Phytophthora palmivora AS THE CAUSATIVE AGENT OF Theobroma cacao

POD ROT DISEASE

Arranged by:

Thalitha Nindika Nathania P. (B1B017009)

Yasmine Aurellia (B1B017019)

PROJECT BASED LEARNING

MINISTRY OF EDUCATION, CULTURE, RESEARCH AND TECHNOLOGY

JENDERAL SOEDIRMAN UNIVERSITY
FACULTY OF BIOLOGY
PURWOKERTO
2021

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Phytophthora palmivora AS THE CAUSATIVE AGENT OF Theobroma cacao
POD ROT DISEASE

Thalitha Nindika Nathania P. (B1B017009)

Yasmine Aurellia (B1B017019)

To fulfill the requirements for the Project Based Learning of the Soil-Borne Plant Pathogen
course at the Faculty of Biology, Jenderal Sudirman University

APPROVED
June .. , 2021

Project Lecturer,

Prof. Endang Sri Purwati, M. P.

NIP 195808141985032001

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 PREFACE

Praise and gratitude the authors to the presence of Allah SWT for blessings and mercy, so that
this structure task can be completed. The title is about Phytophthora palmivora As the Causative Agent
of Theobroma cacao Pod Rot Disease. The author realizes that in the preparation of this structure
assignment can not be separated from the help and support of various parties, therefore on this occasion
the author would like to thank Drs. Aris Mumpuni, M. Phil, Prof. Endang Sri Purwati, M. P. Dr, Juni
Safitri Muljowati, S. Si., M.P. as a lecturer in the course of Soil Borne Plant Pathogens. Hopefully this
structured task is useful for the readers.

Purwokerto, June 2021

Authors

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 TABLE OF CONTENTS

PREFACE........................................................................................................................................ii
TABLE OF CONTENTS...............................................................................................................iii
LIST OF FIGURE..........................................................................................................................iv
SUMMARY.....................................................................................................................................v
I. INTRODUCTION...............................................................................................................1
A. Background.....................................................................................................................1
B. Objectives........................................................................................................................2
II. LITERATURE REVIEW....................................................................................................3
III. RESULT AND DISCUSSION............................................................................................5
IV. CONCLUSION....................................................................................................................8

REFERENCES................................................................................................................................9

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 LIST OF FIGURE

Figure 3.1. T. cacao pod (a) affected by P. palmivora (b) healthy................................................. 5

Figure 3.2. Sporangium P. palmivora............................................................................................. 5
Figure 3.2. T. cacao pod rot disease............................................................................................... 5

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 SUMMARY

Cocoa (Theobroma cacao) is one of the plantation and trade commodities as well as an
important source of foreign exchange earnings in Indonesia. However, cocoa productivity in
Indonesia is still low, only reaching 900 kg, still far below the potential cocoa productivity of 2
tons. One of the factors that is still low in cocoa production in Indonesia is due to disease
attacks on cocoa plants. Important diseases on cocoa plants in Indonesia, among others, fruit rot
disease (Phytophthora palmivora). The selection method that will be carried out refers to the
results of several researchers selecting cocoa that is resistant to fruit rot disease caused by P.
palmivora. P. palmivora is a pathogen causing pod rot disease in cacao plants.
Pod rot disease caused by P. palmivora is one of the important diseases on cocoa crops.
Pod rot is the most important disease because it may cause loss of cocoa pod. Efforts to reduce
losses due to plant disease infection can be controlled with the right targets and methods. Early
observation and identification of the right disease will ensure successful control. Koch's
postulates are a general guide for identifying pathogens that can be isolated.
Keywords: Theobroma cacao, Phytophthora palmivora, Pod rot.

