + Flavobacterium sp., 8- Parafungus (Mancozeb), 9-
Garlic extract (Allium sativum L.), 10- Gawed extract (Piper betle), 11-
Table Salt (.5g/L).

Except for P. betle which was comparable with the untreated control, all
the other treatments gave significantly lower population of the pathogen.
Twenty one (21) days after treatment, the population of F. oxysporum f. sp.
chrysanthemi was lower in the soil applied with mancozeb, garlic extract,
combination of Flavobacterium sp. and Pseudomonas sp., the integration of hot
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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32
water, garlic extract and bacterial antagonists while those treated with hot water,
gawed extract, table salt and Pseudomonas sp. alone did not differ from the
untreated control. Finally, a reduction in the soil population was noted 14 days
after treatment and gradually declined up to the termination of the experiment.
During the last assessment period which was taken 28 days after treatment, all
the treated soil were comparable with the untreated ones. It can also be noted
that a week after treatment, a gradual decline in the population of the pathogen
was observed in the untreated soil. Population decline could be due to the
absence of host or lack of food.
The observed reduction in the pathogen population in the soil treated with
garlic and bacterial antagonists may have an important role in biologically-based
management strategies for Fusarium wilt. One possible management could be the
incorporation into the soil to initially reduce the pathogen population. This would
be followed by application of biological control agent compatible with the extract
three to ten days later to rapidly colonize the treated soil and to further suppress
the development of the pathogen thus achieving sustainable disease control of
Fusarium wilt of chrysanthemum.




Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

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33

Effect of bacterial antagonists and plant extracts
on fusarium wilt infection and
chrysanthemum yield.


Fusarium wilt infection


Figure 3 shows the effect of bacterial antagonists and plant extracts on
Fusarium wilt infection. Apparently, the most effective was thiophanate methyl,
however, it was not significantly different with that of mancozeb and garlic
extract. Although not as effective as the standard fungicides tested, application of
Pseudomonas sp. and Flavobacterium sp. and table salt significantly reduced
Fusarium wilt infection compared with the untreated inoculated plants.

Application of gawed extract and hot water treatment followed by garlic extract
and the combination of bacterial antagonists did not suppress the growth of the
pathogen. The potential of garlic extract and the two bacterial antagonists for the
control of Fusarium wilt in chrysanthemum has been reported by Villanueva et
al. (2004). The efficacy of the extract is attributed to the constituents of garlic
which has a broad spectrum anti-bacterial and anti-fungal activity. This antibiotic
property is due to the presence of allicin, an S-containing compound in the bulb
(Rimando and de Guzman, 1986). Since the inhibition zone obtained in
Pseudomonas sp. and Flavobacterium sp. was not so wide in the bioassay test, it
is presumed that the mode of action of the above antagonists is not antibiosis, but
could be any of the following mechanisms: parasitism, competition or rhizosphere
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

34
competence. On the other hand, the potential of the table salt for the control of
Fusarium wilt was also shown in the experiment. According to Elmer (2000)
chloride nutrition has proven useful in the suppression of Fusarium wilt diseases
in many crops. A single application of salt at the rate of 0.25-.5 g/ liter soil
applied to cyclamen plugs grown in soil infested with F. oxysporum decreased
mortality and increased fresh weight and leaf area. The most noticeable effect
was the ability of salt to postpone the onset of wilt symptoms and delay disease
severity. Plant tissue analysis revealed elevated levels of Na, Cl and Mn. Since
Mn is associated with the defense mechanism in plant tissue, this may be one of
the mechanisms by which table salt suppresses Fusarium wilt. Gawed extract
was not effective against Fusarium wilt of chrysanthemum. This result is similar
to the findings of Villanueva and Lirio (2000) on Fusarium wilt in garden pea.
According to Sullivan (2004) direct inoculation of beneficial organisms like
Trichoderma spp., Flavobacterium spp., Streptomyces sp., Gliocladium spp.,
Bacillus spp. and Pseudomonas spp. could effectively control soil-borne
pathogens.

Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

35
T1
T2
T3
T4
T5
T6
6
T7
T8
T9
a
a
a
5
a
a
a
bc
a
a
4
ab
ab
a
ab
b
b
a
a
ab
b
a
ab
b
e
v
e
r
i
t
y
b
a
b
bc
ab
b
3
a
b
c
e S
ab
b
a
cd
b
c
cd bc
c
a
b
s
e
as
c
c
c
c
c
2
a
cd
d
Di
b
b
b
1
a
e
d
d
c
d
d
0
1
2
3
4
5
6
7
Weeks

Figure 3. Effect of bacterial antagonists and plant extracts on Fusarium wilt
infection of chrysanthemum. Points with a common letter are not
significantly different at 5 % level using DMRT. Note: 1-
Uninoculated/untreated, 2-untreated/inoculated, 3-F.o.c. + Hot H2O +
after 1week add garlic extract + comb. of Pseudomonas sp. +
Flavobacterium sp 17DAT., 4- Pseudomonas sp. + Flavobacterium
sp., 5- Mancozeb (Parafungus), 6- Thiophanate methyl (Fungitox), 7-
Garlic (A. sativum L.), 8- Gawed (P.betle), 9- Table Salt (1g/L H20).


Vascular discoloration

Disease severity in terms of length of vascular discoloration was shortest
in plants applied with thiophanate methyl. However, the effect was not
significantly different from plants applied with mancozeb, combination of
Pseudomonas sp. and Flavobacterium sp., garlic extract, table salt and gawed
extract. On the other hand, application of hot water treatment followed by garlic
extract and the combination of bacterial antagonists was not effective in
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

36
controlling the disease (Table 2 and Plate 8). It is possible that the temperature of
hot water was not high enough to completely kill the pathogen. In addition, there
is possibility that the slight increase in soil temperature may have favored the
growth and multiplication of F. oxysporum f. sp. chrysanthemi.

Table 2. Effect of bacterial antagonists and plant extracts on the vascular
discoloration expressed as disease severity a

TREATMENT



DISEASE SEVERITY
Uninoculated/Untreated
0.00
c
Uninoculated/Treated
8.083
a
F.o.c. + Hot H20 + add garlic extract after a week +
comb. of Pseudomonas sp.+ Flavobacterium sp
after 17days






8.875 a
Pseudomonas sp. + Flavobacterium
sp.
2.878
bc
Mancozeb
(Parafungus
2.210
bc
Thiophanate
methyl
(Fungitox
1.335bc
Garlic
extract
(A.
Sativum
L.) 2.793
bc
Gawed
extract
(P.
Betle)
4.045
b
Table salt (1 g/L H20)
3.020
bc
CV


57.96%
a Data are means of four replications. Means with a common letter are not
significantly different at 1% level using DMRT

Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

37
a
b
c






d
e
f






g
h
i









Plate 8. Vascular discoloration as affected by bacterial antagonists, plant extracts
and fungicides. a=uninoculated –untreated, b=uninoculated-treated, c=
hot h20 treatment + garlic extract (added after 1 week) + combination of
Pseudomonas sp. + Flavobacterium sp (added 17DAT) , d= comb. of
Pseudomonas sp. + Flavobacterium sp., e=mancozeb (parafungus) f=
thiophanate methyl (fungitox), g= garlic extract (Allium sativum L.), h=
gawed extract (Piper betle), i= table salt NaCl=1g/L H20)
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

38
Fresh Weight


Plants treated with the combination of bacterial antagonists gave the
highest fresh top weight with mean of 44.50 g. However, this did not
significantly differ with the top weights of plants treated with thiophanate methyl,
mancozeb, garlic extract and table salt with means of 43.59, 42.84,42.50 and
42.88 g, respectively (Fig.4). The lowest fresh top weight was obtained from
plants treated with hot water + garlic extract introduced one week after and the
combination of the bacterial antagonists with mean of 3.91 g. The same trend was
noted in the fresh root weight: the highest was noted on plants treated with
bacterial antagonists followed closely with thiophanate methyl, garlic extract,
mancozeb, and table salt with respective means of 6.41, 5.84,5.01, 4.89 and 4.66
g. The lowest fresh root weight was again obtained in plants treated with the
integration of hot water, garlic extract and bacterial antagonists with mean of 0.89
g. Although significantly much lower than the above treatments, the fresh top and
root weights of plants applied with gawed extract was significantly higher than
those obtained in the untreated inoculated plants . According to Adams (1990)
susceptible cultivars inoculated with F. oxysporum f. sp. chrysanthemi exhibit
severe symptoms ten days after inoculation and often become severely stunted
and die.

Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

39
Fresh top wt
60
a
Fresh Root wt.
a
50
a
a
a
)

ab
40
b
(g
b
30
g
ht 20
a
a
Wei
c
cd

a
ab
bc
10
ab
ab

0
1
2
3
4
5
6
7
8
9
Treatment


Fig.4. Effect of bacterial antagonists and plant extracts on the fresh weight of
plants. Bars with a common letter are not significantly different at 5 %
level using DMRT. Note: 1-Uninoculated/untreated, 2-
untreated/inoculated, 3-F.o.c. + Hot H2O + garlic extract after 1week +
comb. of Pseudomonas sp. + Flavobacterium sp. after 17days, 4-
Pseudomonas sp. + Flavobacterium sp., 5- Mancozeb (Parafungus), 6-
Thiophanate methyl (Fungitox), 7- Garlic (A. sativum L.), 8- Gawed (P.
betle), 9- Table Salt (.5g/L).


