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Genetic diversity among wild forms and cultivated

varieties of Discus (Symphysodon spp.) as revealed by
Random Amplified Polymorphic DNA....

Article in Aquaculture · March 1999

DOI: 10.1016/S0044-8486(98)00478-5

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 Aquaculture 173 Ž1999. 485–497

Genetic diversity among wild forms and cultivated

varieties of Discus žSymphysodon spp. / as revealed
by Random Amplified Polymorphic DNA žRAPD /
fingerprinting
)
Tieh Ling Koh, Gideon Khoo, Li Qun Fan, Violet Pan Eng Phang
Department of Biological Sciences, National UniÕersity of Singapore, Kent Ridge, Singapore 119260

Abstract

Trial-and-error method has been used extensively in the breeding of Discus. There is limited
knowledge on the genetic structure of its species complex and also the genetic basis of its stock
constitution and management. Random Amplified Polymorphic DNA ŽRAPD. fingerprinting was
used to assess the genetic diversity among four wild forms of Discus: Symphysodon discus
ŽHeckel., S. aequiefasciata aequiefasciata ŽWild green., S. a. axelrodi ŽWild brown. and S. a.
haraldi ŽWild blue. and five cultivated varieties of Discus ŽTurquoise, Pigeon, Ghost, Cobalt and
Solid Red.. The Mann–Whitney U-test used in the comparisons among the inter-wild form,
inter-cultivated variety and between wild form and cultivated variety similarity indices revealed
that the gene pool of the cultivated varieties of Discus is smaller than that of the wild Discus
forms. Unweighted pair–group method with arithmetic means ŽUPGMA. phenogram showed that
the Heckel Discus Ž S. discus . is genetically the most divergent in relation to the other three wild
forms, being 2.89 times further in mean genetic distance from the other three wild forms ŽWild
green, blue and brown. than Wild green to the other two wild forms ŽWild blue and brown.. The
cultivated varieties is 3.18 times genetically closer to the three S. aequiefasciata wild forms ŽWild
green, blue and brown. Žmean genetic distances 0.033. than to S. discus ŽHeckel. Žmean genetic
distances 0.105.. This suggests that the S. aequiefasciata wild form is the more likely genetic
origin of the cultivated varieties. In addition, there is no distinct clustering of individuals from the
same cultivated variety indicating the lack of a genetic basis for the present phenotypic
classification of the cultivated varieties. Outcrossing with the wild forms especially, the Heckel

)
Corresponding author. Tel.: q65-874-2707; Fax: q65-779-2486; E-mail: dbsphang@nus.edu.sg

0044-8486r99r$ - see front matter q 1999 Elsevier Science B.V. All rights reserved.
PII: S 0 0 4 4 - 8 4 8 6 Ž 9 8 . 0 0 4 7 8 - 5
486 T.L. Koh et al.r Aquaculture 173 (1999) 485–497

Discus is recommended to increase the level of genetic variability in the cultivated varieties.
q 1999 Elsevier Science B.V. All rights reserved.

Keywords: Symphysodon discus; S. aequiefasciata; Discus; DNA polymorphisms; Random Amplified Poly-
morphic DNA ŽRAPD.

1. Introduction

The Discus is a popular and expensive aquarium fish. Discus cultivated varieties were
produced from interbreeding and selection among its wild forms. Trial-and-error breed-
ing programs were often implemented to produce Discus of novel characteristics ŽChan,
1991.. For most cultivated varieties, the desirable phenotypic traits have not been fixed
since the genetic basis is still not known. As a result, many of these varieties do not
breed true.
In addition, many cultivated varieties are repeatedly interbred among each other to
produce other new varieties and the genetic background for such crossings had not been
recorded. It is therefore virtually impossible to determine the lineage of most cultivated
Discus varieties ŽBurgess, 1991.. Due to this lack of the knowledge of the genetic
origins of most cultivated varieties, they can only be classified loosely into different
groups based on the phenotypic characteristics of the fish. Such kind of classification is
common in the aquarium literature ŽAxelrod et al., 1991; Yamada and Mori, 1991;
Scamen, 1994. despite the lack of a scientific basis.
Proper management and breeding programs have to be implemented to preserve
genetic variability and prevent inbreeding depression that will probably result from the
present state of unplanned breeding in Discus. However, to carry out such programs
information on the genetic relationships among these cultivated varieties and their
genetic relationships with the wild forms is fundamental. A scientific classification for
all existing Discus varieties that can be accepted worldwide is also necessary as it will

