True bug (Heteroptera) impact on cocoa fruit mortality and productivity.

The real impact of true bug damage on cocoa pods has never been assessed precisely. We conducted a 2-yr study on 1,080 cocoa trees on 36 farms in Cameroon to assess the contribution of true bugs to fruit mortality and production loss. The cocoa fruiting cycle, fruit mortality, and damage caused by true bugs as well as other pests and diseases were monitored on a weekly basis. True bug damage also was described on 2,500 ripe pods per year. Pod weight, bean number, and bean weight were measured and compared for different degrees and types of damage on the ripe pods. Our results showed that true bugs were the main external cause of young fruit abortion. They reduced the abundance of young fruit by up to 10%. In contrast, although one-third of the ripe pods sampled had true bug lesions, only 4% were moderately to heavily damaged. The mean weight of ripe pods was reduced by 12% when there was medium to heavy damage. While the mean weight of wet beans was reduced significantly (by 3-10%), the number of beans per pod was not changed by damage. Despite the reduction in mean weight, the overall weight of beans for the pods sampled was reduced by <2%. Therefore, our study confirmed the common assumption that the economic impact of true bug damage on mature pods is negligible on cocoa farms in Cameroon. However, true bugs have a significant impact on young fruit mortality.

Understanding the recent colonization history of a plant pathogenic fungus using population genetic tools and Approximate Bayesian Computation.

Understanding the processes by which new diseases are introduced in previously healthy areas is of major interest in elaborating prevention and management policies, as well as in understanding the dynamics of pathogen diversity at large spatial scale. In this study, we aimed to decipher the dispersal processes that have led to the emergence of the plant pathogenic fungus Microcyclus ulei, which is responsible for the South American Leaf Blight (SALB). This fungus has devastated rubber tree plantations across Latin America since the beginning of the twentieth century. As only imprecise historical information is available, the study of population evolutionary history based on population genetics appeared most appropriate. The distribution of genetic diversity in a continental sampling of four countries (Brazil, Ecuador, Guatemala and French Guiana) was studied using a set of 16 microsatellite markers developed specifically for this purpose. A very strong genetic structure was found (F(st)=0.70), demonstrating that there has been no regular gene flow between Latin American M. ulei populations. Strong bottlenecks probably occurred at the foundation of each population. The most likely scenario of colonization identified by the Approximate Bayesian Computation (ABC) method implemented in DIYABC suggested two independent sources from the Amazonian endemic area. The Brazilian, Ecuadorian and Guatemalan populations might stem from serial introductions through human-mediated movement of infected plant material from an unsampled source population, whereas the French Guiana population seems to have arisen from an independent colonization event through spore dispersal.

Geographical Distribution of Cacao swollen shoot virus Molecular Variability in Côte d'Ivoire.

The discovery of new outbreaks caused by Cacao swollen shoot virus (CSSV) in Côte d'Ivoire in 2003, when this disease seemed to have been eradicated since the end of the 1950s in that country, casts doubt on the sustainability of Ivorian cocoa production. The aims of this study were, first, to carry out a molecular characterization of CSSV isolates from the main outbreaks in Côte d'Ivoire; second, determine their phylogenetic position in relation to isolates already discovered in Togo and Ghana; and, finally, study their geographical distribution to understand the dispersal of the virus. Additionally, this study was intended to enable the implementation and validation of a polyvalent molecular diagnosis assay for CSSV. Sequences analyses, corresponding to a fragment located at the 5' end of open reading frame (ORF)3 of the CSSV genome, revealed three new CSSV groups (D, E, and F) distinct from the A, B, and C groups already identified in Togo. Only group B was detected in all the outbreaks, whereas groups A and C were not identified in Côte d'Ivoire. In addition, a polymerase chain reaction diagnostic using the ORF3A F/R primer pair was polyvalent, because it enabled the detection of CSSV in 90% of the plots in all the cocoa regions analyzed by this study.

Effect of shade on Arabica coffee berry disease development: Toward an agroforestry system to reduce disease impact.