Cutflower Yield

Plants treated with bacterial antagonists, Flavobacterium sp. and
Pseudomonas sp. produced high quality cutflowers comparable with those treated
with standard fungicides, mancozeb and thiophanate methyl (Table 3.) .
However, the cutflowers were brighter orange than the latter treatment. In
addition, the former treatment produced less non-marketable flowers. On the other
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

40
hand, the highest gross sales from cutflower produced was obtained from the
untreated uninoculated plants, followed closely by those applied with thiophanate
methyl, mancozeb, combination of bacterial antagonists, garlic extract, and table
salt with P60.00, P55.83, P 55.00, P48.34 and P40.84, respectively (Table 4).
Plants applied with the combination of hot water treatment, garlic extract and
bacterial antagonists were not able to produce marketable cutflowers.


Table 3. Effect of bacterial antagonists and plant extracts on the quality of

cutflower yielda
TREATMENT CLASSIFICATION
LONG
MEDIUM
SHORT
CB NON


MARKETABLE
Uninoculated/
0.00b 0.25bc 0.50ab 0.00b
0.00C
Untreated






Uninoculated/Treated
0.00b
0.25c
0.50b
0.00b
2.25ab
F.o.c. + Hot H20 +
0.00b 0.00c 0.00b 0.00b
3.00a
garlic extract +
comb. Of
Pseudomonas sp.+
Flavobacterium sp.
Comb. Of
0.50ab 1.00b 0.50ab 1.00
a
0.00c
Pseudomonas sp. +
Flavobacterium sp.
Mancozeb
1.00a 0.50bc 1.00a 0.50ab
0.00c
(Parafungus)
Thiophanate methyl
1.00a 0.75bc 0.75ab 0.50ab
0.00c
(Fungitox)
Garlic extract (A.
0.25ab 1.00b 0.75ab 0.50ab
0.00c
sativum L.)
Gawed extract (P.
0.00b 0.75bc 0.75ab 0.00b
1.50b
betle)
Table salt (1 g/L H20)
0.25ab 1.00b 0.75ab 0.00b
1.00b
CV


168.32% 56.53%
103.92%
63.94%

a Data are means of four replications. Means followed by a common letter are not
significantly different at 5% level using DMRT
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

41
Table 4. Effect of bacterial antagonists and plant extracts on the gross sales of

cutflowers.

TREATMENT CLASSIFICATION
/ GROSS SALE (PhP)
LONG MEDIUM SHORT CB
NON-
TOTAL
MARKETABLE
Uninoculated/Untreated 0.00 60.00
0.00
0.0 0.00
P60.00
Uninoculated /Treated
0.00
5.00
8.34
0.00
0.00
P13.84
Hot H20 treatment +
0.00 0.00
0.00 0.00 0.00
0.00
garlic extract ( 1 week
after) + comb. of
Pseudomonas ap. &
Flavobacterium sp
after 17 days
Comb. o Pseudomonas 11.66 20.00
8.34 10.00 0.00
50.00
sp. & Flavobacterium
sp.
Mancozeb (Parafungus) 23.32 10.00
16.68
5.00
0.00
55.00
Thiophanate methyl
23.32 15.00
12.51 5.00 0.00
55.83
(Fungitox)
Garlic extract (A.
5.83 20.00 12.51 10.00 0.00
48.34
sativum L)
Gawed extract (P.
0.00 15.00 12.51 10.00 0.00
37.51
betle)
Table Salt (.5g/L H20) 5.83 20.00 12.51 2.50 0.00
40.84




















Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

SUMMARY, CONCLUSION AND RECOMMENDATION

SUMMARY

The study aimed to; 1. determine the effect of selected bacterial
antagonists, plant extracts and fungicides on the growth of Fusarium oxysporum f.
sp. chrysanthemi; 2. determine the compatibility of plant extracts and fungicides
with bacterial antagonists, 3. determine the effectiveness of bacterial antagonists
and plant extracts in reducing the soil population of F. oxysporum f. sp.
chrysanthemi and 4. determine the effect of bacterial antagonists and plant
extracts on Fusarium wilt infection and yield of chrysanthemum.

Results of the in-vitro bioassay test showed that the growth of F.
oxysporum f. sp. chrysanthemi was not significantly inhibited by bacterial
antagonists Flavobacterium sp. and Pseudomonas sp. Garlic extract (Allium
sativum L) gave the widest inhibition zone comparable to the standard fungicides,
mancozeb (Parafungus) and thiophanate methyl (Fungitox).

Mancozeb and garlic extract are not compatible with the bacterial
antagonists while thiophanate methyl and gawed extract did not affect the growth
of Flavobacterium sp. and Pseudomonas sp. Combination of Flavobacterium sp.
and Pseudomonas sp. was comparable with the standard fungicide mancozeb.
Although not as effective as the other treatments, application of garlic extract,
Flavobacterium sp. and Pseudomonas sp. alone and table salt significantly
reduced the soil population of the pathogen compared to the untreated inoculated
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

43
plants. Gawed extract was inferior with the above treatments in suppressing the
pathogen population.

In the greenhouse trial, the lowest fusarium wilt infection was obtained
from plants applied with mancozeb, thiophanate methyl and garlic extract.
Although not as superior as the other treatments, the combination of
Pseudomonas sp and Flavobacterium sp. and application of table salt
significantly reduced the disease infection in chrysanthemum. On the other hand,
integration of hot water treatment, garlic extract and the combination
Pseudomonas sp. and Flavobacterium sp. did not give promising results.
Plants applied with thiophanate methyl and mancozeb gave the best
quality cutflowers comparable with the uninoculated control. However, did not
significantly differ with plants applied with Pseudomonas sp. + Flavobacterium
sp. Application of the bacterial antagonists resulted in more bright and intense
colored flowers which added to the overall quality of the cutflowers. The poorest
quality cutflowers were obtained from plants treated with hot water + garlic
extract + Pseudomonas sp. and Flavobacterium sp. and the untreated-
uninoculated plants.

CONCLUSION
Combination of bacterial antagonists, Pseudomonas sp. & Flavobacterium
sp. and garlic extract could be potential alternatives to soil fungicides in the
management of F. oxysporum f. sp. chrysanthemi in chrysanthemum.
Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

44
RECOMMENDATION

1) Combination of Pseudomonas sp. and Flavobacterium sp. and garlic
extract are recommended for managing fusarium wilt under greenhouse
conditions. However, another trial is necessary , preferably in the open field to
verify the efficacy of the above treatments.

2) For more effective control, the bacterial antagonists should be applied
weekly to give sustainable control of the disease; and

3) For more convenient application, a more rapid and easy technique of
extracting garlic juice should be studied.









Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

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Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in
Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

46
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Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

47
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Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in

Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and
Plant Extracts / Catherine A. Bagsan. 2006

---

APPENDICES


APPENDIX TABLE 1. Inhibition zone (mm) produced by biocontrol agents, plant
extracts and fungicides against Fusarium oxysporum f. sp.
chrysanthemi after 5 days (actual)

REPLICATION

TREATMENT

I II III Total Mean
A. Biocontrol Agents
Sterile Distilled Water (SDW)
0.0 0.0 0.0 0.0
0.0
Isolate 31 (Bacillus sp.)
0.0 0.0 0.0 0.0 0.0

Isolate 131( Bacillus pumilus)
0.0 0.0 0.0 0.0 0.0
Isolate 73 (Bacillus pumilus)
0.0 0.0 0.0 0.0 0.0
Isolate 94 (Flavobacterium sp.) 1.0 0.5 0.5 2.0 0.66
Isolate 158 (Pseudomonas sp.) 1.0 1.0 1.0 3.0 1.0

Verticillium sp.

0.0 0.0 0.0 0.0 0.0
B. Plant Extracts







Garlic (Allium sativum L.) 11.3 9.2 8.43 28.96 9.65
Gawed (Piper betle)
4.0 0.0 2.35 6.35 2.11
Kutsai (Allium schoenoprasum L) 2.5 1.25 0. 0 3.75 1.25
Hot Pepper (Capsicum frutescens)3.0 0.0 2 .0 5.0 1.66
Red Onion (Allium cepa) 0.0 0.0 0.0 0.0 0.0
C. Fungicides
Thiophanate Methyl (Fungitox) 8.25 7.91 6.5 22.66 7.55
Thiophanate Methyl (Topsin–M) 7.33 4.66 3.05 15.04 5.01
Mancozeb (Parafungus) 13.08 9.91 10.05 33.04 11.01
Thiodazole copper (BLB Stopper)0.0 0.0 0.0 0.0 0.0
Captan (Captan)
0.0 5.0 3.75 14.75 4.91
Benomyl (Benlate)
11.16 7.16 8.03 26.35 8.78
Chlorothalonil (Daconil ) 0.0 0.0 0.0 0.0 0.0
Mancozeb (Dithane M-45) 0.0 0.0 0.0 0.0 0.0
D. Others
Table Salt (NaCl= .5g /LH20)
0.0 0.0 0.0 0.0 0.0
GRAND TOTAL





160.9

GRAND MEAN







2.44

---

49
APPENDIX TABLE 2. Inhibition zone (mm) produced by biocontrol
agents, plant extracts and fungicides against Fusarium
after 5 days (transformed)

