The Potential of Malaysian Seaweeds as the Source of Antifungal Compounds for Application - PowerPoint Presentation

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The Potential of Malaysian Seaweeds as the Source of Antifungal Compounds for Application in the Oil Palm Industry

Zetty Norhana Balia Yusof, PhDDepartment of Biochemistry,Faculty of Biotechnology and Biomolecular Sciences,Universiti Putra Malaysia,43400 UPM,Serdang, Selangor,MALAYSIA

Oil Palm

39% palm oil production and 44% export in the worldKingdom: PlantaeFamily: ArecaceaeSubfamily: ArecoideaeTribe: CocoeaeGenus: ElaiesSpecies: E. guineensis

Originated from Africa. Introduced to Malaysia by the British in 1870 and commercial planting started in 1917

G.

boninense

Daily uses

Fusarium wilt

Oil Palm Estates in Malaysia

Basal stem rot disease

Oil palm productivity in Malaysia is threatened by basal stem rot disease caused by Ganoderma boninense

To date, no study is conducted on

the

use of antifungal properties from seaweeds

for the disease control

A more environmental-friendly disease control is needed to replace the current heavy use of fungicides and harmful chemicals

Malaysia is blessed with the abundance of seaweeds which are

underutilised

Problem

Statement

Seaweed as a Source of Natural CompoundsSeveral

reports have shown that compounds deduced from macroalgae are highly bioactive, with properties that have antifungal and antibacterial activities (Oranday MA, MJ Verde, SJ Martínez-Lozano 2004; Freile-Pelegrín & Morales 2004).Bioactive compounds extracted from seaweeds are capable of protecting human and plants from various serious diseases (Paul & Puglisi 2004; Bhadury & Wright 2004).

Antifungal Activity of Seaweeds

Seaweed speciesPlant pathogenEffective extractReferencesAcanthaphora spiciferaMicrosporum gypseumMethanol(Pandian et al., 2011

)

Rhodomela

confervoides

and

Padina

pavonica

Candida

albicans

,

Mucor

ramaniannusMethanol

(

Saidani

et al., 2012)

Alaria

esculenta, Fucus vesiculosus

, Fucus sp., Spirulina platensis, and Ecklonia

maxima

Fusarium

roseum

, F.

oxysporum

,

Alternaria

alternata

, A.

dauci

, A.

longipes

, Trichoderma

viride

, Botrytis

cinerea

, Aspergillus

niger

,

Penicillium

expansum

Ethanol

(

Cosoveanu

et al

., 2010)

Stypopodium

zonale

,

Laurencia

dendroidea

,

Ascophyllum

nodosum

,

Sargassum

muticum

,

Pelvetia

canaliculata

and

Fucus

spiralis.

Colletotrichum

lagenarium

Ethanol

(Peres

et al

., 2012)

Sargassum

vulgare,

Cystoseira

barbata

,

Dictyopteris

membranacea

,

Dictyota

dichotoma

, and

Colpomenia

sinuosa

Alternaria

alternata

,

Fusarium

oxysporum

Cyclohexane

(Am and

Im

, 2015)

Sargassum

myricocystum

Gracilaria

edulis

Colletotrichum

falcatum

Ethanol

(

Ambika

and Sujatha, 2015)

Project AimsTo screen selected

Malaysian seaweeds for antifungal activityTo investigate the potential of bioactive compounds with inhibitory activity against disease-causing fungi in oil palmTo identify the bioactive compounds possessing the antifungal activity against G. boninense

Methodology

Collection of SeaweedsLocation: Teluk Kemang

, Port Dickson, Negeri SembilanThe beach of Teluk Kemang at Port Dickson (2°26′ N,101°51′ E) is located at the West Coast of Peninsular Malaysia, facing the Straits of Malacca.The study sites has vast variety of seaweed species can be found growing on the reefs. SamplingDateTimeLevelTide1stThursday (10th March 2016)2.00 p.m.0.03m

Low tide

2nd

Thursday (2

nd

June

2016)

10.30 a.m.