Table 1
Wild forms and cultivated varieties of Discus studied
Wild forms Discus number Source Cultivated variety Discus number Source
Wild blue L1, L2, L3, L4, L5 4 Turquoise T1, T2, T5 1
T3 2
T4 3
Wild brown R1, R2, R3, R4, R5 1 Pigeon P1, P2 3
P3, P4, P5 1
Wild green N1, N2, N3, N4, N5 4 Ghost G1, G2, G3 1
G4, G5 3
Heckel H1, H2, H3, H4, H5 1 Cobalt C1, C2, C3 1
3 C4, C5 3
Solid Red S1, S5 1
S2, S3, S4 3

1sGan Aquarium Fish Farm ŽSingapore., 2 sGoldwater Aquarium ŽSingapore., 3sSinta Aquarium
ŽSingapore., 4 s International Fisheries ŽSouth America..
 T.L. Koh et al.r Aquaculture 173 (1999) 485–497 487

facilitate the implementation of such programs and the accurate exchange of information
among breeders.
Random Amplified Polymorphic DNA ŽRAPD. fingerprinting was used in our study
to analyse the genetic relationships among four Discus wild forms and five cultivated
varieties. This fingerprinting method was first described by Welsh and McClelland
Ž1990. and Williams et al. Ž1990. in which the amplification of random segments of
DNA in the genome is carried out by polymerase chain reaction ŽPCR. using single
primers of arbitrary nucleotide sequence, typically of a length of 10 nucleotides. It is
able to provide a convenient and rapid assessment of the differences in the genetic
composition of related individuals ŽKazan et al., 1993.. RAPD fingerprinting has been
used recently in many studies for the analysis of phylogenetic and genetic relationships
among organisms ŽStiles et al., 1993; Bardakci and Skibinski, 1994; Orozco-Castillo et
al., 1994; Van Rossum et al., 1995..

2. Materials and methods

2.1. Source of Discus

The wild Discus: Wild blue, Wild brown, Wild green and Heckel, were imported
from their native habitats in the Amazon Basin. The cultivated varieties: Turquoise,
Pigeon, Cobalt, Ghost and Solid Red were either bred in Singapore, Malaysia or
Thailand. Five fish from each of the four wild forms and five cultivated varieties of
Discus were studied giving a total sample size of 45 fish ŽTable 1.. As our aim was to
investigate the overall genetic diversity among the wild forms and cultivated varieties of
Discus, we decided to analyse a small number of fish from all the wild forms and five
cultivated varieties rather than to study a large number of fish from a smaller number of
wild forms and cultivated varieties.

2.2. Extraction of DNA

For DNA extraction, 0.08–0.10 g of tissue was excised from the anal fin of each
Discus fish. The tissue was homogenised in 900 ml of SET buffer Ž50 mM Tris pH 8.0,
100 mM EDTA pH 8.0 and 100 mM NaCl. and 100 ml of 10% sodium dodecyl sulphate
ŽSDS.. It was incubated overnight in 100 ml 10 mgrml proteinase K ŽPromega, USA.,
100 ml 1 M dithiothreitol ŽDTT. ŽSigma, USA. and 40 ml 10 mgrml RNase A
ŽBoehringer Mannheim, Germany. at 558C. The lysate was then centrifuged at 6500 = g
for 15 min to remove cell debris. Phenol–chloroform extraction was used for extracting
the genomic DNA. Genomic DNA was precipitated by the addition of 100 ml of 3 M
sodium acetate followed by 2–3 volumes of ethanol Žy208C. and pelleted down at
6500 = g for 15 min. The DNA pellet was washed once in 3 ml of 70% ethanol and
vacuum-dried for 1–3 h. The dried pellet was then resuspended in 300 ml of TE buffer
Ž10 mM Tris–HCl pH 8.0 with 1 mM EDTA, pH 8.0..
488 T.L. Koh et al.r Aquaculture 173 (1999) 485–497

