Control Measures of Cadang-Cadang Disease on Coconut (Cocos Nucifera) in the

Philippines

Jerrel Ann L. Lagitao1

1
MSc in Plant Pathology, Department of Pest Management – Visayas,

Visayas State University, Visca, Baybay City, Leyte

ABSTRACT

Cadang-cadang disease is widely distributed in the Philippines. It continues to

threaten the coconut (Cocos nucifera L.) industry in the country. Its symptoms develop
slowly but causes premature decline and death of coconut palms. Several approaches were
identified and considered for the management of the disease. But no direct control measures
can be recommended at present. The major objective of research on Cadang-cadang
disease to allow control measures to be developed has been to obtain necessary information
on disease etiology, epidemiology, and host range. The nature of the coconut industry in the
country also requires that control measures should be simple, reliable and cost efficient.

INTRODUCTION
Cadang-cadang is a common viroid disease and the type member of the genus
Cocadviroid within the family Pospiviroidae, the smallest known viroid and infectious
pathogen (Flores et al., 2003; CABI, 2015). The main host of Cadang-cadang is coconut
(Cocos nucifera). The name Cadang-cadang means dead in a local Filipino dialect and is
used to refer to a premature decline and death of coconut palms. Its symptoms develop
slowly and is distinguished initially by the appearance of smaller and rounder nuts with
external scarifications and reduced copra content. Yellow leaf spots develop, new
inflorescences become stunted, partly necrotic, then sterile, and nut production ceases. Leaf
spots enlarge giving fronds a generally chlorotic appearance, crown size declines, and
palms generally die 8 to 16 years after the onset of the disease. Cadang-cadang is usually
observed after palms reach 10 years of age. Incidence then increases linearly for up to 40
years and remains constant for older palms (Zelazny and Pacumbaba, 1982).
Cadang-cadang poses a serious threat in the coconut industry in the country. Its
distribution reaches Bicol Region, Northern Samar, Eastern Samar, Biliran and Quezon.
Approximately 30 million palms are estimated to have been killed by Cadang-cadang during
the last 40 years, but as it seems likely that the disease occurred in the Philippines well
before it was first described, total losses would greatly exceed this estimate. At present, the
disease continues to spread at a gradual rate (Randles et al., 1992). There is no direct
control measure recommended to control Cadang-cadang but several possible strategies
can be considered. More recently, extensive studies were conducted to prove
the viroid nature of the disease and to determine the role in the origins and epidemiology of
CCCVd.

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CADANG-CADANG VIROID
Cadang-cadang was first reported as a lethal disease of coconut palm (Cocos
nucifera) in the central Philippines in 1914 (Ocfemia, 1937). The detection of two small
disease-associated RNAs in 1975 provided the initial clue to the etiology of Cadang-cadang.
Electron microscopy, nucleotide sequencing and transmission experiments that
demonstrated the infectivity of these RNAs finally proved that Cadang-cadang is caused by
a viroid. It is now referred to as the coconut Cadang-cadang viroid (CCCVd). The viroid
causing CCCVd is the smallest known pathogens and they do not have a protein coat, and
consist solely of a small circular, single-stranded infectious RNA molecule that can replicate
in the host cell and be transmitted independently of any other microorganism. It is
transmitted by vegetative multiplication of infected hosts, by seed and pollen and, possibly,
by the action of unknown vector(s) It also has been found only in cultivated plants (e.g.
tomato, coconut, avocado, grapes, hops) and are transmitted mechanically by human
cultural practices.
CONTROL MEASURES OF CADANG-CADANG DISEASE

Necessary and appropriate emergency measures and methodologies in the

treatment of infested coconut trees and other host plants, which may include mechanical,
chemical, and biological measures, as well as the declaration of infested areas to be under
quarantine and the establishment of checkpoints and quarantine stations to prevent the
transportation of unprocessed/untreated parts of coconuts, coconut seedlings and other
host/vector plants from infected areas are the early control measures to mitigate the spread
of such lethal disease.

