Plant Pathology (2006) 55, 671678 Doi: 10.1111/j.1365-3059.2006.01436.

Resistance in water yam (Dioscorea alata) cultivars in

Blackwell Publishing Ltd

the French West Indies to anthracnose disease based on

tissue culture-derived whole-plant assay

T. J. Onyeka*, D. Ptro, G. Ano, S. Etienne and S. Rubens

Unit de Recherche en Productions Vgtales (URPV), Institut National de la Recherche Agronomique (INRA), Centre Antilles-Guyane,
97170 Petit-Bourg, Guadeloupe

Reactions of 60 water yam (Dioscorea alata) cultivars to three isolates of the yam anthracnose fungal pathogen
(Colletotrichum gloeosporioides) were evaluated using tissue culture-derived whole-plant assay. Three disease parameters:
single score on a scale of 06 at the seventh day after inoculation (SD7); area under the disease progress curve (AUDPC);
and disease progress rate (Rd) were compared, and cultivars were classified into disease-response groups using a
rank-sum method based on AUDPC scores for the two most virulent isolates. A wide range of variation in resistance of
the D. alata cultivars, and significant effects of pathogen isolate and isolatecultivar interactions, were observed for all
disease parameters. The three disease parameters were positively correlated; however, four cultivars showed great
dispersions from the regression lines for comparisons of SD7 with the multiple assessments based AUDPC and Rd.
The 60 cultivars were separated into resistant (n = 12), moderately resistant (n = 19), moderately susceptible (n = 18) and
susceptible (n = 11) groups. The potential of the tissue culture-derived whole-plant assay to resistance breeding
programmes and further understanding of the yam anthracnose pathosystem is discussed.

Keywords: anthracnose, Colletotrichum gloeosporioides, food yams, isolate-specific resistance, screening

and where there is potential for development of chemically

Introduction
Yam anthracnose, caused by Colletotrichum gloeosporio-
ides, is the most important foliar disease of the food yams
(Dioscorea spp.), and has been reported on almost all
cultivated species of yam in the humid and subhumid
tropics (Emehute et al., 1998). Water yam (Dioscorea
alata), which is the most popular species in the Caribbean,
is more susceptible than other yam species to anthracnose
(Winch et al., 1984; Mignucci et al., 1988). In the Carib-
bean, the common practice of cultivating very few, often
susceptible, D. alata cultivars on a commercial scale favours
the spread of anthracnose and high yield losses. For
example, in Puerto Rico yield losses of 50 90% due to
anthracnose disease have been common under favourable
conditions (Mignucci et al., 1988).
Chemical control of yam anthracnose requires monthly
or biweekly application of fungicides, which could be
damaging to the environment and increase production costs.
For diseases such as yam anthracnose, which require
sequential fungicide spraying (Hepperly & Vazquez, 1991)
*E-mail: petro@antilles.inra.fr
Accepted 3 March 2006
resistant pathogen strains (Bayart & Pallas, 1994), the use
of host-plant resistance is a more sustainable and environ-
mentally acceptable control option.
The effectiveness of harnessing host-plant resistance for
disease control depends greatly on a good understanding
of the variations within the pathogen and host populations,
and the availability of a reliable method of screening the
host for disease reactions (Parlevliet, 1993; Peever et al.,
2000). Recent studies on C. gloeosporioides isolates asso-
ciated with yam in Nigeria (Abang et al., 2004, 2005) and
in the French West Indies (Frzal et al., 2003; Frzal,
2005) revealed high genetic and pathogenic variations in
between-field and within-field isolates. This high genotypic
diversity in the population of C. gloeosporioides affecting
yam, suggests the pathogen has a strong evolutionary
potential to develop new virulent strains that can break
or erode existing resistance (McDonald & Linde, 2002).
Therefore, there is a need to continuously identify new
sources of resistance that could be used to diversify existing
breeding populations.
The spectrum of resistance to anthracnose in the D. alata
collection in the French West Indies, and in the Caribbean
in general, is not well known. Early efforts at identifying
anthracnose-resistant genotypes resulted in the selection

2006 The Authors

Journal compilation 2006 BSPP 671
672 T. J. Onyeka et al.