REPLICATION

TREATMENT

I II III Total Mean
A. Biocontrol Agents
SDW


0.71 0.71 0.71 2.13 0.71
Isolate 31 (Bacillus sp.) 0.71 0.71 0.71 2.13 0.71
Isolate 131 (Bacillus pumilus) 0.71 0.71 0.71 2.13 0.71
Isolate 73 (Bacillus pumilus.) 0.71 0.71 0.71 2.13 0.71
Isolate 94 (Flavobacterium sp.) 1.22 1.00 1.00 3.22 1.07
Isolate 158 (Pseudomonas sp.) 1.22 1.22 1.00 3.44 1.15
Verticillium
0.71 0.71 0.71 2.13 0.71
B.
Plant
Extracts

Garlic (Allium sativum L.) 3.44 3.11 2.99 9.54 3.18
Gawed (Piper betle)
2.12 0.71 1.69 4.52 1.51
Kutsai (Allium schoenoprasum L.)1.73 1.32 0.71 3.76 1.25
Hot Pepper(Capsicum frutescens) 1.87 0.71 1.58 4.16 1.39
Red Onion (Allium cepa) 0.71 0.71 0.71 2.13 0.71
C. Fungicides
Thiophanate Methyl (Fungitox) 2.55 2.35 2.06 6.96 2.32
Thiophanate Methyl(Topsin–M) 2.80 2.27 3.05 5.32 1.77
Mancozeb (Parafungus) 3.69 3.23 3.25 10.17 3.39
Thiodazole Copper(BLB Stopper)0.71 0.71 0.71 2.13 0.71
Captan (Captan) 2.96 2.90 2.65 5.61 1.87
Benomyl (Benlate) 3.41 2.77 2.92 9.10 3.03
Chlorothalonil (Daconil) 0.71 0.71 0.71 2.13 0.71
Mancozeb (Dithane M-45) 0.71 0.71 0.71 2.13 0.71
D. Others
Table Salt (.5g/L H20)
0.71 0.71 0.71 2.13 0.71

ANOVA TABLE
Source of Degree of
Sum of
Mean of
Computed Prob.
Variation Freedom
Square Squares
F

Model
20
813.82 40.69 34.57
0.0001
Treatments
20
813.82 40.69 34.57
0.0001
Error
42
49.43 1.18
Total
62
863.25
* - Significant at 5% level using DMRT


CV = 42.48%

---

50
APPENDIX TABLE 3.
Inhibition
zone
(mm)
produced
by
plant
extracts and fungicides against bacterial antagonists;
Pseudomonas sp. and Flavobacterium sp. after 24
hours (actual)

REPLICATION

TREATMENT

I II III Total Mean
Flavobacterium sp. + SDW 0.0
0.0 0.0
0.0 0.0
Flavobacterium sp. +
Mancozeb (Parafungus) 5.17 5.0 4.0
14.17 4.72
Flavobacterium sp. +
Thiophanate Methyl (Fungitox)
0.0
0.0
0.0
0.0 0.0
Flavobacterium sp. +
Garlic (Allium sativum L.) 11.75 6.5 8.75
27 9.0
Flavobacterium sp. +
Gawed (Piper betle) 0.0
0.0 0.0
0.0 0.0
Flavobacterium sp. +
.5 g Table salt/li H20 0.0
0.0 0.0
0.0 0.0
Pseudomonas sp. + SDW 0.0
0.0 0.0
0.0 0.0
Pseudomonas sp +
Mancozeb (Parafungus) 4.5
1.0 5.0
10.5 3.5
Pseudomonas sp. +
Thiophanate Methyl (Fungitox) 0.0
0.0 0.0
0.0
0.0
Pseudomonas sp. +
Garlic (Allium sativum L.) 4.25
3.0 4.75 12.0
4.0
Pseudomonas sp. +
Gawed (Piper betle) 0.0 0.0 0.0
0.0 0.0
Pseudomonas sp. +
Table salt ( .5g/L H20) 0.0
0.0 0.0 0.0 0.0
GRAND TOTAL





63.67

GRAND MEAN







1.93

---

51
APPENDIX TABLE 4. Inhibition zone (mm) produced by plant


extracts and fungicides against bacterial
antagonists; Pseudomonas sp. and Flavobacterium sp.
after 24 hours (transformed)

REPLICATION

TREATMENT

I II III Total Mean
Flavobacterium sp. + SDW 0.71 0.71 0.71 2.13
0.71
Flavobacterium sp. +
Mancozeb (Parafungus) 2.83 2.34 2.12
7.29
2.43
Flavobacterium sp. + Thiophanate Methyl
(Fungitox )
0.71 0.71 0.71 2.13
0.71
Flavobacterium sp. +
Garlic (Allium sativum L.) 3.50 2.64 3.04 9.18 3.06
Flavobacterium sp. +
Gawed (Piper betle) 0.71 0.71 0.71 2.13 0.71
Flavobacterium sp. +
.5 g Table salt/li H20 0.71 0.71 0.71 2.13 0.71
Pseudomonas sp. + SDW 0.71 0.71 0.71 2.13
0.71
Pseudomonas sp +
Mancozeb (Parafungus) 2.24 1.22 2.35
5.81
1.94
Pseudomonas sp. + Thiophanate Methyl
(Fungitox)



0.71 0.71 0.71 2.13 0.71
Pseudomonas sp. +
Garlic (Allium sativum L.) 1.18 1.87 2.29
5.34 1.78
Pseudomonas sp. +
Gawed (Piper betle) 0.71 0.71 0.71 2.13
0.71
Pseudomonas sp. +
.5 g Table salt/li H20
0.71 0.71 0.71 2.13
0.71
GRAND TOTAL





63.67

GRAND
MEAN



1.93

ANOVA TABLE
Source of
Degree of Sum of
Mean of
Computed
Prob.

Variation
Freedom
Square Squares
F

Treatment
12
290.73
24.23
24.42
0.0001
Error
26
25.80
0.99
Total
38
316.53
** - highly significant at 1% CV = 61.01%

---

52
APPENDIX TABLE 5. Effect of bacterial antagonists and plant extracts on soil



Population count of F. oxysporum f. sp. chrysanthemi ( 0 day



(before treatment) (actual)

REPLICATION

TREATMENT

I II III Total Mean

Uninoculated-Untreated
0.0
0.0 0.0
0.0 0.0
Uninoculated-Untreated
200000 600000 500000 1300000 433333.3
F.o.c. + Hot H20
67000 300000 100000 467000 155666.7
F. o. c. + Hot H20, after 1
week add garlic, after 10
days add Pseudomonas sp
+ Flavobacterium sp.
200000 100000 100000 400000 133333.3

F. o.c + Flavobacterium sp. 100000 100000 200000 400000 133333.3

F.o.c + Pseudomonas sp. 600000 100000 300000 1000000 333333.3

F.o.c. + Flavobacterium sp. 200000 67000 200000 467000 155666.7
& Pseudomonas sp.

Mancozeb (Parafungus)
200000 100000 200000 500000 166666.7

Garlic (Allium sativum L.) 200000 300000 100000 600000 200000.0
Gawed (Piper betle) 233000 400000 667000 1300000 433333.3

Table salt (.5 g/L H20)
200000 100000 900000 1200000 400000
GRAND TOTAL





7634000
GRAND MEAN






231333.3

---

53
APPENDIX TABLE 6. Effect of bacterial antagonists and plant extracts on the soil
Population count of F. oxysporum f. sp. chrysanthemi (0 day
before treatment) (transformed)

REPLICATION

TREATMENT

I II III Total Mean






Uninoculated-Untreated
0.0
0.0
0.0
0.0
0.0
Uninoculated-Treated
2.0x105
6.0x105 5.0x105 13.0x105
4.3x105
F.o.c. + Hot H20





Treatment
6.7x104
3.0x105 1.0x105 4.6x105
1.5x105
F. o. c. + Hot H20, after





1 week add garlic, after





10 days add





Pseudomonas sp.&





Flavobacterium sp.
2.0x105
1.0x105 1.0x105 4.0x105
1.3x105
F. o.c + Flavobacterium





sp.
1.0x105
1.0x105 2.0x105 4.0x105
1.3x105
F.o.c + Pseudomonas sp.
6.0x105
1.0x105 3.0x105 10.0x105
3.3x105
F.o.c. + Flavobacterium





Sp. & Pseudomonas sp.
2.0x105
6.7x104 2.0x105 4.6x105
1.5x105
Mancozeb (Parafungus)

2.0x105 1.0x105 2.0x105 5.0x105
1.6x105
Garlic (Allium sativum L.)
2.0x105 3.0x105 1.0x105 6.0x105
2.0x105
Gawed (Piper betle)
2.3x105
4.0x105 6.6x105 13.0x105
4.3x105
Table salt (NaCl = .5g/L)
2.0x105 1.0x105 9.0x105 12.0x105
4.0x105
GRAND
TOTAL
76.3X105


GRAND
MEAN
2.3X105



ANOVA
TABLE

Source of
Degree of
Sum of
Mean of


Variation Freedom Squares Squares
F
Value
Probability

Treatment
10
77.48
7.74
100.63
0.0001
Error
22
1.69
0.08


Total
32
76.89



** - highly significant at 1% level using DMRT CV= 5.77%

---

54
APPENDIX TABLE 7. Effect of bacterial antagonists and plant extracts on the soil



population count of F. oxysporum f. sp. chrysanthemi 1 day



after treatment (actual)

REPLICATION

TREATMENT
I II III Total Mean

Uninoculated-Untreated

0.0
0.0 0.0
0.0 0.0

Uninoculated-Treated 300000 500000 600000 1400000 466666.7

F.o.c. + Hot H20 100000 200000 100000 400000 133333.3

F. o. c. + Hot H20, after 1
week add garlic, after
10 days add
Pseudomonas sp
+Flavobacterium sp. 200000 100000 133000 433000 144333.3

F. o.c +Flavobacterium sp. 100000 100000 167000 367000 122333.3

F.o.c + Pseodomonas sp. 500000 100000 200000 800000 2666666.7

F.o.c. + Flavobacterium sp. 100000 100000 200000 400000 133333.3
& Pseudomonas sp.