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 I. INTRODUCTION

A. Background
Theobroma cacao (cocoa) belongs to the kingdom Plantae, division
Magnoliophyta, class Magnoliopsida, order Malvales, family Malvaceae, genus
Theobroma and species Theobroma cacao. T. cacao is one of the plantation
commodities that has a fairly high economic value among other plantation crops and
plays an important role as a source of foreign exchange through exports and encourages
the regional economy, especially in rural areas (Defitri, 2017). However, the cultivation
of T. cacao faces many obstacles, including the attack of disease-causing pests that can
reduce crop production (Pratama et al., 2017).
The biotic environment of the soil consists of living things, both animals (fauna)
and plants (flora) which all or part of their life phases are in the soil system. Plants are
said to be sick if they experience changes in their body's physiological processes caused
by factors that cause disease so that clear symptoms appear with the factors causing the
disease, including abiotic factors consisting of weather, temperature, minerals and toxic
compounds and biotic factors consisting of fungi, bacteria, viruses, mycoplasmas,
nematodes and higher plants (Agrios, 1997).
Phytophthora spp. is an oomycete type of pathogen that causes cocoa pod (fruit)
rot. Some species that have been reported can infect T. cacao were Phytophthora
palmivora, Phytophthora citrophthora, Phytophthora megakarya and Phytophthora
tropicalis (Maharaj et al. 2016). Of the diverse Phytophthora species, P. palmivora has
the broadest distribution in infecting T. cacao trees in tropical plantations. P. palmivora
is a great destroyer. This pathogen favored moisture conditions in soil and T. cacao
trees. The ability to reproduce asexually with coenocytic mycelium will address it as the
soil-borne pathogen in a favorable condition (Edy et al., 2019).
P. palmivora belongs to the kingdom Stramenopila, class Oomycetes, order
Peronosporales, family Pythiaceae, genus Phytophthora and species Phytophthora
palmivora. Within the order Peronosporales, the largest genus with over 120 described
species, Phytophthora sp. is a hemibiotrophic pathogen capable of infecting a wide
range of hosts, including many agricultural crops, worldwide. Hemibiotrophic are the
spectrum of plant pathogens, including bacteria, oomycete and a group of plant

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 pathogenic fungi that keep its host alive while establishing itself within the host tissue,
taking up the nutrients. Some rot pod-causing Phytophthora genus have distinct
geographical distributions. P. palmivora originally isolated from Palmyra palm
(Borassus flabellifer) in 1907, has a pantropical (all tropics) geographical distribution
and is found in virtually all cocoa production areas (Walker, 2020).
Symptoms are abnormalities or deviations from the normal state shown by plants
or plants. A disease can cause different symptoms or can be the same from different
plants. Symptoms of diseased plants can be divided based on the nature of the
symptoms that arise (local and systemic), direct and indirect effects (primary and
secondary), based on symptom size (microscopic and macroscopic), as well as
morphologically and anatomically (hyperplasia, hypoplasia, discoloration, dryness or
wilting, necrosis and growth of fungi) (Sutarman, 2017). In fruit or pod rot disease,
symptoms can occur at various ages of pod. The color of the pod begins to change from
the tip of the pod or near the stalk, then quickly spreads throughout the pod and finally
the pod turns black. On the surface of the diseased pod and turns black a powdery layer
appears, consisting of secondary fungi that form many spores (Semangun, 1990).
Symptoms are often very specific depending on the species that infects it, so that the
symptoms of the disease can be used to identify the type of pathogen that infects in the
field (Agrios, 1999).
Plant diseases can cause losses both in quantity and quality of crop yields.
Efforts to reduce losses due to plant disease infection can be controlled with the right
targets and methods. Early observation and identification of the right disease will ensure
successful control (Abadi, 2005). Koch's Postulates or Henle-Koch's Postulates consist
of four criteria formulated by Robert Koch in 1884 and published in 1890. Koch's
postulates developed in the 19th century as a general guide for identifying pathogens
that can be isolated by certain techniques (Suada & Suniti, 2014).
B. Objectives
The objectives to be achieved from this project are:
1. To understand the type and development of T. cacao pod rot disease symptoms.
2. To understand the stages to find out the cause of disease in T. cacao pod.

