Yam Anthracnose Research Articles

Rapid assessment of resistance of tissue-cultured water yam (Dioscorea alata) and white guinea yam (Dioscorea rotundata) to anthracnose (Colletotrichum gloeosporioides Penz.)

Yam anthracnose is caused by the pathogen Colletotrichum gloeosporioides Penz. and has been identified as the most important biotic constraint to yam production worldwide. Rapid assessment of the disease is vital to its effective diagnosis and management. In this study, tissue-cultured yam plantlets of five lines of Dioscorea alata and nine of D. rotundata were rapidly assessed for their reactions to two isolates of yam anthracnose. The plantlets, obtained from meristem of the nodal cuttings, were grown for 8 weeks on Murashige and Skoog (MS) basal medium, acclimatised for 3 weeks, hardened for an additional 3 weeks, arranged in screen house in completely randomised design and sprayed with spore inocula prepared from 7 day-old culture of the two strains of Colletotrichum gloeosporioidies Penz. The relative resistance of the different Dioscorea spp. was evaluated using three disease indices – severity at seventh day after inoculation, SD7; area under disease progress curve, AUDPC; and disease severity rate, Rd. A modified rank-sum classification method put TDa 1425 and TDr 2040, with rank sum of 2.0 each, as resistant. TDr 2121, TDr 2287 and TDr 2048 were susceptible with rank sum of 27.50, 25.50 and 24.50, respectively. Dioscorea alata TDa 1425 and Dioscorea rotundata TDr 2040 were recommended in areas endemic with yam anthracnose, and also as parent lines while breeding for resistance to anthracnose.

Modelling the effects of temperature and leaf wetness on monocyclic infection in a tropical fungal pathosystem

Modelling the epidemiology of water yam anthracnose (Dioscorea alata) caused by the fungus Colletotrichum gloeosporioides is an important research goal, as it will allow the investigation of a wide range of scenarios of new practices to reduce the disease impact before experimentation in the field. Developing such a model requires a prior knowledge of the fungus’s response to the environmental conditions, which will be affected by pest management. In this work, we first measured the response of the fungus to the main physical environmental factors controlling its development, namely temperature (ranging from 18 °C to 36 °C) and wetness duration (from 2 h to 72 h). As response variables, we measured the percentage of formed appressoria (relative to the total number of spores), the length of the latent period (time lag between inoculation and first symptoms observed), and the rate of necrotic lesion extension (percentage of diseased leaf surface at different time steps). These variables allow us to estimate the effects of temperature and wetness duration on the success of infection (appressoria formation) and the subsequent rate of disease development (latent period length and lesion extension rate). The data were fitted to non-linear models chosen for their ability to describe the observed patterns. From our data and model analyses, we were able to estimate parameters such as the optimal and maximal temperatures (25–28 °C and 36 °C, respectively), the required wetness duration to reach 20 % of infection success and the time to reach 5 % disease severity as a function of temperature.

Quantification of the aggressiveness of a foliar pathogen, Colletotrichum gloeosporioides, responsible for water yam (Dioscorea alata) anthracnose

In the absence of a gene-for-gene relationship between a pathogen and its host, knowledge about aggressiveness is crucial to characterize novel pathogen populations that potentially emerge in agricultural pathosystems. Information about pathogen aggressiveness is also critical when establishing representative panels of pathogen isolates to test host resistance and in mapping quantitative trait loci involved in the host resistance. In this study, we focused on the fungus C. gloeosporioides that causes necrosis on the aerial part of one of its host plants, Dioscorea alata, and identified the in vitro conditions required to assess fungal aggressiveness on this host. Our main purpose was to convert the necrosis area development into a unique index for quantifying pathogen aggressiveness. The ‘Ag’ index described here has two advantages. First, it integrates the variance of symptom evolution curves to estimate the lesion development rates (initial and secondary) and the maximal necrosis area. Secondly, the new index takes two different symptoms commonly observed when inoculating D. alata leaves with C. gloeosporioides into account, one correlated with high leaf colonisation efficiency and the other with low colonisation efficiency. The weights accorded to each symptom in the index were proportional to leaf colonisation efficiency. We propose a framework for the acquisition of this index that has been designed to be conveniently combined with the routine bioassays required to establish representative panels of pathogen isolates. The general framework for the construction of this index can be broadly applied to diseases with necrotic symptoms.

