VIRUS DISEASES OF CACAO IN WEST AFRICA

Of the tested plants which are indigenous to West Africa, three species in the Bombacaceae and four in the Steruliaceae are susceptible to one or more of four viruses from cacao, three occurring in the Gold Coast and one in Nigeria. These species are less affected than cacao by the viruses; some show transient leaf symptoms and others are symptomless carriers. The development of spines on the stems of Ceiba pentandra seedlings is suppressed by infection with virus 1 A.
In general, the indigenous species are more difficult to infect than cacao, and mealybugs do not become infective as readily when feeding on them as when feeding on infected cacao. The availability of the viruses to vectors seems to be correlated with severity of symptoms, and transmission from infected plants to cacao becomes less frequent with increasing duration of infection.
Ceiba pentandra trees were found naturally infected in the Gold Coast and Nigeria. In the Western Province of the Gold Coast, Cola chlamydantha trees in cacao farms and forests were naturally infected with viruses apparently identical with those causing swollen shoot of cacao there.
There is little doubt that C. chlamydantha trees are an important source of virus for cacao trees. Whenever possible these and other alternative hosts growing near to cacao should be destroyed.     

VIRUS DISEASES OF CACAO IN WEST AFRICA VIII. THE SEARCH FOR VIRUS-RESISTANT CACAO

The methods used and the results of 10 years' search for cacao resistant to swollen-shoot disease are described. Selection among the trees surviving in farms devastated by this virus disease led to the discovery of mild virus strains which can protect trees against virulent strains.
When tested by graft inoculation with virulent virus, none of the selections showed any immunity or resistance save that conferred by previous mild-strain infection. A low degree of tolerance was found in some selections.
Local selections and a range of new introductions were tested by mealybug infection, and only types from the Upper Amazon region of Ecuador were consistently resistant to infection. This genetical resistance seems to be strongest in cacao from the Nanay peninsula, near Iquitos.     

FURTHER OBSERVATIONS ON THE PATHOGENICITY OF CALONECTRIA RIGIDIUSCULA (BERK. & BR.) SACC. TO THEOBROMA CACAO L

Quantitative data are provided for the occurrence of Calonectria rigidiuscula and other fungi in occluded Mirid lesions and other sites in shoots of cacao plants. Mirid lesions appear to be particularly favourable for the development of C. rigidiuscula. The fungus also occurs as a saprophyte on cacao pods, and as a wound parasite in woody plants other than cacao; it was successfully introduced into plants of varied affinities by inoculation. C. rigidiuscula spreads from inoculations in cacao stems much more rapidly than other fungi. The results confirm that it is the most important fungus infecting Mirid lesions and causing dieback of cacao in West Africa.
Inoculation tests with a range of Amelonado and Trinitario clones suggested that the clones vary in their susceptibility to C. rigidiuscula , but the plant-to-plant variation was too great to conclude that any one is highly resistant. Various types of introduced cacao were also tested; preliminary experiments indicate that certain types of Upper Amazon cacao may be resistant, but they need further investigation.     

STUDIES ON THE HOST RANGES OF VIRUSES FROM THEOBROMA CACAO L.

Only plants in the natural order Tiliales were susceptible to experimental infection by viruses found infecting cacao. Of sixty-three species in the Tiliales colonized with infective mealybugs, twenty-eight became infected with the New Juaben virus. The ease with which different species contracted infection and the severity of symptoms varied considerably. The host ranges of seven other viruses were all contained in that of the New Juaben virus. The viruses could be divided into three groups according to their host ranges, and it is suggested that host ranges may prove helpful in attempts to classify cacao viruses. Four species of Corchorus were all rapidly killed by any of the eight viruses.     

SOME HISTOLOGICAL OBSERVATIONS ON VIRUS-INFECTED THEOBROMA CACAO L.

Although the five cacao viruses studied produce different external symptoms in infected plants, they affect internal anatomy similarly. Symptoms on leaves occur only if these are still developing when they become infected, and the viruses seem to produce their effects usually by preventing differentiation of the cells. The tissues of chlorotic areas of infected leaves are undifferentiated and similar in structure to young unexpanded leaves. In stem and root swellings xylem and phloem are both increased, but they occur in the same proportions as in normal secondary thickening. The anatomical effects of infection seem insufficient alone to account for the death of cacao trees, but they may well be complementary to the serious necrosis of the root system which results from virus infections.     