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 II. REVIEW OF LITERATURE

In Theobroma cacao, the infection of Phytophthora palmivora could occur in almost

whole parts of the plant such as the stem, flower, leaf surface and pod. T. cacao pod was the
most susceptible part against the invasion of P. palmivora. Pathogens could infect at all stages
of pod development and immature pod was the most susceptible phage toward pathogen attack
(Komalasari et al., 2018). P. palmivora, as an oomycete, is part of a distinct group of fungus-
like eukaryotic microbes. It shares a range of morphological features with fungi, but it
possesses other features unique to plants, such as the major component of its cell wall being
cellulose, unlike true fungi, which consists mainly of chitin. Another feature is that its
mycelium is composed of hyaline, branched and non-septate filaments, while fungal hyphae
have septate. The dispersal of P. palmivora by wind or water is achieved by asexual sporangia
which develop at the ends of specialized hyphal tips (Walker, 2020). According to Purwantara
(2008), soil from T. cacao plantations in West Java is a consistent source of large and consistent
P. palmivora inocula and that P. palmivora infection from soil to T. cacao pods appears to be
mainly through contact or splashing of rainwater.
Pod infection leads to pod rot (black pod) and any stages of pod development and parts
of all the pods are susceptible to infection. Immature pods between 10 and 20 weeks were
reported to have the highest disease incidence when pod production dynamics. According to
Hebbar (2007), such infected immature pods are rendered useless, while for ripe pods, it results
in a reduction in bean quality. The initial symptom observed for all Phytophthora genus on T.
cacao pods is the appearance of a small translucent spot. The appearance of the spot takes about
2-3 days after infection, to manifest. The spot then turns brown and eventually darkens. Under
humid conditions, the spot spreads rapidly to cover the entire pod in 7-14 days. Three to five
days after the appearance of the first symptom, whitish spores are produced. Black pod disease
incidence in the field is influenced by environmental conditions. Numerous studies have
established the role of climatic factors on the incidence of black pod disease, caused by
Phytophthora spp. Rainfall, high relative humidity, and low temperature are known to create
favourable humid conditions for the development of the disease (Akrovi, 2015).
Studies by Ali et al. (2015) and Tey (1982) demonstrated that P. palmivora cysts
formed germ tubes and appressoria on T. cacao pod husks and leaf tissues. P. palmivora causes

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two main types of disease on T. cacao trees, which are black pod and stem canker. In black
pods, pods or cherelles (immature pods) can be infected at any place on the surface. Studies in
12 diverse T. cacao genotypes demonstrated that germinating zoospores of P. palmivora could
penetrate through stomata, epidermal hair base, scar and by direct penetration of pods.
Symptoms are a brown or black spot on the pod, which spreads to cover the whole pod
(Walker, 2020).
Correct identification of plant pathogens is critical and fundamental to population
genetics, epidemiological studies and the development of disease control strategies (Akrofi,
2015). The Koch postulate developed in the 19th century as a general guide for identifying
pathogens that can be isolated by certain techniques. In 1880, Koch took advantage of advances
in laboratory methods and determined the criteria needed to prove that a specific microbe was
the cause of a particular disease. These criteria are known as Koch's Postulates. According to
Neville et al. (2018), Koch's original postulates consist of four criteria. First, microorganisms
must be present in every case of disease. Second, microorganisms must be isolated from
diseased hosts and grown as pure cultures in the laboratory. Third, microorganisms must have
caused the same disease when introduced to a new host. Fourth, microorganisms must be
recovered from a new host. The four microorganisms can be isolated again from the infected
plants. All four criteria must be met to determine causation between parasites and disease.

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 III. RESULT AND DISCUSSION

The results of a review of several references (Figures 3.1, 3.2 and 3.3) macroscopically
infected cocoa pods show symptoms of blackish chocolate, microscopically Phytophthora
palmivora sporangium and its secondary disease (stem canker).

