Frosty pod of cacao: a disease with a limited geographic range but unlimited potential for damage

ABSTRACT Moniliophthora roreri, the cause of frosty pod rot (FP), is a specialized fungal pathogen (family Marasmiaceae) that invades only actively growing pods of cacao, Theobroma cacao, and related species of Theobroma and Herrania. FP damages pods and the commercially important seeds that some of these species produce. M. roreri was confined to northwestern South America until the 1950s. Its appearance in Panama in 1956 signaled a change in its geographic distribution. Now, it is found in 11 countries in tropical America. The fungus is currently in an active dispersal phase, possibly due to an increase in human-mediated spread. FP is more destructive than black pod (Phytophthora spp.) and more dangerous and difficult to control than witches' broom, caused by Moniliophthora (Crinipellis) perniciosa. The aggressiveness of M. roreri, its capacity to survive different environmental conditions, its rapid natural dispersal, its propensity for man-mediated dispersal, and the susceptibility of most commercial cacao genotypes, all indicate that FP presents a substantial threat to cacao cultivation worldwide.     

Development of a marker assisted selection program for cacao

ABSTRACT Production of cacao in tropical America has been severely affected by fungal pathogens causing diseases known as witches' broom (WB, caused by Moniliophthora perniciosa), frosty pod (FP, caused by M. roreri) and black pod (BP, caused by Phytophthora spp.). BP is pan-tropical and causes losses in all producing areas. WB is found in South America and parts of the Caribbean, while FP is found in Central America and parts of South America. Together, these diseases were responsible for over 700 million US dollars in losses in 2001 (4). Commercial cacao production in West Africa and South Asia are not yet affected by WB and FP, but cacao grown in these regions is susceptible to both. With the goal of providing new disease resistant cultivars the USDA-ARS and Mars, Inc. have developed a marker assisted selection (MAS) program. Quantitative trait loci have been identified for resistance to WB, FP, and BP. The potential usefulness of these markers in identifying resistant individuals has been confirmed in an experimental F(1) family in Ecuador.     

Cacao diseases: important threats to chocolate production worldwide

ABSTRACT Theobroma cacao, cacao, is an ancient, neotropical domesticate. It is now grown throughout the humid, lowland tropics and is the basis of a multibillion dollar confectionary trade. Diverse diseases impact production of the crop. They reduce yields by ca. 20%, but could cause far greater losses if certain highly damaging diseases were to become more widely distributed. Among the most potentially dangerous of these diseases are frosty pod, caused by Moniliophthora roreri, and witches' broom, caused by M. perniciosa (previously Crinipellis perniciosa). These two diseases occur only in the Western Hemisphere, and severe losses would follow their introduction to West Africa and Asia, where ca. 86% of all cacao production occurs. Elsewhere, Cacao swollen shoot virus and the damaging black pod agent, Phytophthora megakarya, are found in Western Africa; whereas vascular streak dieback, caused by Oncobasidium theobromae, is present only in Asia. Breeding programs are challenged by minimal resistance to some of the diseases. Progress that has been made is threatened by the "emergence" of other serious diseases, such as Ceratocystis wilt (Ceratocystis cacaofunesta). During this symposium, new insights are discussed on the biology, origins, pathology and phylogeny of the pathogens; as well as the biological, chemical and genetic management of the diseases that they cause.     

Cacao diseases: a global perspective from an industry point of view

ABSTRACT Diseases of cacao, Theobroma cacao, account for losses of more than 30% of the potential crop. These losses have caused a steady decline in production and a reduction in bean quality in almost all the cacao-producing areas in the world, especially in small-holder farms in Latin America and West Africa. The most significant diseases are witches' broom, caused by Moniliophthora perniciosa, which occurs mainly in South America; frosty pod rot, caused by M. roreri, which occurs mainly in Central and northern South America; and black pod disease, caused by several species of Phytophthora, which are distributed throughout the tropics. In view of the threat that these diseases pose to the sustainability of the cacao crop, Mars Inc. and their industry partners have funded collaborative research involving cacao research institutes and governmental and nongovernmental agencies. The objective of this global initiative is to develop short- to medium-term, low-cost, environmentally friendly disease-management strategies until disease tolerant varieties are widely available. These include good farming practices, biological control and the rational or minimal use of chemicals that could be used for integrated pest management (IPM). Farmer field schools are used to get these technologies to growers. This paper describes some of the key collaborative partners and projects that are underway in South America and West Africa.     

