Phytochemistry 114 (2015) 168–177

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Phytochemistry
journal homepage: www.elsevier.com/locate/phytochem

Review

Molecular defense response of oil palm to Ganoderma infection

C.-L. Ho a,b,⇑, Y.-C. Tan b,1
a
Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM-Serdang, Selangor, Malaysia
b
Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400 UPM-Serdang, Selangor, Malaysia

a r t i c l e i n f o a b s t r a c t

Article history: Basal stem rot (BSR) of oil palm roots is due to the invasion of fungal mycelia of Ganoderma species which
Available online 12 November 2014 spreads to the bole of the stem. In addition to root contact, BSR can also spread by airborne basidiospores.
These fungi are able to break down cell wall components including lignin. BSR not only decreases oil
Keywords: yield, it also causes the stands to collapse thus causing severe economic loss to the oil palm industry.
Ganoderma The transmission and mode of action of Ganoderma, its interactions with oil palm as a hemibiotroph,
Oil palm and the molecular defence responses of oil palm to the infection of Ganoderma boninense in BSR are
Elaeis guineensis Jacq.
reviewed, based on the transcript profiles of infected oil palms. The knowledge gaps that need to be filled
Basal stem rot
Molecular defence
in oil palm–Ganoderma molecular interactions i.e. the associations of hypersensitive reaction
Lignin (HR)-induced cell death and reactive oxygen species (ROS) kinetics to the susceptibility of oil palm to
Ganoderma spp., the interactions of phytohormones (salicylate, jasmonate and ethylene) at early and late
stages of BSR, and cell wall strengthening through increased production of guaiacyl (G)-type lignin, are
also discussed.
Ó 2014 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168
2. Ganoderma spp. reported in BSR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
2.1. Life cycle of Ganoderma spp. and its relation to BSR. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
2.2. Transmission of BSR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
2.3. Development and progress of BSR in oil palm roots . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170
3. Plant-pathogenic interactions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170
4. Gene expression of oil palm roots in response to Ganoderma infection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172
5. Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 176

1. Introduction
Oil palm (Elaeis guineensis Jacq.) is one of the main sources of
edible oil in the world with Indonesia and Malaysia contributing
about 85% of the world’s palm oil production. Total export revenue
⇑ Corresponding author at: Department of Cell and Molecular Biology, Faculty of
Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM-
Serdang, Selangor, Malaysia. Tel.: +60 3 89467475; fax: +60 3 89467510.
E-mail address: clho@upm.edu.my (C.-L. Ho).
1
Present address: Codon Genomics Sdn. Bhd., No. 11/A-2, Jalan Bandar Lapan Belas,
Pusat Bandar Puchong, 47160 Puchong, Selangor, Malaysia.
of oil palm products (palm oil, palm kernel oil, palm kernel cake,
oleo chemicals, biodiesel and finished products) for Malaysia and
Indonesia in 2012 was approximately USD 22 billion (bepi.mpob.-
gov.my/; www.indonesia-investments.com/), respectively. In order
to meet the high demand for palm oil which increases annually,
the oil palm plantation areas in these two main palm oil producing
countries in South East Asia have expanded. Over the last three
decades, the oil palm plantation area in Malaysia has expanded
from 641,791 hectares in 1975 to 5.23 million hectares in 2013
(bepi.mpob.gov.my), while the total oil palm plantation area in
Indonesia has reached 8 million hectares in 2010 (Rianto et al.,
2011). Meanwhile, breeding and selection of palms with high yield