of some less-susceptible cultivars such as Belep, Plimbite, Table 1 Sources of Dioscorea alata cultivars evaluated for anthracnose
Kinabayo and Oriental, which were recommended in differ- disease resistance
ent Caribbean countries and in the Pacific Islands (Green
& Simons, 1992; Plumbley & Sweetmore, 1994; Wright & Genotype Country of collection Region

Peters, 2002). Some of these previously resistant cultivars, Oriental Barbados Caribbean
however, have been reported to be susceptible in some DA28 Cuba Caribbean
farmers fields. This has been attributed to the presence of DA26 French Guiana South America
new or previously unknown virulent strains of C. gloe- DA27 French Guiana South America
osporioides (Ano et al., 2005). It could also be that these Divin 2 French Guiana South America
Fenakue French Guiana South America
varieties were selected under suboptimal conditions for
Pacala Guyane French Guiana South America
disease development, as early work on identification of
Cinq Guadeloupe Caribbean
resistance was conducted under natural field infection. Cuella Largo Guadeloupe Caribbean
Screening of yam in a controlled environment that Divin 1 Guadeloupe Caribbean
favours both disease development and plant growth is Florido Guadeloupe Caribbean
required to evaluate effectively the reactions of different Kinabayo Guadeloupe Caribbean
yam cultivars to anthracnose disease. The lack of standard Pacala station Guadeloupe Caribbean
screening procedures for yam anthracnose resistance limited St Sauveur 1 Guadeloupe Caribbean
progress in resistance breeding until recently, when Abang Ste Catherine Guadeloupe Caribbean
et al. (2001b) proposed a tissue culture-based in vitro Tahiti French Guadeloupe Caribbean
Tahiti Messaien Guadeloupe Caribbean
method for rapid assessment of yam genotypes under a
65 IRAT Guadeloupe/IRAT Caribbean
controlled environment. This method, which involves
Pacala Cacao Guadeloupe/French Guiana Caribbean
inoculation of tissue culture-derived whole plants, has 76 IRAT Guadeloupe/IRAT Caribbean
been optimized for the evaluation of levels of anthracnose Asmhore Guadeloupe/IRAT Caribbean
resistance in yam accessions (Onyeka et al., 2005a, 2006). Igname serpent Guadeloupe/Martinique Caribbean
This study used the optimized tissue culture-derived Jardin Hatien Guadeloupe/Martinique Caribbean
whole-plant assay to evaluate the D. alata collection of the Brazzo fuerte Haiti Caribbean
Institut National de la Recherche Agronomique (INRA), Caillade 2 Haiti Caribbean
Guadeloupe, French West Indies, for varying levels of Plimbite Haiti Caribbean
resistance to anthracnose. Toro Haiti Caribbean
Pyramide Puerto Rico Caribbean
Renta Yam Jamaica Caribbean
Materials and methods Igname deau Martinique Caribbean
St Martin Martinique Caribbean
Dioscorea alata cultivars and C. gloeosporioides St Vincent Blanc 1 Martinique Caribbean
St Vincent Violet Martinique Caribbean
isolates
Sassa 1 Martinique Caribbean
A total of 60 D. alata cultivars from the yam germplasm Telemaque Martinique Caribbean
collection of INRA were evaluated for anthracnose resist- Belep New Caledonia Pacific
ance. These consisted of tetraploid, hexaploid and octoploid Caplaou New Caledonia Pacific
D. alata cultivars obtained from various yam programmes Goana New Caledonia Pacific
Gordito New Caledonia Pacific
in different Caribbean countries, South America and the
Morado New Caledonia Pacific
Pacific (Table 1).
Nouma New Caledonia Pacific
All the cultivars were evaluated for response to three Pakutrany New Caledonia Pacific
isolates of C. gloeosporioides (Cg78, Cg118 and Cg206) Tana New Caledonia Pacific
with varying degrees of aggressiveness. The three isolates Wabe New Caledonia Pacific
were selected based on prescreening of 66 isolates from the Wenefela bis New Caledonia Pacific
INRA culture collections of C. gloeosporioides from the Kokoeta Philippines Pacific
French West Indies, using detached-leaf bioassay (Jacqua Lupias Philippines Pacific
et al., 2005), with two D. alata cultivars (Plimbite and St Smooth statia Puerto Rico Caribbean
Catherine). Two of the isolates (Cg78 and Cg118) originated Buet Puerto Rico Caribbean
Cuba 1 Puerto Rico Caribbean
from Grande-Terre, while Cg206 originated from the Base-
DA32 Puerto Rico Caribbean
Terre region of Guadeloupe (G. Jacqua, personal communi-
De Agua Puerto Rico Caribbean
cation). Initial AFLP analysis showed that the three isolates Feo Puerto Rico Caribbean
represent distinct genetic groupings (Degueret, 2001). Grand Etang 1 Puerto Rico Caribbean
Hawai branched Puerto Rico Caribbean
Igname rouge Puerto Rico Caribbean
Preparation of tissue culture-derived whole plants
Purple Lisbon Puerto Rico Caribbean
Tissue culture-derived whole plants were raised from Vino purple form Puerto Rico Caribbean
in vitro plantlets obtained by nodal explant cultures. The Vino white form Puerto Rico Caribbean
in vitro plantlets were grown for 5 weeks on modified solid White Lisbon Puerto Rico Caribbean