Molecular characterization of fungal endophytic morphospecies isolated from stems and pods of Theobroma cacao

Endophytic fungi were isolated from healthy stems and pods of cacao ( Theobroma cacao ) trees in natural forest ecosystems and agroecosystems in Latin America and West Africa. These fungi were collected for screening as a potential source of biocontrol agents for the basidiomycetous pathogens of cacao in South and Central America, Moniliophthora roreri (frosty pod rot) and Moniliophthora perniciosa (witches' broom). Many of these isolates were morphologically unidentifiable as they failed to form fruiting structures in culture, or only produced arthrosporic stages. Affinities with basidiomycetes were suspected for many of these based on colony morphology. Fifty-nine of these morphologically unidentifiable isolates were selected for molecular identification by DNA extraction and sequence analysis of nuclear ribosomal DNA (rDNA). The large subunit (LSU) was chosen for initial sequencing because this region has been used most often for molecular systematics of basidiomycete fungi, and comprehensive LSU datasets were already available for sequence analyses. Results confirmed that the majority of the isolates tested belonged to the Basidiomycota, particularly to corticoid and polyporoid taxa. With LSU data alone, identification of the isolates was resolved at varying taxonomic levels (all to order, most to family, and many to genus). Some of the isolates came from rarely isolated genera, such as Byssomerulius , whilst the most commonly isolated basidiomycetous endophyte was a member of the cosmopolitan genus Coprinellus (Agaricales). The role of these fungi within the host and their potential as biological control agents are discussed.     

Phylogeny of the frosty pod rot pathogen of cocoa

Morphological, cytological and molecular evidence is presented which confirms that the frosty pod rot pathogen of cocoa, formerly classified as the mitosporic fungus Moniliophthora roreri (Deuteromycota), belongs to the hymenomycetous genus Crinipellis (Basidiomycota) and that two varieties should now be recognized: Crinipellis roreri var. roreri and the new variety C. roreri var. gileri . The latter was collected on Theobroma gileri , an endemic tree of submontane forests in north-west Ecuador, and can be distinguished from Ecuadorian and Peruvian isolates from cocoa ( T. cacao ) on the basis of spore morphology, incompatibility and nucleotide sequence data. As with var. roreri , meiosis is shown to occur within the dispersive and infective spore stage of var. gileri and these meiospores are interpreted to represent a much modified probasidium. In addition, in a field inoculation experiment, an isolate from T. gileri proved to be noninfective to cocoa pods when compared with positive control strains isolated from T. cacao in western Ecuador and T. bicolor in eastern Ecuador. It is concluded that var. gileri is the vestigial progenitor of the frosty pod rot pathogen of cocoa, with a host range and distribution restricted to T. gileri in the mesic forests of north-west South America.     

Molecular and metabolic changes of cherelle wilt of cacao and its effect on Moniliophthora roreri

Young Theobroma cacao pods, known as cherelles, are commonly lost to physiological thinning known as cherelle wilt. Cherelles are susceptible to frosty pod rot caused by Moniliophthora roreri. We studied the cherelle wilt process and its impact on M. roreri infection using microscopic, metabolite, and gene expression analyses. Wilt was associated with increased levels of tricarboxylic acid cycle intermediaries and decreased levels of major metabolites. Expression changes of cacao ESTs in response to wilt suggest induction of the polyamine, ethylene, and jasmonic acid biosynthetic pathways and regulation of abscisic acid and cytokinin levels. M. roreriinfection caused little alteration of cherelle physiology. M. roreri responded to the late stage of wilt by altering the expression of M. roreri ESTs associated with metabolite detoxification and host tissue degradation. The environment of the wilting cherelles may truncate the disease cycle of frosty pod rot, by limiting M. roreri sporulation and stopping the lifecycle.     