CONTAINMENT PROGRAM FOR CADANG-CADANG DISEASE

In 2004, the PCA implemented a comprehensive Cadang-Cadang Disease

Containment Program which also resulted to the issuance of a BPI Quarantine
Administrative Order No. 13. Series of 2004 with the SUBJECT: Rules and Regulations
Further Modifying BPI Administrative Order No. 15. Series of 1988 Entitled "Declaring the
Cadang-cadang as a Manageable Disease and Providing for a Containment and Quarantine
Control Program in Affected Areas." The Cadang-cadang Disease Containment Program
has 3 components geared towards increasing coconut production and resumption of coconut
exports: The identification and accreditation of disease-free areas assure importers that
coconut will be sourced from disease free areas that have been subjected to highly sensitive
detection techniques. Establishment of buffer zones in Luzon and Visayas will prevent
further spread of the disease and confine it to Cadang-cadang-affected areas of the country.
Eradication of Cadang-cadang and rehabilitation of affected areas will remove diseased
palms and other possible sources of inocula in the field and replant with certified disease-
free high yielding planting materials.
In addition, no coconut leaves or fronds, young coconut and other raw or
unprocessed or untreated coconut products, coco seedlings, and seedlings of host or vector
plants of Cadang-cadang shall be transported outside of barangays, municipalities or
provinces declared under quarantine especially in Bicol Region, Northern Samar, Eastern
Samar, Biliran and Quezon. With this action this will help to prevent the spread of a pest
from the said areas.

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REPLANTING

The replacement of infected palms, or replanting of infected plantations, appears to

be the earliest recommended means of reducing the losses due to Cadang-cadang (Bigornia
1977). This recommendation was based on the observation that the rate of spread in new
plantings was not influenced by the proximity of infected palms. Although having an
economic cost in lost production and cost of planting, this practice has allowed production to
continue in the Cadang-cadang area.

ERADICATION

Control by removal of diseased palms has been attempted in two sets of trials. In
1952–55, diseased palms were removed every 3 months; 2 years later, the rate of spread
was reported to be about 1/10 that in the untreated area (Zelazny et al. 1982). A trial that
began in 1979 on an isolated island with about 300 000 palms, in which all palms with
disease symptoms were cut annually, showed that there was a marked decline in new cases
of disease during the first two years, but in the third year, as many new cases occurred as at
the beginning of the trial. This trial is continuing, but it is noteworthy that early attempts to
eradicate Cadang-cadang from apparently new outbreak sites have not succeeded—
infected palms still appear in these areas. Control by eradication of all palms with viroid
detectable by molecular methods should therefore be tested.

USE OF RESISTANT VARIETIES

The development of field resistance relies on selecting and breeding from survivor
palms in high-incidence areas. Evaluation of populations exposed to natural Cadang-cadang
infection at several field sites indicated that after 10 years, palms in the Laguna Tall, Rennel
Island Tall, Paulog and Tambolilid populations contained infected individuals, whereas no
palms in the Catigan Dwarf, Malayan Red Dwarf, Malayan Yellow Dwarf and West African
Tall populations were infected (Orolfo et al. 2000). Such trials may reduce the risk of field
infection by selecting parent lines cither for direct planting or for producing FI hybrid seed in
current attempts to exploit heterosis.

VECTOR CONTROL

Although the search for a vector has been unsuccessful, the probability that one or
more exists means that control measures may eventually be directed towards vectors.

USE OF ADVANCED TECHNIQUE FOR DETECTION, IDENTIFICATION AND

DIAGNOSIS FOR CONTROL

Identification and fine characterization of the genetic basis CCCVd diseases using
high-throughput methods, such as genomic biomarkers, next-generation sequencing or
genome-wide association studies, provide invaluable insights into the mechanisms of
disease, and allow physicians to not only assess disease predisposition but also to design

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and implement accurate diagnostic methods and to develop a cooperative management
program.