Mancozeb (Parafungus ) 100000 100000 167000 367000 122333.3

Garlic (Allium sativum L.) 200000 200000 200000 600000 200000.0

Gawed (Piper betle) 200000 300000 600000 1100000 366666.7

Table salt (.5g/L H20) 300000 100000 667000 1067000 355666.7
GRAND TOTAL





6934000


GRAND MEAN





210121.2

---

55
APPENDIX TABLE 8. Effect of bacterial antagonists and plant extracts on the soil



Population count of F. oxysporum f. sp. chrysanthemi 1 day



after treatment (transformed)

REPLICATION

TREATMENT
I II III Total Mean

Uninoculated-Untreated 0.0
0.0
0.0
0.0
0.0
Uninoculated-Treated 3.0x105
5.0x105 6.0x105
4.0x105
4.6x105
F.o.c. + Hot H20





Treatment
1.0x105
2.0x105
1.0x105
4.0x105
1.3x105
F. o. c. + Hot H20,





after 1 week add





garlic, after 10 days





add Pseudomonas sp.&





Flavobacterium sp.
2.0x105
1.0x105
1.3x105
4.3x105
1.4x105
F. o.c + Flavobacterium





sp.
1.0x105
1.0x105
1.6x105
3.6x105
1.2x105
F.o.c + Pseudomonas





sp.
5.0x105
1.0x105
2.0x105
8.0x105
2.6x105
F.o.c. +





Flavobacterium sp.





& Pseudomonas sp.
1.0x105
1.0x105
2.0x105
4.0x105
1.3x105
Mancozeb (Parafungus)







1.0x105
1.0x105
1.6x105
3.6x105
1.2x105
Garlic (Allium sativum





L.)
2.0x105
2.0x105
2.0x105
6.0x105
2.0x105
Gawed (Mancozeb)





2.0x105
3.0x105
6.0x105
11.0x105
3.6x105
Table salt (NaCl





=.5g/L)
3.0x105
1.0x105
6.6x105
10.6x105
3.5x105






GRAND TOTAL
69.3X105







GRAND MEAN
2.1 X105







ANOVA
TABLE

Source of
Degree of
Sum of
Mean of

Probability
Variation Freedom
Squares Squares F
Value

Treatment
10 76.05 7.60 197.74 0.0001
Error 22 0.85
0.04

Total
32
76.89



** - highly significant at 1% level using DMRT CV= 4.13%

---

56
APPENDIX TABLE 9. Effect of bacterial antagonists and plant extracts on the soil



population count of F. oxysporum f. sp. chrysanthemi 3 days



after treatment (actual)


REPLICATION
TREATMENT
I
II
III
TOTAL
MEAN

Uninoculated-Untreated
0.0 0.0
0.0 0.0
0.0
Uninoculated-Treated 400000 600000 500000 1500000 500000.0
F.o.c. + Hot H20
67000 167000 100000 334000 111333.3

F. o. c. + Hot H20,
after 1 week add garlic,
after 10 days add
Pseudomonas sp +
Flavobacterium sp. 100000 200000 100000 400000 133333.3

F. o.c + Flavobacterium
sp. 100000 100000 133000 333000 111000.0

F.o.c + Pseudomonas
sp. 400000 100000 200000 700000 233333.3

F.o.c. + Flavobacterium
sp. & Pseudomonas sp. 67000 100000 200000 367000 122333.3

Mancozeb (Parafungus) 67000 67000 100000 234000 78000.0

Garlic (Allium sativum L.) 200000 100000 200000 500000 166666.7

Gawed (Piper betle) 200000 200000 600000 1000000 333333.3

Table salt (.5g/L H20) 200000 200000 600000 1000000 333333.3
GRAND TOTAL




6368000
GRAND MEAN





192969.7

---

57
APPENDIX TABLE 10. Effect of bacterial antagonists and plant extracts on the soil
population count of F. oxysporum f. sp. chrysanthemi 3 days
after treatment (Transformed)


REPLICATION
TREATMENT
I
II
II
TOTAL
MEAN

Uninoculated-Untreated 0.0 0.0 0.0 0.0 0.0
Uninoculated-Treated
4.0x 105 6.0
x105
5.0 x 105 15.0 x105 5.0 x 105
F.o.c. + Hot H20
Treatment
6.7 x 104
1.6 x 105
1.0 x 105 3.3 x 105 1.1x105
F. o. c. + Hot H20, after 1
week add garlic, after
10 days add
Pseudomonas sp.&
Flavobacterium sp.
1.0x 105
2.0 x 105
1.0 x 105 4.0 x 105
1.3 x 105
F.o.c + Flavobacterium
sp.
1.0 x 105
1.0 x 105
1.3 x 105 3.33 x 105 1.1x
105
F.o.c + Pseudomonas sp. 4.0x105
1.0 x 105
2.0x 105 7.0 x 105 2.3x
105
F.o.c. + Flavobacterium
sp. & Pseudomonas sp. 6.7x
104
1.0 x 105
2.0x 105 3.6 x 105 1.2x
105
Mancozeb (Parafungus)
6.7x 104
6.7 x 104
1.0 x 105 2.3 x 104 7.8x
104
Garlic (Allium sativum L.)
2.0x 105
1.0 x 105
2.0x 105 5.0 x 105
1.6 x 105
Gawed (Piper betle)
2.0 x 105 2.0x
105 6.0x105
10.0 x 105
3.3 x 105
Table salt (NaCl=5g/L)
2.0x 105 2.0x
105 6.0x105
10.0x 105
3.3 x 105
GRAND TOTAL 63.68 X 105
GRAND MEAN 2.10 X 105

ANOVA TABLE

Source of
Degree of
Sum of
Mean of

Variation
Freedom
Squares
Squares
F Value
Probability
Treatment
10
87.62
8.76
164.12
0.0001
Error
22
1.17
0.05


Total 32
88.79

** - highly significant at 1% level using DMRT CV= 4.59%

---

58
APPENDIX TABLE 11. Effect of bacterial antagonists and plant extracts on the soil



population count of F. oxysporum f. sp. chrysanthemi 7 days



after treatment (actual)


REPLICATION
TREATMENT
I
II
III
TOTAL
MEAN

Uninoculated-Untreated
0.0
0.0
0.0

0.0

0.0
Uninoculated-Treated 800000 1633000 933000 3366000 1122000.0
F.o.c. + Hot H20
1100000 833000 1933000 3866000 1288667.0

F. o. c. + Hot H20,
after 1 week add garlic,
after 10 days add
Pseudomonas sp +
Flavobacterium sp. 500000 267000 100000 867000 289000.0

F. o.c + Flavobacterium
sp. 200000 600000 967000 1767000 589000.0

F.o.c + Pseodomonas
sp. 400000 200000 267000 867000 289000.0

F.o.c. + Flavobacterium
sp. & Pseudomonas sp. 167000 133000 133000 433000 144333.3

Mancozeb (Parafungus 67000 33000 100000 200000 66666.7

Garlic (Allium sativum L.) 600000 300000 900000 1800000 600000.0

Gawed (Piper betle) 200000 300000 600000 1100000 366666.7

Table salt (.5g/L H20) 300000 200000 700000 1200000 400000.0

GRAND TOTAL




15466000
GRAND MEAN





468666.7

---

59
APPENDIX TABLE 12. Effect of bacterial antagonists and plant extracts on the soil



population count of F. oxysporum f. sp. chrysanthemi 7 days



after treatment (Transformed)


REPLICATION
TREATMENT
I
II
III
TOTAL MEAN
Uninoculated-Untreated
0.0
0.0
0.0 0.0 0.0
Uninoculated-Treated 8.0x 105 16.33x105 9.33x 105 33.66x 105 11.22x 105

F.o.c. + Hot H20
11.0x 104 8.33x 105 19.3x105 38.66x 103 12.88x 105

F. o. c. + Hot H20, after
1 week add garlic,
after 10 days add
Pseudomonas sp +
Flavobacterium sp. 5.0x 105 2.67x 105 1.0x 105 8.67x 105 58.9x 105


F. o.c + Flavobacterium
sp. 2.0x 105 6.0x 105 9.67x 105 17.6x105 5.89x 105

F.o.c + Pseudomonas sp. 4.0x 105 2.0x 105 2.67x 105 8.67x105 2.89x 105

F.o.c. + Flavobacterium
sp. & Pseudomonas sp. 1.67x 105 1.33x 105 1.33x 105 4.33x 105 1.44x 105

Mancozeb (Parafungus) 6.7x 104 3.3x 104 1.0x 105 2.0x105 6.66x 104

Garlic (Allium sativum L.) 6.0x 105 3.0x 105 9.0x 105 18.0x 105 6.0x 105

Gawed (Piper betle) 2.0x 104 3.0x 105 6.0x 105 11.0x 105 3.66x 105


Table salt (.5g/L H20) 3.0x 105 2.0x 105 6.0x 105 12.0x 105 4.0x 105
GRAND TOTAL





154.66 x 105
GRAND MEAN







46.86x105

ANOVA TABLE

Source of
Degree of
Sum of
Mean of

Variation
Freedom
Squares
Squares
F Value
Probability
Treatment
10
87.62
8.76
164.12
0.0001
Error
22
1.17
0.05


Total 32
88.79

** - highly significant at 1% level using DMRT CV= 4.59%

---

60
APPENDIX TABLE 13 . Effect of bacterial antagonists and plant extracts on the soil



population count of F. oxysporum f. sp. chrysanthemi 14 days
after treatment ((actual)

REPLICATION
TREATMENT
I
II
III
TOTAL MEAN

Uninoculated-Untreated
0.0 0.0
0.0 0.0 0.0
Uninoculated-Treated
600000 800000 500000 1900000 633333.3
F.o.c. + Hot H20
400000 200000 167000 767000 255666.7

F. o. c. + Hot H20, after 1
week add garlic, after 10
days add Pseudomonas
sp +Flavobacterium sp. 100000 200000 167000 467000 155666.7

F. o.c + Flavobacterium sp. 133000 100000 300000 533000 177666.7

F.o.c + Pseudomonas sp. 267000 133000 100000 500000 166666.7

F.o.c. + Flavobacterium sp. 133000 67000 100000 300000 100000.0
& Pseudomonas sp.