Figure 3.1. T. cacao pod (a) pod affected by P. palmivora infection symptoms (b) healthy
………...….pod without symptoms of infection. Source: Nurfianti, 2019.

Figure 3.2. Sporangium which were directly isolated from the surface of infected T. cacao
pods. ..….Source: Komalasari et al., 2018.

Figure 3.3. T. cacao pod rot disease (down arrow) followed by stem canker (up arrow). Source:
...Edy et al., 2019.
The disease observed was from the pathogen species Phytophthora palmivora. According
to Sutarman (2017), the types of symptoms that appear on diseased plants can be divided based
on the nature of the symptoms that arise, direct and indirect effects, symptom size, as well as

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morphology and anatomy. Based on the nature of the symptoms that arise, P. palmivora is
included in the systemic symptoms. Systemic symptoms are symptoms that arise due to diseases
that attack all parts of the plant. According to (Komalasari et al., 2018), P. palmivora can occur
in almost all parts of the plant such as stems, flowers, leaf surfaces and pods, this makes P.
palmivora included in the type of systemic symptoms. Based on direct and indirect effects, P.
palmivora is included in the primary symptom. Primary symptoms are symptoms that arise
directly on the part of the plant where it is infected. Pod rot caused by P. palmivora according to
Akrovi (2015), is characterized by early symptoms in the form of the appearance of spots on the
pod surface. The appearance of these spots takes about 2-3 days after infection, to manifest on
the pod that will be infected.
Based on the size of the symptoms, P. palmivora included in the macroscopic symptoms.
Macroscopic symptoms are symptoms of a disease that can be seen with the naked eye.
Symptoms of pod rot due to P. palmivora can be observed directly marked by brown to black
spots located at the bottom of the pod (Venegtern et al., 2018). Based on morphology and
anatomy, P. palmivora is included in the symptoms of discoloration. According to Hebbar
(2007), this color change begins with the appearance of small transparent spots on the pod. Then
the spots will be brown, then turn black and expand rapidly until the whole pod is covered.
The development of P. palmivora infection symptoms on T. cacao pods with the initial
time of infection symptoms appearing on the 3rd to 7th day after inoculation, characterized by
the appearance of a blackish brown color on the surface of the T. cacao pod (Fig.3.1) then
becoming wet rot and then the symptoms spread to cover the entire pod surface. On the
blackened part, a white powdery layer will appear which is the spore of P. palmivora. According
to Afriyeni (2013), the symptoms start from the tip of the pod, there is a collection of mycelium
which is white, yellow and brown spots and there is a spiral-shaped circle on the surface of the
pod skin. In addition, P. palmivora was also found in pods which have the characteristics of a
soft, black base that extends almost to cover the entire surface of the pod skin, covered by a
collection of white mycelium like flour and brown spots on the skin surface (Nurfianti, 2019).
According to Komalasari et al. (2018), the initial symptom of P. palmivora infection
was spot on T. cacao pod which would quickly develop within 14 days and then could extend
covering the whole pod’s surface. Such symptoms were various on pods, from the tip of pod,
base of pod close to stalk and irregular pattern. Further infection was indicated with the

