The Effect of Basil Leaves Extract (Ocimum Sanctum L.) on Mycelial

Growth of Postharvest Fungi

Kiki Nurtjahja

, Albert Pasaribu

and Roslindawati

Department of BiologyUniversitas Sumatera Utara, Medan, Indonesia

Department of Chemistry, Universitas Sumatera Utara, Medan, Indonesia

Keywords: Antifungal, Basil leaves, Postharvest fungi, Minimum inhibitory concentration

Abstract: Basil (Ocimum sanctum L.) contain secondary metabolites which inhibit fungal grow. This research aims to

investigate methanolic extract of basil leaves against postharvest fungi. Dried basil leaves were extracted and

macerated using methanol. The extract was stored for 3×24 hours at room temperature and homogenized for

24 hours. The homogenise was filtered using whatman filter paper and evaporated at 50 ºC. The thick extract

was made a serial concentration 100, 80, 60, 40 and 20 % by dimethylsulfoxide 10%. The postharvest fungi

used were Aspergillus oryzae, Aspergillus flavus, Aspergillus fumigatus, Aspergillus tamarii, Aspergillus

niger, Aspergillus candidus, and Penicillium sp. The minimum inhibitory concentration was determined by

liquid dilution. Ketoconazole 2% and dimethyl sulfoxide were used as positive and negative control

respectively. Results showed that basil leaves with concentration 100, 80, 60, 40 and 20% inhibit fungal

growth, the highest inhibition occurred on Aspergillus tamarii with inhibition 8.7; 8.0; 7.2; 6.0; 5.0 mm

respectively. The lowest inhibition occurred on Penicillium sp. with inhibition 2.5; 2.0, 1.8; 0.0 mm

respectively.

1 INTRODUCTION

Basil (Ocimum sanctum L.) is one of the traditional

herbs that can be used as antifungi. Solikhah (2015)

reported that the antifungi as secondary metabolites

were accumulated on leave. Sopianti and Sary (2018)

stated that secondary metabolites on basil were

alkaloid, saponin, tannin, flavonoid, steroid and eteris

oil. Previous study by Omay et al. (2017) reported the

potential of basil extract to inhibit Candida albicans.

Whereas, Berlian et al. (2016) used the leave extract

to inhibit Fusarium oxysporum. Basil leave extract in

inhibiting postharvest fungi such as Aspergillus

niger, Fusarium solani, Penicillium funicolusum,

Trichoderma reesei was reported by Dharmagadda et

al. (2005) and Bansod and Rai (2008). Postharvest

fungi commonly infect crops and dried stored spices.

The used of chemical compounds such as fungicide

or fumigants to control fungal infection on

agricultural products was expensive and the chemical

residues were harmful for human health. The aim of

the recent study was to determine the potential of

leave basil extract to inhibit mycelial growth of

postharvest fungi.

2 MATERIALS AND METHOD

2.1 Preparation of Fungal Isolates

Postharvest fungi used in this experiment were

culture collection of Microbiology and

Biotechnology Laboratory, Faculty of Mathematics

and Natural Sciences, Universitas Sumatera Utara.

All of the storage fungi were isolated from dried-

stored spices. The fungal isolates were subcultured in

potato dextrose agar (PDA) and incubated at 29

C for

5 days.

2.2 Extraction of Basil Leaves

Ten kilogram of fresh basil leaves were air dried for

4 days and groud using RT 04 Mill Powder Tech. Co

LTD Taiwan at 25.000 rpm for 30 second, and 756 g

of the simplicia were extracted in 6 L methanol for

3×24 hours. The homogenate was filtered using

Whatman filter paper no.1, the filtrate then was

evaporated by rotavapor 100 rpm at 50

C. The extract

concentrations used were 100, 80, 60, 40 and 20%,

dimethyl sulfoxide 10% used as a solvent.

542

Nurtjahja, K., Pasaribu, A. and Roslindawati, .

The Effect of Basil Leaves Extract (Ocimum sanctum L.) on Mycelial Growth of Postharvest Fungi.

DOI: 10.5220/0010612700002775

In Proceedings of the 1st International MIPAnet Conference on Science and Mathematics (IMC-SciMath 2019), pages 542-544

ISBN: 978-989-758-556-2

2.3 Determination of Basil Leaves

Extract in Inhibiting Fungal

Growth

Paper disc (5 mm in diameter) containing 10 µl of

extract in petri dish (9 cm in diameter) containing 10

mL potato dextrose plate agar. Each fungal isolate

was tested by inoculating agar plug containing fungal

mycelia. Ketoconazole 2% and dimethyl sulfoxide

10% were used as positive and negative control

respectively. All plates were incubated at 27°C for 5

days. Three replicates were used for each

concentration.