Mancozeb (Parafungus)
67000 33000 33000 133000 44333.30

Garlic (Allium sativum L.) 300000 200000 100000 600000 200000.0

Gawed (Piper betle) 900000 200000 333000 1433000 477666.7

Table salt (.5g/L H20) 100000 200000 600000 900000 300000.0
Grand Total





7533000
Grand Mean






228272.7


APPENDIX TABLE 14. Effect of bacterial antagonists and plant extracts on the soil



population count of. F. oxysporum f. sp. chrysanthemi 14 days
after treatment (transformed)

REPLICATION

TREATMENT

I II III Total Mean






Uninoculated-Untreated
0.0
0.0
0.0
0.0
0.0

---

61
Uninoculated-Treated
6.0x105
8.0x105 5.0x105 19.0x105
6.3x105
F.o.c. + Hot H20





Treatment
4.0x104
2.0x105 1.6x105 7.6x105
2.5x105
F. o. c. + Hot H20, after





1 week add garlic, after





10 days add





Pseudomonas sp.&





Flavobacterium sp.
1.0x105
2.0x105 1.6x105 4.6x105
1.5x105
F. o.c + Flavobacterium





sp.
1.3x105
1.0x105 3.0x105 5.3x105
1.7x105
F.o.c + Pseudomonas sp.
2.6x105
1.3x105 1.0x105 5.0x105
1.6x105
F.o.c. + Flavobacterium





Sp. & Pseudomonas sp.
1.3x105
6.7x104 1.0x105 3.0x105
1.0x105
Mancozeb (Parafungus)

6.7x104 3.3x104 3.3x104 1.3x105
4.4x104
Garlic (Allium sativum L.)
3.0x105 2.0x105 1.0x105 6.0x105
2.0x105
Gawed (Piper betle)
9.0x105
2.0x105 3.3x105 14.3x105
4.7x105
Table salt (NaCl =.5g/L)
1.0x105 2.0x105 6.0x105 9.0x105
3.0x105
GRAND
TOTAL
75.3X105


GRAND
MEAN
2.2X105


ANOVA TABLE

Source of
Degree of
Sum of
Mean of


Variation Freedom Squares Squares
F
Value
Probability

Treatment
10
78.05
7.8
153.11
0.0001
Error
22
1.12
0.05


Total
32
79.17



** - highly significant at 1% level using DMRT CV= 4.73
APPENDIX TABLE 15. Effect of bacterial antagonists and plant extracts on the soil
Population count of. F. oxysporum f. sp. chrysanthemi 21
days after treatment (actual)

REPLICATION

TREATMENT

I II III Total Mean

Uninoculated-Untreated 0.0 0.0 0.0 0.0

---

62
Uninoculated-Treated 400000 300000 300000 1000000 333333.3
F.o.c. + Hot H20
300000 100000 100000 500000 166666.7
F. o. c. + Hot H20, after 1
week add garlic, after 10
days add Pseudomonas sp
+ Flavobacterium sp.
33000 67000 100000 200000 66666.7

F. o.c + Flavobacterium sp. 100000 100000 100000 300000 100000.0

F.o.c + Pseodomonas sp. 267000 300000 67000 634000 211333.3

F.o.c. + Flavobacterium sp. 100000 67000 67000 234000 78000.0
& Pseudomonas sp.

Mancozeb (Parafungus)
33000 33000 33000 99000 33000.0

Garlic (Allium sativum L.) 67000 33000 33000 133000 44333.3

Gawed (Piper betle) 300000 167000 100000 567000 189000.0

.5g Table salt/li H20 100000 100000 300000 500000 166666.7
GRAND TOTAL





4167000
GRAND MEAN






126272.7







APPENDIX TABLE 16. Effect of bacterial antagonists and plant extracts on the soil



population count of. F. oxysporum f. sp. chrysanthemi 21 days



after treatment (transformed)


REPLICATION

TREATMENT

I II III Total Mean






Uninoculated-Untreated
0.0
0.0
0.0
0.0
0.0
Uninoculated-Treated
4.0x105
3.0x105 3.0x105 10.0x105
3.3x105

---

63
F.o.c. + Hot H20





Treatment
3.0x105
1.0x105 1.0x105 5.0x105
1.6x105
F. o. c. + Hot H20, after





1 week add garlic, after





10 days add





Pseudomonas sp.&





Flavobacterium sp.
3.3x104
6.7x104 1.0x105 2.0x105
6.6x104
F. o.c + Flavobacterium





sp.
1.0x105
1.0x105 1.0x105 3.0x105
1.0x105
F.o.c + Pseudomonas sp.
2.6x105
3.0x105 6.7x104 2.3x105
2.1x105
F.o.c. + Flavobacterium





Sp. & Pseudomonas sp.
1.0x105
6.7x104 6.7x104 2.3x105
7.8x104
Mancozeb (Parafungus)

3.3x104 3.3x104 3.3x104 9.9x104
3.3x104
Garlic (Allium sativum L.)
6.7x104 3.3x104 3.3x104 1.3x105
4.4x104
Gawed (Piper betle)
3.0x105
1.6x105 1.0x105 5.6x105
1.8x105
Table salt (NaCl =.5g/L)
1.0x105 1.0x105 3.0x105 5.0x105
1.6x105
GRAND
TOTAL
41.6X105


GRAND
MEAN
1.2X105




ANOVA TABLE

Source of
Degree of
Sum of
Mean of


Variation Freedom Squares Squares
F
Value
Probability

Treatment
10
71.11
7.11
175.41
0.0001
Error
22
0.89
0.04


Total
32
72.00



** - highly significant at 1% level using DMRT CV= 4.41%



APPENDIX TABLE 17. Effect of bacterial antagonists and plant extracts on the soil
population count of. F. oxysporum f. sp. chrysanthemi 28 days
after treatment (actual)

REPLICATION

TREATMENT

I II III Total Mean

Uninoculated-Untreated
0.0 0.0 0.0 0.0 0.0
Uninoculated-Treated 100000 67000 133000 300000 100000.0

---

64
F.o.c. + Hot H20
33000 33000 33000 99000 33000.0
F. o. c. + Hot H20, after 1
week add garlic, after 10
days add Pseudomonas sp
+ Flavobacterium sp.
33000 33000 0 66000 22000.0

F. o.c + Flavobacterium sp. 33000 33000 33000 99000 33000.0

F.o.c + Pseodomonas sp. 167000 33000 33000 233000 77666.7

F.o.c. + Flavobacterium sp. 33000 33000 33000 99000 33000.0
& Pseudomonas sp.

Mancozeb (Parafungus)
33000 0 0 33000 11000.0

Garlic (Allium sativum L.) 67000 0 0 67000 22333.3

Gawed (Piper betle) 33000 33000 0 66000 22000.0

Table salt (.5g/L H20 33000 0 100000 133000 44333.3
GRAND TOTAL





1195000
GRAND MEAN






36212.12











APPENDIX TABLE 18. Effect of bacteria l antagonists and plant extracts on the soil



population count of F. oxysporum f. sp. chrysanthemi 28
days after treatment (transformed)

REPLICATION

TREATMENT

I II III Total Mean






Uninoculated-Untreated
0.0
0.0
0.0
0.0
0.0
Uninoculated-Treated
1.0x105
6.7x104 1.3x105 3.0x105
1.0x105
F.o.c. + Hot H20

---

65
Treatment





3.3x104
3.3x104 3.3x104 9.9x104
3.3.x104
F. o. c. + Hot H20, after





1 week add garlic, after





10 days add





Pseudomonas sp.&





Flavobacterium sp.
3.3x104
3.3x104 0.0
6.6x104
2.2x104
F. o.c + Flavobacterium





sp.
3.3x104
3.3x104 3.3x104 9.9x104
3.3x104
F.o.c + Pseudomonas sp.
1.6x105
3.3x104 3.3x104 2.3x105
7.7x104
F.o.c. + Flavobacterium





Sp. & Pseudomonas sp.
3.3x104
3.3x104 3.3x104 9.9x104
3.3x104
Mancozeb (Parafungus)