2.3. Amplification of DNA

PCR was carried out in 80 ml reactions containing 0.5 mg DNA sample, 16 ml 4 =

PCR buffer with 15 mM MgCl 2 ŽPromega, USA., 0.1 mM primer, 200 mM dNTPs
ŽPromega, USA., 2U Taq polymerase ŽPromega, USA.. Five arbitrary primers were used
in this study Žsee Table 2.. The three primers: NUSZG4, 6 and 8 were provided by
Dinesh et al. Ž1993. and the other two primers: Operon A3 ŽOPA-3. and Operon A4
ŽOPA-4. were obtained from Operon Technologies ŽUSA.. PCR was performed using
the GeneAmp PCR system 9600 ŽPerkin Elmer, USA. programmed for an initial
denaturation step of 5 min at 988C followed by 30 cycles, each consisting of 948C
denaturation for 1 min, 328C annealing for 1 min and 728C extension for 1 min. A final
extension for 10 min was performed at 728C and the PCR products were then held
indefinitely at 48C. A negative control, consisting of all the reaction components except
template DNA, was also included for each amplification.

2.4. Detection of amplification products

PCR products were resolved by urea-polyacrylamide gel electrophoresis Ž8% resolv-

ing and 3% stacking. at 110 V for 14 h using the Protean II xi cell ŽBio-Rad, USA..
Twelve microliters of PCR products were loaded together with 2 ml of loading buffer
Ž50 mM EDTA, 30% glycerol, 0.25% xylene cyanol and 0.25% bromophenol blue. into
each lane. The amplification products were visualised by silver staining following the
procedure by Dinesh et al. Ž1993.. A 100 bp DNA ladder ŽGibco BRL, USA. was used.

2.5. Data analysis

The RAPD fingerprints generated for all samples were scored in the following way:
presence of a band was scored as 1 while absence of the band was scored as 0 Žraw
data.. Only bands between 200 and 1500 bp were scored. Comparisons were carried out
between samples amplified by the same primer in a pairwise manner.
Nei and Li’s genetic similarity index ŽSI. ŽNei and Li, 1979. was used in this study. It
exhibits the least percent bias for closely related organisms as compared to two other
similarity coefficients, i.e., the simple matching coefficient and the Jaccard’s coefficient
ŽLamboy, 1994.. SI values are presented as mean values for pairwise comparisons
between all individuals in a pair of wild forms or cultivated varieties being compared.

Table 2
The five arbitrary primers used for RAPD fingerprinting of Discus
X X
Name of primer Nucleotide length Sequence Ž5 –3 . ŽGqC.% Tm Ž8C.
NUSZG4 ŽDinesh et al., 1993. 9-mer GGAGCTGGC 77.7 32
NUSZG8 ŽDinesh et al., 1993. 10-mer TGCCGAGCTG 70.0 34
NUSZG6 ŽDinesh et al., 1993. 10-mer CGGTCACTGT 60.0 32
OPA-3 ŽOperon Technologies, USA. 10-mer AGTCAGCCAC 60.0 32
OPA-4 ŽOperon Technologies, USA. 10-mer AATCGGGCTG 60.0 32
 T.L. Koh et al.r Aquaculture 173 (1999) 485–497 489