Biodiversity and biogeography of the cacao (Theobroma cacao) pathogen Moniliophthora roreri in tropical America

Moniliophthora roreri , the cause of moniliasis or frosty pod rot, occurs on the neotropical rainforest genera Theobroma and Herrania . While this basidiomycete has had devastating effects on the cacao tree ( T. cacao ) in tropical America, where it is confined, little is known of its biogeography and intraspecific genetic variability. Here, AFLP and ISSR profiles of 94 isolates of M. roreri from across its geographic range in Central/South America were analyzed. The study provided limited evidence to support the hypothesis that M. roreri is capable of sexual reproduction. The highest levels of genetic diversity occurred in Colombia and not in Ecuador as originally believed. The fungus was broadly divided into five genetic groups. Two of these have a wide geographic range: Bolívar group (north of Santander in Colombia, eastern Venezuela, peripheral Ecuador, Peru), and Co-West group (western Colombia, central Ecuador, Central America). The other groups are all apparently endemic to Colombia (Co-East and Co-Central groups) or north-western Ecuador ( Gileri group). We speculate that central/north-eastern Colombia may represent the centre of origin for M. roreri . Sequence data from the internal transcribed spacer region of the nuclear rDNA repeat were congruent with the AFLP/ISSR results, dividing M. roreri into two broad groups: the Orientalis group, comprising most isolates from the Co-East, Co-Central and Bolívar groups, and the Occidentalis group, comprising isolates from the Co-West and Gileri groups. The spread of M. roreri into new areas and countries mediated by human activity is discussed.     

A disease and production index (DPI) for selection of cacao (Theobroma cacao) clones highly productive and tolerant to pod rot diseases

Frosty pod rot (FPR) (Moniliophthora roreri), along with black pod rot (Phytophthora species) and witches’ broom disease (Moniliophthora perniciosa) constitute the main phytosanitary problems limiting cacao (Theobroma cacao) production causing severe yield losses. One of the main sought after methods of pod rot management is the selection of tolerant cacao genotypes. Typically, the selection is carried out through the quantification of the percentage of diseased pods (PDP). However, PDP does not consider the relative productivity, or production potential (PT) of the genotype. Production potential can vary among cacao genotypes. Consequently, genotypes with similar PT can have similar or vastly different disease tolerance levels as measured by PDP. The disease and production index (DPI) was developed to integrate a genotype's tolerance to M. roreri and other diseases as measured by PDP, with its PT. Here, we evaluated the number of healthy pods, number of diseased pods, and weight of fresh seed for 29 clones grown in replicated five-tree plots over 4 years. The data obtained was used to calculate PDP and DPI for each clone for three different disease combinations: frosty pod rot alone, pod rots other than frosty pod rot, and the combination of all pod rots. Multivariate analysis verified that DPI discriminated between clones based on productivity and disease tolerance. Surprisingly, there was a close ranking of clones between resistance to FPR and resistance to all other pod rots. The DPI can be used in breeding programmes focused on the selection of high yielding disease-tolerant cacao genotypes.     

Of mushrooms and chocolate trees: aetiology and phylogeny of witches' broom and frosty pod diseases of cacao

The troubled history of the two major diseases of the chocolate tree (Theobroma cacao) in South America, witches' broom and frosty pod, is reviewed, concentrating on critical aspects of the aetiology as well as the phylogeny of the causal agents. Both diseases are caused by sister species within the genus Moniliophthora, belonging to the Marasmiaceae family of mushrooms. The witches' broom pathogen, Moniliophthora perniciosa, evolved on the Amazonian side of the Andes and induces brooms not only in cacao and its relatives in the genera Theobroma and Herrania (Malvaceae), but also in species in the plant families Bignoniaceae, Malpighiaceae and Solanaceae, on which the mushrooms (basidiomata) are produced. Moniliophthora roreri, the type species of the genus, evolved as a pod pathogen on endemic Theobroma species on the western side of the northern Andean Cordillera. Because Moniliophthora was described originally as the asexual form of an unknown basidiomycete, the generic diagnosis is amended here to accommodate species with agaricoid basidiomata. In addition, the new variety M. roreri var. gileri is designated for the morphotype occurring on Theobroma gileri, in northwest Ecuador. Cytology studies indicate that the supposed conidia of M. roreri are, in fact, sexual spores (meiospores) and it is posited that the fruiting structure represents a much‐modified mushroom. Finally, based on preliminary data from pathogenicity testing, it is hypothesized that the true causal agent of both diseases is an as yet unidentified infectious agent vectored into the host by the fungus.     