http://dx.doi.org/10.1016/j.phytochem.2014.10.016
0031-9422/Ó 2014 Elsevier Ltd. All rights reserved.
 C.-L. Ho, Y.-C. Tan / Phytochemistry 114 (2015) 168–177
potentials and disease tolerance (thus reducing yield loss due to
diseased palms) are crucial for sustainable production of palm oil
without further expansion of plantation area. Hence, an under-
standing of oil palm disease infection process and host defence
mechanisms at the molecular level are required.
Oil palm is susceptible to a number of fungal diseases, including
Phytophthora, Fusarium wilt, upper stem rot (USR) and basal stem
rot (BSR) (Turner and Bull, 1967; Aderungboye, 1977). Among
these, USR and BSR are caused by pathogenic fungi belonging to
the genus Ganoderma and BSR is considered to be one of the most
devastating oil palm diseases. BSR is an oil palm root disease which
manifests itself as decay of roots and lower stem, and can be
spread by direct root contact and basidiospores by still unknown
means (Pilotti et al., 2002; Rees et al., 2007, 2009, 2012). Ganoder-
ma spp. are white rot fungi that are capable of degrading compo-
nents of the plant cell wall including lignin (Paterson, 2007). The
fungal pathogen disrupts water and nutrient transport to the upper
part of the palm thus causing frond wilting, yellowing of fronds,
unopened spear leafs, reduced and ‘‘one-sided mottling’’ of canopy,
and emergence of basidiocarps on the lower stem (Turner, 1981;
Chung, 2011). Infected oil palms were reported to have a lower
oil yield before the stands collapsed eventually (Singh, 1991). To
make the condition worse, BSR which was initially thought to
infect only mature palms over 25 years of age (Thompson, 1931),
was found to infect also young palms (10–15 years) (Turner,
1981) and seedlings less than 5 years old (Singh, 1991). In this
review, the focus is mainly on BSR since it is more prevalent in
most of the oil palm plantations in South East Asia.
BSR occurs frequently in coastal marine clay areas which were
previously planted with coconut trees in western Peninsular
Malaysia (Navaratnam, 1964; Lim et al., 1992). However, it was
also reported to happen in peat soil situated inland (Ariffin et al.,
1989; Rao et al., 2003). In order to control and suppress the spread
of the disease, several approaches were explored. Practices that
involved fungicides and burning of infected oil palms are unfavor-
able to the environment. Furthermore, most fungicides are not
effective or specific against BSR (Idris and Arifurrahman, 2008).
Meanwhile, conventional cultural practices, such as elimination
of infected palms and improved sanitation processes, can only
delay the spread of the disease (Breton et al., 2006), while
extensive physical clearing of infected palms is not economically
feasible. Biological controls of BSR could be the most suitable
approach, as it is safe for the users, and poses little threat to
non-target organisms. However, to develop a highly selective bio-
logical control for BSR is difficult, expensive, time consuming, and
less effective when applied in the field (Damon, 2000).
Host resistance can be used in preventing and controlling plant
diseases. Oil palms from different genetic origins with differences
in disease susceptibility to BSR have been reported (Idris et al.,
2004; Durand-Gasselin et al., 2005). They could serve as useful
genetic resources for the improvement of oil palm tolerance or
resistance to Ganoderma spp., employing a similar strategy which
was used to improve the resistance of oil palm to Fusarium wilt
(Cochard et al., 2005).
2. Ganoderma spp. reported in BSR
The fruiting body of Ganoderma lucidum has gained wide popu-
larity as a dietary supplement especially in China, Taiwan and,
Japan for its perceived health benefits in preventing immunological
diseases, such as hypertension and tumorigenesis (Liu et al., 2002).
Many Ganoderma species are also plant pathogens to khair, grape-
vines, betel palm, rubber, tea, and oil palm (Turner, 1965; Bakshi
et al., 1976; Adaskaveg and Gilbertson, 1987). G. lucidum was first
reported as a causal agent of BSR in oil palm in 1930 (Thompson,
169
1931; Utomo et al., 2005). Subsequently, six additional species
were reported to be associated with BSR in Malaysia and Indonesia,
including Ganoderma boninense, Ganoderma tornatum, Ganoderma
chalceum, Ganoderma zonatum, and Ganoderma xylonoides
(Steyaert, 1967). To date, a total of 15 Ganoderma species have
been detected in oil palms (Turner, 1981). Although G. boninense
was identified as the most virulent species that caused BSR, multi-
ple species could be responsible for the disease on individual trees
(Ho and Nawawi, 1985).
2.1. Life cycle of Ganoderma spp. and its relation to BSR
Ganoderma spp. are classified as basidiomycetes. The life cycle
of G. boninense in relation to BSR has been studied by Hasan and
Flood (2003). Generally, each basidiospore germinates into a
genetically unique monokaryotic hypha which is saprophytic and
able to colonize dead palm wood. Ganoderma spp. have a tetrapolar
mating system which favors outcrossing. They are heterothallic
with two pairs of alleles at two mating loci, thus ensuring
maximum genetic diversity by restricting inbreeding to 25%
(Rees et al., 2009). Hyphae of compatible mating type anastomose
to produce a dikaryotic mycelium which could be potentially inva-
sive. This heterokaryon proliferates and grows, with two haploid
nuclei dividing and multiplying independently in each septated
unit until the life cycle of Ganoderma is completed when the dik-
aryotic hyphae produce a fruiting body known as a basidiocarp
(Hasan and Flood, 2003). The basidiocarp bears specialized cells
called basidia which resemble little clubs where karyogamy occurs.
The basidia then divide meiotically to produce genetically unique
basidiospores (Campbell et al., 2008).
2.2. Transmission of BSR
Various sources of infection and modes of transmission, have
been proposed for BSR. Basidiospores have been proposed as a
source of BSR infection (Pilotti et al., 2003; Rees et al., 2012). Bas-
idiocarps are able to release a high number of basidiospores which
can travel a long distance (Sanderson, 2005) and become a source
of infection for a wounded palm surface created during plantation
harvesting and management (Rees et al., 2012). Anastomosis of
basidiospore germlings could occur on palm surface, debris or
fallen palms in soil causing direct infection via cut fronds or
indirect infection through roots (Rees et al., 2012). Spread through
basidiospores must be accountable for from the genetic diversity of
Ganoderma isolates from the field (Miller et al., 1999; Pilotti et al.,
2003). Root invasion was considered as the primary route of trans-
mission of BSR (Rees et al., 2009). Since Ganoderma spp. have poor
competitive saprophytic capability in soil, colonized debris left in
the field by infested palms from previous planting of coconut or
oil palm have been proposed as a very important source that pro-
vides the substantial amount of inocula required for root infection
(Hasan and Turner, 1998; Rees et al., 2007). The infection of oil
palm is likely to occur when the roots come into contact with inoc-
ula from the debris left in the ground (Flood et al., 2005), or from
roots of neighboring infected palms.
The tetrapolar heterothallism of Ganoderma spp. also explains a
number of phenomena:
1. Genetic diversity of isolates within a plantation was as
great as between plantations whereby the vast genetic
diversity of fungi observed on the infected palms could be
a result of plasmogamy of genetically different mycelia
derived from basidiospores;
2. BSR takes a long time to be evident in the field due to the
requirement of plasmogamy or anastomosis of compatible
170 C.-L. Ho, Y.-C. Tan / Phytochemistry 114 (2015) 168–177
mating types to form virulent dikaryotic mycelium, and
they have weak competitive ability in soil or on organic
debris (Rees et al., 2007);
3. BSR symptoms may not be detectable even though Gano-
derma spp. can be detected on palms whereby the fungus