They are also the mean values for the five primers used. The mean SI for the inter-wild
form, inter-cultivated variety and also for between wild form and cultivated variety
pairwise comparisons were corrected for their respective mean intra-wild form and
intra-cultivated variety SI values based on the equation given by Lynch Ž1990.;
SXi j s 1 q Si j y 0.5Ž Si q S j . where Si and S j are the intra-population SI, Si j is the
inter-population SI and SXi j is the corrected inter-population SI.
The Mann–Whitney U-test Žalso known as the Wilcoxon Rank Sum test. was
performed for the pairwise comparisons among the inter-wild form, inter-cultivated
variety and between wild form and cultivated variety SI values using Statistical Analysis
System ŽSAS. version 6.12 ŽSAS Institute, USA.. SI values for all pairwise comparisons
between individuals were used to calculate genetic distances between them by using the
formula: 1 y SI. The average genetic distances among individuals calculated for the five
primers were then used to construct a phenogram based on unweighted pair–group
method with arithmetic means ŽUPGMA. ŽSneath and Sokal, 1973. using the PHYLIP
Žver. 3.57c. program ŽFelsenstein, 1995..

3. Results

3.1. Comparisons of SI Õalues using Mann–Whitney U-test

Fig. 1 shows the RAPD fingerprints of one fish from each of the wild forms and
cultivated varieties generated for the five primers used. The mean SI values calculated
for all pairwise comparisons among the wild-forms and cultivated varieties are shown in
Table 3. The results of the Mann–Whitney U-test performed on these SI values are
recorded in Table 4. These results indicate that individuals of the wild forms of Discus
are as genetically similar to one another as to individuals of the cultivated varieties.
However, the converse is not true, i.e., individuals of the cultivated varieties are closer
to one another than with individuals of the wild forms, even though the genetic
similarity among cultivated varieties is not significantly different from that among the
wild forms.

3.2. UPGMA phenogram based on SI Õalues of indiÕidual pairwise comparisons

In the phenogram ŽFig. 2. based on the mean genetic distances among all 45
individuals for the five primers used Ždata not shown., the cluster of Heckel individuals
lies furthest from the rest of the wild forms and cultivated varieties. It exists as a distinct
cluster, 0.131 Žmean genetic distance., away from the cluster of individuals belonging to
the other three wild forms and 0.121 Žmean genetic distance. from the main cluster of
cultivated varieties. The five Heckel individuals are clustered together, indicating a high
genetic similarity among them. This is only seen in one of the other three wild forms:
Wild green. Wild blue and Wild brown individuals are intermingled loosely with each
other in a group distinct Žmean genetic distance of 0.045. from Wild green individuals.
 490
T.L. Koh et al.r Aquaculture 173 (1999) 485–497
Fig. 1. RAPD fingerprints generated for one fish of each of the four wild forms and five cultivated varieties of Discus using the five primers ŽNUSZG4, NUSZG6,
NUSZG8, OPA-3 and OPA-4.. T s Turquoise, P s Pigeon, G sGhost, C sCobalt, SsSolid Red, L sWild blue, R sWild brown, NsWild green, H s Heckel,
E s negative control and M s100 bp DNA marker ŽGibco BRL, USA..
 T.L. Koh et al.r Aquaculture 173 (1999) 485–497 491

Table 3
Mean similarity indices Žcorrected for intra-wild form or intra-cultivated variety similarity after Lynch Ž1990..
for the pairwise comparisons among the four wild forms and the five cultivated varieties of Discus for the five
primers
L R N H T P G C S
L 0.906 0.972 0.957 0.872 0.904 0.883 0.914 0.925 0.907
R 0.860 0.982 0.894 0.907 0.901 0.919 0.932 0.934
N 0.838 0.860 0.893 0.877 0.906 0.930 0.911
H 0.910 0.821 0.807 0.799 0.825 0.834
T 0.820 0.975 0.953 0.966 0.946
P 0.834 0.971 0.953 0.957
G 0.868 0.983 0.970
C 0.790 0.985
S 0.834

L sWild blue, R sWild brown, NsWild green, H s Heckel, T s Turquoise, P s Pigeon, G sGhost,
C sCobalt and SsSolid Red.