Coffee berry disease (CBD), caused by Colletotrichum kahawae, is a major constraint for Arabica coffee cultivation in Africa. The disease is specific to green berries and can lead to 60% harvest losses. In Cameroon, mixed cropping systems of coffee with other crops, such as fruit trees, are very widespread agricultural practices. Fruit trees are commonly planted at random on coffee farms, providing a heterogeneous shading pattern for coffee trees growing underneath. Based on a recent study of CBD, it is known that those plants can reduce disease incidence. To assess the specific effect of shade, in situ and in vitro disease development was compared between coffee trees shaded artificially by a net and trees located in full sunlight. In the field, assessments confirmed a reduction in CBD on trees grown under shade compared with those grown in full sunlight. Artificial inoculations in the laboratory showed that shade did not have any effect on the intrinsic susceptibility of coffee berries to CBD. Coffee shading mainly acts on environmental parameters in limiting disease incidence. In addition to reducing yield losses, agroforestry system may also be helpful in reducing chemical control of the disease and in diversifying coffee growers' incomes.

Spatio-Temporal Dynamics of Arabica Coffee Berry Disease Caused by Colletotrichum kahawae on a Plot Scale.

Coffee berry disease (CBD) is caused by Colletotrichum kahawae. This pathogen only attacks green berries; it causes cherry rot and premature fruit fall. The disease leads to major harvest losses in the western highland region of Cameroon. The origin of the primary inoculum and the beginning of epidemics are unknown. The interactions between the pathogen and its host were studied at locations where CBD was known to cause severe disease. The disease was monitored weekly in uniform plots of adjacent coffee trees at Santa (1,750 m) in 2003 and 2004 and Bafou (1,820 m) in 2004 and 2005. The logistic model provided good fit of the epidemic's temporal dynamics. The spatial distribution of CBD over time indicated that plants in a plot were contaminated stepwise from the first infected coffee tree. An analysis of semi-variograms and the disease dispersal maps obtained by kriging revealed primary infection foci at both sites. They were observed from the 8th to the 10th week after flowering at Bafou and from the 11th to the 13th week at Santa. CBD affected the entire plots 3 weeks after the foci first appeared. These results suggest that inoculum from previous epidemics survives at points in the initial foci in a coffee plantation.

Relationships Between Black Pod and Witches'-Broom Diseases in Theobroma cacao.

ABSTRACT Field observations were conducted from 1998 to 2001 at the International Cocoa Genebank, Trinidad, to evaluate 57 cacao clones for resistance to black pod (BP) and witches'-broom (WB) diseases (caused by Phytophthora sp. and Crinipellis perniciosa, respectively). Each month ripe pods were harvested and the number of healthy and diseased was recorded. The number of brooms on vegetative shoots was recorded three times a year on selected branches. Twenty-three clones showed less than 10% of infection for both BP and WB on pods. Among those, eight clones showed an absence of brooms on the observed branches: IMC 6, MAN 15/60 [BRA], PA 67 [PER], PA 195 [PER], PA 218 [PER], PA 296 [PER], PA 303 [PER], and POUND 32/A [POU]. Broad-sense heritability was estimated at 0.38 and 0.57 for WB disease on pods and shoots, respectively, and at 0.51 for BP disease. Genetic correlation between WB disease on pods and on shoots was low and estimated at 0.39, whereas the correlation between WB and BP diseases on pods was 0.48. To choose putative parents for breeding schemes, it is suggested that clones are first assessed for their level of resistance to WB on shoots, and the most promising individuals are screened for BP with a detached pods test. Further studies are needed to confirm whether the level of resistance to WB on pods can be predicted using an early test on seedlings.

Occurrence of Cacao swollen shoot virus in Litimé, the Main Cocoa-Producing Area of Togo.

Cacao swollen shoot disease, first described from Ghana on Theobroma cacao in 1922, was identified in Togo in 1955 (4) in Kloto, a minor cocoa-producing area. Litimé is the main production zone in the country. However, its share of production has fallen from approximately 75% of national production in the 1980s to 55% currently because of aging plantations and new swollen shoot foci in this region that had previously been free of the disease. This disease is caused by Cacao swollen shoot virus (CSSV), of which the first isolate to be studied molecularly (Agoul) came from Kloto (1). Since then, the different cocoa-producing regions have been surveyed on several occasions and swollen shoot foci were detected for the first time in Litimé at the end of the 1990s. During 2000, symptomatic leaves were taken from trees which exhibited characteristic symptoms of the disease including swellings on fan branches and chupons, and red vein banding on young flush leaves. The existence of the virus was confirmed using immunosorbent electron microscopy (2) with a composite polyclonal Banana streak virus/Sugarcane bacilliform virus antiserum supplied by B. Lockhart and using polymerase chain reaction amplification with CSSV-specific primers (3). Degenerate primers for CSSV detection were designed in the 5' region (the first 350 amino acids) of open reading frame (ORF3) because this region of the CSSV genome was found to be highly conserved among available full-length CSSV sequences. An isolate from the Litimé area (Wobe 12) was sequenced completely (Genbank Accession No. AJ781003), revealing that it shared a nucleotide sequence identity of only 77% with Agou1. Since the initial observations, the disease has spread rapidly and approximately 60% of the plots in Litimé contain infected trees. Hence, there is an urgent need for a program to rogue infected trees and replant with tolerant material. Moreover, this situation is a threat to cocoa plantations in neighboring Ghana, where numerous eradication operations to control this disease have been launched. References: (1) L. Hagen et al. Virology 196:619, 1993. (2) B. E. L. Lockhart et al. Phytopathology 82:691, 1992. (3) E. Muller et al. J. Virol. Methods 93:15, 2001. (4) M. Partiot et al. Café Cacao Thé 22:217, 1978.