Genetic diversity of Colletotrichum gloeosporioides in Nigeria using amplified fragment length polymorphism (AFLP) markers

Colletotrichum gloeosporioides is the causal agent of yam anthracnose disease in Nigeria. Differential cultivars and amplified fragment length polymorphic DNA markers were used to assess the extent of genetic diversity among 39 isolates of the pathogen. Fourteen (14) pathotypes of the pathogen were identified based on inoculation of a differential set of Dioscorea alata genotypes of which pathotype P11 was the most virulent attacking all differential D. alata host genotypes, while pathotype P8 was the least virulent. 52% of the isolates were avirulent, 23% were slightly virulent and 25% were virulent. Amplified fragment length polymorphism (AFLP) analysis confirmed genetic variation among the C. gloeosporioides isolates and was effective in establishing genetic relationships between them. However, the grouping of the isolates based on AFLP analysis was not directly related to virulence groups or geographical origin of the isolates. The genetic variation in C. gloeosporioides is important in choosing strategies to develop durable resistance Keywords: Anthracnose disease, pathotypes, genetic diversity, amplified fragment length polymorphism (AFLP) markers, yam

Effects of long-term salinity on the growth of the halophyte Spartina alterniflora Loisel

Colletotrichum gloeosporioides is the causal agent of yam anthracnose disease in Nigeria. Differential cultivars and amplified fragment length polymorphic DNA markers were used to assess the extent of genetic diversity among 39 isolates of the pathogen. Fourteen (14) pathotypes of the pathogen were identified based on inoculation of a differential set of Dioscorea alata genotypes of which pathotype P11 was the most virulent attacking all differential D. alata host genotypes, while pathotype P8 was the least virulent. 52% of the isolates were avirulent, 23% were slightly virulent and 25% were virulent. Amplified fragment length polymorphism (AFLP) analysis confirmed genetic variation among the C. gloeosporioides isolates and was effective in establishing genetic relationships between them. However, the grouping of the isolates based on AFLP analysis was not directly related to virulence groups or geographical origin of the isolates. The genetic variation in C. gloeosporioides is important in choosing strategies to develop durable resistance Key words: Anthracnose disease, pathotypes, genetic diversity, amplified fragment length polymorphism (AFLP) markers, yam.

Biological control of anthracnose (Colletotrichum gloeosporioides) in yam by Streptomyces sp.MJM5763

To find a suitable biocontrol agent for yam anthracnose caused by Colletotrichum gloeosporioides. An actinobacterial strain, MJM5763, showing strong antifungal activity, multiple biocontrol and plant growth-promoting traits was isolated from a yam cultivation field in Yeoju, South Korea. Based on morphological and physiological characteristics and analysis of the 16S rDNA sequence, strain MJM5763 was identified as a novel strain of Streptomyces and was designated as Streptomyces sp. MJM5763. Treatment with MJM5763 and the crude culture filtrate extract (CCFE) was effective in suppressing anthracnose in detached yam leaves in vitro and reduced incidence and severity of anthracnose in yam plants under greenhouse conditions. The CCFE treatment was the most effective of all the treatments and reduced the anthracnose severity by 85-88% and the incidence by 79-81%, 90 days after inoculation with the pathogen. CCFE treatment was also effective under field conditions and showed a reduction of 86 and 75% of anthracnose severity and incidence, respectively. Streptomyces sp. strain MJM5763 was effective in biocontrolling anthracnose in yam caused by C. gloeosporioides. Streptomyces sp. MJM5763 is a potential alternative to chemical fungicides for reducing yield losses to anthracnose in yam.