In conclusion, Heckel Discus is 2.89 times Žin terms of mean genetic distance. further
from the other wild forms ŽWild green, blue and brown. than Wild green to the other
two wild forms ŽWild blue and brown..
The individuals of each of the five cultivated varieties are not tightly clustered with
other individuals of the same variety ŽFig. 2.. This indicates the lack of a genetic basis
for the present classification of the cultivated varieties which based solely on pheno-
types. Most of the individuals of the cultivated varieties lie in the same cluster distinct
from the wild forms. This cluster of cultivated varieties is 3.18 times genetically closer
to the cluster of the three Symphysodon aequiefasciata wild forms ŽWild green, blue and
brown. Žmean genetic distances 0.033. than to S. discus ŽHeckel. Žmean genetic
distances 0.105.. However, individuals such as Turquoise 4 ŽT4., Pigeon 4 ŽP4.,
Pigeon 5 ŽP5. and Cobalt 3 ŽC3. lie out of this main cluster and are genetically closer to
Heckel than to the other three wild forms.

Table 4
Probabilities Ž P . obtained for comparisons among Ž1. inter-wild form, Ž2. inter-cultivated variety and Ž3.
between wild form and cultivated variety similarity indices using the Mann–Whitney U-test ŽWilcoxon Rank
Sum test. by Statistical Analysis System ŽSAS.
Similarity indices Ž1. Inter-wild form Ž2. Inter-cultivated Ž3. Between wild
variety form and
cultivated variety
Ž1. Inter-wild form P s 0.2321 n.s. P s 0.2868 n.s.
Ž2. Inter-cultivated P s 0.0001 sig.
variety
Ž3. Inter-wild form and
cultivated variety

sig.ssignificant Ž P - 0.05. and n.s.s not significant Ž P ) 0.05..

 492
T.L. Koh et al.r Aquaculture 173 (1999) 485–497
Fig. 2. UPGMA phenogram showing the genetic relationships among all individuals of the four wild forms and five cultivated varieties of Discus constructed using
PHYLIP Žver. 3.57c.. One unit of the scale represents 0.01 mean genetic distance. T s Turquoise, P s Pigeon, G sGhost, C sCobalt, SsSolid Red, L sWild blue,
R sWild brown, NsWild green and H s Heckel.
 T.L. Koh et al.r Aquaculture 173 (1999) 485–497 493

4. Discussion

4.1. Comparisons of SI Õalues using Mann–Whitney U-test

The intra-wild form and intra-cultivated variety SI values may be overestimated due
to the small sample size analysed. However, all following conclusions based on the
inter-wild form, inter-cultivated variety and between wild form and cultivated variety SI
Žcorrected for their respective intra-wild formr-cultivated variety SI values and there-
fore dependent on those values. could only be further strengthened if a higher sample
size was available. This is because these conclusions are concerned with the relative
comparisons among the inter-wild form, inter-cultivated variety and the between wild
form and cultivated variety genetic diversity and not the absolute genetic variation in the
wild forms or the cultivated varieties.
From our results, we postulate that the gene pool of the cultivated varieties consists
only a portion of the gene pool of the wild forms of Discus. The genetic variability
among the cultivated varieties is comparable although not as high as that within wild
forms. This suggests that the genetic diversity in the wild Discus is still not being fully
exploited in the present culture of Discus varieties. In view of this, there will be a higher
chance of generating new fin and colour morphs of Discus by frequent outcrossing of
the cultivated Discus with the wild Discus rather than by selective breeding followed by
inbreeding among cultivated varieties. In this way, the genetic variability among the
cultivated varieties would also be enhanced thus preventing the occurrence of inbreeding
that will probably take place if the present situation of unplanned breeding persists.
Therefore, for high sustainability of the culture of Discus, proper breeding programs
must be implemented with careful management and monitoring such that there is
frequent outcrossing with the wild forms as well as maintenance of any newly emerged
traits by inbreeding.