NOTES ON A VIRUS DISEASE OF CACAO

A virus disease of cacao, discovered in Trinidad towards the end of 1943, is apparently confined to the north-western corner of the is land and does not occur in the main cacaogrowing areas of eastern, central and southern Trinidad; nor has it been found in Venezuela, Tobago or the Windward Islands of Grenada, St Vincent, St Lucia and Dominica. The symptoms resemble those of West African swollen shoot disease, consisting of vein-clearing or mosaic of the leaves and red-mottle of the leaves and pods. No swellings are formed, but recent observations indicate that the disease may cause both dieback and reduction in yield. There are two strains closely resembling each other. The spread of the disease under field conditions has been demonstrated, and although the rate of spread has varied somewhat, a mean increase of 41 % over the original number of infected trees has been found over a 10-month period. As with swollen shoot disease, existing diseased areas have increased in size and new outbreaks at short distances have occurred. Both strains were readily transmitted by budding, the incubation period varying from 34 to 136 days, with a mean around 90 days, but no insect vectors have been found. There is at present little reason to believe that any varieties of cacao are immune to the disease or are even symptomless carriers. The question of eradicating this virus, while its distribution remains limited, should be seriously considered.     

VIRUS DISEASES OF ADANSONIA DIGITATA L. (BOMBACACEAE) AND THEIR RELATION TO CACAO IN GHANA

Virus-infected Adansonia digitata L. were found in the Guinea-savannah woodland forming the Northern Territories and on the Accra plains of Ghana, but those tested in the rain forest were not infected. Three viruses isolated from infected trees resembled the Kpeve cacao virus, but no definite relationship could be established. It is unlikely that the original outbreaks of swollen shoot in cacao came from A. digitata.     

VIRUS DISEASES OF CACAO IN WEST AFRICA IX. STRAIN VARIATION AND INTERFERENCE IN VIRUS 1A

Cacao virus iA, the most important and prevalent of the viruses that attack cacao in the Gold Coast, occurs in strains that differ widely in their virulence towards cacao. Outbreaks usually contain trees infected with different strains and individual trees are often infected simultaneously with more than one strain; this can be demonstrated by coppicing the trees, and by inoculating sets of test plants with grafts from different parts of one tree. Neither mild nor virulent strains seemed to be consistently dominant in roots or in other parts of cacao trees.
Cacao plants infected with mild strains were nearly always protected against the effects of infection by virulent strains; however, virulent strains entered hosts already infected with mild strains, but usually without causing any symptoms unless the plants were coppiced. The severe symptoms that developed on new growth from such coppiced plants were seldom repeated in later growth. Mealybugs transmitted the virulent strains from leaves with symptoms characteristic of infection by the latter, but not from leaves free from such symptoms. These results suggest that the multiplication of a virulent strain is impeded in plants infected with a mild strain.
In the field, infection with a mild strain protected mature trees against the effects of virulent strains spread by mealybugs. During 3 years in which 273 out of 387 previously uninfected trees became severely diseased, only 35 out of 416 infected with themildstrain developed symptoms of infectionwiththevirulent strain. Five years after infection with the mild strain, trees were yielding I pod per tree more than in the year they were infected, whereas the decrease on trees infected with the virulent strain was 16 pods per tree. Some limitations in the practical application of protection by mild strains, and objections to its use as a control measure, are discussed.     

STRUCTURE AND DEVELOPMENT OF THE DIMORPHIC BRANCH SYSTEM OF THEOBROMA CACAO

The vegetative morphology of Theobroma cacao, the cacao tree, was studied in order to provide a foundation for further investigations on the morphogenesis of the cacao dimorphic shoot system. The seedling of cacao has a determinate orthotropic shoot with a (2+3) phyllotaxis. Branch dimorphism is initiated after 1 to 2 years of growth at which time the apical meristem of the orthotropic shoot aborts and a pseudowhorl of plagiotropic branches is initiated from axillary positions in the shoot tip. The plagiotropic branches are characterized by a distichous phyllotaxis and indeterminate growth. Subsequently an axillary bud below the pseudowhorl develops into a new orthotropic shoot. The apical meristem of this shoot eventually aborts and another pseudowhorl is formed. The apical anatomy of the two types of shoots is similar. The developmental potentiality of the orthotropic shoot axillary buds to form one or the other type of shoot was investigated. The phyllotaxis of the axillary buds of the orthotropic shoot is spiral and that of the axillary buds of the plagiotropic branch is distichous. Pruning and apical puncture experiments showed that the axillary buds of a plagiotropic branch, and of an orthotropic seedling shoot which has not yet formed a pseudowhorl, always give rise to the parent type of shoot. However, the axillary buds of an orthotropic shoot which already bears a pseudowhorl give rise to either type of shoot for several nodes below the point of origin of the pseudowhorl. The type of shoot has no influence on the form of branch which develops from an axillary bud grafted to it. This evidence supports the hypothesis that the axillary buds are initiated as one or the other type of shoot, i.e., once initiated they are predestined.     