3.3x104
0.0
0.0
3.3x104
1.1x104
Garlic (Allium sativum L.)
6.7x104
0.0
0.0
6.7x104
2.2x104
Gawed (Piper betle)
3.3x104
3.3x104
0.0
6.6x104
228x104
Table salt (NaCl =.5g/L)
3.3x104
0.0 1.0x105 1.3x105
4.4x104
GRAND
TOTAL
11.9X105


GRAND
MEAN
3.6X104




ANOVA TABLE

Source of
Degree of
Sum of
Mean of


Variation Freedom Squares Squares
F
Value
Probability

Treatment
10
79.53
7.95
2.43
0.0396
Error
22
71.96
3.29


Total
32
151.49



* - significant at 5% level using DMRT CV= 55.87%

---

66


APPENDIX TABLE 19. Effect of bacterial antagonists and plant extracts on
Fusarium wilt infection of chrysanthemum (1st week)

REPLICATION
TREATMENT
I II III I
TO
V TAL MEAN
Uninoculated – Untreated
1.00
1.00
1.00
1.00
4.00
1.00







Uninoculated – Treated
2.33
2.00
2.33
2.00
8.66
2.17







F.o.c + Hot H20 + garlic
extract (1 week after) +
comb. of Pseudomonas
sp. and Flavobacterium
sp. after 17 days
1.67
1.67
1.33
1.33
6.00
1.50







Comb. of Flavobacterium
sp. and Pseudomonas sp.
2.00
2.00
1.67
2.00
7.67
1.92







Mancozeb (Parafungus)
1.67
1.33
1.33
1.33
5.66
1.42







Thiopanate methyl
(Fungitox)
1.33
1.67
1.00
1.00
5.00
1.25







Garlic (Allium sativum L.)
2.33
2.00
1.67
1.67
7.67
1.92







Gawed (Piperbetle)
2.33
2.00
2.33
2.33
8.99
2.25







Table salt (1g/l H20)
1.67
2.00
1.67
2.33
7.67
1.92







GRAND
TOTAL 66.32

GRAND
MEAN
1.51

ANOVA

Source of
Degree of
Sum of
Mean of

Variation
Freedom
Squares
Squares
F Value Probability






Model 35
21.85
0.62
3.06
0.0001
Treatment 8
17.69
2.21
14.33
0.0001
Rep(TRT) 27
4.17
0.15
0.76
0.7875
Error 72
14.67
0.2


TOTAL 107
36.52









** Highly significant at 1% level using DMRT CV= 26.49%

---

67
APPENDIX TABLE 20. Effect of bacterial antagonists and plant extracts on
the fusarium wilt infection of chrysanthemum (2nd week)


REPLICATION
TREATMENT
I
II
III
IV
TOTAL
MEAN







Uninoculated – Untreated
1.00
1.00
1.00
1.00
4.00





1.00
Uninoculated – Treated
3.33
3.00
3.00
3.00
12.33





3.082
F.o.c + Hot H20 + garlic
extract (1 week after) +
comb. of Pseudomonas
sp. and Flavobacterium
sp. after 17 days
2.67
3.00
3.00
3.00
11.67





2.92
Comb. of Flavobacterium
sp. and Pseudomonas sp.
2.00
1.67
2.00
2.00
7.67





2.33
Mancozeb (Parafungus)
1.67
2.33
1.33
1.67
7.00





1.92
Thiopanate methyl
(Fungitox)
2.00
2.33
1.67
2.67
8.67





1.75
Garlic (Allium sativum L.)
2.67
2.67
3.33
1.67
10.34





2.17
Gawed (Piperbetle)
2.33
2.00
2.33
2.33
8.99





2.59
Table salt (1g/l H20)
2.00
2.00
2.00
2.67
8.67





2.25
GRAND TOTAL
2.17
GRAND MEAN




2.23








ANOVA

Source of
Degree of
Sum of
Mean of

Variation
Freedom
Squares
Squares
F Value Probability
Model 35
48.55
1.39
6.81
0.0001
Treatment
8
39.96 4.99
15.71 0.0001
Rep(TRT)
27
8.58 0.32 1.56 0.0696
Error 72
14.67
0.2


TOTAL
107 63.21

** Highly significant at 1% level using DMRT CV= 20.22%

---

68


APPENDIX TABLE 21. Effect of bacterial antagonist and plant extracts
on fusarium wilt infection of chrysanthemum (3rd week)


REPLICATION
TREATMENT
I
II
III
IV
TOTAL
MEAN







Uninoculated – Untreated
1.00
1.00
1.00
1.00
4.00
1.00







Uninoculated – Treated
3.67
4.00
3.67
3.67
15.01
3.7525







F.o.c + Hot H20 + garlic
extract (1 week after) +
comb. of Pseudomonas sp.
and Flavobacterium sp.
after 17 days
3.67
3.33
4.00
4.00
15.00
3.75







Comb. of Flavobacterium
sp. and Pseudomonas sp.
3.00
3.33
4.00
2.33
12.66
3.165







Mancozeb (Parafungus)
2.00
2.67
2.00
2.00
8.67
2.1675







Thiopanate methyl
(Fungitox)
2.00
2.33
2.00
2.00
8.33
2.0825







Garlic (Allium sativum L.)
2.00
3.33
2.67
3.33
11.33
2.8325







Gawed (Piperbetle)
3.00
3.33
3.67
3.67
13.67
3.4175







Table salt (1g/l H20)
3.00
3.00
2.00
3.33
11.33
2.8325







GRAND TOTAL
18.67 21.32 20.34 20.66 80.99 20.2475
GRAND MEAN







ANOVA

Source of
Degree of
Sum of
Mean of

Probability
Variation Freedom Squares Squares F
Value









Model 35
92 2.63 7.1 0.0001

Treatment 8
77.67
9.71
18.29 0.0001

Rep
(TRT) 27
14.33 0.53 1.43 0.1153

Error 72
26.67
0.37


TOTAL 107
118.67



** Highly significant at 1% level using DMRT CV= 21.91%

---

69
APPENDIX TABLE 22. Effect of bacterial antagonist and plant extracts on
fusarium wilt infection of chrysanthemum (4th week)


REPLICATION
TREATMENT
I
II
III
IV
TOTAL
MEAN







Uninoculated – Untreated
1.00
1.00


1.00 1.00 4.00 1.00
Uninoculated – Treated
4.00
4.67


4.00
4.33
17.00
4.25
F.o.c + Hot H20 + garlic
extract (1 week after) +
comb. of Pseudomonas
sp. and Flavobacterium
sp. after 17 days
3.67
4.00
4.33
4.00
16.00





4.00
Comb. of Flavobacterium
sp. and Pseudomonas sp.
3.33
3.33
4.33
2.33
13.32
3.33







Mancozeb (Parafungus)
2.33
2.67
2.00
2.00
9.00
2.25







Thiopanate methyl
(Fungitox)
2.00
2.33
2.00
2.00
8.66
2.17







Garlic (Allium sativum L.)
2.33
4.00
2.67
4.00
12.67
3.17







Gawed (Piperbetle)
2.00
3.67
4.00
3.67
14.67
3.67







Table salt (1g/l H20)
3.33
3.33
2.00
4.33
12.99
3.25







GRAND
TOTAL

3.33

GRAND
MEAN


3.01

ANOVA

Source of
Degree of
Sum of
Mean of

Variation Freedom Squares Squares F
Value Probability
Model 35
128.32
3.67
6.49
0.0001
Treatment 8 101.57
12.7 12.82 0.0001
Rep(TRT) 27 26.75
0.99 1.75 0.031
Error 72
40.67
0.56

TOTAL 107
168.99



** Highly significant at 1% level using DMRT CV= 24.97%

---

70
APPENDIX TABLE 23. Effect of bacterial antagonist and plant extracts on
fusarium wilt infection of chrysanthemum (5th week)


REPLICATION
TREATMENT
I
II
III
IV
TOTAL
MEAN







Uninoculated – Untreated
1.00
1.00
1.00
1.00
4.00
1.00







Uninoculated – Treated
4.00
4.67
4.33
4.67
17.67
4.42







F.o.c + Hot H20 + garlic
extract (1 week after) +
comb. of Pseudomonas
sp. and Flavobacterium
sp. after 17 days
4.00
4.00
4.67
4.67
17.34
4.36







Comb. of Flavobacterium
sp. and Pseudomonas sp.
3.33
5.00
2.33
2.33
12.99
3.25







Mancozeb (Parafungus)
2.33
2.67
2.00
2.33
9.33
2.33







Thiopanate methyl
(Fungitox)
2.33
2.67
2.00
2.00
9.00
2.25







Garlic (Allium sativum L.)
2.00
4.67
2.67
4.33
13.67
3.42







Gawed (Piperbetle)
4.00
4.33
4.33
3.67
16.33
4.08







Table salt (1g/l H20)
3.67
3.67
2.00
4.67
14.01
3.50







GRAND TOTAL






GRAND MEAN







ANOVA

Source of
Degree of
Sum of
Mean of

Variation Freedom Squares Squares F
Value Probability
Model
35
164.85
4.71
5.98
0.0001
Treatment
8
123.18
15.4
9.98
0.0001
Rep(TRT)
27
41.67
1.54
1.96
0.0125
Error
72
56.67
0.78


TOTAL 107 221.52


** Highly significant CV= 27.69%

---

71

APPENDIX TABLE 24. Effect of bacterial antagonist and plant extracts on
fusarium wilt infection of chrysanthemum (6th week)