4.2. UPGMA phenogram based on SI Õalues of indiÕidual pairwise comparisons

The controversy over the number of subspecies and species in the genus Sym-
physodon has persisted since it was first described by Heckel Ž1840.. Schultz Ž1960.
proposed the presence of two species of Discus: S. discus ŽHeckel. and S. aquiefasciata
with three subspecies, namely, S. a. aquiefasciata ŽWild green., S. a. haraldi ŽWild
blue. and S. a. axeroldi ŽWild brown.. Teton and Allgayer Ž1986., however, considered
only one species of Discus, S. discus, with two subspecies: S. d. discus and S. d.
aquiefasciata to be present. In this classification, Wild green, Wild blue and Wild brown
are not considered as subspecies. The later classification is supported by the fact that
there are more than one natural hybrid of S. discus ŽHeckel. and S. aquiefasciata,
namely, S. d. willischwartzi and ŽBurgess, 1991. Jordan’s Brown Heckel ŽJordan, 1993..
There is also difficulty in distinguishing subspecies of Schultz Ž1960. both morphologi-
cally and geographically ŽAxelrod et al., 1991; Yamada and Mori, 1991..
Our results showed that the Heckel Discus appeared to be the most divergent in
relation to the other wild forms ŽFig. 2.. In addition, since Wild blue and Wild brown
494 T.L. Koh et al.r Aquaculture 173 (1999) 485–497

individuals do not cluster together with individuals of the same wild form as in the case
of the Heckel and Wild green Discus, it is very likely that Wild blue and Wild brown are
not distinct subspecies. Nevertheless, the possibility of these S. aquiefasciata wild forms
existing as a clinal populations has not been ruled out. Therefore, although S. discus and
the S. aquiefasciata wild forms are genetically distinct, the debate over their exact
species status has yet to be resolved. This would require additional investigations
involving geographical, morphological as well as genetic approaches.
The distinct dichotomy between the main cluster of cultivated varieties and S.
aquiefasciata is indicative of the genetic effect of more than 50 years of artificial
selection in Discus breeding for strong colours and desirable body and fin shapes ŽFig.
2.. The phenogram also shows that this main cluster of cultivated varieties is in fact
genetically closer to S. aquiefasciata than to S. discus. This agrees with the aquarium
literature which mentioned that most of the cultivated varieties are derived from either
Wild green or Wild blue. The Turquoise variety was derived from two Wild green
Discus according to Wattley Ž1985.. The Pigeon variety was reported to be produced
from Red Royal Blue ŽWeiss, 1993. which had been bred from Turquoise ŽYamada and
Mori, 1991.. Cobalt was known to be derived also from Turquoise ŽYamada and Mori,
1991.. The Solid Red variety was reported to be bred either from Wild green or Wild
brown Discus ŽSchmidt-Focke, 1990.. There is no record on the genetic background of
the Ghost variety.
In conclusion, S. aquiefasciata is more likely to be the genetic origin of the cultivated
varieties than S. discus ŽHeckel.. Thus, in order to enhance the genetic variability of the
cultivated varieties, they should be outcrossed not only to the wild forms Žas explained
in the previous section. but as far as possible also to the Heckel Discus, although the
rate of success is lower in the latter case ŽWattley, 1985.. Two potential problems may
arise in such cross-breeding if there are in fact two species of Discus. However, both of
these problems are not entirely insurmountable. The first one is the possible meiotic
problem in the subsequent generations of such interspecific cross-breeding. This can be
solved by the production of allotetraploids through the induction of endomitosis so that
the successful meiotic pairing of their chromosomes can occur between the duplicated
ones ŽChevassus, 1983.. In this way, fertile and stable ‘new species’ Žhybrids. can be
created.
It is obvious that such interspecific hybridisation may also raise questions on issues
regarding the conservation of wild Discus natural genetic resources as well as the
preservation of the genetic distinctness of the two Discus species if they truly exist. The
solution to the first aspect of the issue lies in the setting up of a large captive breeding
facility in situ, i.e., at the Amazon basin where rare and distant wild Discus forms can be
caught and tank-bred to produce progeny that will be exported for interbreeding with
cultivated varieties. Wild caught material will only be introduced occasionally for the
strengthening of genetic lines. In this way, the original wild caught stocks need not be
exported at all and would most likely not suffer from severe exploitation that might lead
to the gradual extinction of the species. This has, in fact, been carried out by an African
rift lake exporter ŽGrant, 1995..
The answer to the second aspect of the issue lies in the use of chromosome
manipulation methods to stock new hybrids as sterile triploid fish as in the case of
 T.L. Koh et al.r Aquaculture 173 (1999) 485–497 495