Plant Pathology (2006) 55, 671678

 Anthracnose resistance in water yam
Murashige and Skoog medium under 16-h photoperiods
at 24 2C (Onyeka et al., 2006). Plantlets were then
transferred from the tubes into propagation trays containing
sterilized soil amended with organic manure, and acclima-
tized in a bioclimatic chamber for 3 weeks at 28/25C day/
night temperature and a 16-h photoperiod. Acclimatized
plants were transferred into plastic pots and grown for an
additional 3 weeks in the bioclimatic chamber before inoc-
ulation. Prior to inoculation, potted plants were transferred
into a miniserre (BHR serre, France). This mini-glasshouse-
like containment facility enabled maintenance of the high
relative humidity (>80%) required for disease development.
The miniserre also prevented cross-contamination of the
different pathogen isolates.
Plant inoculation and disease assessment
Inoculation and disease-assessment procedures were carried
out according to the modified tissue culture-derived whole-
plant assay (Onyeka et al., 2005a, 2006). Potted plants were
spray-inoculated with a spore suspension (105 conidia
mL1) in the miniserre using approximately 1 mL inoculum
per plant. The inoculated plants were scored for disease
severity at 3, 5 and 7 days after inoculation (DAI), on a
scale of 06 using the percentage whole plant-area scoring
method (Simons & Green, 1994) with slight modification,
where 0 = 0%, 1 = 1%, 2 = 2%, 3 = 39%, 4 = 1024%,
5 = 2550% and 6 > 50% affected plant tissue.
The evaluation of the 60 cultivars was conducted in three
different batches using a randomized complete block design
with two blocks of four replicates per treatment combina-
tion. Two of the cultivars were included in all the batches
as internal checks, and the study was repeated once. The
disease score (DS) of each cultivar was adjusted for varia-
tion between experimental batches based on the reactions
of the internal checks as: DScor = DS (CHKn CHKN),
where DS = disease score of the cultivar; CHKn = average
of checks in a particular batch; and CHKN = total mean of
checks in all batches.
Data analysis
Variation in resistance of yam cultivars was evaluated
based on a single disease score at the seventh day after
inoculation (SD7), the cumulative area under the disease
progress curve (AUDPC) as described by Shaner & Finney
(1977), and the disease progress rate (Rd) taken as the
slope of regression line for disease severity against time.
All statistical analyses were carried out using the sas
statistical package, ver. 612 (SAS, 1989). As both random
and fixed effects were involved in the experimental designs,
treatment effects were compared using the linear mixed
model procedure. Trial, isolate within trial, and block
within trial were fitted as random effects, while cultivar,
isolate, and interaction between cultivar and isolate were
fitted as fixed effects. Relationships among the disease
parameters were tested by simple linear regression.
The relative resistance of the different D. alata cultivars
was evaluated using a modified rank-sum classification
Plant Pathology (2006) 55, 671678
673
Table 2 Summary of generalized linear mixed modelling of the
reactions of tissue culture-derived whole plants of 60 Dioscorea alata
cultivars to anthracnose disease severity evaluation
SD7a AUDPCb Rdc
Fixed term df rdfd Fe Pf F P F P
Genotype 59 1331 1402 *** 1655 *** 1212 ***
Isolate 2 1331 5856 *** 6741 *** 6578 ***
Genotype isolate 118 1331 2469 *** 2848 *** 3005 ***
a
Disease severity was evaluated as a single disease score on a scale
of 06 at the seventh day after inoculation (SD7).
b
Area under the disease progress curve.
c
Rd, disease progress rate.
d
rdf, Residual degrees of freedom.
e
Type 3 F statistics.
f
Probability of F statistics: ***, P < 0001.
method (Onyeka et al., 2005b) based on the mean AUDPC
scores of each cultivar for the two most virulent pathogen
isolates. To calculate the rank sum, the AUDPC scores of the
cultivars averaged across the two trials for each pathogen
isolate were assigned ranks in ascending order using the
RANK procedure of sas with option average for handling
ties. The sum of ranks (rank sum) was computed for each
cultivar and compared with the grand mean of the rank
sums across all cultivars. Deviation of each cultivar from the
grand mean was calculated, and cultivars were classified
based on deviation from the grand mean as follows: 20
to 11 SD of grand mean = resistant; 10 to 01 SD =
moderately resistant; 00 to 10 SD = moderately susceptible;
11 to 20 SD = susceptible.
Results
Variation in anthracnose resistance among
D. alata cultivars
All three isolates were capable of initiating anthracnose
symptoms on most of the yam cultivars. The three disease
parameters revealed wide variations among yam cultivars
and pathogen isolates (Table 2). The isolatecultivar
interaction term was significant and relatively large for all
parameters, indicating that specific resistance or suscepti-
bility accounted for a high proportion of the variations
observed. Disease score 0, which indicates complete resist-