CCCVd can be identified by analyzing leaf tips using polyacrylamide gel

electrophoresis (PAGE) followed by silver staining, dot-blot or gel electroblot hybridization
(Hanold and Randles, 1991; Imperial et al., 1985; Mohamed & Imperial, 1984) or RT-PCRs
(Rodriguez and Randles, 1993). Improved extraction and RT-PCR protocols are available for
the sensitive detection of CCCVd and its variants and its reliable discrimination from other
viroid such as Coconut tinangaja viroid (CTiVd) that produces similar symptoms (Hodgson et
al., 1998; Vadamalai et al., 2006). In addition, a ribonuclease protection assay (RPA)
(Vadamalai et al., 2009) and a reverse transcription loop-mediated isothermal amplification
(RT-LAMP) (Thanarajoo et al., 2014) may also efficiently detect low amounts of CCCVd-
related RNAs. Symptoms are not reliable for the detection of CCCVd as they appear years
after the initial infection (up to 6 years in the field) and they may resemble those caused by
CTiVd (tinangaja disease) or physiological changes due to other biotic (insect, microbes) or
abiotic stresses.

Consequently, a continuing evaluation of disease epidemiology is required to clarify

the pattern of spread of CCCVd and to deduce its source and possible mode of spread. Host
range studies are particularly important for identifying possible virus sources, and common
plants in six monocotyledon and six dicotyledon families have been inoculated or collected
from high-incidence areas, and assayed for CCCVd. The detection of non-palm host(s)
would open up a new dimension in studies.

In a different study, mutants of CCCVd associated with a 'brooming' effect in the

crown of the coconut (where the leaflet lamina is greatly reduced) have been identified. The
increased pathogenicity was shown to be associated with no more than three point
mutations (Rodriguez, 1993; Rodriguez and Randles, 1993; Randles and Rodriguez, 2003).
A range of molecules related to CCCVd in structure and nucleotide sequence has been
detected in coconut palm and in several other tropical monocotyledon families in many
locations both inside and outside the Philippines by hybridization assay (Hanold and
Randles, 1991). They are defined as CCCVd-related RNAs because their degree of
relatedness to CCCVd, and their importance as pathogens, still needs to be investigated. A
study of CCCVd-related RNA in a commercial oil palm identified viroid molecules of various
sizes that contained the minimal CCCVd sequence (Vadamalai, 2005; Vadamalai et al.,
2006). Similar sequences have been detected in oil palm in a Botanical Garden in Java
(Hanold and Randles, 1998). The presence of a CCCVd-related RNA has also been
detected in a limited sample of coconuts in Sri Lanka (Vadamalai et al., 2009). The exact
significance of these findings requires further research. Sequencing of further isolates will
help to provide answers to questions about origin, pathogenicity, epidemiology and
management of CCCVd and its variants. All commercial varieties cultivated in the Philippines
are susceptible with this disease.

MILD STRAIN PROTECTION

Mild strains of viroid may cross-protect against severe strains (Horst 1975; Niblett et
al. 1978), at least in the short term. The lack of other control strategies could favor the use of
mild strain protection. The mild strain of PSTVd differs only slightly in its nucleotide
sequence from the severe strain (Gross et al. 1981), and it seems likely that eventually the
sites on viroid molecules where changes in sequence can be induced so as to modify
pathogenicity will be defined (Keese and Symons 1985). Research on the forms of CCCVd
has shown that considerable variation at the right end of the molecule apparently does not
affect their ability to infect. Variation at the left end of the molecule has not been reported by

4
Haseloff et al. (1982), but Rodriguez and Randles (1993) have described mutations in the
pathogenicity and central conserved domain of CCCVd that are associated with the severe
‘brooming’ form of the disease. Experiments need to be done to determine the stability and
pathogenicity of mutants. Evidence from other viroid has shown that mutation is frequent,
and that passage through different plant species can be associated with selection of viroid
mutants (Fagoaga et al. 1995).