striped bass = white bass hybrids ŽTave, 1993.. This approach will ensure that in the
case of accidental escape of hybrids into the Discus native habitats, introgressive
hybridisation and consequent contamination of the wild Discus gene pool will not occur.
This has actually been done fairly routinely and is the basis for certain industries ŽTave,
1993.. Triploid fish can be produced by mating tetraploids with diploids ŽTave, 1993.
Žtetraploids can be produced in the same way as mentioned before.. The tetraploids
would be mated for self-perpetuation and maintained as a broodstock. In this way, the
genetic distinctness of the two species of Discus can be preserved.
Moreover, such interspecific hybridisation could have been responsible for the vivid
hues achieved in aquarium species although systematic study of such case is not
reported ŽPurdom, 1993.. The major colour morphs of goldfish and koi are certainly
believed to relate to hybridisation in their distant pasts ŽPurdom, 1993.. Therefore, such
interspecific hybridisation should be integrated into the more general genetic broodstock
management and improvement program Žlike the increase of the effective breeding
number, Ne , through optimising the total number of breeding individuals, sex ratio,
mating system and variance of family size. with due consideration of the potential
problems and the adoption of preventive measures mentioned in the previous few
paragraphs. This will definitely be able to enhance the genetic variance for selection
programs in the cultivated varieties. Nevertheless, additional genetic analyses regarding
this enhancement must be carried out to allow the assessment of the effectiveness of
such an aquacultural practice in Discus breeding before its large-scale and long-term
implementation.
At present, there are three attempts at the classification of cultivated Discus varieties.
Yamada and Mori Ž1991. provided a system to classify the cultivated discus varieties by
grouping them into three categories: Turquoise, Wattley and Red Royal Blue. However,
similar varieties Že.g., Cobalt. are grouped in both the Turquoise and the Wattley
categories. In addition, the basis of classifying the varieties under the third group, Red
Royal Blue, was not explained. Axelrod et al. Ž1991. did not give any description for his
classification and only photographs of the varieties in each category were given. The
classification given by Scamen Ž1994. utilises the typing of the background colours of
Discus to separate them into different races. Although, this method allows easy
communications among breeders due to the usage of easily understood terms ŽScamen,
1994., it consists of insufficient descriptions of the fish with important phenotypes like
body patterning being excluded. In addition, there is also subjectivity involved in
deciding the background colour. Moreover, since it is based entirely on the physical
appearance of the Discus, it is sometimes difficult to determine the breed of the fish as it
may possess intermediate morphological characteristics.
A widely accepted standard classification and nomenclature system for the cultivated
Discus should be formulated. This is to facilitate the communicating of the fish among
breeders so that industry-wide breeding programs can be properly implemented to
produce higher quality fish. The present phenotypic classification and nomenclature
system used in the industry is not unified and confusion may arise due to the fact that
the same term is being used for different varieties and vice versa. The present system
also lacks genetic basis since the individuals categorised and named under the same
variety possessing the same set of phenotypes do not cluster together in the UPGMA
496 T.L. Koh et al.r Aquaculture 173 (1999) 485–497

phenogram. It would be more useful if the classification of cultivated Discus can be

standardised to one where the present cultivated Discus are grouped according to their
genetic similarities. In this way, interbreeding among the varieties could be planned on
the basis of this classification system and information can be conveyed more accurately
using the associated nomenclature. In view of this, there should be a more extensive
study using molecular methodologies to clarify the genetic relationships among all of the
existing cultivated Discus so that such a classification system can be formulated.

Acknowledgements

This work was supported by a grant from the National University of Singapore and
an EC-Singapore grant Žcontract number: C11)CT94-0021..

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