ance with no anthracnose symptoms throughout the duration
of the study, was obtained in 10, eight and one cultivars
for isolates Cg78, Cg206 and Cg118, respectively.
Resistance reactions to isolate Cg78 (scores 0 4) were
obtained for 57 (95%) of the cultivars, while 29 (43%) and
37 (61%) of the cultivars showed resistance reactions to
isolates Cg118 and Cg206, respectively. Resistance reac-
tions to all the three isolates were obtained in 24 cultivars.
Isolate-specific reactions
Isolate Cg78 was generally the least virulent, and only
three cultivars (Ashmore, Jardin Haitien and Pacala Guyane)
674 T. J. Onyeka et al.
Figure 1 Scatter plot showing the variation in responses of 60
Dioscorea alata cultivars to three isolates of Colletotrichum
gloeosporioides (Cg78, Cg118 and Cg206) on a scale of 0 6, where
scores 0 4 correspond to <25% of whole plant affected = resistance
and scores >4 = susceptible.
were susceptible to this isolate (score >4). All three cultivars
were also susceptible to the more widely virulent isolate
Cg118, but all were resistant to isolate Cg206 (Fig. 1a,b).
This is an indication that these three cultivars have specific
resistance to isolate Cg206. More isolate-specific reactions
were obtained when the cultivars were compared based
on their responses to isolates Cg118 and Cg206 (Fig. 1c).
A total of 13 cultivars were resistant to isolate Cg206 but
susceptible to Cg118, while five cultivars were resistant to
Cg118 but susceptible to Cg206.
Relationship between disease parameters
The regression coefficients generally indicated strong linear
relationships between disease parameters (Fig. 2). However,
four cultivars showed large deviations from the regression
line for the relationship between disease severity 7 days
after inoculation (SD7) and the rate of disease progress
(Rd) (Fig. 2a). These are cultivars that had early but slow
development of the disease (St Martin and St Vincent
Blanc), or cultivars with late but rapid development of the
disease (St Catherine and St Sauveur). The same trend was
observed for the relationship between SD7 and AUDPC
(Fig. 2b). Comparison of Rd and AUDPC showed a stronger
linear relationship (r2 = 087; Fig. 2c), indicating that the
cultivars maintained their disease ranking based on these
two parameters.
Relative resistance among D. alata cultivars
The relative resistance among the D. alata cultivars was
compared on the basis of the cumulative AUDPC, as there
were good correlations between disease parameters.
The rank-sum classification of relative resistance using the
two most widely virulent isolates identified 12 cultivars
(20%) as resistant; 19 (317%) moderately resistant; 18
(30%) moderately susceptible; and 11 (183%) susceptible
(Table 3). Cultivars in the resistant and susceptible groups
showed no isolate-specific reaction as they were, respec-
tively, resistant and susceptible to the two isolates. Isolate-
specific reactions were observed for some of the cultivars in
the moderately resistant and moderately susceptible groups.
A total of seven moderately resistant and 11 moderately
susceptible cultivars were specifically resistant to either
Cg118 or Cg206.
Discussion
All three parameters considered in this study revealed a
wide range of reactions among D. alata cultivars. Any of
these parameters could therefore be used for assessment of
anthracnose resistance in the tissue culture-derived whole-
plant assay. However, the presence of cultivars with early
but slow development of disease, or with late but fast
development of disease, showed that a single disease score
may not compare resistance responses among cultivars
adequately. Du Toit et al. (1997) noted that the construc-
tion of disease progress curves based on multiple disease
assessments has the advantage of showing how early
resistance to maize fusarium seedling blight was expressed.
Sweetmore et al. (1994) showed that yam anthracnose
symptoms in the field occurred mainly on younger leaves.
Therefore cultivars with delayed disease onset are likely to
be valuable sources of field resistance.
Variations among pathogen isolates and their interactions
with host cultivars are important considerations in tests
to identify host-plant resistance to diseases. The results
obtained in this study showed that variation among isolates
of C. gloeosporioides should be adequately considered in
resistance screening. The use of a single but highly aggressive
isolate in screening for host resistance has been recommended
for some plant-disease systems (Arseniuk & Czembor,
1999). The identification of some cultivars that were
resistant to the highly aggressive Cg206 but susceptible to
the less aggressive Cg78 in this study indicates that the use of
a single aggressive isolate for screening may not ade-
quately reveal the full spectrum of anthracnose resistance/
susceptibility in yam germplasm. It is important, therefore,
to subject the resistant materials to a wide variety of pathogen
Plant Pathology (2006) 55, 671678
 Anthracnose resistance in water yam 675