could be in the form of monokaryotic mycelia living sapro-
phytically on dead tissues on the palm surface since
monokaryotic mycelium are non-infective (Goh, 2005;
Rees et al., 2007).
2.3. Development and progress of BSR in oil palm roots
The development and progress of BSR in oil palm roots have
been studied by Rees et al. (2009) in great detail. Briefly,
infection by Ganoderma spp. is initiated by the penetration of oil
palm root surface (epidermis and exodermis) by fungal mycelia,
followed by a longitudinal progression of hyphae through inner
and thin-walled cortex, and colonization of the lower stem (bole)
eventually. Host cells in newly colonized tissue were shown to
be colonized by intracellular hyphae and contained intact cell wall,
intact cytoplasm, and organelles (Rees et al., 2009). At this stage,
depletion of starch grains in the cytoplasm was observed in host
cells in advance of invasion and in the lower stem of infected oil
palm (Rees et al., 2009). Ganoderma spp. may behave as hemi-bio-
trophs in newly colonized tissues before turning into necrotrophic
pathogens as implicated by extensive degradation of host cell
walls. Cell wall degrading enzymes (CWDEs) such as cellulase,
manganese peroxidases and laccases that are involved in the deg-
radation of cellulose and lignins, are expected to be released by the
fungus. At this stage, the host cells were colonized by hyphae intra-
, intercellularly and intramurally. Rees et al. (2009) also suggested
that the defense response of oil palm may rely on production and
release of antimicrobial compounds rather than on cell wall
strengthening. In the subsequent stage, the oil palm roots are sur-
rounded by a tough and melanised mycelium (pseudo-sclerotium)
with thin-walled hyphae encased by many thick-walled cells (Rees
et al., 2009). This leads to massive hyphal aggregations outside the
oil palm roots, culminating in the formation of basidiocarps and
release of basidiospores (Rees et al., 2009). An infected oil palm
tree may or may not bear any fruiting bodies. The presence of a
fruiting body on an infected palm normally shows that the fungus
has been in the wood for at least several years, and an extensive
decay could have taken place in the stem (Najmie et al., 2011).
3. Plant-pathogenic interactions
The genetics of a host plant and its pathogen were first
explained by the gene-for-gene hypothesis developed by Flor
(1942). According to this hypothesis, the inheritance of both resis-
tance in the host plant and ability of pathogen to cause disease is
controlled by a pair of matching genes, namely the resistance (R)
gene in the plant host, and the avirulence (Avr) gene in the patho-
gen. When a pathogen carrying a single dominant Avr gene is rec-
ognized by a single dominant R gene from a plant host, it fails to
cause any disease. It is thus called an avirulent pathogen and the
host is classified to be resistant, with their interaction known as
being an incompatible interaction (Prell and Day, 2001). In the
absence of an Avr gene in the pathogen and/or an R gene in the
host, the pathogen is considered to be virulent and the host is sus-
ceptible, with their interaction known as a compatible interaction.
In the latter, the pathogen is able to form a parasitic relationship
with the host plant. R gene products may not bind to Avr gene
products directly, but rather detect alterations in host proteins that
are caused by the gene products from the pathogens, as explained
by the guard model (van der Biezen and Jones, 1998). Other models
for R activation including the switch model (Takken et al., 2006)
and decoy model (van der Hoorn and Kamoun, 2008) have been
described.
The interactions of plants and microbial pathogens are among
the most complex phenomena in nature, in which an unlimited
variety of pathogenic molecules can interact with the cellular
component of host plants (Schneider and Collmer, 2010). In brief,
common microbe-associated molecular patterns (MAMPs) or path-
ogen-associated molecular patterns (PAMPs) from microbes
including non-pathogens such as flagellin, lipopolysaccharide, fun-
gal chitin and b-glucans are detected by host plants (Nurnberger
et al., 2004; Zipfel and Felix, 2005). Some pathogens may secrete
CWDEs that can degrade host cell wall producing fragments of
structural polysaccharides known as damage associated molecular
pattern (DAMPs). The recognition of these patterns by transmem-
brane pattern recognition receptors (PRRs) activates the primary
defense response in host plants (PAMP-triggered immunity, PTI)
through signal transduction of a few signalling pathways (de Wit,
2007). PTI could negatively affect colonization of the pathogens
through production of reactive oxygen species (ROS) and phytoal-
exins, cell wall alterations, deposition of callose, and accumulation
of defence-related or pathogenesis-related (PR) proteins (van Loon
et al., 2006; de Wit, 2007).
In addition to PTI, plants have a secondary defense system
(effector-triggered immunity, ETI) whereby the plant resistance
proteins (RPs) encoded by R genes often with characteristic nucle-
otide binding (NB) and leucine rich repeats (LRRs), could monitor
the presence of effectors or their perturbations and trigger RP-
mediated secondary responses. ETI and PTI respond to different
pathogen-derived molecules and they differ by the intensity of
their immune responses. The recognition of effectors can be direct
(Dodds et al., 2006), or indirect (van der Biezen and Jones, 1998). In
order to avoid recognition by PRP, biotrophic pathogens may
produce multiple effectors that suppress PTI. The ETI induced by
a biotrophic pathogen often results in a hypersensitive response
(HR) and other locally induced defence responses that block fur-
ther invasion of the pathogens. HR-associated cell death which
confines pathogens at the infection site and limits their growth
and supply of nutrients, is a typical indicator of host resistance
to biotrophic pathogen.
Over time, pathogens may evolve under selection pressure and
acquire ability to overcome ETI and evade defense responses of
host plants through complete removal or subtle changes of amino
acids of effectors (Chisholm et al., 2006; Jones and Dangl, 2006).
The functions of effectors include suppression of PTI; suppression
of HR (Janjusevic et al., 2006); induction of abscisic acid (ABA)
pathway to facilitate disease (de Torres-Zabala et al., 2007);
production of metabolites that suppress salicylic acid (SA)- and
jasmonic acid (JA)-induced defence responses; as well as establish-
ment of disease symptoms (Zhao et al., 2003; Brooks et al., 2005).
When an effector can no longer be detected by the host plants,
fewer RPs will be induced. The failure of host plants to trigger
ETI leads to susceptibility to pathogen.
The presence of R gene or RP which is specific against BSR in oil
palm has not been investigated. However, sequencing of a partial-
length disease resistance gene homologue encoding NBS–LRR type
R protein in oil palm has been reported (Chin et al., 2012). Many
sequences related to R genes have also been recently identified
from the transcriptomes of oil palm (Low et al., 2014). Despite this,
the existence and involvement of the R gene in BSR which is caused
by a hemi-biotroph, remain a speculation without evidence
showing that these putative R proteins are involved in disease
resistance. So far the R genes associated with plant resistance to
necrotrophs have not been reported, except for a Toll/interleukin
1 receptor domain R protein in Arabidopsis which conferred resis-
tance to Leptosphaeria maculans (Staal et al., 2008).
 C.-L. Ho, Y.-C. Tan / Phytochemistry 114 (2015) 168–177
The heterogeneity of commercial oil palm populations derived
from crosses of dura and pisifera could create a selection pressure
for Ganoderma spp. that favor outcrossing. If the R protein for
BSR does exist, tetrapolar heterothallism may allow Ganoderma
spp. to segregate for more aggressive pathogenicity to overcome
host defense response. Given the way Ganoderma spp. reproduce