VIRUS DISEASES OF CACAO IN WEST AFRICA

Increasing the number of Pseudococcus njalensis per test plant increased the infection rate with cacao virus 1A and 1M to 95 % with twenty-five insects. Young leaves with the red vein-banding symptoms were better sources of virus than mature leaves, and there was evidence of virus localization in the plant. It was estimated that Ps. njalensis nymphs took about 16 min. to penetrate plant tissues with their stylets.
Infection rates increased with time of test feeding up to 50 min. The virus was non-persistent in the feeding insect, but persisted up to 36 hr. in insects starved after the infection feed. Starvation before the infection feed increased the infection rate with infection feeds up to 10 hrs., after which the infection rate with pre-starved insects declined to the level attained by unstarved insects.     

VIRUS DISEASES OF CACAO IN WEST AFRICAL: II. CROSS-IMMUNITY EXPERIMENTS WITH VIRUSES 1A, 1B AND 1C

Experiments on cross-immunity reactions between three viruses attacking Theobroma cacao L. on the Gold Coast are described. A field trial involving 3 acres of graft-inoculated trees revealed some degree of protection afforded by Theobroma virus 1B against infection with virus 1A. The protection appeared to be more effective against insect inoculation than against graft transmission, being only temporary in the latter. Virus 1C (probably unrelated but not to be called Theobroma virus 2 until more evidence is available) conferred no protection against virus 1A.
The latent periods for these viruses were calculated from this experiment, which also provided data on their effects on yield. Virus 1A reduced yield by 50% in the first year after inoculation and killed the trees in the second. Virus 1B had no appreciable effect on yield; virus 1C reduced yield by 50 % in the third year after inoculation but there was no further decline in the fourth.
The rates of spread of these viruses were compared and significant differences demonstrated.     

OBSERVATIONS ON THE PATHOGENICITY OF CALONECTRIA RIGIDIUSCULA (BERK. & BR.) SACC. ON THEOBROMA CACAO L

Calonectria rigidiuscula has been found associated with an acute and a chronic form of dieback of cacao. It can also infect cankers caused primarily by Phytophthora palmivora , and the lesions following attack by the capsids Sahlbergella singidaris and Distantiella theobroma. The association with the capsid lesions is of great economic importance since it appears that capsids alone are capable of killing only green shoots, and that the severe damage caused to woody shoots follows C. rigidiuscula infection of the capsid lesions. Acute dieback occurs only in certain districts after exceptional drought and may be considered primarily the result of environmental conditions. The main importance of chronic dieback is that it prevents the recovery of cacao which has been seriously weakened from other causes. C. rigidiuscula infection retards the healing of P. palmivora cankers and also produces a lesion in the xylem tissue which may be extensive.
C. rigidiuscula has been established in wounded cacao stems of all ages without difficulty. The spread of the fungus is slow, but is more rapid in the xylem than in the cortical tissues. Spread is greatest in the unlignified tissues.     

COPPICING EXPERIMENTS ON THE SPREAD AND CONTROL OF CACAO SWOLLEN-SHOOT DISEASE IN NIGERIA

A coppicing technique was used to determine the incidence and distribution of latent and unrecognized infection around naturally occurring outbreaks of cacao swollen-shoot virus disease in Western Nigeria. All the apparently healthy trees within 30 yards of eighty outbreaks of various sizes were coppiced and most of the infected stumps which regenerated showed symptoms within a year.
The distribution of the infected stumps around thirty-five outbreaks studied in detail is expressed by the equation log₁₀ I = a + bx. I is the estimated intensity of infection in the coppiced stumps at distance x from the nearest infected tree removed at the time of coppicing. The constant a determines the height of the peak of the infection gradient and increases with outbreak size, whereas the slope of the gradient, determined by the negative constant b , is similar around all outbreaks.
The results are consistent with information on the movement of the mealybugs and spread of cacao swollen-shoot virus in outbreaks. Moreover, they indicate that outbreaks are controlled most economically by removing all obviously infected trees and adjacent apparently healthy ones. Control does not require the destruction of all the trees around outbreaks up to a distance of 30 yards as done previously, but can be achieved by felling fewer trees, the actual number depending on the size of the outbreak.     