Cocoa production in Ecuador in relation to dry-season escape from pod rot caused by Crinipellis perniciosa and Moniliophthora roreri

Cropping and disease patterns were observed over a period of 4 years in individual cocoa trees at three contrasting sites near Quevedo, Ecuador, with the aim of evaluating the importance of dry season production and the consequent avoidance of pod disease caused by Crinipellis perniciosa and Moniliophthora roreri. Distinction of the diseases on necrotic pods was aided by splitting open pods after removal from the tree. The year was divided into wet season (April to September) and dry season (October to March) harvest periods. The 3-month lag between the actual wet and dry seasons and these harvest periods allowed time for symptom development. Combined disease losses ranged from 19·0% to 62·1 % at the various sites. At all sites and in each year, more ripe healthy pods were harvested in the dry season harvest period than in the wet season harvest period. In general, pod losses caused by C, perniciosa were curtailed more sharply by the dry season than those caused by M. roreri , which at one site caused as much damage in the dry season as in the wet season. The proportion of the annual production falling in the dry season harvest period varied among sites and between years at a given site. A comparison of trees from two progenies growing side by side showed consistent differences in cropping patterns, and identified certain trees that were productive, yielded consistently in the dry season harvest period, and were little affected by M, roreri. Analysis of long-term rainfall data for the Quevedo region indicated that years which lack a normal dry season do occur, but only once every 10 years on average. Further exploitation of disease escape through selection and breeding appears to be both feasible and appropriate.     

A qualitative host-pathogen interaction in the Theobroma cacao-Moniliophthora perniciosa pathosystem

The aim of this study was to test whether resistance of clones of Theobroma cacao (cocoa) varied between isolates of Moniliophthora (formerly Crinipellis ) perniciosa , the cause of witches' broom disease. Developing buds of vegetatively propagated T. cacao grown in greenhouses in the UK were inoculated with 16 000 spores of M. perniciosa per meristem in water, under conditions where water condensed on the inoculated shoot for at least 12 h after inoculation. The proportion of successful inoculations varied between clones and was inversely correlated with time to symptom production or broom formation. A specific interaction was demonstrated among three single-spore isolates of M. perniciosa and the clone Scavina 6 (SCA 6) and a variety of susceptible clones. Isolates Castenhal-I and APC3 were equally likely to infect SCA 6 and the other clones, but isolate Gran Couva A9 never infected SCA 6, although it was as virulent on the other clones. The interaction was maintained when the wetness period was extended to 70 h. Offspring of SCA 6 × Amelonado matings were all susceptible to both Castenhal-I and GC-A5, with no evidence of greater variability in susceptibility to GC-A5 than Castanhal-I. This suggests recessive inheritance of a single homozygous factor conferring resistance to GC-A5, from SCA 6. The progenies were slightly more susceptible to Castanhal-I than GC-A5. The implications for managing the disease are discussed.     

Induction of resistance in cocoa against Crinipellis perniciosa and Verticillium dahliae by acibenzolar- S-methyl (ASM)

The benzothiadiazole compound acibenzolar- S -methyl (ASM) was assessed as an inducer of resistance against Crinipellis perniciosa , agent of witches' broom, and Verticillium dahliae , agent of vascular wilt, both on cocoa. ASM induced a reduction in incidence of witches' broom of up to 84·5% when sprayed 30 days before inoculation on cocoa seedlings of cv. Catongo. ASM also induced a reduction in severity of Verticillium wilt to 55·4% on cv. Theobahia. For both pathosystems, effects of dose on disease were not clearly observed. The efficacy of the inducer increased with the interval between sprayings and the respective inoculations with the pathogens. In another experiment, the effect of ASM on the control of witches' broom on cocoa seedlings was compared with that of cuprous oxide and tebuconazole, all sprayed 15 days before inoculation. ASM reduced disease incidence by 60·1% compared with the inoculated control. ASM was superior to tebuconazole, and there was also a tendency for ASM to be better than cuprous oxide. To understand the mechanism of action of ASM as an inducer of resistance, alterations in the levels of total phenolics, polyphenol oxidases and peroxidases were evaluated 3, 15 and 30 days after spraying of seedlings of cv. Catongo. Enzyme activities from seedlings of cv. Theobahia were evaluated 30 days after spraying. On cv. Catongo, no significant differences in total phenolic content and polyphenol oxidase activity were detected after spraying. However, an increase in peroxidase activity was detected at all times of evaluation. On cv. Theobahia, significant increases in activities of peroxidase and polyphenol oxidase were detected, indicating that defence responses due to ASM were dependent on host genotype.     