REPLICATION
TREATMENT
I
II
III
IV
TOTAL
MEAN







Uninoculated – Untreated
1.00
1.00
1.00
1.00
4.00
1.00







Uninoculated – Treated
5.33
5.00
4.67
4.67
19.67
4.92







F.o.c + Hot H20 + garlic
extract (1 week after) +
comb. of Pseudomonas
sp. and Flavobacterium
sp. after 17 days
5.00
4.67
5.33
5.00
20.00
5.00







Comb. of Flavobacterium
sp. and Pseudomonas sp.
4.33
4.00
5.00
2.33
15.66
3.92







Mancozeb (Parafungus)
2.33
2.67
2.00
2.00
9.00
2.25







Thiopanate methyl
(Fungitox)
2.33
2.67
2.00
2.00
9.00
2.25







Garlic (Allium sativum L.)
2.00
4.67
2.67
4.33
13.67
3.42







Gawed (Piperbetle)
4.00
4.00
5.00
3.67
16.67
4.17







Table salt (1g/l H20)
3.67
3.67
2.00
4.67
14.01
3.50







GRAND
TOTAL
GRAND
MEAN
3.38

ANOVA

Source of
Degree of
Sum of
Mean of

Variation Freedom Squares Squares F
Value Probability
Model
35
213.43
6.10
4.88
0.0001
Treatment
8
169.52
21.19
13.03
0.0001
Rep(TRT)
27
43.92
1.63
1.30
0.1882
Error
72
90.00
1.25


TOTAL 107
303.43


** Highly significant at 1% level using DMRT CV= 33.08%

---

72
APPENDIX TABLE 25. Effect of bacterial antagonists and plant extracts on
Fusarium wilt infection of chrysanthemum (7th week)

REPLICATION
TREATMENT
I II III I
TO
V TAL MEAN
Uninoculated – Untreated
1.00
1.00
1.00
1.00
4.00
1.00







Uninoculated – Treated
5.33
5.33
5.67
4.67
21.00
5.25







F.o.c + Hot H20 + garlic

extract (1 week after) +


comb. of Pseudomonas


sp. and Flavobacterium


sp. after 17 days
6.00
6.00
5.33
5.00
22.33
5.58







Comb. of Flavobacterium
sp. and Pseudomonas sp.
4.33
4.00
5.33
2.33
15.99
4.00







Mancozeb (Parafungus)
2.33
2.67
2.00
2.67
9.67
2.42







Thiopanate methyl
(Fungitox)
3.33
2.67
2.00
2.00
10.00
2.50







Garlic (Allium sativum L.)
2.00
2.67
4.33
13.67
3.42

4.00



Gawed (Piperbetle)
4.67
5.00
4.67
18.34
4.59







Table salt (1g/l H20)
4.00
4.00
2.00
5.00
15.00
3.75







GRAND
TOTAL 130.00

GRAND
MEAN
3.61

ANOVA

Source of
Degree of
Sum of
Mean of

Variation
Freedom
Squares
Squares
F Value Probability






Rep(TRT) 8
68.82
8.60
13.11
0.0001
Error 27
17.72
0.66


TOTAL 35
86.54









** Highly significant at 1% level using DMRT CV= 22.43%

---

73
APPENDIX TABLE 26. Effect of bacterial antagonists and plant extracts



on Vascular discoloration (cm) on chrysanthemum

TREATMENTS REPLICATION
I
II
III
IV
TOTAL
MEAN







Uninoculated/Untreated
0.00 0.00 0.00 0.00 0.00 0.00
Uninoculated /Treated
5.00
5.33
8.17
13.83
32.33
8.08
Hot H20 treatment +
7.17 12.67 10.33 5.33 35.50 8.88
garlic extract ( 1 week
after) + comb. Of
Pseudomonas ap. &
Flavobacterium sp
after 17 days
Comb. of Pseudomonas
2.67 3.50 3.67 1.67 11.51 2.88
sp. & Flavobacterium
sp.
Mancozeb (Parafungus)
3.67
0.67
2.17
2.33
8.84
2.21
Thiophanate methyl
0.67 0.17 3.50 1.00 5.34 1.34
(Fungitox)
Garlic extract (Allium
1.33 3.67 3.50 2.67 11.17 2.79
sativum L)
Gawed extract (Piper
2.67 4.17 5.67 3.67 16.18 4.05
betle)
Table Salt (1g/L H20) 0.67
2.67
2.07
6.67
12.08 3.02







GRAND TOTAL




132.95

GRAND MEAN





3.69


ANOVA

Sources of
Degree of
Sum of
Mean of
Computed
Probability
Variation
Freedom
Squares
Squares
F
Treatment 8
278.29
34.79
7.59 0.0001
Error 27
123.73
4.58


TOTAL 35
402.02


** -highly significant at 1% level using DMRT
CV
57.96%

---

74
APPENDIX TABLE 27. Effect of bacterial antagonists and plant extracts on
fresh top weight (g) of chrysanthemum plant

TREATMENTS REPLICATION
I
II
III
IV
TOTAL
MEAN







Uninoculated/Untreated 34.00 34.33 34.00 34.33 136.66 34.17
Uninoculated
/Treated 18.33 19.67 38.33 36.33 112.66 28.17
Hot H20 treatment +
5.00 3.00 0.00 7.67 15.67 3.92
garlic extract ( 1 week
after) + comb. Of
Pseudomonas ap. &
Flavobacterium sp
after 17 days
Comb. of Pseudomonas
37.33 43.67 49.67 47.33 178.00 44.50
sp. & Flavobacterium
sp.
Mancozeb
(Parafungus) 45.33 41.67 41.67 42.67 171.34 42.84
Thiophanate methyl
26.67 46 58.00 43.67 174.34 43.59
(Fungitox )

Garlic extract (Allium
41.67 48.67 35.00 44.67 170.01 42.50
sativum L)
Gawed extract (Piper
30.67 33.00 37.33 6.00 107.00 26.75
betle)
Table Salt (.5g/L H20) 41.33 45.33 50.67 31.00 168.33 42.08
GRAND
TOTAL

1234.01
GRAND MEAN





34.28


ANOVA

Sources of
Degree of
Sum of
Mean of
Computed
Probability
Variation
Freedom
Squares
Squares
F
Treatment 8
5634.79
704.35
10.17
0.0001
Error 27
1870.31
69.27


TOTAL 35
7505.10


** -highly significant at 1% level using DMRT
CV
24.28%

---

75
APPENDIX TABLE 28. Effect of bacterial antagonists and plant extracts



on the Fresh root weight (g) of chrysanthemum plant


TREATMENTS REPLICATION
I
II
III
IV
TOTAL
MEAN
Uninoculated/Untreated 5.73 6.50 6.70
4.77 23.70 5.93
Uninoculated /Treated
2.40
2.63
1.23
4.23
10.49
2.62
Hot H20 treatment +
1.50 0.73 0.67
0.67 3.57 0.89
garlic extract ( 1 week
after) + comb. Of
Pseudomonas ap. &
Flavobacterium sp.
after 17 days)
Comb. of Pseudomonas
6.37 7.03 7.13
5.10 25.63 6.41
sp. & Flavobacterium
sp.
Mancozeb (Parafungus)
4.10
5.23
6.63
3.63
19.59
4.90
Thiophanate methyl
4.33 6.03 5.53
7.47 23.36 5.84
(Fungitox )
Garlic extract (Allium
5.23 5.63 4.47
4.73 20.06 5.02
sativum L)
Gawed extract (Piper
4.23 5.50 3.57
2.80 16.10 4.03
betle)
Table Salt (.5g/L H20) 4.01
5.83
6.03
2.67
118.63
4.66
Grand Total




130

Grand Mean





3.61


ANOVA

Sources of
Degree of
Sum of
Mean of
Computed
Probability
Variation
Freedom
Squares
Squares
F
Treatment 8
98.69
12.34
10.20
0.0001
Error 27
32.66
1.21


Total 35
131.35



** -highly significant at 1% level using DMRT
CV
24.57%

---

76
APPENDIX TABLE 29. Quality cutflower (Long)

TREATMENTS REPLICATION
I
II
III
IV
TOTAL
MEAN
Uninoculated/Untreated 0.00 0.00 0.00
0.00 0.00 0.00
Uninoculated /Treated
0.00
0.00
0.00
0.00
0.00
0.00
Hot H20 treatment +
0.00 0.00 0.00
0.00 0.00 0.00
garlic extract ( 1 week
after) + comb. of
Pseudomonas ap. &
Flavobacterium sp
after 17 DAT)
Comb. of Pseudomonas
0.00 1.00 0.00
1.00 2.00 0.50
sp. & Flavobacterium
sp.
Mancozeb (Parafungus)
0.00
0.00
2.00
2.00
4.00
1.00
Thiophanate methyl
0.00 1.00 2.00
1.00 4.00 1.00
(Fungitox )

Garlic extract (Allium
0.00 0.00 1.00
0.00 1.00 0.25
sativum L)
Gawed extract (Piper
0.00 0.00 0.00
0.00 0.00 0.00
betle)
Table Salt (.5g/L H20) 0.00
0.00
1.00
0.00
1.00 0.25
Grand Total




12.00

Grand Mean





0.33


ANOVA

Sources of
Degree of
Sum of
Mean of
Computed
Probability
Variation
Freedom
Squares
Squares
F
Treatment 8
5.50
0.09
2.18
0.0618
Error 27
8.50
0.32