and the genetic divergence of their basidiospores, Durand-
Gasselin et al. (2005) have suggested breeding for partial resistance
as a means for durable and long term solution to the BSR problem
in oil palm. This can be achieved by integrating many genes of
minor effect to provide reasonable levels of resistance in some
oil palm lines. Furthermore, oil palm with an ‘‘all-or-nothing type’’
of resistance has not been reported.
Biotrophs and necrotrophs are fundamentally different in their
infection processes, the nature of effector proteins, and the elicited
host-defence responses (Laluk and Mengiste, 2010). Biotrophs
grow between host cells and invade relatively few host cells, as
well as secrete a limited amount of CWDEs and generally lack
phytotoxic compounds (Mendgen and Hahn, 2002). Through this
feeding activity, biotrophs establish a long-term feeding relation-
ship with the living cells of their hosts. Attempted infection by bio-
trophic pathogens would trigger plant immune responses,
including HR which causes cell death at the site of infection. HR
confines the spread of a pathogen by abolishing its nutrient supply,
thereby limiting the growth of biotrophic pathogen. Conversely,
cell death is an indicator of successful infection of necrotrophs
which extract nutrients from dead cells (Govrin and Levine,
2000). In contrast to biotrophs, nectrophs secrete diverse phyto-
toxic compounds and CWDEs to induce cell necrosis and cause
leakage of nutrients. Plant mutants with enhanced cell death
showed increased resistance to biotrophic pathogens but were
susceptible to necrotrophs (Veronese et al., 2004). Generally, cell
death promotes susceptibility of plant host to necrotrophs; how-
ever, it is unknown whether it also applies to all necrotrophs
(Laluk and Mengiste, 2010).
Many fungi that are considered as necrotrophs may be hemi-
biotrophs, as they have biotrophic stage early in the infection pro-
cess, including Ganoderma spp. are amongst this group (Rees et al.,
2009). Infected oil palm may respond to the invasion of Ganoderma
spp. (which behave as biotrophs) at the initial stage, by inducing
HR. The induction of HR, which is in favor of necrotroph growth
may trigger the switch of Ganoderma spp. from biotrophs to necro-
trophs, and render the oil palm susceptible to BSR. Nevertheless,
HR was reported to be associated with the resistance of plant hosts
to two hemi-biotrophs Magnaporthe oryzae (Jia et al., 2000) and
Phytophthora infestans (Vleeshouwers et al., 2000). In addition,
ROS was found to have contrasting roles in plant defense depend-
ing on the lifestyle of the pathogens and the kinetics of the oxida-
tive burst (Lamb and Dixon, 1997). At early stages of infection, ROS
may act as signalling molecules that activate various immune
responses in plants, resulting in resistance to both biotrophs and
necrotrophs. Upon the establishment of infection, ROS may
promote cell death causing HR-associated cell death in biotrophs,
and also susceptibility to necrotrophs. The associations of HR-
associated cell death and the kinetics of the oxidative burst to
the susceptibility of oil palm to Ganoderma spp. await further
investigation. Knowledge on the ability of oil palm cells to attenu-
ate necrosis or to alleviate the effects of necrosis is lacking.
The interplay of hormones such as JA, SA and ethylene and their
related pathways also contribute to the resistance and susceptibil-
ity of plants to pathogens. JA signalling and ethylene-response
pathway are important to the resistance of plant hosts to necro-
trophic pathogens (Glazebrook, 2005; Laluk et al., 2011), whereas
the accumulation of SA increases the resistance of plant host to
hemibiotrophic pathogens but promotes the susceptibility to
necrotrophic pathogens (Veronese et al., 2004, 2006). The roles of
171
different phytohormones during BSR have not been investigated.
The question as to whether oil palm can also switch its defense
responses rapidly through the interplay of hormones according
to the switch in the feeding modes or life styles of Ganoderma
spp. remains unanswered.
The ability of necrotrophs to induce necrosis is central to their
successful invasion, while the ability of host plants to counter fun-
gal toxins (or other virulence factors) or their effects on necrosis
are major factors in host resistance (Mengiste, 2012). Although
the production of necrosis and ethylene inducing proteins which
are able to cause cell death in dicotyledonous plants has been
reported in necrotrophic fungal and bacterial species (Staats
et al., 2007), little is known about production of phytotoxic com-
pounds in Ganoderma spp., including their targets and ability to
cause cell death to monotyledonous plants. The published genome
of G. lucidum (Liu et al., 2012) may provide insights into the pro-
duction of phytotoxic compounds and wood degradation by fungi
belonging to the same genus. Although there are efforts by the
Malaysian Palm Oil Board (MPOB; www.mpob.gov.my/Ganoder-
ma/), ACGT Sdn. Bhd. (www.acgt.asia/press/pdf/02Nov2010.pdf),
Malaysian Genomics Resource Centre (MGRC, (www.mgrc.
com.my/)) and Felda Agricultural Services Sdn. Bhd. to sequence
the genome of G. boninense in recent years, the sequences have
not been made available in the public domain. The availability of
this data will enhance our understanding of the potential virulence
genes involved in Ganoderma infection of oil palm and enable us to
develop better methods for BSR detection and to devise more effec-
tive strategies to prevent BSR.
As white rot fungi, Ganoderma spp. are able to degrade the
lignin component in host cell walls through the production and
secretion of CWDEs such as lignin peroxidases, manganese perox-
idases and laccases (Paterson et al., 2009; Liu et al., 2002). In gen-
eral, lignin protects cell wall polysaccharides from microbial
degradation. The lignin fraction in the oil palm trunk predomi-
nantly comprised syringyl unit (S-type lignin) (Sun et al., 1999),
which is presumably more susceptible to degradation than the
guaiacyl unit (G-type lignin). Paterson et al. (2009) have proposed
improvement of oil palm resistance to Ganoderma spp. through
genetic manipulation of lignin biosynthesis in oil palm. Techni-
cally, this can be achieved by producing transgenic oil palms with
lignin which are rich in guaiacyl unit through genetic manipulation
of enzymes/pathway involved in its biosynthesis. The enzymes and
pathway involved in lignin biosynthesis have been well character-
ized in higher plants (Vanholme et al., 2010). The newly available
oil palm genome (Singh et al., 2013) can be referred for relevant
gene sequences to be transformed, and using well established oil
palm transformation methods (Parveez et al., 2000; Abdullah
et al., 2005), genetic engineering of oil palm for disease resistance
can be applied. Overexpression of transcripts encoding key
enzymes that produce the precursors of lignin units in the phenyl-
propanoid pathway, and polymerization of lignin, together with
the suppression of enzyme that produce the S-type lignin such as
coniferaldehyde (ferulate) 5-hydroxylase by the RNAi approach,
could be attempted for this purpose. Nonetheless, the manipula-
tion of enzymes involved in phenylpropanoid pathway may affect
other related pathways such as the biosynthesis of flavonones/
flavonols which share the same precursors i.e. p-coumaric, cin-
namic and caffeic acids. The partition of these precursors to differ-
ent pathways justifies further research because the production of
other important secondary metabolites such as phytoalexin isofl-
avonoids, which are important anti-microbial compounds, could
be reduced at the expense of increasing lignin biosynthesis. Fur-
thermore, Skyba et al. (2013) have shown that transgenic poplars
with syringyl-rich lignin are more resistant to degradation by
wood decay fungi including white rot fungi, suggesting that
elevated guaiacyl content does not necessary improved decay
172 C.-L. Ho, Y.-C. Tan / Phytochemistry 114 (2015) 168–177
resistance of wood. The biochemistry of lignolytic enzymes from
