Development of Biopesticide Beauveria bassiana Bals. (Vuill.) Fungi

for Pest Control Vegetable in Berastagi

Sularno

and Edi Azwar

Lecturer of Faculty of Teacher Training and Education Islamic University of Sumatera Utara.

Keywords: Bioinsectisida, Beauveria bassiana Bals. (Vuill.), Chemical pesticides, vegetables

Abstract: The abstract should summarize the contents of the paper and should contain at least 70 and at most 200 words.

It should be set in 9-point font size, justified and should have a hanging indent of 2-centimenter. There should

be a space before of 12-point and after of 30-point. This research was conducted in two stages. The research

method is the isolation and application of biopesticide. In the first stage aims to isolate entomopathogenic

fungi Beauveria bassiana around Berastagi vegetable farms and grow on agar and corn mashed artificial

medium. Testing the virulence of Beauveria bassiana in vitro in several types of vegetable pests before it is

applied on agricultural land. In phase II aims to application B. bassiana in the vegetable patch. In the in vitro

treatment, the ability to infect insects beauvria average reached 75%. , Controls on crop lands, in the third

week began their Crocidolomia pests that attack the growing point so that the cabbage plants fail to bloom.

Average cropland applied biopesticides B. bassiana has been no attack. At week 7 attacks on control plants

reach 13 plants or 17.3%. In field applied while Beauveria bassiana attack reached six plants, or about 8%.

1 INTRODUCTION

Be advised that papers in a technically unsuitable

form will be returned for retyping. After returned the

manuscript must be appropriately modified.

Pesticides can cause acute and long-term effects

for farm workers who are exposed. Exposure to

pesticides can cause varying effects ranging from

irritation to the skin and eyes to a more deadly effect

that affects the nerve, disrupting the reproductive

hormone system and cause cancer. In 2007,a study in

non-Hodgkin's lymphoma and leukemia showed a

positive association with exposure to pesticides.

Strong evidence also shows that the negative effects

of pesticide exposure include neurological damage,

congenital abnormalities, fetal death and

developmental disorders of the nervous system. The

American Medical Association recommends limiting

exposure to pesticides and starts using safer

alternatives (Jakubowski,. 2011).

The use of pesticides increases the number of

issues on the environment. More than 90%

insecticides and 95% herbicides are sprayed toward

the place that was not targeted. Flow pesticides occur

when pesticides suspended in the air as particles

carried by the wind to other areas, potentially causing

pollution. Pesticides are the main problems of water

pollution, and some pesticides are persistent organic

pollutants that cause ground contamination.

Pesticides also reduce agricultural biodiversity in

the soil, thereby reducing the rate of nitrogen fixation.

loss of pollinators, destroy habitat (especially bird

habitats), and harm endangered animals. Over time,

the pest species can develop resistance to pesticides

so it takes research to develop new pesticides.

Of the approximately one million species of

insects are known, approximately 15,000 species

known as pests. Along with the increase in

agricultural activity, interference also increases insect

pests that cause damage or destruction of food crops

(Bale, et al, 2008). Fortunately, most insect pests have

the potential pathogenic microorganisms associated

with it. One of these microorganisms is an

entomopathogenic fungus.

Entomopathogenic fungi are microorganisms that

are used first as a biological pest control agent. More

than 700 species from 90 genera of fungi are

pathogenic to insects (Khan, et al, 2012). Most

species of entomopathogenic fungi derived from the

division Ascomycota and Zygomycota. Generally

fungal pathogens of insects include Metarhizium

anisopliae, Beauveria bassiana, Nomuraea rileyi,

Paecilomyces farinosus and Paecilomyces

fumosoroseus (Takhur and Sandhu, 2010),

442

Sularno, . and Azwar, E.

Development of Biopesticide Beauveria bassiana Bals. (Vuill.) Fungi for Pest Control Vegetable in Berastagi.

DOI: 10.5220/0010044504420445

In Proceedings of the 3rd International Conference of Computer, Environment, Agriculture, Social Science, Health Science, Engineering and Technology (ICEST 2018), pages 442-445

ISBN: 978-989-758-496-1

Lagenidium, Coelomomyces, Conidiobolus,

Entomophaga, Entomophthora, Erynia, Neozygites,

Pandora, Zoophthora, Cordyceps, Hypocrella,

Torrubiella, Aschersonia, Hirsutella, Tolypocladium

and Verticillium (Pell and Shah, 2003).

