Molecular Surveillance, Prevalence, and Distribution of Cacao Infecting Badnavirus Species in Côte d'Ivoire and Ghana.

The cacao swollen shoot disease (CSSD) caused by a complex of badnavirus species presents a major challenge for cacao production in West Africa, especially Ghana and Côte d'Ivoire. In this study, CSSD species detection efficiency, diversity, and geographic distribution patterns in cacao plantations in Ghana and Côte d'Ivoire were investigated through field surveillance, PCR detection assays, sequencing of positive amplicons, and phylogeographic clustering. Cumulatively, the detection efficiency of the tested CSSD primer sets that were targeting the movement protein domain of the virus ranged from 0.15% (CSSD-3 primer) to 66.91% (CSSD-1 primer) on all the symptomatic cacao leaf samples assessed. The identified CSSD species differed phylogenetically and overlapped in distribution, with the cacao swollen shoot Togo B virus (CSSTBV) (n = 588 sequences) being the most prevalent and widely distributed compared to the other CSSD species that were encountered in both countries. Geographically, the cacao swollen shoot CE virus (CSSCEV) species (n = 124 sequences) that was identified was largely restricted to the bordering regions of Ghana and Côte d'Ivoire. These results provide updated knowledge of the geographic distribution of the key CSSD species and their diagnostic efficiency and, thus, provide guidance in identifying locations for structured testing of cacao germplasm and optimal diagnostics for the predominant CSSD species in Ghana and Côte d'Ivoire.

Cacao Swollen Shoot Viruses in Ghana.

Cacao swollen shoot virus causes cacao swollen shoot disease of Theobroma cacao (cacao) plants. At least six cacao-infecting Badnavirus species-Cacao swollen shoot Togo A virus, Cacao swollen shoot Togo B virus (previously known as Cacao swollen shoot virus), Cacao swollen shoot CE virus, Cacao swollen shoot Ghana M virus, Cacao swollen shoot Ghana N virus, and Cacao swollen shoot Ghana Q virus-are responsible for the swollen shoot disease of cacao in Ghana. Each of these species consists of a multiplicity of strains. The New Juaben strain, the most virulent cacao swollen shoot virus strain in Ghana, belongs to the Cacao swollen shoot Togo B virus species, and is a commonly used strain in laboratory transmission assays. Infection of cacao trees with multiple strains of the virus is common and new evidence suggests that these coinfections may have resulted in the emergence of recombinant strains of the virus. The impact of these emerging recombinant strains on disease severity is uncertain. This review focuses largely on the discovery of cacao swollen shoot virus in Ghana, diversity of the virus strains, molecular characterization, propagation of virus infection in cacao plants, emergence of recombinant virus strains, vector-mediated transmission of the virus, and the management of the cacao swollen shoot disease in Ghana. It also contains sections on the botany and origin of the cacao tree, its introduction to Ghana, the role of cacao swollen shoot disease in facilitating Ghana's independence from Britain, and a brief history of chocolate.

Epidemiology and Diagnostics of Cacao Swollen Shoot Disease in Ghana: Past Research Achievements and Knowledge Gaps to Guide Future Research.

Cacao swollen shoot disease (CSSD) caused by complexes of cacao swollen shoot badnaviruses (family Caulimoviridae, genus Badnavirus) remains highly prevalent and devastating in West Africa. The disease continues to impact substantially on cacao yield loss, cacao tree mortality, and decline in foreign exchange income from cacao bean sales. Currently, the disease is estimated to have a prevalence rate of over 30% in Ghana, as assessed in the ongoing third country-wide surveillance program. Although achievements from past research interventions have greatly elucidated the etiology, biology, epidemiology, diagnostics, and management of the disease, there are some outstanding knowledge gaps. The role of these information gaps and their effect on CSSD epidemiology and prevalence remain unanswered. This paper summarizes existing scientific knowledge from past research achievements that have provided elucidation on CSSD epidemiology, management options, and guided future research. The discussion highlights the need for multidisciplinary research with modern tools and institutional collaborators to holistically bring clarity on knowledge gaps on pathogen biology, virus-host--vector interactions, role of environmental and soil nutrient effects on CSSD severity, evolution pattern, role of alternative hosts on virus species diversity, vector population dynamics, and their overall impact on CSSD prevalence and integrated management in cacao plantations.