Ganoderma spp. should be investigated to shed light on the types
of lignin monomers preferred by these enzymes.
Paterson et al. (2009) also suggested the assessment of the
lignin content of oil palm genetically transformed with Bacillus
thuringiensis toxin genes and their resistance to Ganoderma spp.
based on reports that corns transformed with B. thuringiensis toxin
genes have increased lignin contents (Saxena and Stotzky, 2001;
Poerschmann et al., 2005). However, Bt-corns were not found to
have a significant higher content of lignin by Jung and Sheaffer
(2004) and Poerschmann et al. (2008). Furthermore, the increase
in total lignin in these plants could be due to random and uninten-
tional insertion of B. thuringiensis toxin gene into genes that control
lignin biosynthesis as explained by Paterson et al. (2009). If this is
the case, it is unlikely that the oil palm genetically transformed
with B. thuringiensis toxin genes will have a higher content of lig-
nin. In regard to transgenic oil palms, the acceptability of palm
oil from transgenic oil palms by consumers especially from the
European market remains the main concern of palm oil producers
and oil palm planters (Paterson et al., 2009), and should not be
taken lightly. In light of that, screening and breeding for oil palm
with cell walls enriched in the presumably more resistant G-type
lignin could be performed despite its lengthy process.
4. Gene expression of oil palm roots in response to Ganoderma
infection
Disease resistant oil palms are crucial for sustainable produc-
tion of palm oil. DNA markers that are linked to disease resistance
in oil palm are required for screening and marker-assisted breed-
ing of disease resistant oil palms but have not been reported so
far. An in depth understanding of oil palm defence mechanisms
in response to colonization of Ganoderma spp. at various stages
of infection, are necessary at the transcript level to identify poly-
genic genes that contribute to a reasonable and durable disease
resistance to Ganoderma spp. Knowledge on the molecular interac-
tions between oil palm–Ganoderma may also offer alternatives to
management of BSR disease.
The oil palm molecular defence mechanisms in response to
invasion of Ganoderma spp. can be elucidated by analysing the
expression of individual genes that are homologous to genes
reported as involved in defense pathway in other plants, or by ana-
lysing the global gene expression of the total mRNA populations.
The former approach involves fewer genes, is less costly, less time
consuming and facilitates the profiling of targeted transcripts only;
while the latter approach involves a collection of transcripts that
represent the total mRNA, is in large scale, costly, needs develop-
ment of tools/facilities and is able to provide global information
on gene or protein networks in the plant materials. Both
approaches have been used by researchers to study the gene
expression of oil palm in response to controlled inoculation with
Ganoderma spp.
Prior to the genome sequencing of oil palm and its deposition in
the public database, global analysis of oil palm gene expression
relied on information generated by expressed sequence tags (ESTs)
which are short cDNA sequences generated and sequenced from
the mRNA of oil palm. In the last decade, a total number of
40,809 ESTs from oil palm have been generated from various oil
palm tissues and deposited at the EST database by various groups
of researchers (www.ncbi.nlm.nih.gov/genbank/dbest/, Jouannic
et al., 2005; Ho et al., 2007; Low et al., 2008). The generation of
ESTs has facilitated the development and fabrication of a cDNA
microarray consisting of more than 3700 cDNA probes for gene
expression study of oil palm (Lim et al., 2010; Tee et al., 2013).
In the preparation of cDNA microarray probes, cDNA sequences
amplified by PCR are fixed onto a microscopic slide with a
chemically treated surface. Fluorescent-labelled mRNA samples
extracted from un-inoculated oil palm seedlings and oil palm
seedlings inoculated with Ganoderma, respectively, can then hybri-
dise with the cDNA microarray. By measuring and analysing the
fluorescence emitted by mRNAs that are complementary to indi-
vidual cDNA probes, the gene expression level of individual
sequences can be profiled and compared between un-inoculated
oil palm seedlings and oil palm seedlings inoculated with Ganoder-
ma. Using this approach, Tee et al. (2013) reported a total of 61
transcripts being differentially expressed (up-regulated more than
twofold or down-regulated less than twofold) in oil palm roots that
have been artificially inoculated with G. boninense for 3 and
6 weeks, respectively, compared to those from un-inoculated roots.
Upon Ganoderma inoculation, oil palm genes could either be up-
regulated in infected oil palms to strengthen the host defense
against fungal invasion including genes encoding pathogenesis-
related protein 1, heat-shock protein-70 cofactor, isoflavone
reductase, early-methionine-labelled polypeptides and early nodu-
lin-20); or be down-regulated due to the suppression of the host
defense system by the pathogen such as genes encoding vicilin-like
antimicrobial peptide, pecanex-like protein and extension-1 (Tee
et al., 2013). Some of these genes that were differentially expressed
in inoculated oil palms are summarized in Fig. 1.
Similar to other plants, oil palm can activate inducible defence
response to pathogenic invasion; however, the effectiveness of
these responses against Ganoderma is unknown. Although global
gene expression analysis has been initiated to analyse the defence
response of oil palm to Ganoderma, the study by Tee et al. (2013)
suffered a major limitation whereby only a small number of gene
sequences (approximately 3000 genes) were included in the cDNA
microarray, and therefore could not provide a complete picture of
the molecular defence response of oil palm against Ganoderma
infection. We still do not have answers to many questions, for
example, what are the oil palm defence genes and mechanisms
besides those already known and reviewed here? Although the
general defence pathways are well characterized in higher plants,
the applicability of those models developed for disease resistance
in model plants has to be evaluated in oil palm. In addition, there
are specific questions that need to be answered; for example, can
oil palm switch its defense responses rapidly to Ganoderma spp.
at biotrophic and necrotrophic phases, through the interplay of
hormones? What are the associations of HR-associated cell death
and the kinetics of oxidative burst to the susceptibility of oil palm
to Ganoderma spp.? Can oil palm produce antimicrobial com-
pounds at the site of infection to suppress the spread of Ganoderma
spp. (Fig. 1)? The availability of the oil palm genome information,
cheaper and faster sequencing service will contribute to the global
gene expression profiling of oil palm genes in response to the path-
ogen soon.
In addition, oil palm ESTs have also provided information for
molecular cloning and characterization of targeted genes. A total
of 22 oil palm genes including those encoding PR proteins i.e., glu-
canases, chitinases, proteinase-inhibitors, defensin and isoflavone
reductase have been identified from the oil palm ESTs for sequence
analysis and gene expression profiling (Naher et al., 2011; Yeoh
et al., 2012, 2013; Tan et al., 2013; Tee et al., 2013). In this
approach, the gene expression level of individual sequences can
be profiled and compared between un-inoculated oil palm
seedlings and oil palm seedlings inoculated with Ganoderma by
quantitative reverse-transcription (qRT)-PCR (Naher et al., 2011;
Yeoh et al., 2012, 2013; Tan et al., 2013; Tee et al., 2013) or
semi-quantitative RT-PCR (Alizadeh et al., 2011).
Induction of pathogenesis-related proteins (PRs) has been found
in many plant species belonging to various families (van Loon,
1999; van Loon and van Strien, 1999). Currently, there are 17
 C.-L. Ho, Y.-C. Tan / Phytochemistry 114 (2015) 168–177 173