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emergence of white powder which was the sporangium of P. palmivora on the surface of
diseased pod (Fig.3.2). At the same time, neighboring pods either on the same or different trees
would express varying symptoms.
According to Edy et al. (2019), the symptoms of T. cacao black pod were commonly
wet brown spots on the pod surface and in some cases followed by stem canker (Fig.3.3). In
stem canker, P. palmivora mycelia spread from infected pods (Walker, 2020). Usually brown to
black spots located at the base port of the pods, especially at the pods that were hanging below
part of the T. cacao tree which was adjacent to the soil. The brown spot subsequently spread on
the pod surface. Once the spot covers the entire pod surface, the infection may continue to the
beans. The pathogen attacked both young and old pods. However, farmers can still harvest T.
cacao beans before more damage occurs. Occasionally, infected pods also white mycelium
presence on infected trees. The pathogen can penetrate the wax layer before the cuticle of the T.
cacao pod through the epidermis (Vanegtern et al., 2018).
Based on Nurfianti's research (2019), the technique used to test Koch's postulates on T.
cacao pods begins with isolation, namely taking the source of P. palmivora inoculum from T.
cacao pods infected with pod rot disease, then inoculating them into healthy T. cacao pods. This
is in accordance with the statement of Hafsah & Zuyana (2013), Koch's Postulate Koch's
Postulate technique begins with transmitting pathogens to healthy cocoa pods until they show the
same symptoms.

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 IV. CONCLUSION

Based on the result and discussion, we conclude that:

1. Based on the type of disease symptoms, P. palmivora included systemic symptoms, primary
symptoms, macroscopic symptoms and discoloration. Based on the development of the
disease, the initial symptoms are characterized by the appearance of a blackish brown color
on the surface of the pod and eventually covers the entire surface of the pod.
2. Koch's postulate test started with the isolation of P. palmivora from infected T. cacao pods
and inoculated to healthy T. cacao pods.

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Abadi, A., 2005. Ilmu Penyakit Tumbuhan. Jakarta: Bayu Media.

Afriyeni, Y. N., 2013. Jenis-Jenis Jamur pada Pembusukan Buah Kakao (Theobroma cacao L.)
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Agrios, G., 1997. Plant Pathology, 4th ed. San Diego: Academic Press.

Agrios, G., 1999. Ilmu Penyakit Tumbuhan. Yogyakarta: Gajah Mada University Press.

Akrovi, A. Y., 2015. Phytophthora megakarya: A Review on its Status as a Pathogen on Cacao
in West Africa. African Crop Science Journal, 23(1), pp.67-87.

Ali, S., Shao, J., Lary, D., Strem, M., Meinhardt, L. & Bailey, B., 2017. Phytophthora
megakarya and P. palmivora, Causal Agents of Black Pod Rot, Induce Similar Plant
Defense Responses Late during Infection of Susceptible Cacao Pods. Front Plant Sci, 8,
pp.1-10.

Defitri, Y., 2017. Penyakit Busuk Buah Tanaman Kakao (Theobroma cacao l.) serta Persentase
Serangannya di Desa Betung Kecamatan Kumpeh Ilir Kabupaten Muaro Jambi. Jurnal
Media Pertanian, 2(2), pp.98-103.

Edy, N., Angelia, M., Lakani, I. & Panggeso, J., 2019. Current Status on Cacao Disease
Incidence in Central Sulawesi. AGROLAND The Agricultural Sciences Journal (e-
Journal), 6(1), pp.57-62.

Hafsah, S. & Zuyasna, Z., 2013. Uji Patogenisitas Beberapa Isolat Penyakit Busuk Buah Kakao
Asal Aceh dan Evaluasi Efektivitas Metode Inokulasi. Jurnal Agrista, 17(1), pp.42-48.

Hebbar, P. K., 2007. Cacao Diseases a Global Perspective from an Industry Point of View.
Phytopathology, 97, pp.1658-1663.

Herliani, H., Sulaiman, A., Hidayat, M. I. & Hidayat, A. M., 2019. Pathogenicity Test Bacteria
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Komalasari, I., Suryanti, S. & Hadisutrisno, B., 2018. Identification of the Causal Agent of
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Maharaj, S., Sreenivasan, T. N., Motilal, L. A. & Umaharan, P., 2016. Black Pod and Other
Phytophthora Induced Diseases of Cacao: History, Biology and Control, in: Cacao
Diseases. Springer International Publishing, pp. 213-266.

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Neville, B. A., Forster, S. C. & Lawley, T. D., 2018. Commensal Koch's Postulates: Establishing
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