Evaluation of cacao (Theobroma cacao) clones against seven Colombian isolates of Moniliophthora roreri from four pathogen genetic groups

Artificial pod inoculation was used to compare the relative aggressiveness of seven Colombian isolates of Moniliophthora roreri (the causal agent of moniliasis or frosty pod disease), representing four major genetic groupings of the pathogen in cacao (cocoa), when applied to five diverse cacao genotypes (ICS-1, ICS-95, TSH-565, SCC-61 and CAP-34) at La Suiza Experimental Farm, Santander Department, Colombia. The following variables were evaluated 9 weeks after inoculation of 2- to 3-month-old pods with spore suspensions (1·2 × 10⁵ spores mL⁻¹): (i) disease incidence (DI); (ii) external severity (ES); and (iii) internal severity (IS). IS was found to be of greatest value in classifying the reaction of the host genotype against M. roreri . Genetic variation reported between isolates and cacao genotypes was not matched by similar diversity in their aggressiveness. All isolates were generally highly aggressive against most cacao genotypes, with only two isolates showing reduced IS and ES reactions. There was considerable variation between clones in the IS and ES scores, but one cultivated clone (ICS-95) displayed a significant level of resistance against all seven isolates. This clone may be useful in cacao breeding initiatives for resistance to moniliasis of cacao.     

Pathogenic variability amongst isolates of Crinipellis perniciosa from cocoa (Theobroma cacao)

Dry witches' brooms from cocoa were imported from various areas within Bolivia, Brazil, Colombia, Ecuador, Trinidad and Venezuela. Basidiocarps of Crinipellis perniciosa were induced to form on these brooms and seedlings of different types of cocoa were inoculated with basidiospores either on the hypocotyl or cotyledon bud. Host reactions were assessed mainly by recording stem base swelling and broom development at the cotyledon node (hypocotyl inoculations) or the extent of swelling and branching of shoots (cotyledon bud inoculations).
Results from 30 experiments indicated considerable diversity amongst isolates in inducing disease symptoms, but suggested that two groups or populations of C. perniciosa exist on cultivated cocoa. One group (A), comprising isolates from Bolivia and Pichilingue (Ecuador) and most isolates tested from Colombia, induced severe symptoms on cocoa with Scavina 6 as one parent; the other group (B), comprising isolates from Brazil, Trinidad and Venezuela, did not. Within these groupings variants could be further distinguished by particular host reactions. Isolates from Ecuador, especially from the Oriente, a centre of diversity for Theobroma cacao, showed a range of pathogenicity comparable to that found amongst isolates from cultivated cocoa over a much wider area.     

An optimized screening method for identifying levels of resistance to Crinipellis perniciosa in cocoa (Theobroma cacao)

The effects of host age, leaf number, host type (clone or seedling), pathogen spore concentration and incubation time on inoculation with Crinipellis perniciosa (witches' broom disease of cocoa) were studied in greenhouse experiments using susceptible cocoa genotypes. Three methods of inoculation (agar-drop, water-drop and spray) were also tested. An optimized inoculation method was selected and tested for its repeatability as well as its ability to discriminate between various levels of resistance to C. perniciosa in cocoa. The optimized method (350 000 viable basidiospores per mL, 60 h incubation, agar-drop technique) produced 100% infection repeatedly, on both clonal and seedling plants of a susceptible genotype. Seedling age (2–12 months) and leaf number did not significantly affect the percentage of plants with symptoms or broom characteristics. This method discriminated effectively between the various levels of resistance in 14 cocoa genotypes and is recommended as an inoculation method to identify levels of resistance in germplasm collections. Symptom severity was shown to be a better measure of resistance than infection success.     