Field evaluation of the impact of cocoa swollen shoot virus disease infection on yield traits of different cocoa (Theobroma cacao L.) clones in Ghana.

Cocoa swollen shoot virus disease (CSSVD) is a major disease of cacao (Theobroma cacao L.) in Ghana and other West African countries that grow the crop. Attempts to develop resistant varieties since the discovery of the disease in 1936 have yielded little success. Recently, planting materials that are tolerant to the disease have been recommended for planting in areas with high CSSVD prevalence. However, the effect of CSSVD on yield component traits of most cacao clones including the tolerant varieties has not been well studied. To investigate the impact of CSSVD on these traits, reduction in bean weight (BW), number of beans per pod (NoBP) and dry bean yield (DBY) between symptomless and symptomatic trees, and disease incidence (DI) among 210 different cacao clones were evaluated. The clones were transplanted in June 2010 following a randomised complete block design with four replications consisting of three trees per clone per block. Response of the clones to CSSVD incidence had 180 of the genotypes having < 50% symptoms. Clones of Contanama, Iquitos, Marañon, Nanay and unknown derived from Upper Amazon parentage had less DI at the end of the study. The CSSVD effect (symptomless vs. symptomatic) was significant (p < 0.05) for DBY and NoBP, and the symptomless trees surpassed on average their symptomatic trees by 21.17% for DBY. Some of the best-performing clones identified under natural CSSVD infection were; COCA 3348/52 and GU 219/V among the underrepresented clones, B 36, ICS 40, NA 33 x IMC 67, T30/628, T60/887, T63/971, T 81/1879 and T 82/503 among those that combine high DBY with low yield reduction, and NA 124, T17/358, T35/78, T57/305, T63/971 x SCA 6, T65/239, T76/1835 and T82/2294 among those that combine high DBY with no disease incidence. Their inclusion in breeding programs that seek to develop resistant CSSVD varieties or deployment as planting materials in endemic areas to improve yield production in Ghana is recommended on the basis of the present observations.

The proposed new species, cacao red vein virus, and three previously recognized badnavirus species are associated with cacao swollen shoot disease.

BACKGROUND: Cacao swollen shoot virus (CSSV), Cacao swollen shoot CD virus (CSSCDV), and Cacao swollen shoot Togo A virus (CSSTAV) cause cacao swollen shoot disease (CSSD) in West Africa. During 2000-2003, leaf and shoot-swelling symptoms and rapid tree death were observed in cacao in Cote d'Ivoire and Ghana. Molecular tests showed positive infection in only ~50-60% of symptomatic trees, suggesting the possible emergence of an unknown badnavirus. METHODS: The DNA virome was determined from symptomatic cacao samples using Illumina-Hi Seq, and sequence accuracy was verified by Sanger sequencing. The resultant 14, and seven previously known, full-length badnaviral genomic and RT-RNase H sequences were analyzed by pairwise distance analysis to resolve species relationships, and by Maximum likelihood (ML) to reconstruct phylogenetic relationships. The viral coding and non-coding sequences, genome organization, and predicted conserved protein domains (CPDs) were identified and characterized at the species level. RESULTS: The 21 CSSD-badnaviral genomes and RT-RNase H sequences shared 70-100% and 72-100% identity, respectively. The RT-RNase H analysis predicted four species, based on an ≥80% species cutoff. The ML genome sequence tree resolved three well-supported clades, with ≥70% bootstrap, whereas, the RT-RNase H phylogeny was poorly resolved, however, both trees grouped CSSD isolates within one large clade, including the newly discovered Cacao red vein virus (CRVV) proposed species. The genome arrangement of the four species consists of four, five, or six predicted open reading frames (ORFs), and the CPDs have similar architectures. By comparison, two New World cacao-infecting badnaviruses encode four ORFs, and harbor CPDs like the West African species. CONCLUSIONS: Three previously recognized West African cacao-infecting badnaviral species were identified, and a fourth, previously unidentified species, CRVV, is described for the first time. The CRVV is a suspect causal agent of the rapid decline phenotype, however Koch's Postulates have not been proven. To reconcile viral evolutionary with epidemiology considerations, more detailed information about CSSD-genomic variability is essential. Also, the functional basis for the multiple genome arrangements and subtly distinct CPD architectures among cacao-infecting badnaviruses is poorly understood. New knowledge about functional relationships may help explain the diverse symptomatologies observed in affected cacao trees.