pathogen CWDEs from pathogen

pathogen

ROS Strengthening PAMPs DAMPs

of cell wall effector Cell wall
SOD
Cell membrane
Rboh R protein?
? Phytotoxic PRR
compounds? JA
?
?
? ?
ROS Ethylene SA Phe
? PAL
CA
Disease- and HR-
IFR
associated cell
death
Phenolic Phytoalexin
accumulation accumulation

Regulation of gene expression

PR proteins: Stress-related proteins Other proteins

Proteinase inhibitors, Transcription factor UNE10, Auxin-responsive protein IAA7,
defensins, HSP-70 cofactor, wall-associated receptor kinase,
pathogenesis related-1 metallothionien-like, sedoheptulose-1,7-biphosphate,
glucanases Em protein H2, EMZ08, early nodulin, IFR
chitinases Extensin , extracellular ribonuclease, YABBY 2, pecanex-like protein, alpha
SPX-domain containing protein, ketoglutarate-dependent dioxygenase,
vicilin-like antimicrobial peptidase PAL

Fig. 1. Proposed oil palm–Ganoderma molecular interactions and summary of oil palm genes that respond to the infection by Ganoderma boninense. At an early stage of
interaction, fungal pathogen may release pathogen associated molecular patterns (PAMPs) which are recognized by the pattern recognition receptors (PRRs) residing in the
cell membrane. In the necrotrophic phase, the fungal pathogen secretes cell wall degrading enzymes (CWDEs) that degrade cell wall of plant host (including lignin) producing
fragments of structural polysaccharides known as damage associated molecular patterns (DAMPs). The recognition of P/DAMPs by PRRs in the host plant converges into a few
signalling pathways that regulate transcription of defense genes in oil palm including those encoding pathogenesis-related (PR) proteins, stress proteins and other proteins in
the cell. It is unknown whether an effector is released by Ganoderma spp. during its biotrophic phase, which could possibly interact with R protein and elicit HR-associated cell
death via the production of reactive oxygen species (ROS). ROS could also induce disease-associated cell death during necrotrophic phase. The ability of Ganoderma spp. to
produce phytotoxin is unknown. Blue arrows in broken line indicate multiple steps in a pathway. Brown arrows in broken line represent the possible signalling pathways
while the black arrows in broken line represent the roles of protein encoded by some of the up-regulated or down-regulated genes. The blue dotted line arrows show the
possible defence responses in oil palm, whereas the black dotted lines arrows indicate possible interactions (synergy between SA and ethylene, and antagonism between SA
and JA) between hormones. The question marks highlight the gaps that need to be filled in oil palm molecular defences responses to Ganoderma spp. The words in red and
green indicate plant genes that were up-regulated and down-regulated in Ganoderma-inoculated oil palm in comparison to the un-inoculated oil palm root tissues,
respectively. CA, cinnamate; HR, hypersensitive reaction; IFR, isoflavone reductase; JA, jasmonate; PAL, phenylalanine ammonia-lyase; Phe, phenylalanine; ROS, reactive
oxygen species; SA, salicylate; SOD, superoxide dismutase. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this
article.)

families of PR proteins: PR-1 (antifungal), PR-2 (b-1,3-glucanase),
PR-3 (chitinase type I, II, IV, V, VI, VII), PR-4 (chitinase type I, II),
PR-5 (thaumatin-like), PR-6 (proteinase-inhibitor), PR-7 (endopro-
teinase), PR-8 (chitinase type III), PR-9 (peroxidase), PR-10 (ribonu-
clease-like), PR-11 (chitinase type I), PR-12 (defensin), PR-13
(thionin), PR-14 (lipid-transfer protein), PR-15 (oxalate oxidase),
PR-16 (‘oxalate oxidase-like’), PR-17 (unknown), with most of their
activities known (van Loon and van Strien, 1999). The structural
and functional characteristics of each class were well reviewed
(Selitrennikoff, 2001) and will not be discussed here. In this review,
the expression of genes encoding PR proteins and stress related
proteins in Ganoderma-inoculated oil palm tissues, and their possi-
ble roles are summarized (Table 1 and Fig. 1). Only the changes in
gene expression level in Ganoderma-infected oil palm tissues in
comparison to un-inoculated oil palm tissues that are equal or
more than 2-fold will be considered. The genes were up-regulated
possibly as part of plant defence against Ganoderma spp. whereas
the down-regulated genes in oil palm roots were possibly sup-
pressed by the pathogen. Discussion on individual gene expression
profiles has been detailed by individual references and thus will
not be repeated here. Many of these genes (Table 1) display tran-
sient up- or down-regulation at different stages of infection
thereby complicating the analysis of gene expression profiles. Nev-
ertheless, transcript profiling of some of these genes belonging to a
big family, especially genes encoding chitinases and glucanases
from oil palm, has enabled identification of specific isoforms
related to pathogenesis (Naher et al., 2011; Yeoh et al., 2012,
2013). This is especially important for isozymes that may have
many functions other than that in plant defense.
Expression analyses of genes encoding enzymes related to phe-
nylpropanoid and isoflavonoid phytoalexin pathways (Table 1) are
important to understand the molecular defense of oil palm against
Ganoderma spp. since many flavonoids and isoflavonoid are impor-
tant antimicrobial compounds (Kramr et al., 1984; Cruickshank,
1962). The expression of an oil palm gene encoding isoflavone
reductase (EgIFR) was higher in roots inoculated with G. boninense
compared to that of the un-inoculated roots (Tee et al., 2013) sug-
gesting that the biosynthesis isoflavonoid phytoalexin could be
induced by this pathogen in oil palm. Furthermore, the transcript
profiles of EgPAL and EgC4H which encode L-phenylalanine lyase
and cinnamate 4-hydroxylase, respectively, may provide more
information on the biosynthesis of lignin precursors i.e. cinnamate
and p-coumarate. The transcript abundance of EgPAL and EgC4H
was down-regulated in oil palm roots at an early stage of G. bonin-
ense inoculation, supporting the finding of Rees et al. (2009) that
oil palm may not rely on wall-associated responses against inva-
sion of Ganoderma spp. Instead, oil palm may be more reliant on
the production of antimicrobial compounds against the fungal