Table 3 Rank-sum classification of anthracnose severity of 60 Dioscorea alata cultivars inoculated with isolates of Colletotrichum gloeosporioides
in tissue culture-derived whole-plant assay

Cg118 Cg206 Rank-sum classification

Cultivar SD7 AUPDC1 a SD7 AUDPC2 b c d Group

Renta Yam R 000 100 R 000 450 550 176 R

Igname deau R 075 250 R 000 450 700 171 R
DA27 R 113 450 R 000 450 900 165 R
Telemaque R 113 450 R 000 450 900 165 R
Caillade 2 R 075 250 R 038 900 1150 157 R
Sassa 1 R 238 700 R 000 450 1150 157 R
65 IRAT R 288 1000 R 000 450 1450 147 R
Toro R 175 600 R 063 1000 1600 142 R
De Agua R 331 1300 R 075 1100 2400 117 R
Tahiti French R 250 800 R 306 1700 2500 114 R
DA26 R 350 1450 R 150 1300 2750 106 R
Tahiti Messaien R 269 900 R 369 2000 2900 101 R
St Martin R 294 1100 R 400 2150 3250 090 MR
Pacala station S 888 3100 R 000 450 3550 081 MR
Belep R 300 1200 R 525 2550 3750 074 MR
Tana R 700 2650 R 088 1200 3850 071 MR
Wabe R 694 2500 R 263 1600 4100 063 MR
Feo R 513 2000 R 400 2150 4150 062 MR
Fenakue R 500 1900 R 575 2800 4700 044 MR
76 IRAT S 1300 4300 R 000 450 4750 043 MR
Asmhore S 1025 3500 R 163 1400 4900 038 MR
St Vincent Violet R 369 1600 S 788 3500 5100 032 MR
Igname rouge S 863 3000 R 488 2300 5300 025 MR
Smooth statia R 569 2300 R 638 3000 5300 025 MR
St Vincent Blanc 1 R 400 1700 R 825 3700 5400 022 MR
Buet S 1216 4000 R 189 1500 5500 019 MR
Purple Lisbon R 350 1450 R 956 4100 5550 017 MR
Morado R 663 2400 R 681 3200 5600 016 MR
Hawa branched S 919 3300 R 538 2700 6000 003 MR
Nouma R 850 2900 R 676 3100 6000 003 MR
Cuella Largo R 531 2100 R 938 3950 6050 002 MR
DA32 S 963 3400 R 588 2900 6300 006 MS
Florido S 1150 3850 R 525 2550 6400 009 MS
Plimbite S 1227 4100 R 497 2400 6500 013 MS
Kokoeta R 550 2200 S 1113 4400 6600 016 MS
Oriental R 700 2650 S 938 3950 6600 016 MS
Pacala Cacao S 1373 4900 R 315 1800 6700 019 MS
Gordito S 913 3200 S 869 3800 7000 028 MS
Vino white form R 706 2800 S 1038 4300 7100 032 MS
Igname serpent R 494 1800 S 1750 5400 7200 035 MS
Divin 2 S 1113 3600 S 1014 4200 7800 054 MS
Jardin Hatien S 1556 5900 R 344 1900 7800 054 MS
Grand Etang 1 S 1369 4800 R 738 3300 8100 063 MS
Pakutrany S 1325 4600 S 800 3600 8200 066 MS
White Lisbon S 1125 3700 S 1226 4700 8400 073 MS
Pacala Guyane S 1401 5200 R 776 3400 8600 079 MS
St Catherine S 1306 4500 S 1169 4500 9000 092 MS
Divin 1 S 1301 4400 S 1226 4700 9100 095 MS
Goana S 1288 4200 S 1394 4900 9100 095 MS
Lupias S 1329 4700 S 1411 5000 9700 114 S
Wenefela bis S 1388 5000 S 1226 4700 9700 114 S
Kinabayo S 1150 3850 S 1922 6000 9850 119 S
Pyramide S 1439 5400 S 1658 5300 10700 146 S
Brazzo fuerte S 1394 5100 S 1850 5800 10900 152 S
Cinq S 1500 5800 S 1414 5100 10900 152 S
DA28 S 1416 5300 S 1843 5700 11000 155 S
Vino purple form S 1448 5550 S 1780 5600 11150 160 S
Cuba 1 S 1576 6000 S 1539 5200 11200 161 S
St Sauveur 1 S 1463 5700 S 1775 5500 11200 161 S
Caplaou S 1448 5550 S 1905 5900 11450 169 S
6100

SD7 = Resistance assessment on the basis of a single disease score at the seventh day after inoculation on a scale of 0 6, where scores 4 are
considered resistance reactions and scores >4 are considered susceptible reactions.
AUDPC1 = cumulative area under the disease progress curve with Cg118; AUDPC2 = cumulative area under the disease progress curve with Cg206.
Grand mean of rank sums, a = cultivar ranking on the basis of AUDPC1; b = cultivar ranking on the basis of AUDPC2; c = rank sum (a + b) for
each cultivar; d = deviation from grand mean (G) of rank sums [d = (c G)/SD].
R = resistant; MR = moderately resistant; MS = moderately susceptible; S = susceptible.