ARREST OF THE SWOLLEN-SHOOT VIRUS DISEASE OF CACAO IN THE GOLD COAST BY CONTROLLING THE MEALYBUG VECTORS WITH THE SYSTEMIC INSECTICIDE, DIMEFOX

A 13-acre plot of cacao, in an area devastated by swollen shoot, was treated by dimefox applied to the soil nine times. The plot was examined every month and trees recorded as they showed symptoms. The number of such trees decreased very rapidly a year after the first treatment and remained low for the 3 years that the experiment was continued. This suggests that the time between infection and symptom development can extend to about I year. The cause of the continued appearance of a few diseased trees could not be determined.
As cacao farms differ from one another, it was difficult to find suitable control plots. Three farms in the neighbourhood of the chemically treated area were selected. The initial percentage of diseased trees in each was lower than on the chemically treated plot. The three farms were inspected every month and newly discovered trees with symptoms were felled. The number of such trees decreased 12 months after the initial cutting out and then increased abruptly until it again reached a high level.
Chemical treatment combined with cutting out was more efficient than cutting out alone in arresting the advance of the disease.     

Intersterility, morphology and taxonomy of Ceratocystis fimbriata on sweet potato, cacao and sycamore

Ceratocystis fimbriata is a large, diverse complex of species that cause wilt-type diseases of many economically important plants. Previous studies have shown that isolates in three monophyletic lineages within the Latin American clade of C. fimbriata are host-specialized to cacao (Theobroma cacao), sweet potato (Ipomoea batatas) and sycamore (Platanus spp.), respectively. We paired testers of opposite mating type from isolates of these lineages to find intersterility groups. Two intersterility groups corresponded to the sweet potato and sycamore lineages, respectively. The cacao lineage contained two intersterility groups, corresponding to two genetic sublineages centered in western Ecuador and Brazil/Costa Rica/Colombia. Six isolates from cacao that were not members of the cacao lineage and were not pathogenic to cacao in an earlier study also were intersterile with members of the two cacao intersterility groups. Some pairings between testers from different lineages or sublineages yielded perithecia from which a few abnormal progeny could be recovered, typical of interspecific hybrids. These progeny showed abnormal segregation of the MAT-2 gene and mycelial morphology, showing that they were indeed the result of crosses. Isolates of the sweet potato, cacao, and sycamore lineages were indistinguishable morphologically except for the presence or absence of a doliform (barrel-shaped) conidial state and minor differences in size of perithecial bases and necks and ascospores. C. fimbriata originally was described from sweet potato. We describe the cacao pathogen as a new species, Ceratocystis cacaofunesta and we raise the sycamore pathogen from a form to species Ceratocystis platani.     

Effects of an experimental drought on the functioning of a cacao agroforestry system,Sulawesi, Indonesia

Agroforestry systems may play a critical role in reducing the vulnerability of farmers' livelihood to droughts as tree‐based systems provide several mechanisms that can mitigate the impacts from extreme weather events. Here, we use a replicated throughfall reduction experiment to study the drought response of a cacao/Gliricidia stand over a 13‐month period. Soil water content was successfully reduced down to a soil depth of at least 2.5 m. Contrary to our expectations we measured only relatively small nonsignificant changes in cacao (?11%) and Gliricidia (?12%) sap flux densities, cacao leaf litterfall (+8%), Gliricidia leaf litterfall (?2%), soil carbon dioxide efflux (?14%), and cacao yield (?10%) during roof closure. However, cacao bean yield in roof plots was substantially lower (?45%) compared with control plots during the main harvest following the period when soil water content was lowest. This indicates that cacao bean yield was more sensitive to drought than other ecosystem functions. We found evidence in this agroforest that there is complementary use of soil water resources through vertical partitioning of water uptake between cacao and Gliricidia. This, in combination with acclimation may have helped cacao trees to cope with the induced drought. Cacao agroforests may thus play an important role as a drought‐tolerant land use in those (sub‐) tropical regions where the frequency and severity of droughts is projected to increase.     