Table 1
A summary of gene expression profiles of individual defense related genes in inoculated oil palm seedlings compared with un-inoculated oil palm seedlings.
Transcripts
Glucanases
b-D-glucan exohydrolase
Glucan endo-1,3-D-
glucosidase
Chitinases/chitinase-like
Class I chitinase
Class II chitinase
Class III chitinase
Class V chitinase
Proteinase-inhibitor
Bowman–Birk serine
protease inhibitor
Defensin
Pathogenesis-related 1
protein
Ribosome-inactivating
protein
Metallothioneins
Late embryogenesis associated proteins
Early methionine-labeled
polypeptide (group 1
LEA)
Dehydrin (group 2 LEA)
174
Putative functions Abbreviation Gene expression profiles in inoculated oil palm seedlings compared with un- References
inoculated oil palm seedlings
PR-2; degrade fungal cell wall component by hydrolyzing b-1,3- EgGlc1-1 Up-regulated in the leaves of inoculated oil palm seedlings at 3 and 12 wpi Yeoh et al. (2012)
glucosidic linkages and promote the release of cell-wall derived fungal EgGlc5-1 Down-regulated in the roots of inoculated oil palm seedlings from 3, 6
elicitors and12 wpi, and leaves of inoculated oil palm seedlings at 6 wpi
EgGlc5-2 Down-regulated in the roots of inoculated oil palm seedlings from 3, 6
and12 wpi, suppressed in the leaves of inoculated oil palm seedlings at 6
and12 wpi; up-regulated in the leaves of inoculated oil palm seedlings at
3 wpi
C.-L. Ho, Y.-C. Tan / Phytochemistry 114 (2015) 168–177
PR-3, PR-4, PR-8 and PR-11; cleave the b-1,4-glycosidic linkages EgCHI1 Up-regulated in the roots of inoculated oil palm seedlings at 5 wpi Naher et al. (2011)
between N-acetylglucosamine residues in fungal chitin to chitin EgChit1-1 Up-regulated in the leaves of inoculated oil palm seedlings at 3 wpi; Yeoh et al. (2013)
oligosaccharides suppressed in the roots and leaves of inoculated oil palm seedlings at 6 and
12 wpi
EgCHI2 Up-regulated in the roots of inoculated oil palm seedlings at 2 and 5 wpi; Naher et al. (2011)
EgCHI3 Up-regulated in the roots of inoculated oil palm seedlings at 5 wpi Naher et al. (2011)
EgChit3-1 Up-regulated in the roots of inoculated oil palm seedlings at 12 wpi, and Yeoh et al. (2013)
leaves of inoculated oil palm seedlings at 6 and 12 wpi; down-regulated in the
roots and leaves of inoculated oil palm seedlings at 3 and 6 wpi
EgChit5-1 Up-regulated in the leaves of inoculated oil palm seedlings at 3 wpi, down- Yeoh et al. (2013)
regulated in the roots at 6 wpi and leaves of inoculated oil palm seedlings at
3 wpi
PR-6; serine proteinase inhibitors that specifically regulate proteinases EgBBI1 Up-regulated in the roots of inoculated oil palm seedlings at 12 wpi and Tan et al. (2013)
belonging to serine classes leaves of inoculated oil palm seedlings at 3 wpi
EgBBI2 Up-regulated in the roots of inoculated oil palm seedlings at 3 and 6 wpi and
leaves of inoculated oil palm seedlings at 6 wpi
PR-12; cysteine-rich proteins with many different modes of actions, EgDFS Up-regulated in the roots of inoculated oil palm seedlings at 12 wpi; down- Tan et al. (2013)
including anti-microbial, anti-fungal, insecticidal, protease inhibiting, regulated in the roots of inoculated oil palm seedlings at 6 wpi
and a-amylase inhibiting activities (Stotz et al., 2009)
PR-1; antifungal with unknown modes of action, cellular and EgPRP Down-regulated in the roots of inoculated oil palm seedlings at 12 wpi and Tan et al. (2013)
molecular targets leaves at 6 and 12 wpi; up-regulated in the leaves of inoculated oil palm
seedlings at 3 wpi
Ribotoxins, type 2 ribosome-inactivating protein EgT2RIP Up-regulated in the roots of inoculated oil palm seedlings at 12 wpi but Tan et al. (2013)
down-regulated in roots at 3 wpi; up-regulated in the leaves of inoculated oil
palm seedlings at 3 wpi
Intracellular cysteine-rich metal-binding proteins. with reactive MT3-A Up-regulated in the roots of inoculated oil palm seedlings at 21 dpi; induced Alizadeh et al. (2011)
oxygen species scavenging activities in the leaves of inoculated oil palm seedlings at 3 dpi
MT3-B Induced in the leaves of inoculated oil palm seedlings at 7 dpi Alizadeh et al. (2011)
EgMT Up-regulated in the roots of inoculated oil palm seedlings at 12 wpi; down- Tan et al. (2013)
regulated in the roots of inoculated oil palm seedlings at 3 and 6 wpi
Related to development and abiotic stresses EgEMLP1 Up-regulated in the roots of inoculated oil palm seedlings at 6 wpi, down- Tan et al. (2013)
regulated in the leaves of inoculated oil palm seedlings at 3,6 and12 wpi
EgEMLP2 Up-and down-regulation less than 2-fold in all tissues examined
EgDHN Up-regulated in the roots of inoculated oil palm seedlings at 3, 6 and 12 wpi Tan et al. (2013)
and leaves at 3 wpi
 C.-L. Ho, Y.-C. Tan / Phytochemistry 114 (2015) 168–177 175

invasion, which happens at a later stage as evident by the gene

Alizadeh et al. (2011)