Plant Pathology (2006) 55, 671678

676 T. J. Onyeka et al.
isolates. The generally strong isolatecultivar interactions
mean that we should be cautious about identifying a
particular cultivar as resistant without defining the isolate.
This study provides the first detailed report of the
spectrum of anthracnose resistance in the collection of
D. alata in the French West Indies. There are previous
reports of the disease reactions of some of the cultivars
tested under field conditions in Puerto Rico (Hepperly &
Vazquez, 1991); Barbados (McDonald et al., 1998); and
Guadeloupe (Ano et al., 2005), but field identification of
resistance to yam anthracnose could be of limited value
because the pathogen population at the screening sites
is not necessarily representative of pathogenic variation
elsewhere, especially as high temporal and spatial variation
exists in C. gloeosporioides isolates (Chakraborty et al.,
2002). The potential problems in field screening of yam
for resistance to unknown isolates of the anthracnose
pathogen are demonstrated by the seemingly inconsistent
disease reactions of some cultivars in previous reports, for
example cv. Kinabayo was reported as moderately suscep-
tible in Puerto Rico (Hepperly & Vazquez, 1991), but highly
Figure 2 Relationships between anthracnose
disease parameters on inoculated tissue
culture-derived whole plants of Dioscorea
alata.
resistant in Guadeloupe (Ano et al., 2005). Also, cvs
Plimbite, Belep and Pacala were reported as more resistant
than Oriental in Barbados (McDonald et al., 1998), while
Plimbite was reported as susceptible and Oriental as highly
resistant in Guadeloupe (Ano et al., 2005). Using conditions
favourable to both plant and pathogen development, as
described here, demonstrated that cv. Belep was resistant
to all the isolates tested, in agreement with previous
reports on the reaction of this cultivar (McDonald et al.,
1998; Ano et al., 2005), while cv. Plimbite was resistant to
isolate Cg206 but susceptible to Cg118, and cv. Oriental
was resistant to Cg118 but susceptible to Cg206. These
isolate-specific reactions of cvs Plimbite and Oriental may
partly explain the variations in their field reactions in
different countries. Although cv. Kinabayo is resistant to
anthracnose in the field (Ano et al., 2002, 2005), this
cultivar was susceptible to direct inoculation with both
isolates Cg118 and Cg206 used in this study, suggesting
isolate-specific resistance in this cultivar.
Another important outcome of this study is the
separation of the various D. alata cultivars into different
Plant Pathology (2006) 55, 671678
 Anthracnose resistance in water yam
anthracnose-response groups. The modified rank-sum
classification method used in this study separated the
cultivars into different compact groups based on their
reactions across two different isolates. The use of ranks is
preferred to the means of the observed data because
rank sum is less sensitive to outliers (Deng et al., 2004;
Onyeka et al., 2005b). The 60 D. alata cultivars were
separated into four response groups (resistant, moderately
resistant, moderately susceptible and susceptible). A total