2 LIFECYCLE FUNGI

ENTOMOPATHOGENIC

The life cycle starts from the entomopathogenic fungi

spores germinate to form a mycelium, the mycelium

is then reshaped spores. Entomopathogenic fungi life

cycles generally consist of two phases. The first phase

is normal mycelial growth that occurs outside the host

body and budding yeast-like phase occurs within the

host homeosol.

3 ENZYMES OF VIRULENT

ENTOMOPATHOGENIC FUNGI

Initial interaction in the pathogenicity is mediated by

a mechanical force, enzymatic process and the

strength of certain acids. These enzymes responsible

for the interactions of fungi with their hosts and

environment. The enzymes involved in the

pathogenicity of a group of insects generally protease,

peptidase, chitinase, and lipase. Its main constituent

insect cuticle chitin and protein degradation by

protease and a peptidase enzyme. Another role as a

growth factor saprophytic fungi, propanol oxidase

activation in the hemolymph and as a virulence factor

of fungi (Khan, et al, 2012).

The main component of insect cuticle is the chitin

substance. Therefore both endo and Ekto khitinase

plays an important role in breaking down the N-

acetylglucosamine (NAGA) the polymer of insect

cuticle becomes smaller monomer units. So that the

enzyme chitinolytic also be a virulence determinant

factor of fungi. While the lipase enzyme plays a role

in epicuticular parse the outer surface of insects

composed of a mixture of non-polar lipid complex

(Blomquist and Vogt, 2003).

4 HOST RANGE

ENTOMOPATHOGENIC FUNGI

Entomopathogenic fungi have a very broad host range

in the class Insecta. Insect pests are hosts from

Lepidoptera, coleopteran, Diptera, Orthoptera and

other insect orders. Entomopathogenic fungi,

Verticillium lecanii, Beauveria bassiana and

Metarhizium anisopliae intensively studied for use as

the main natural enemies of the pest aphid and other

major agricultural pest (Roberts and St. Leger, 2004).

Beauveria bassiana (Bals.) Vuill. Has been developed

as a microbial insecticide to eradicate many major

pests, including lepidopterans and orthopterans.

approximately 33.9% of mycoinsecticide made of B.

bassiana, Metarhizium anisopliae followed by

(33.9%), Isaria fumosorosea (5.8%) and Beauveria

brongniartii (4.1%) (Wraight and Carruthers, 1999).

Therefore, the presence of fungi, these fungi play a

major role in overcoming the vegetable pests in

vegetable organic Brastagih as centers in North

Sumatra.

5 MATERIALS AND METHODS

Step 1. Exploration research includes fungi present

on the ground around the roots of plants with a depth

of 5 to 15 cm. Soil samples were collected in

accordance with the determination of sampling. Soil

is then inserted into the plastic and mixed until

homogenous 2. Soil samples were obtained is put into

a plastic container, filled with about half of the

volume of the container. 3. Before larvae Tenebrio

molitor inserted, the soil in the container moistened

with enough water added. 4. Putting worm Tenebrio

molitor at ground level in the container, which

included caterpillar is a caterpillar newly molted

(molting) is white 5. Further containers are closed

using gauze that caterpillars are not out of the

container, then incubated for 1 to 2 weeks in a dark

place in order to trap caterpillars move on, so it's easy

contact with entomopathogenic fungi that are in the

soil samples.

6 ISOLATION STAGE

The isolate phase includes 1. Tenebrio molitor

caterpillars that infected Beauveria bassiana fungi

were isolated by instilling sample of infected tissue

on the medium Potato dextrose Agar (PDA) and

incubated for 5 to 7 days 2. Insulation is done by

dipping the infected tissue samples (worm Tenebrio

molitor) while (± 3 minutes) to a solution of clorox,

alcohol, and then rinsed with sterile distilled water.

3.fungi growing on media is identified and

transmitted back (reinokulate) on the test insects.

Virulence test is done by calculating lethal

Development of Biopesticide Beauveria bassiana Bals. (Vuill.) Fungi for Pest Control Vegetable in Berastagi

443

concentration 50% (LC50) and 50% lethal dose

(LD50). 4. virulent entomopathogenic fungi

reproduced and developed to control the fi

Isolation of Beauveria bassiana

Stages isolation covers 1. Tenebrio molitor larvae

infected with Beauveria bassiana Fungi were isolated

by instilling sample of infected tissue on the medium

Potato dextrose Agar (PDA) and incubated for 5 to 7

days 2. Insulation is done by dipping the infected

tissue samples ( Tenebrio molitor) while (± 3

minutes) to a solution of clorox, alcohol, and then

rinsed with sterile distilled water. 3.fungi growing on

media is identified and transmitted back (reinoculate)

on the test insects Sometimes the isolation process

had to be repeated because of contamination by other

bacteria or fungi. Kesabarabn and accuracy needed to

obtain pure isolates before propagated in the

propagation medium (corn).