Alizadeh et al. (2011)

expression profile of EgIFR (Table 1).
So far, molecular identification and characterization of oil palm
Tan et al. (2013)
Tee et al. (2013)

Tee et al. (2013)

Tee et al. (2013)

Tee et al. (2013) defense genes have been carried out on oil palm seedlings derived
from tenera seeds with different degrees of susceptibility to Gano-
derma spp., and thus may limit the identification of major genes
that contribute to resistance/tolerance. Expansion of these studies
to clonal oil palms that have been proven to be tolerant/resistant
to Ganoderma spp. may provide more consistent and remarkable
Up-regulated in the roots of inoculated oil palm seedlings at 21 dpi; down-
Down-regulated in the roots of inoculated oil palm seedlings at 3 wpi; up-

differences in gene expression profiles that make the identification

regulated in the leaves of inoculated oil palm seedlings at 42 and 63 dpi
Down-regulated in the roots of inoculated oil palm seedlings at 3 wpi

Down-regulated in the roots of inoculated oil palm seedlings at 3 wpi

of resistance genes easier. However, oil palm with an ‘‘all-or-

Up-regulated in the leaves of inoculated oil palm seedlings at 21 dpi
Up-regulated in the roots of inoculated oil palm seedlings at 6 wpi

nothing type’’ of resistance has not been reported. In addition, the

Up-and down-regulation less than 2-fold in all tissues examined

regulated in the roots of inoculated oil palm seedlings at 12 wpi

reproducibility of artificial inoculations of oil palm materials with

Ganoderma spp. is important for gene expression analysis. Reliable
methods which can detect infection stages of oil palm roots without
uplifting the plants from the soil are required. In most of the previ-
ous studies, destructive sampling has been practised and the oil
palm seedlings were harvested according to the duration of inocu-
lation instead of the stage of infection. The duration of inoculation is
not a good indicator for infection stage because the levels of
infection can be easily affected by the age of the plants, size and vir-
ulence of inoculum, shading and temperature (Rees et al., 2007).
Due to the non-synchronous nature of infection of Ganoderma
spp., the tissues collected were not specific to a particular stage of
infection alone, in fact they were either a mixture of healthy and
necrotic tissues, or a mixture of tissues at various infection stages
most of the time. The difficulties encountered in correlating tempo-
ral gene expression to infection stages have to be overcome.

5. Conclusions

In recent years, comparative analyses of global gene expression

and transcript profiles of oil palm PR proteins in inoculated and un-
EgC4H
EgPAL

EgCHI
EgIFR

SAD1

SAD1

inoculated oil palm seedlings have started to reveal the molecular

defence mechanisms of oil palm against Ganoderma spp. Findings
Catalyzes conversion of saturated stearic acid to mono-saturated oleic

inducing salicyclic acid and jasmonic acid-mediated defence response

on the gene regulation of oil palm defence pathways although frag-

mentary have brought us a step closer toward the understanding of
acid; regulates the defence response of plants to pathogens by

polygenic resistance of oil palm against Ganoderma spp. Polygenic

resistance comprising many genes of minor effect may provide
reasonable levels of resistance in oil palm. Identification and char-
acterization of genes that contribute to this polygenic resistance
could be useful for screening for disease resistant oil palm and
marker-assisted breeding programme. Knowledge on associations
Enzymes related to phenylpropanoid and isoflavonoid phytoalexin pathways

of HR-induced cell death and ROS kinetics, to the susceptibility

Biosynthesis of isoflavonoid phytoalexin

Catalyzes the biosynthesis of cinnamate

of oil palm to Ganoderma spp.; the interactions of phytohormones

Catalyzes cinnamate to p-coumarate

(salicylate, jasmonate and ethylene) at early and late stages of BSR;

and cell wall strengthening through increased production of G-
Catalyzes chalcone to flavone

type lignin; are required to understand the molecular interactions

between oil palm and Ganoderma spp. The advance of new gener-
wpi, weeks post inoculation; dpi, days post inoculation.

ation of sequencing technologies and availability of oil palm gen-

ome information will hasten the pace of gene expression
profiling in oil palm and Ganoderma spp. in the near future. The
lignolytic activities of plant CWDEs from Ganoderma spp. on differ-
D9 stearoyl-acyl carrier protein desaturase

ent types of lignin are obscure, and await further investigations.

The breeding of disease resistant oil palms should be considered
despite the lengthy process required. The issues of acceptability
of consumers to genetically manipulated oil palm have to be
Chalcone flavone isomerase

Oil palm D9 stearoyl-acyl

considered prior to genetic modification of oil palm with higher

Cinnamate 4-hydrolase
L-Phenylalanine lyase

resistance to Ganoderma spp.

Isoflavone reductase

carrier protein
desaturase

Acknowledgements

This project was funded by RUGS Initiative 6 UPM (05-02-11-

1408RU) and Putra Grant (GP-IPB/2013/9413601). Tan Y.-C. was
supported by Malaysia Ministry of Science, Technology and
176 C.-L. Ho, Y.-C. Tan / Phytochemistry 114 (2015) 168–177
Innovation under National Science Fellowship (NSF). We thank the
reviewers for their constructive comments.
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C.-L. Ho is currently an associate professor at the Fac-
ulty of Biotechnology and Biomolecular Sciences in
University Putra Malaysia. She obtained her PhD from
the Faculty of Pharmaceutical Sciences, Chiba Univer-
sity, Japan. Her current research is focused on oil palm
defense responses to Ganoderma sp. and transcriptomes
of an agarophyte, Gracilaria changii. She has published
more than 60 peer-reviewed research and review
papers.
Y.-C. Tan graduated in 2008 from Universiti Malaysia
Sabah with a BSc. degree in Biotechnology. After grad-
uation, he worked in Nimura Genetic Solutions (M) Sdn.
Bhd., on bioprospecting and drugs discovery from soil
bacteria (actinomycetes) and fungi. Then, he pursued
his MSc degree in the field of Molecular Biology at
Universiti Putra Malaysia and graduated in 2013. He
studied some of the host defence-related genes in oil
palm when exposed to pathogenic fungus Ganoderma
sp. He is currently working as a genome informaticist in
Codon Genomics Sdn. Bhd. analyzing big data from next
generation sequencing.