Conservation Value of Cacao Agroforestry Systems for Terrestrial Herbaceous Species in Central Sulawesi,Indonesia

Tropical secondary forest and agroforestry systems have been identified as important refuges for the local species diversity of birds and other animal groups, but little is known about the importance of these systems for terrestrial herbs. In particular, few studies report how the conversion from tropical forest to technified cacao plantation affects the species richness and the community structure of herbs. We conducted surveys in 43 cacao plantations along the border of the Lore Lindu National Park in Central Sulawesi, ranging from agroforests to technified cacao, categorizing the plantations as rustic cacao, planted shade cacao, and technified cacao. We recorded 91 herb species. Of the 74 species determined to species level, 21 were also found in natural forests, while 53 were recorded only in agricultural habitats. Araceae was the most forest‐dependent plant family while Asteraceae included the highest number of nonforest species. Overall, the presence of forest species was confined to moderately intensively managed rustic and planted shaded plantations. Distance from the forest, which has been identified as a crucial parameter for the diversity and composition of other taxa in cacao agroforests, only played a minimal role for herbs. Our study suggests that native forest herbs maybe more vulnerable to forest conversion than animal groups. The intensification of cacao plantation management increases the presence of weedy species to the detriment of native forest species.     

Genetic Structure and Molecular Diversity of Cacao Plants Established as Local Varieties for More than Two Centuries: The Genetic History of Cacao Plantations in Bahia,Brazil

Bahia is the most important cacao-producing state in Brazil, which is currently the sixth-largest country worldwide to produce cacao seeds. In the eighteenth century, the Comum, Pará and Maranhão varieties of cacao were introduced into southern Bahia, and their descendants, which are called ‘Bahian cacao’ or local Bahian varieties, have been cultivated for over 200 years. Comum plants have been used to start plantations in African countries and extended as far as countries in South Asia and Oceania. In Brazil, two sets of clones selected from Bahian varieties and their mutants, the Agronomic Institute of East (SIAL) and Bahian Cacao Institute (SIC) series, represent the diversity of Bahian cacao in germplasm banks. Because the genetic diversity of Bahian varieties, which is essential for breeding programs, remains unknown, the objective of this work was to assess the genetic structure and diversity of local Bahian varieties collected from farms and germplasm banks. To this end, 30 simple sequence repeat (SSR) markers were used to genotype 279 cacao plants from germplasm and local farms. The results facilitated the identification of 219 cacao plants of Bahian origin, and 51 of these were SIAL or SIC clones. Bahian cacao showed low genetic diversity. It could be verified that SIC and SIAL clones do not represent the true diversity of Bahian cacao, with the greatest amount of diversity found in cacao trees on the farms. Thus, a core collection to aid in prioritizing the plants to be sampled for Bahian cacao diversity is suggested. These results provide information that can be used to conserve Bahian cacao plants and applied in breeding programs to obtain more productive Bahian cacao with superior quality and tolerance to major diseases in tropical cacao plantations worldwide.     

Analyses of polyphenols in cacao liquor, cocoa, and chocolate by normal-phase and reversed-phase HPLC

The antioxidant polyphenols in cacao liquor, a major ingredient of chocolate and cocoa, have been characterized as flavan-3-ols and proanthocyanidin oligomers. In this study, various cacao products were analyzed by normal-phase HPLC, and the profiles and quantities of the polyphenols present, grouped by molecular size (monomers to approximately oligomers), were compared. Individual cacao polyphenols, flavan-3-ols (catechin and epicatechin), and dimeric (procyanidin B2), trimeric (procyanidin C1), and tetrameric (cinnamtannin A2) proanthocyanidins, and galactopyranosyl-ent-(-)-epicatechin (2alpha-->7, 4alpha-->8)-(-)-epicatechin (Gal-EC-EC), were analyzed by reversed-phase HPLC and/or HPLC/MS. The profile of monomers (catechins) and proanthocyanidin in dark chocolate was similar to that of cacao liquor, while the ratio of flavan-3-ols to the total amount of monomeric and oligomeric polyphenols in the case of pure cocoa powder was higher than that in the case of cacao liquor or chocolate.     

The status of cacao (Theobroma cacao,sterculiaceae) in the western hemisphere

There are, at the present time, effectively no long-range, ongoing programs in any tropical country of the western hemisphere dedicated to the improvement of cacao (Theobroma cacao, Sterculiaceae). While some effort is currently made to obtain new acquisitions of cacao cultivars exhibiting desirable characteristics and to maintain genepools of these trees, there are few data from field trials to prove and substantiate these qualities. In addition, there is a growing concern regarding the disparities between predicted yields of cacao trees through the use of “hybrid” seed and from actual production under field conditions. This has stimulated an awareness of the current inadequate understanding of the genetics of cacao and the lack of comprehension as to which cultivars, under distinct ecological conditions, are precocious, resistant to disease, or heavy bearing, or indeed demonstrate those traits vital to the success of farming programs adapted to today’s market conditions. This paper examines the events that have led to the current status of selection, development, and breeding of cacao. Alternative approaches are suggested.