Evaluation of Water Yam (Dioscorea alata L.) Germplasm for Reaction to Yam Anthracnose and Virus Diseases and their Effect on Yield

AbstractUse of genetic resistance is the most practical and economic way to manage major diseases of yams. In a search for sources of resistance, 40 water yam (Dioscorea alata L.) accessions from Benin, Ghana, Nigeria and Puerto Rico were screened under natural disease infection conditions in Ibadan, Nigeria. The accessions were evaluated at 1, 3 and 6 months after planting (MAP) for severity of yam anthracnose and viral diseases. The effect of the pathogens on yield was also evaluated at harvest 9 MAP. There were significant differences (P < 0.001) between accessions for severities of anthracnose and viral diseases. Eight (20%) of them had lower anthracnose area under disease progress curves (AUDPC) values than the resistant check while 10 (25%) had AUDPC values below the trial mean. There were significant variations (P < 0.001) in yield components among the accessions. There was significant negative correlation of anthracnose severity with fresh tuber yield (r = −0.51) and with number of tubers per plot (r = −0.40). Similarly, significant negative correlations were observed of virus disease severity with fresh tuber yield (r = −0.78) and number of tubers per plot (r = −0.65). Linear regression models also showed that the fresh yield had significant negative relationships with anthracnose (R2 = 0.26) and viral (R2 = 0.62) diseases. The accessions identified as resistant constitute a valuable resource for breeding of resistant germplasm.

Resistance in water yam (Dioscorea alata) cultivars in the French West Indies to anthracnose disease based on tissue culture‐derived whole‐plant assay

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 0–6 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 isolate–cultivar 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.

Optimizing Controlled Environment Assessment of Levels of Resistance to Yam Anthracnose Disease Using Tissue Culture‐derived Whole Plants

AbstractStudies were conducted to determine the timing and frequency of disease assessment required to effectively identify levels of resistance to yam anthracnose using tissue culture‐derived whole plant inoculation assay. The effects of inoculation methods (paint brush and spray), and disease scoring methods [individual leaf area (ILA) and whole plant area (WPA)] were also assessed. Spray inoculation resulted in rapid infection and higher variations among yam genotypes, leading to earlier discrimination of genotypes than with the paintbrush method. Both the ILA and WPA scoring methods showed variation among yam genotypes, and association between the two methods gave a high positive correlation (r > 0.90). However, the WPA was faster and had the advantage of detecting differences in reactions of yam genotypes to less aggressive pathogen isolates to which the ILA method showed no variation. A single disease evaluation at 7 days after inoculation was as good as the area under the disease progress curve (AUDPC) and the disease progress rate (Rd) derived from multiple evaluations. However, a significant time–genotype interaction, suggests a need for more than a single assessment for effective comparison of genotypes. AUDPC derived from two assessments (5 and 7 DAI) was better than AUDPC from three assessments (5, 7 and 9 DAI) in separating genotypes reactions to a less aggressive pathogen isolate. This study showed that the use of spray inoculation method, the WPA scoring method, and AUDPC derived from two assessments (5 and 7 DAI) provided best conditions for evaluating yam genotypes for levels of anthracnose resistance with the tissue culture‐derived whole plant assay.

Two Genetically Distinct Populations of Colletotrichum gloeosporioides Penz. Causing Anthracnose Disease of Yam (Dioscorea spp.)

AbstractVariation within Colletotrichum gloeosporioides, the causal agent of yam anthracnose disease, is still poorly defined and this hinders breeding for resistance. Two morphotypes of C. gloeosporioides, designated slow‐growing grey (SGG) and fast‐growing salmon (FGS), are associated with anthracnose disease of yam in Nigeria. The morphotypes are distinguishable based on colony and conidial morphology, growth rate, virulence, as well as vegetative compatibility, but molecular differentiation of SGG and FGS strains is needed to facilitate epidemiological studies. Denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR)‐amplified small subunit (18S) rDNA fragments, and microsatellite‐primed PCR (MP‐PCR) genomic fingerprinting were employed to provide a basis for molecular differentiation of the morphotypes. DGGE analysis revealed patterns that clearly differentiated isolates of the aggressive defoliating SGG from the moderately virulent non‐defoliating FGS strains. Genetic analysis based on 52 MP‐PCR markers revealed highly significant differentiation between the SGG and FGS populations on yam (GST = 0.22; Nei's genetic identity = 0.85; θ = 0.28, P < 0.001), indicating that the SGG and FGS morphotypes represent genetically differentiated populations. The results of the molecular typing using DGGE and MP‐PCR analyses were consistent with the disease phenotype caused by the two morphotypes. Consequently, these molecular techniques might be used, at least partly, to replace time‐consuming virulence studies on yam.