of 12 cultivars with high nonisolate-specific resistance were
identified. All cultivars exhibiting isolate-specific reactions
were grouped as either moderately resistant or moderately
susceptible. The various cultivar groupings, particularly
groups with isolate-specific reactions, can serve as study
subsets to further understand the basis of hostpathogen
interactions for yam anthracnose. Subject to further
validation of the present results, by inoculation with more
C. gloeosporioides isolates from other locations, the resistant
groups with near-complete resistance, as well as the
moderately resistant group with reduced levels of disease,
can serve as potential sources of resistance in various yam-
breeding programmes towards development of D. alata
varieties with durable anthracnose resistance. This study
has shown that both isolate-specific as well as good partial
resistance to the three isolates of C. gloeosporioides tested
are present in the French West Indies collection of D. alata
cultivars. Similar results were obtained by Mignouna
et al. (2001), who identified both race-specific and race-
nonspecific resistance to two strains of C. gloeosporioides
from Nigeria. It is expected that gene pyramids would be
efficient when breeding for anthracnose resistance.
The tissue culture-derived whole-plant assay offers a
rapid, reliable and robust method for evaluating a large
number of yam germplasms, to select a subset that can
be evaluated further in field tests. The technique reduces
the number of lines that need to be evaluated in expensive
field trials. The results obtained in this and the earlier
study by Mignouna et al. (2001) indicated that both isolate-
specific and isolate-nonspecific reactions could be involved
in yam anthracnose resistance. Initial inheritance studies
(Mignouna et al., 2001, 2003) suggest that resistance is
dominant, but inherited quantitatively. The ability to evalu-
ate parental and progeny lines rapidly under reproducible
conditions using the tissue culture-derived whole-plant
assay will facilitate better understanding of the genetics of
yam anthracnose resistance. Segregating populations can
be characterized rapidly and useful phenotypes utilized in
quantitative trait analyses using marker-assisted selection.
In addition to its potential for rapid screening of a large
number of genotypes, the tissue culture-based assay could
be used for precise analysis of the virulence and aggressive-
ness of C. gloeosporioides isolates (Abang et al., 2001a),
studies of cultivarisolate interactions (Green et al., 2000;
Onyeka et al., 2006), and studies on the infection process.
Furthermore, the assay could be used to evaluate cultivars
for response to specific isolates of interest. This will enable
screening of exotic isolates of C. gloeosporioides and
assessment of yam accessions in international exchange
programmes.
Plant Pathology (2006) 55, 671678
677
Acknowledgements
The postdoctoral position of T.J.O. was supported by
INRA Dpartement Sant des Plantes et Environnement.
We thank R. Renac and J. Gelabale for their technical
contributions.
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