At present, little is known about the natural occurrence of mild strains of CCCVd.
CTiVd shows too much difference in sequence from CCCVd to allow regions of molecular
variation to be defined that may be responsible for the particular symptoms of each disease.
The detection of CCCVd-like sequences in coconut and oil palms in Solomon Islands
(Hanold and Randles 1991) should allow research to begin on the relationship between
sequence and pathogenicity, and initiate the collection of a range of naturally occurring
variants for evaluation in mild strain protection. Preliminary results that have shown that
ginger (Zingiberaceae) and arrowroot (Marantaceae) collected in the Philippines contain
sequences partially homologous with CCCVd (Rodriguez 1993) suggest that other naturally
occurring variants of CCCVd will be found as molecular hybridisation is more widely used in
surveys for the distribution of CCCVd.

CONCLUSION
At present, there is no direct control measure that can be recommended to control
Cadang-cadang but several possible strategies can be considered. Rules and regulations
have been made by the Philippine government to control the spread of this lethal disease.
Exclusion is the only method considered to be effective in controlling the spread of Cadang-
cadang. No genetically resistant or tolerant coconut (Cocos nucifera) cultivars have been
identified (Randles, 1985), but field resistance may be available (Randles and Rodriguez,
2003). On the other hand, replacing infected palms may not decrease disease incidence, but
has allowed reduced production to continue in affected areas (Randles, 1987). Moreover,
research on Cadang-cadang has been to obtain sufficient information on disease aetiology,
epidemiology and host range to allow control measures to be developed.
Thus, as a plant breeder, plant pathology student and at the same time working in a
coconut research center, it is a privilege and great challenge to protect the Philippine
coconut industry and further contributes to the development of simple, reliable and cost-
efficient control measures against CCCVd. Though there are no direct effective control can
be recommended at present, but several possible avenues for future development can be
considered. Work is in progress to improve the sensitivity, rapidity, simplicity and portability
of diagnostic procedures for CCCVd using advanced technologies; continued monitoring,
detection, and survey of CCCVd of their distribution throughout the country; and following
the employed regulations and protocol employed by the Philippine government are essential
prerequisite to a rational management program.

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REFERENCES

Bigornia, A. E. 1977. Evaluation and trends of researches on the coconut cadang-

cadang disease. Philipp. J. Coconut Studies 2:5-34. 

CABI. 2015. Coconut cadang-cadang viroid (cadang cadang disease), Invasive Species

Compendium. CAB International, Wallingford, UK. Available online: www.cabi.org/isc

Fagoaga, C. 1994. Occurrence of Small RNAs in Severely Diseased Vegetable Crops. Plant
Disease. 78. 749. 10.1094/PD-78-0749.

Flores, R., J.W., Randles and R.A.,Owens. 2003. Classification. In: A Hadidi, R.,

Flores, J.W., Randles, J.S., Semancik (eds.). Viroids. CSIRO Publishing, Collingwood,
Australia, pp. 71– 75.

Gross, Hans & Liebl, Ursula & Alberty, Heidemarie & Krupp, Guido & Domdey, Horst &
Ramm, Karla & Sänger, Heinz. 1981. A severe and a mild potato spindle tuber viroid isolate
differ in three nucleotide exchanges only. Bioscience reports. 1. 235-41.
10.1007/BF01114910.

Hanold, D., and J.W. Randles. 1991. Coconut cadang-cadang disease and its viroid agent.
Plant Disease, 75(4):330-335

Hanold, D., and J.W. Randles. 1998. Report on ACIAR-funded research on viroids and
viruses of coconut palm and other tropical monocotyledons 1985-1993. ACIAR Monograph
no. 45. Canberra, Australia: Australian Centre for International Agricultural Research, 222
pp. http://www.agwine.adelaide.edu.au/plant/plant_protection/pv/aciar_viroids.pdf

Haseloff, J., N.A. Mohamed, and R.H. Symons. 1982. Viroid RNAs of cadang-cadang
disease of coconuts. Nature, UK, 299(5881):316-321

Hodgson, R. A. J., G.C. Wall, J.W. Randles. 1998. Specific identification of coconut
tinangaja viroid for differential field diagnosis of viroids in coconut palm. Phytopathology,
88(8):774-781; 19 ref.

Horst, R. 1975. Detection of a Latent Infectious Agent That Protects Against Infection by
Chrysanthemum Chlorotic Mottle Viroid. Phytopathology. 65. 10.1094/Phyto-65-1000.

