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Abstract

This study aimed to determine the effectiveness of Bacillus subtilis in


controlling ascospore development of Aspergillus niger, in vitro.
A. niger ascospores were discharged into petri dishes with water agar (T 1)
and into petri dishes with water agar and B. subtilis (T2). The lengths of the germ
tubes emerging from the ascospores were measured after forty-eight (48) hours.
Results of the in vitro test showed that the length of the germ tubes of the
ascospores of M. fijiensis in T2 (________ m) was significantly shorter than that
observed in T1-control (________ m).
It was concluded that B. subtilis is effective in controlling ascospore
development of A. niger.

Introduction

Bacillus subtilis(Bacillaceae)is a widespread bacterium found in soil, water,


and air. It controls the growth of certain harmful bacteria and fungi, presumably by
competing for nutrients, growth sites on plants, and by directly colonizing and
attaching to fungal pathogens. When applied to seeds, it colonizes the developing
root system of the plants and continues to live on the root system and provides
protection throughout the growing season.
Aspergillus niger (Trichocomaceae)is a fungi thatcan cause the rotting of
numerous fruits, vegetables, and other food products. It causes root curling and top
deformation in plants, thus causing substantial economic losses due to spoilage. B.
subtilis has the capability to shorten the germ tubes of mycotoxins, which A. niger is
one. The aim of this study is to test the effectivity of B. subtilis to reduce or remove
the effect of A. niger on Mangifera indica (Anacardiaceae), particularly black molds.
M. indica (Mango) one of the commercially produced in the world, but it is
also very susceptible to diseases which A. niger is one, with the help of the B.
subtilis the amount of damage A. niger can cause to mangoes and other plants, will
be drastically reduced, increasing the harvest and production of mangoes.

Chapter 3
METHODOLOGY

Phase I: In Vitro test


Gathering of specimens: Five (5) leaves from the
Cavendish Banana variety with black Sigatoka, as indicated by
necrotic burnt tissue, were collected from Bureau of Plant
Indusstry (note: the leaves have not experienced rain for three
days). The leaves were cleaned with dry tissue paper and it was
made sure that the leaves did not get wet. The samples were
incubated in a plastic bag with a moist towel for 48 hours at room
temperature. The bag was sealed in order to maintain the
moisture in the bag.

Preparation of the Broth Culture of Bacillus Subtilis: One


hundred ml of distilled water was mixed with 2.8 grams of
nutrient broth in Erlenmeyer flask. The mixture was heated and
stirred until the solution is clear. The broth was then sterilized in
the oven for 15 minutes at 121 C. The flask was soaked in the
water bath until the broth cooled. Each 20-ml sterilized test tube
was poured with 15 ml of broth. Bacillus subtilis was then
inoculated into the test tube and were incubated at 30 C for 24
hours.

Preparation of Treatments: Water Agar, a culture


medium of 2% water agad formulation, was prepared, For every

1000 ml of water, there was 10 g of agar. For T1, 5 petri dishes


each with 25 ml 2% water agar were prepared. For T 2, 5 petri
dishes with 25 ml 2% water agar were poured plated with 2 ml of
broth culture containing Bacillus subtilis.

Bacillus subtilis as a biological control agent of


Mycosphaerella Fijiensis causing Black Sigatoka in
Bananas.
This study aimed to determine the effectiveness of Bacillus
subtilis in controlling Development of Mycosphaerella fijiensis
in vitro, and to compare its effectiveness with that of different
fungicides.
M. fijiensis ascopores were discharged into petri dishes
with water agar (T1) and into petri dishes with water agar and B.
Subtilis (T2). The lengths of the green tubes emerging from
ascopores were measured after forty-eight (48) hours. In the field,
Bacillus subtilis at 80 ml and 40 ml/lt water, Dithane, Daconil,
bumper and Bankit were sprayed on different leaves.
Effectiveness was measured by the number of days after treating
the symptoms of the disease first appeared.
Results of the in vitro test showed that the length of the
germ tubes of the ascopores of M. Fijiensis in T2 (0.3066672)
was significantly shorter than that observed in T 1-control. In the
field test, the appearance of the symptoms were most delayed by
Bankit and Bumper while there was no significant difference
between the number of days the symptoms appeared in the
leaves sprayed with B. Subtilis at 80 ml and 40 ml/lt water,
Dithane and Daconil.
It was concluded that B. Subtilis is effective in controlling
ascospore development of M. Fijiensi and that bumper and
bankit are the most effective in delaying the appearance of the

symptoms of M. Fijiensi while Bacillus subtilis at 80 ml and 40


ml/lt water, Dithane and Daconil are equally effective in delaying
the appearance of the symptoms of M. fijiensi