Here is a picture Tenebrio molitor larvae infected

with Beauveria bassiana

Figure 1. Tenebrio molitor larvae infected with beauveria

bassiana and metarhizium anisopliae

7 STAGE PURIFICATION AND

THE MULTIPLICATION OF

BEAUVERIA BASSIANA

Beauveria bassiana purification method is done by

streak plate method or by pour plate method on PDA

media. Beauveria bassiana already growing on the

PDA medium and then reproduced in corn medium.

Mashed corn steamed later washed Clean Up nearly

ripe. Enter Into plastic glass hearts and sterilized

using an autoclave. Corn medium cold ready for

review inoculated with Beauveria bassiana as a

propagation medium. Beauveria bassiana is well

propagated used to review applications Pest control

Plants flower cabbage Brassica oleracea.

From in vitro experiments on the data obtained as

follows:

Table. Number of Tenebrio molitor larvae infected and died

after treatment with Beauveria bassiana

Repeat of

Treatment

with 10

T.molitor

larvae

Number larvae T.molitor

infected at day ....

4 5 6 7

I 1 3 3 8

II 0 1 2 7

III 1 2 5 7

IV 0 2 4 6

V 1 3 5 8

VI 2 5 6 9

From the table above it can be seen that the

highest number of deaths occurred on the seventh-day

average larvae infected and die reached 75%. This

data will be compared with the treatment of

Beauveria bassiana in the field.

8 APPLICATIONS OF

B.BASSIANA IN THE FIELDS

OF RESEARCH

Applications Beauveria bassiana conidia done by

mixing with water

Application spraying is done in the afternoon with

the intensity 3 times in one week. Observations were

made each week cover the range of the pest

population, the diversity of types of pests, damage

leaves and pest infestation levels in both fields of

research.

Observations in the first week, second and third

after the second application of Beauveria bassiana on

land not seen a significant difference. The types of

pests of vegetables such as cauliflower B. oleracea

snails, grasshoppers leaf, ladybugs, Plutella and

Crossidolomia. In qualitative damage to the leaves of

plants by insects or slightly higher in the control

plants, the damage is massive because the pests only

attack leaves lateral. Group grasshopper populations

are low and do not cause severe damage was Plutella

xylostella the solitary life also does not cause severe

damage to both research fields. The leaves of the

cabbage flowers on the lateral are perforated, not

sporadic pest attack and not to attack the growing

point (crop) cabbage plants harvested so that interest

will continue to grow well.

ICEST 2018 - 3rd International Conference of Computer, Environment, Agriculture, Social Science, Health Science, Engineering and

Technology

444

Observations on the third week, on land that is

applied with Beauveria bassiana, the pest population

remains relatively low, while on land without any

treatment or control Crocidolomia binotalis

populations have started to emerge. Population

growth of Crocidolomia binotalis is quite rapid and

sporadic, larvae are green with light green stripes

back and on the left and right sides with the older

color yellow on the the abdomen. After hatching, the

larvae feed on leaves immediately with very

voracious, especially the leaves inside covered by the

outer leaves for larvae afraid of sunlight. The larvae

live in groups, if attacks increase, the larvae will reach

the point of growing crops or crop cauliflower. As a

result of this attack,, cabbage plants failed to form

interest and result in loss of harvest (yield loss). Of

the 75 plants on land control, attack the crop reaches

comparatively quite high, reaching at week 3

contained 5 plants damaged or approximately 6.7% in

the five weeks to 9 plants, or about 12%, and at week

7 reaches 13 plants or 17.3%. In field applied while

Beauveria bassiana, Crocidolomia binotalis attacks

are relatively smaller. At three weeks there have been

no plants are attacked, at week five there are three

affected plants to the growing point and at week 7

there are 6 plants or about 8% that was attacked.

Figure 2. Damage to the leaves of the growing points or

crop in cauliflower, cabbage crop failure flowering due to

pests attacking at the growing point

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