English

Water yam (Dioscorea alata L.) is the most widely cultivated yam species globally. The major limitation to the profitable and sustainable production of D. alata is its susceptibility to anthracnose disease. The availability of resistant varieties could potentially form the cornerstone of an integrated management strategy for yam anthracnose; however, anthracnose resistance breeding is hampered by the dearth of knowledge on pathogen identity and diversity. Four forms of Colletotrichum are now known to be associated with foliar anthracnose of yam: the slow-growing grey (SGG), the fast-growing salmon (FGS), the fast-growing olive (FGO), and the fast-growing grey (FGG) forms. The close phylogenetic relationship of the first three forms to reference isolates of Colletotrichum gloeosporioides, and the fact that only strains of these forms have been observed to induce typical anthracnose symptoms on D. alata, recently confirmed that C. gloeosporioides is the causal agent of yam anthracnose disease. The FGG form possibly represents a distinct, endophytic, species as indicated by morphological, biological and molecular criteria. Previous research emphasized epidemiology and control but limited progress was made in understanding yam anthracnose disease based on this classical approach. Molecular approaches have started to unravel the systematics and ecology of Colletotrichum strains associated with yam anthracnose, as well the population biology of C. gloeosporioides on yam. Sexual recombination is a likely mechanism contributing to the high genetic diversity of C. gloeosporioides in yam-based cropping systems. Studies have been initiated to understand the mechanisms that generate genetic variation in C. gloeosporioides, and to gain some insight into the biochemistry of the interactions between the pathogen and yam. Our thesis in this article is that integrating traditional and molecular approaches to understanding C. gloeosporioides systematics, epidemiology and population genetics will lead to a much better understanding of yam anthracnose disease, and thus to the development of effective and sustainable control measures. Research successes and challenges are discussed, as well as their implications for future studies on pathogen evolutionary potential, anthracnose resistance breeding, and the deployment of resistance genes. Key words : Anthracnose, Colletotrichum gloeosporioides, Dioscorea spp., molecular markers, molecular systematics, population biology, resistance breeding, yam. African Journal of Biotechnology Vol. 2 (12), pp. 486-496, December 2003

Identification and application of RAPD markers for anthracnose resistance in water yam (Dioscorea alata)

SummaryAnthracnose, caused by Colletotrichum gloeosporioides, is the most severe foliar disease of water yam (Dioscorea alata) worldwide. The tetraploid breeding line, TDa 95/00328, is a source of dominant genetic resistance to the moderately virulent fast growing salmon (FGS) strain of C. gloeosporioides. Bulked segregant analysis was used to search for random amplified polymorphic DNA (RAPD) markers linked to anthracnose resistance in F1 progeny derived from a cross between TDa 95/00328 and the susceptible male parent, TDa 95–310. Two hundred and eighty decamer primers were screened using bulks obtained from pooled DNA of individuals comprising each extreme of the disease phenotype distribution. A single locus that contributes to anthracnose resistance in TDa 95/00328 was identified and tentatively named Dcg‐1. We found two RAPD markers closely linked in coupling phase with Dcg‐1, named OPI71700 and OPE6950, both of which were mapped on the same linkage group. OPI71700 appeared tightly linked to the Dcg‐1 locus; it was present in all the 58 resistant F1 individuals and absent in all but one of the 13 susceptible genotypes (genetic distance of 2.3 cM). OPE6950 was present in 56 of the 58 resistant progeny and only one susceptible F1 plant showed this marker (6.8 cM). Both markers successfully identified Dcg‐1 in resistant D. alata genotypes among 34 breeding lines, indicating their potential for use in marker‐assisted selection. OPI71700 and OPE6950 are the first DNA markers for yam anthracnose resistance. The use of molecular markers presents a valuable strategy for selection and pyramiding of anthracnose resistance genes in yam improvement.