Imperial, J.S., R.M. Bautista, and J.W. Randles. 1985. Transmission of the coconut cadang-
cadang viroid to six species of palm by inoculation with nucleic acid extracts. Plant
Pathology, 34(3):391-401

Keese, P., M. Osorio-Keese, and R. Symons. 1988. Coconut tinangaja viroid: Sequence
homology with coconut cadang-cadang viroid and other potato spindle tuber viroid related
RNAs. Virology. 1988;162:508–510. doi: 10.1016/0042-6822(88)90497-7.
Mohamed, N. A., and J.S. Imperial. 1984. Detection and concentration of coconut cadang-
cadang viroid in coconut leaf extracts. Phytopathology. 74 (2), 165-169. DOI:10.1094/Phyto-
74-165

Niblett, C., E. Dickson, F. Elizabeth, K. H. Fernow, R. K. Horst, and M. Zaitlin. 1978. Cross
protection among four viroids. Virology. 91. 198-203. 10.1016/0042-6822(78)90368-9.

Ocfemia, G.O. 1937. The probable nature of cadang-cadang disease of coconuts. Phil Agr.
26:338–340.

7
Orolfo, M.B., L.P. Estioko, and M.J.B. Rodriguez. 2000. Screening of coconut populations for
resistance to coconut cadang-cadang viroid (CCCVd). PCA-ARDB Annual Report.

Randies, J.W., D. Hanold, E.P. Pacumbaba and M.J.B. Rodriguez. 1992. Cadang-cadang
disease of coconut palm. In: AN Mukhopadhyay, J Kumar, HS Chaube and US Singh, eds.
Plant Diseases of International Importance, pp 277–295. Prentice Hall Inc., New Jersey.

Randles, J.W., and M.J.B. Rodriguez. 2003. Coconut cadang-cadang viroid. In: Viroids [ed.
by Hadidi, A.\Flores, R.\Randles, J. W.\Semancik, J. S.]. Collingwood, Australia: CSIRO
Publishing, 233-241

Rodriguez, M.J.B. 1993. Molecular variations in coconut cadang-cadang viroid (CCCVd).

PhD Thesis, University of Adelaide, South Australia.

Rodriguez, M.J.B., and J.W. Randles. 1993. Coconut cadang-cadang viroid (CCCVd)
mutants associated with severe disease vary in both the pathogenicity domain and the
central conserved region. Nucleic Acids Research, 21(11):2771

Sathis Sri Thanarajoo, Kong LihLing, Jugah Kadir, Lau WeiHongi, Ganesan Vadamalai.
2014. Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse
transcription loop-mediated isothermal amplification (RT-LAMP).Journal of Virological
Methods, 20219-23.

Vadamalai, G. 2005. PhD Thesis, University of Adelaide. Adelaide, Australia: University of

Adelaide.

Vadamalai, G., D. Hanold, M.A. Rezaian, and J.W. Randles. 2006. Variants of Coconut
cadang-cadang viroid isolated from an African oil palm (Elaeis guineensis Jacq.) in Malaysia.
Archives of Virology, 151(7):1447-1456.
http://springerlink.metapress.com/content/309455v362247577/?
p=ae735118762a404baf8306d8ef463410&pi=17

Vadamalai, G., A.A.F.L.K. Perera, D. Hanold, M.A. Rezaian and, J.W. Randles. 2009.
Detection of Coconut cadang-cadang viroid sequences in oil and coconut palm by
ribonuclease protection assay. Annals of Applied Biology, 154(1):117-125.
http://www.blackwell-synergy.com/loi/aab

Zelazny, B., J.W. Randies, G. Boccardo and J.S. Imperial. 1982. The viroid nature of the
cadang-cadang disease of coconut palm. Scientia Filipina 2:46–63.

Zelazny, B., and E.E. Pacumbaba. 1982. Phytophagous insects associated with cadang‐
cadang infected and healthy coconut palms in South‐eastern Luzon, Philippines. Ecological
Entomology, 7(1), 113-120.