Random amplified polymorphic DNA (RAPD) analysis of Crinipellis perniciosa isolates from different hosts

DNA sequence polymorphisms among isolates of Crinipellis perniciosa , causing witches' broom disease in several genera and species in the Sterculiaceae, Solanaceae and Bixaceae, were revealed by random amplified polymorphic DNA (RAPD) banding patterns. Comparisons were made among isolates from Theobroma cacao (cultivars SCA6 and SVB), T. grandiflorum, T. obovatum, T. subincanum, Herrania sp. (all collected near Belém, Brazil, at three plots within two kilometres of each other), and Bixa orellana and Solanum rugosum (150 and 1440 km from Belém, respectively). Genetic similarities, as determined by Sorensen's coefficient ( Sc_ij ; fraction of shared bands), ranged from 1·00 ( T. cacao SCA6 and T. grandiflorum ) to 0·56 ( S. rugosum and T. subincanum). Results indicate that proximity is more important than host species for determining the genetic relationships among isolates, that there were two independent sources of inoculum at the site near Belém, and that the isolate from B. orellana is most closely related to the isolate from T. cacao SVB. Comparisons were also made among multispore cultures from different basidiocarps on the same T. cacao SVB broom, as well as monospore cultures from the same S. rugosum basidiocarp. Banding patterns were similar among basidiocarps on the same broom, but differences were detected among monospore cultures from the same basidiocarp.     

泡桐丛枝病病原MLO在寄主离体培养组织中的保存与增殖

 感染丛枝病病原MLO的泡桐幼苗茎梢,经表面灭菌后,置于附加0~3mg/1 6-BA的MS培养基中进行离体培养,由腋芽或顶芽直接伸长生长所形成的试管苗或经不定芽发生途径所形成的试管苗在1个月内均表现出典型的丛枝症状。在高浓度6-BA(2~3mg/1)处理培养基中,丛枝试管苗1月左右病死。当外植体为0.5mm长的微茎尖时,试管苗的丛枝症状有所减轻。DAPI荧光染色镜检结果表明,这种已表现出丛枝的试管苗的根、茎、叶柄的韧皮部筛管,以及已具维管束分化的愈伤组织的筛管部位发出很强的MLO特异荧光。超薄切片电镜观察发现,离体培养组织中的MLO形态具有较多处于活跃增殖期的哑铃形和芽殖形。以30天为一个继代周期,在无激素培养基或低浓度6-BA(lmg/1)与无激素培养基交替继代培养1年之后,来源于染病愈伤组织和试管苗中的MLO深度一直维持在相当高的水平。试验表明,利用寄王组织进行离体培养是保存、繁殖并进一步深入研究泡桐丛枝病病原MLO的一条良好途径。     

Detection of a mycoplasma-like organism in peanut plants with witches' broom using indirect enzyme-linked immunosorbent assay (ELISA)

The mycoplasma-like organism (MLO) associated with peanut (groundnut) witches' broom (PWB) from India was partially purified and an antiserum produced against it. Using a protein A indirect enzyme-linked immunosorbent assay (ELISA) procedure, PWB MLO was detected in crude extracts of leaves. stems and pegs of infected peanut plants, although stems were a better source than leaves and pegs. Extracts of infected tissues of three diseases of assumed MLO etiology in India, little leaf of brinjal (eggplant). I med rosed witches "broom, and Daturd sp. witches' broom, failed to react with the PWB MLO antiserum.     

Molecular Fingerprinting Suggests Two Primary Outbreaks of Witches' Broom Disease (Crinipellis perniciosa) of Theobroma cacao in Bahia, Brazil

Crinipellis perniciosa (Stahel) Singer is the causal agent of witches' broom disease in the Sterculiaceae, Solanaceae, and Bixaceae families. The disease is endemic to the Brazilian Amazon, and was first reported infecting Theobroma cacao (cocoa) in the State of Bahia, Brazil, in 1989. Random amplified polymorphic DNA (RAPD) analyses were performed on 46 isolates of C. perniciosa from cocoa that were collected from 15 counties in Bahia and the Brazilian Amazon. A total of 258 RAPD loci from 20 primers and three mixed primers were analyzed. Of these loci, 108 (42%) were polymorphic, with an average of 4.7 polymorphic loci per primer produced. Genetic similarities were estimated using Nei and Li's index and UPGMA clustering. Bootstrap analysis divided the phenogram into four significantly different clusters: two groups contained isolates from Ariquemes and from Ouro Preto, Rondônia, and the other two separated the isolates from Bahia into two major groups of C. perniciosa, classified as Group 1 (G1) and Group 2 (G2). The two groups of isolates from Bahia differed for their genetic similarity with the isolates from the Brazilian Amazon. The geographic distribution of the groups in Bahia suggests two independent focal points of introduction. Ongoing programs to screen for resistant cocoa genotypes should consider both groups of isolates.