Yield stability in Coffea canephora from diallel mating designs monitored for 14 years.

The genetic parameters of several agronomic traits were estimated in Coffea canephora in a triangular diallel with six parents and in a partial diallel with 18 parents. The yield data came from harvests obtained over 14 years divided into an initial 6-year cycle, a second cycle of 5 years after cutting back and a third cycle of 3 years after topping. Risk incidence was also observed over several years. The general combining abilities (GCA) were the principal sources of variation for the various traits considered. Parents derived from the Yangambi selections (Democratic Republic of Congo) proved to be best for yields in the three cropping cycles. The rank correlations between the GCA of the various traits indicated that first-cycle production was still not enough to predict the GCA for yield over 14 years. Susceptibility to leaf anthracnose and to drought could lead to modifications in successive classifications, with a more significant impact on production during the first cycle. The differences in susceptibility to Coffee Berry Borers had an impact on yield, which increased in line with coffee tree age. There was a rather good relation between inherent values and values in parent combinations and it was thus possible to choose parents on their own performance.

Durability of Field Resistance to Black Pod Disease of Cacao in Papua New Guinea.

Black pod disease of cacao, caused by various species of Phytophthora, is one of the most serious causes of cacao yield losses worldwide. In Papua New Guinea, such losses on local smallholdings are estimated at more than 40% of annual production. Genetic improvement of cacao resistance is the most promising way of reducing the impact of these pathogens. Field infection of 25 selected Trinitario × Upper Amazonian hybrid progenies in a factorial design (5 by 5) was monitored for five consecutive years at the Cocoa and Coconut Research Institute in Papua New Guinea. The resistance durability of the various hybrids was analyzed using a factorial split-plot model with years as the main factor. The reaction of the hybrid progenies remained stable and similar throughout the 5 years. Similarly, the general combining ability ranking of the parents was also the same for each year. Moreover, data accumulated over several years provided a clearer distinction between the parents. The analyses clearly showed female inheritance under field conditions, and the progenies from what are believed to be resistant females showed good resistance.

Genetic mapping of resistance factors to Phytophthora palmivora in cocoa.

Phytophthora palmivora causes pod rot, a serious disease on cocoa widespread throughout the producing regions. In order to ascertain the genetic determination of cocoa resistance to P. palmivora, a study was carried out on two progenies derived from crosses between a heterozygous, moderately resistant Forastero clone, T60/887, and two closely related and highly susceptible Forastero clones, one completely homozygous, IFC2, and one partially heterozygous, IFC5. The cumulative size of both progenies was 112 individuals. Plants were subjected to natural and artificial inoculation of P. palmivora in C te d'Ivoire. The genetic maps of T60/887 and of IFC5 were constructed using amplified fragment length polymorphism (AFLP) markers and microsatellites. The map of T60/887 comprised 198 markers assembled in 11 linkage groups and representing a total length of 793 cM. The map of IFC5 comprised 55 AFLP markers that were assembled into six linkage groups for a total length of 244 cM. Ratio of rotten over total number of fruit under natural infection was measured for each tree over two harvests. Artificial inoculations were performed on leaves and pods. These tests were weakly correlated with the pod rot rate in the field. Five quantitative trait loci (QTLs) of resistance were detected for T60/887 but none were common between the three traits measured. Stability and reliability of the experimental procedures are discussed and revealed the difficult use of these artificial tests on adult trees for a good prediction of field resistance.