2.4 Determination of Minimum

Inhibitory Concentration of Basil

Leaves Extract

Concentration extract used for minimum inhibitory

experiment was 20, 40, 60, 80, 85, 90, 95 and 100%.

Ketoconazole 2% and dimethylsulfoxide 10% were

used as positive and negative control respectively.,

One mililiter of fungal spore suspension (10

/ml) then

was mixed with 1 ml of each extract concentration,

the mixture thes was.homogenized and dilute by a

serial dilution until the concentration 10

cfu/ml.

Each 1 ml thes was pour plate in petri dish containing.

PDA. Each treatment was replicate 3 times All plates

were incubated at 27°C for 5 days.

3 RESULTS AND DISCUSSION

3.1 Inhibition Zone of Basil Leaves

Methanolic Extract on Mycelial

Growth of Postharvest Fungi

Leave extract of basil leaves potential to inhibit

mycelial growth of postharvest fungi (Table 1). As

shown in Table 1 all fungal species were affected by

the presence of extract, however, each of the species

has different inhibition zone. The higher extract

concentration is followed by the increasing of

inhibition. Among the postharvest fungi, Aspergillus

tamarii was the most affected by basil extract (6.98

mm) followed by A. candidus (6.48 mm). Whereas,

Penicillium sp. was the less affected (1.26) followed

by A. oryzae and unknown yeast (2.58 mm).

Fardiaz (1989) stated that the ability of antifungal

compounds in inhibiting mycelial growth was

determined by concentration of the compound and

fungal species. The range of inhibition zone indicate

that basil leave extract has mild to low potential in

inhibiting storage fungi. We assumed that the use of

crude extract lowering antifungal compounds. Davis

and Stout (1971) reported that the effect of antifungi

on mycelial growth as indicated by inhibition zone

with mild (5 and 10 mm in diameter) and low

inhibition (0 to 4 mm in diameter). The presence of

chemical compounds in the basil extract might inhibit

mycelial growth. Previously studied by Kadian and

Parle (2012) reported that eteris oil in basil leaves

contain methyl chavicol and linalool that has

antifungal activity, the compounds affect on

ergosterol in fungal cell membrane.

Table 1: Inhibition zone (mm) of basil leaves methanolic extract (%) againts postharvest fungi

Fungal isolates

Extract concentration (%) / inhibition zone (mm)

K- K+ 20 40 60 80 100 Average

Aspergillus oryzae 0 5.2 1.8 2.1 2.6 3.0 3,4 2.58

A. flavus 0 7.0 4.0 5.0 5.3 5.7 6,6 5.32

A. niger 0 5.5 2.9 3.0 3.7 4.0 4,2 3.56

A. candidus 0 8.0 5.5 5.8 6.6 7.0 7,5 6.48

A. fumigatus 0 6.0 2.8 3.0 3.2 3.6 4,0 3.32

A. tamarii 0 9.5 5.0 6.0 7.2 8.0 8,7 6.98

Penicillium sp. 0 3.0 0.0 0.0 1.8 2.0 2,5 1.26

The Effect of Basil Leaves Extract (Ocimum sanctum L.) on Mycelial Growth of Postharvest Fungi

543

Table 2: Minimum inhibitory concentration methanolic extract of basil leave on the growth of postharvest fungi

Extract

(%)

Fungal species/mycelial growth

A. oryzae A. flavus A. niger A. candidus A fumigatus A. tamarii Penicillium sp.

K- + + + + + + +

K+ - - - - - - -

100 + - + - + - +

95 + - + - + - +

90 + - + - + - +

85 + + + - + - +

80 + + + + + + +

60 + + + + + + +

40 + + + + + + +

20 + + + + + + +

Ruijter et al. (2004) reported that xerophilic fungi

accumulate polyol such as glycerol, eryhritol, and

arabitol to maintain their grow in low water activity.

Whereas, the effect of tannin in basil leaves was

reported by Ajizah (2004), the compound reduce

membrane permeability.

3.2 Determination of Minimum

Inhibitory Concentration of Basil

Leaves Extract

Minimum inhibitory concentration of the extract to all

fungi was shown in Table 2. Basil leave with

concentration lower than 85 % has no effect on all

fungi tested. The minimum inhibitory of the extract

begin to inhibit fungal mycelia at concentration 85%

particularly on Aspergillus candidus and Aspergillus

tamarii. Some species of fungi such as A. oryzae, A.

niger, A. fumigatus and Penicillium have no effect,

even at the highest concentration (100%).

4 CONCLUSION

Basil leaves extract reduce mycelial growth of

postharvest fungi, however, the potential of the

extract in inhibiting the fungal growth was different

for each species.

ACKNOWLEDGEMENT

The research was supported by TALENTA-Sumatera

Utara University, Grant no. 322/UN5.2.3.1/PPM/KP-

TALENTA USU/2019.

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