Total 35
14.00



* - significant at 5% level using DMRT
CV
168.327%

---

77
APPENDIX TABLE 30. Quality cutflower (Medium)

TREATMENTS REPLICATION
I
II
III
IV
TOTAL
MEAN
Uninoculated/Untreated 3.00 3.00 3.00 3.00 12.00 3.00
Uninoculated /Treated
0.00
0.00
0.00
1.00
1.00
0.25
Hot H20 treatment +
0.00 0.00 0.00
0.00
0.00 0.00
garlic extract ( 1 week
after) + comb. of
Pseudomonas ap. &
Flavobacterium sp
after 17 days
Comb. of Pseudomonas
1.00 1.00 1.00
1.00
4.00 1.00
sp. & Flavobacterium
sp.
Mancozeb (Parafungus)
0.00
0.00
1.00
1.00
2.00
0.50
Thiophanate methyl
1.00 0.00 1.00
1.00
3.00 0.75
(Fungitox )
Garlic extract (Allium
2.00 0.00 1.00
1.00
4.00 1.00
sativum L)
Gawed extract (Piper
1.00 1.00 1.00
0.00
3.00 0.75
betle)
Table Salt (.5g/L H20) 2.00 1.00 1.00
0.00
4.00 1.00
Grand Total




33.00

Grand Mean





0.92


ANOVA

Sources of
Degree of
Sum of
Mean of
Computed
Probability
Variation
Freedom
Squares
Squares
F
Treatment 8
23.50
2.94
10.94 0.0001
Error
27
7.25 0.27

Total 35
30.75


** -highly significant at 1% level using DMRT
CV
56.53%

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78
APPENDIX TABLE 31. Quality cutflower (Short)

TREATMENTS REPLICATION
I
II
III
IV
TOTAL
MEAN
Uninoculated/Untreated 0.00 0.00 0.00
0.00 0.00 0.00
Uninoculated /Treated
1.00
1.00
0.00
0.00
2.00
0.50
Hot H20 treatment +
0.00 0.00 0.00
0.00 0.00 0.00
garlic extract ( 1 week
after) + comb. of
Pseudomonas ap. &
Flavobacterium sp
after 17 days
Comb. of Pseudomonas
0.00 1.00 1.00
0.00 2.00 0.50
sp. & Flavobacterium
sp.
Mancozeb (Parafungus)
2.00
2.00
0.00
0.00
4.00
1.00
Thiophanate methyl
1.00 1.00 0.00
1.00 3.00 0.75
(Fungitox)
Garlic extract (Allium
1.00 1.00 1.00
0.00 3.00 0.75
sativum L)
Gawed extract (Piper
0.00 1.00 1.00
1.00 3.00 0.75
betle)
Table Salt (.5g/L H20) 1.00
1.00
1.00
0.00
3.00 0.75
Grand Total




20.00

Grand Mean





0.56


ANOVA

Sources of
Degree of Sum of
Mean of Computed
Probability
Variation
Freedom
Squares Squares F
Treatment 8
3.89
0.49
1.46
0.2156
Error 27
9.00
0.33


Total 35
12.89



* - significant at 5% level using DMRT
CV
103.92%

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79
APPENDIX TABLE 32. Quality cutflower (CB/Reject)

TREATMENTS REPLICATION
I
II
III
IV
TOTAL
MEAN
Uninoculated/Untreated 0.00 0.00 0.00
0.00 0.00 0.00
Uninoculated /Treated
2.00
2.00
3.00
2.00
9.00
2.25
Hot H20 treatment +
3.00 3.00 3.00
3.00 12.00 3.00
garlic extract ( 1 week
after) + comb. of
Pseudomonas ap. &
Flavobacterium sp
after 17days
Comb. of Pseudomonas
2.00 0.00 1.00
1.00 4.00 1.00
sp. & Flavobacterium
sp.
Mancozeb (Parafungus)
1.00
1.00
0.00
0.00
2.00
0.50
Thiophanate methyl
1.00 1.00 0.00
0.00 2.00 0.50
Fungitox)
Garlic extract (Allium
0.00 2.00 0.00
2.00 4.00 1.00
sativum L)
Gawed extract (Piper
2.00 1.00 1.00
2.00 6.00 1.50
betle)
Table Salt (.5g/L H20) 0.00
1.00
0.00
3.00
4.00 1.00
GRAND TOTAL




43.00

GRAND MEAN





1.19

ANOVA

Sources of
Degree of
Sum of
Mean of
Computed
Probability
Variation
Freedom
Squares
Squares
F
Treatment 8
27.89
3.49
5.98
0.0002
Error 27
15.75
0.58


TOTAL 35
43.64


** - highly significant at 1% level using DMRT
CV
27.89%

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80
APPENDIX TABLE 33. Effect of bacterial antagonists and plant extracts on



cutflower yield.a


TREATMENT CLASSIFI
CATION
LONG
MEDIU
SHORT CB NM
M
Uninoculated/Untreated
0.00b 3.00a 0.00b 0.00b 0.00c
Uninoculated
/Treated
0.00b 0.25bc 0.50ab 0.00b 2.25ab
Hot H20 treatment + garlic
0.00b 0.00c 0.00b 0.00b 3.00a
extract ( 1 week after) +
comb. Of Pseudomonas
ap. & Flavobacterium sp
(after 17 DAT)
Comb. Of Pseudomonas sp.
0.50b 0.00c 0.00b 0.50a 0.50bc
& Flavobacterium sp.
Parafungus
(Mancozeb)
1.00a 0.50bc 1.00a 0.50a 0.00c
Fungitox (Thiophanate
1.00a 0.75bc 0.75ab 0.50a 0.00c
methyl)
Garlic extract (Allium
0.25ab 1.00b 0.75ab 0.50a 0.00c
sativum L)
Gawed extract (Piper betle) 0.00b 0.75bc 0.75ab 0.50a 0.50bc
Table Salt (.5g/L H20)
0.25ab 1.00b 0.75ab 0.25ab 0.75b
Grand
Total



CV

168.32% 56.53% 103.92% 63.94 % 63.94%
aData are means of four replications. Means followed by a common letter are not significantly
different at 5% level using DMRT

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81
APPENDIX TABLE 34 . Effect of bacterial antagonists and plant extracts on



cutflower yield.a


TREATMENT
CLASSIFICATION/GROSS SALE (PhP)
Long
Medium
Short
CB
Reject
Total







Uninoculated/Untreated 0.00
60.00
0.00
0.0
0.00
P60.00
Uninoculated /Treated
0.00
5.00
8.34
0.00
0.00
P13.84
Hot H20 treatment + garlic
0.00 0.00 0.00 0.00 0.00 0.00
extract ( 1 week after) +
comb. Of Pseudomonas
ap. & Flavobacterium sp
( after 17 DAT)
Comb. Of Pseudomonas
11.66 20.00 8.34 10.00 0.00 50.00
sp. & Flavobacterium sp.
Parafungus (Mancozeb)
23.32
10.00
16.68
5.00
0.00
55.00
Fungitox (Thiophanate
23.32 15.00 12.51 5.00 0.00 55.83
methyl)
Garlic extract (Allium
5.83 20.00 12.51
10.00 0.00 48.34
sativum L)
Gawed extract (Piper
0.00 15.00 12.51
10.00 0.00 37.51
betle)
Table Salt (.5g/L H20) 5.83
20.00
12.51
2.50
0.00
40.84







GRAND
TOTAL

GRAND
MEAN

aData are means of four replications.

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Document Outline

• Management of Fusarium Wilt (Fusarium oxysporum f. sp. chrysanthemi) in Chrysanthemum (Dendranthema grandiflora T. Zveler) Using Bacterial Antagonists and Plant Extracts
  • BIBLIOGRAPHY
  • ABSTRACT
  • TABLE OF CONTENTS
  • INTRODUCTION
    • Background of the Study
    • Importance of the Study
    • Objectives of the Study
    • Time and Place of the Study
  • REVIEW OF RELATED LITERATURE
    • The Disease
    • Symptoms
    • Sign
    • Survival
    • Management
    • Biological Control
    • Plant Extracts
    • Chloride and Soil pH.
  • MATERIALS AND METHODS
    • Collection and Isolation of the Pathogen
    • Laboratory Experiment
      • Bioassay Test
      • Preparation of bacterial suspension
      • Preparation of Plant Extracts
      • Preparation of Fungicides
      • Compatibility Test
    • Greenhouse Experiments
      • Effect of bacterial antagonists and plant extractson the soil population of Fusarium oxysporumf.sp. chrysanthemi
      • Effect of bacterial antagonists and plant extracts onFusarium wilt infection and chrysanthemum yield
      • Data Gathered
  • RESULTS AND DISCUSSION
    • Laboratory Experiment
      • Effect of bacterial antagonists, plant extractsand fungicides on the growth of the pathogen
      • Compatibility Test
    • Greenhouse Experiment
      • Effect of Bacterial Antagonists and Plant Extractson the Soil Population of F.oxysporumf. sp. chrysanthemi
      • Effect of bacterial antagonists and plant extractson fusarium wilt infection andchrysanthemum yield.
        • Fusarium wilt infection
        • Vascular discoloration
      • Fresh Weight
      • Cutflower Yield
  • SUMMARY, CONCLUSION AND RECOMMENDATION
    • SUMMARY
    • CONCLUSION
    • RECOMMENDATION
  • LITERATURE CITED
  • APPENDICES

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