0.63m

Low tide

Tide

schedule of

Teluk

Kemang, Port Dickson during sampling.Source : http://tides.mobilegeographics.com/locations/4893.html?y=2016&m=3&d=10

Collected seaweeds and their potentials

Seaweeds / BioactivityAntifungal

Antibacterial

Antioxidant

Red seaweeds

(Rhodophyta)

Kappaphycus

alvarezii

√

√

√

Brown seaweeds

(Phaeophyta)

Sargassum

duplicatum

NI

NI

√

Green

seaweeds

(

Chlorophyta

)

Halimeda opuntia

√

NI

√

Caulerpa

racemosa

√

√

√

Caulerpa

microphysa

√

√

NI

Extract preparation

VolatileDissolve wide range of organic compoundsEfficient for lipid extractionInexpensiveRecent publications reported a higher extraction yield in water for seaweeds identified with high levels of water-soluble components, such as soluble polysaccharides, protein and peptides, revealing the hydrophilic nature of the majority of the seaweed components (Farvin and Jacobsen 2013; Tierney et al. 2013).InexpensiveVolatileCan dissolve many compoundsLow boiling point

Extraction Yield

SeaweedsPowder weight (g)Extraction yield (g)Methanol

Dichloromethane

Chloroform

Aqueous

Sargassum

duplicatum

5.0

1.13

0.289

0.048

0.501

Caulerpa

microphysa

5.0

NA

0.331

0.421

0.832

Caulerpa

racemosa

5.0

0.862

0.159

0.417

0.789

Halimeda

opuntia

5.0

NA

0.785

0.533

0.748

Antifungal Assay - Poisoned Food Technique(Bussaman et al., 2012; Schmitz, 1930)

Ganoderma boninenseBasidiomycetesReproduce through spores and mycelia

Spreads in the soil through roots and also through the airG. boninense causes basal stem rot disease in oil palmFusarium oxysporum

Ascomycetes

Soilborne

pathogen

P

roduces

macroconidia, microconidia

, and

chlamydospores

Chlamydospores

allow survival in plant debris and

soil

F.

oxysporum

causes fusarium wilt in oil palm

Attack oil palm at all ages from seedling to mature palms (Prendergast, 1957)

Ganoderma boninense

Culture of G. boninenseMicroscopic identification of G. boninense with Lactophenol cotton blue staining. Right :Image observed under 40x100 magnification, Left: Image adapted from Rahamah et al., 2012

Fusarium oxysporum

Culture of F. oxysporumMicroscopic identification of F. oxysporum with Lactophenol cotton blue staining. Right :Image observed under 40x100 magnification, Left: Image adapted from http://thunderhouse4-yuri.blogspot.my/2012/06/fusarium-oxysporum.html

Antifungal Assay (Bussaman et al., 2012; Schmitz, 1930)

Antifungal Assay

G.b.= Ganoderma boninense, F.o.= Fusarium oxysporum, X=contaminated Seaweed

Extract concentration

(µl/ml)

Mycelial

growth (mm

)

% reduction over control

Control (mm)

G.b

.

F.o

.

G.b

.

F.o

.

G.b

.

F.o

.

Sargassum

duplicatum

5

18.7

26.0

63.1

46.9

50.7

49.0

10

16.0

20.0

68.4

59.2

20

10.0

19.7

80.3

59.8

Caulerpa

microphysa

5

X

21.0

-

57.1

10

14.5

26.0

71.4

46.9

20

X

20.7

-

57.8

SummaryMethanol solvent gave the most extract yield compared to other

solventsA significant amount of reduction in the mycelial growth were observed in extract-added PDAThe seaweed extracts showed high levels of activities against the tested fungus G. boninense is more sensitive towards the seaweeds extracts compared to F. oxysporum

GC-MS,HPLC – for identification of compounds (

phenolics

and fatty acids)

Testing of seaweed extracts on fungi-infected oil palm seedlings

Compounds pathway analysis for overexpression studies

Future Work

Project BenefitThis

project will pave the way for the production of a more natural and environmental friendly method to control the disease caused by G. boninense in oil palm The utilization of the readily abundant and naturally occurring seaweeds as a source of potential bioactive compounds with antifungal activityThis project will be crucial for the sustainability of palm oil production in the oil palm industry of Malaysia

AcknowledgementsSyamimi Diyana Abdul Aziz

Nur Sakinah GhazaliAtiqah SubkiAisamuddin Ardi Zainal AbidinLee Li FernNur Husna AzimLab 230, Department of Biochemistry, FBSB, UPMGeran Putra IPM Universiti Putra Malaysia (9425900)

Thank youZetty Norhana Balia Yusof, PhD (Cambridge, UK)

Senior LecturerDepartment of Biochemistry,Faculty of Biotechnology & Biomolecular Sciences,Universiti Putra Malaysia,43400 UPM, Serdang,Selangor, MALAYSIA+60389466712 / +60123307339zettynorhana@upm.edu.my / zettybyusof@gmail.com