Molecular identification of Colletotrichum gloeosporioides causing yam anthracnose in Nigeria

Four forms of Colletotrichum representing three distinct virulence phenotypes were found associated with foliar anthracnose of yam in Nigeria: the highly virulent (= severity of disease) slow‐growing grey (SGG); the moderately virulent fast‐growing salmon (FGS); the weakly virulent fast‐growing grey (FGG); and the moderately virulent fast‐growing olive (FGO) morphotype. Isolates of the four forms were identified as C. gloeosporioides, based on morphology. The reaction of monoconidial cultures on casein hydrolysis medium (CHM), PCR‐RFLP and sequence analysis of the internal transcribed spacer region of the ribosomal DNA (ITS1‐5·8S‐ITS2) were used to establish the identity of the yam anthracnose pathogen(s). All yam isolates were distinguished from C. acutatum by the absence of protease activity on CHM. On ITS PCR and enzymatic digestion of PCR products, all FGS, FGO and SGG isolates produced RFLP patterns identical to those of C. gloeosporioides reference isolates, while FGG isolates revealed unique ITS RFLP banding patterns. Sequence analysis of the ITS1 region and of the entire ITS region revealed that SGG, FGS and FGO isolates were highly similar (98–99% nucleotide identity) and showed 97–100% identity to C. gloeosporioides. Less than 93% similarity of these fungal isolates to reference C. acutatum and C. lindemuthianum isolates was observed. The molecular study confirmed that foliar anthracnose of yam is caused by C. gloeosporioides. While a high similarity was found among most C. gloeosporioides fungi from yam, isolates of the FGG form did not cluster with any previously described Colletotrichum species, and probably represent a distinct species.

Screening of cassava and yam cultivars for resistance to anthracnose using toxic metabolites of colletotrichum species.

Collectotrichum gloeosporioides f. sp. manihotis and C. gloeosporioides, causal agents of cassava (Manihot spp.) and yam (Dioscorea spp.) anthracnose diseases, respectively, produce toxic metabolites in culture that fluoresce at 254 nm and 366 nm, producing bands with Rf of 0.65 and 7.0, respectively. Symptoms induced on yam and cassava by the extracted metabolites were similar to those induced by the pathogens. Twenty-four clones of tropical D. rotundata (TDr), D. alata (TDa), D. esculenta (TDe), and D. cavenensis (TDc) were screened by applying toxic metabolites of C. gloeosporioides to their leaves and stems. Only TDr131, TDe179 and TDc750 were resistant. Other clones were susceptible to varying degrees. Nineteen of the 45 clones of M. esculenta were resistant to varying degrees of toxic metabolites of C. gloeosporioides f. sp. manihotis. Results from in vitro screening of' cassava and yam clones using toxic metabolites compared favourably with field screening based on natural epidemics. Using toxic metabolites appears to be a more effective technique for screening for disease resistance than conventional inoculation with plant pathogens.

SEMI-SELECTIVE MEDIUM FOR ISOLATION OF COLLETOTRICHUM GLOEOSPORIOIDES FROM SOIL

Isolation of Colletotrichum gloeosporioides (causal agent of yam anthracnose) from soil is difficult because of the presence of fast growing micro-organisms like Rhizoctonia spp., Rhizopus spp., Penicillium spp. and bacteria. Therefore, a semi-selective medium consisting of antimicrobial agents such as pencycuron 50 mg L−1, tolclofos-methyl 10 mg L−1, streptomycin sulphate 100 mg L−1, chloramphenicol 100 mg L−1 and chlortetracycline 100 mg L−1 was developed using potato dextrose agar as the basal medium. The procedure involved screening of candidate fungicides, developing a suitable combination of antimicrobial agents, assessing the proposed semi-selective medium and adjustment of pH of the medium to 5. The effectiveness of the medium is due to selective inhibition of micro-organisms by the antimicrobial agents and the development of distinct salmon-pink colonies of C. gloeosporioides.

Comparison of disease progress curves for yam anthracnose (Colletotrichum gloeosporioides)

The incidence and severity of foliar anthracnose, caused by Colletotrichum gloeosporioides, were recorded for seven fields of yams (Dioscorea alata var. White Lisbon) in Barbados over a complete growing season. A scoring system was developed to estimate severity, placing the emphasis on the earliest, least visible stages of the disease. Disease progress curves were described in terms of the logistic model and the effects of site and leaf age on the parameters as well as on disease at tuber bulking were analysed. Differences between sites indicated that disease was more severe in areas of higher rainfall. In severe epidemics, disease predominated on young leaves. Fungicides applied sufficiently early can delay the onset of disease and slow the rate of disease progress.