Chemical control of vascular-streak dieback disease of cocoa in Papua New Guinea

High-volume foliage sprays of 18 compounds including 13 systemic fungicides in field trials at 4-week intervals did not control vascular-streak dieback (VSD) of cocoa caused by Oncobasidium theobromae in Papua New Guinea. Soil drenching with carbendazim and other systemics was also ineffective, possibly because insufficient carbendazim may have been taken up into the xylem to inhibit the fungus. Stem paints with four systemic fungicides in aqueous formulations were also ineffective, but 4-weekly stem painting of the hardened bark of young seedlings with 10 g/l propiconazole in 25% white oil caused a highly significant reduction in VSD incidence under field conditions. The treatment had no curative effect. Some phytotoxicity was observed but this was not apparent when the concentration of propiconazole was doubled and the treatment interval lengthened to 8 weeks. The ineffectiveness of systemic fungicides as soil drenches and foliage applications was probably due to failure of these compounds to accumulate at the shoot apex where infection occurs. A stable and non-toxic formulation containing surfactants was developed which may improve control by aiding bark penetration by propiconazole or related compounds.     

Black pod: diverse pathogens with a global impact on cocoa yield

ABSTRACT Pathogens of the Straminipile genus Phytophthora cause significant disease losses to global cocoa production. P. megakarya causes significant pod rot and losses due to canker in West Africa, whereas P. capsici and P. citrophthora cause pod rots in Central and South America. The global and highly damaging P. palmivora attacks all parts of the cocoa tree at all stages of the growing cycle. This pathogen causes 20 to 30% pod losses through black pod rot, and kills up to 10% of trees annually through stem cankers. P. palmivora has a complex disease cycle involving several sources of primary inoculum and several modes of dissemination of secondary inoculum. This results in explosive epidemics during favorable environmental conditions. The spread of regional pathogens must be prevented by effective quarantine barriers. Resistance to all these Phytophthora species is typically low in commercial cocoa genotypes. Disease losses can be reduced through integrated management practices that include pruning and shade management, leaf mulching, regular and complete harvesting, sanitation and pod case disposal, appropriate fertilizer application and targeted fungicide use. Packaging these options to improve uptake by smallholders presents a major challenge for the industry.     

On-farm selection for quality and resistance to pest/diseases of cocoa in Sulawesi: (ii) quality and performance of selections against Phytophthora pod rot and vascular-streak dieback

The cocoa industry in Sulawesi, the main region of cocoa production in Indonesia, is threatened by destructive diseases, including vascular-streak dieback (VSD) caused by the basidiomycete Oncobasidium theobromae and stem canker and Phytophthora pod rot (PPR) or black pod, caused by Phytophthora palmivora. Using the considerable genetic diversity of cocoa on farms, host resistance was identified and tested with the participation of farmers. Forty-nine local and international cocoa selections with promising resistance characteristics (as well as susceptible controls) were side-grafted onto mature cocoa in a replicated trial with single-tree plots. Developing grafts were assessed in the dry season for severity of VSD infection, scored from 0 (no infection) to 4 (graft death). All of the 49 clones in the trial became infected with VSD in at least some replicates. Average severity varied from 0.2 to 1.6. Potential VSD-resistance was found in eight clones, including DRC 15, KA2 106 and a local Sulawesi selection, VSD2Ldg. Some of the most susceptible clones were local Sulawesi selections from areas with a history of little or no VSD. Thirty-four pod-bearing clones were evaluated over a 2-year period for yield, quality and resistance to natural infections of PPR. Cumulative PPR incidence for all clones was 22% but varied from 8.6 to 43% among clones. Clones with less than 15% PPR incidence were designated as resistant, including DRC 16 and local Sulawesi selections, Aryadi 1, Aryadi 3 and VSD1Ldg. Scavina 12 was moderately resistant in the trial with a PPR incidence of 23%. Cumulative incidences of the mirid, Helopeltis spp., determined in the same evaluation period, indicated that DRC16 was the most susceptible clone with an incidence of 52% in ripe pods and 23% in immature pods. In comparison, KKM4 showed evidence of resistance to Helopeltis spp., with incidences of 34 and 0.8% in ripe and immature pods, respectively. The impact of diseases and pests (including cocoa pod borer) on bean losses and bean quality varied between clones but generally the bean size (or bean count) was affected more than the fat content or shell content.     

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.     

The effects of mild cacao swollen shoot virus strains N1 and SS365B on growth and yield of cacao

A field experiment started at the Cocoa Research Institute of Ghana in 2007 to investigate the effects of the cacao swollen shoot virus (CSSV) mild stains N1 and SS365B on growth and yield of mixed hybrid cacao (Theobroma cacao L.). Growth was significantly influenced by the duration of infection with the two CSSV mild strains. Slicing analyses showed no significant differences in stem circumference among treatments up to the third year of field data collection. However, from the fourth to the seventh year, growth of N1-inoculated cacao plants was significantly higher than that of SS365B-inoculated plants. Overall, growth did not differ significantly between cacao plants inoculated with mild strain and the non-inoculated plants. Contrary to some previous reports, yield of mild strain-inoculated cacao plants was significantly lower than that of the non-inoculated plants. Polymerase chain reaction was used to detect the presence of CSSV in randomly selected field samples. The implications of these results (based on field data collected from 2008 to 2014) in the management of cocoa swollen shoot virus disease in Ghana are discussed.     

Limited morphological,physiological and genetic diversity of Phytophthora palmivora from cocoa in Papua New Guinea

In Papua New Guinea (PNG) cocoa (Theobroma cacao) is one of the most important cash crops grown in the tropical lowland and island regions. As in most cocoa‐growing areas, phytophthora black pod and canker cause significant yield losses. Cocoa breeding activities in PNG are focused in East New Britain province where disease control recommendations are also developed. This study tested the hypothesis that there was no diversity in the Phytophthora palmivora population causing black pod on cocoa by characterizing the variation in pathogen populations within and between the five major cocoa‐growing areas. Diseased pods were sampled hierarchically from the five locations and additional isolates were collected from soil, stem and leaf lesions, or retrieved from culture collections. Morphological characters showed continuous variation within the range described for P. palmivora. Genetic analysis revealed that the isolates belonged to one dominant clonal lineage, with restricted distributions of several other subpopulations. Lowest diversities were found in the geographically isolated Karkar Island and East Sepik province. Soil isolates showed greater genetic diversity than isolates from cocoa lesions. Intra‐farm variation was as much as inter‐farm or inter‐province variation. Both mating types were detected, although no strong evidence of sexual recombination was observed. The analysis revealed limited geographic, temporal or host specialization, suggesting continuous selection for pathogenicity from a genetic pool of P. palmivora. These findings have significant implications on the deployment of cocoa genotypes, enforcement of inter‐province quarantine and sustainable disease management strategies.     

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.     

香蕉黑腐病菌(Botryodiplodia theobromae)的PCR检测

 根据香蕉黑腐病菌可可球二孢菌(Botryodiplodia theobromae)与其它香蕉病原真菌核糖体基因转录间隔区(rDNA-ITS)ITS1和ITS2间序列差异,设计了特异引物Bth-S(5'-TCTCCCACCCTTTGTGAAC-3')和Bth-A(5'-AAAAGT-TCAGAAGGTTCGTC-3'),利用此引物对包括可可球二孢菌在内的21个菌株基因组DNA进行PCR扩增,结果只有4个可可球二孢菌菌株扩增到422bp特异带,其它17个菌株无扩增产物。灵敏度测试结果表明此特异引物能对1pg的可可球二孢菌基因组DNA进行扩增。对自然感染黑腐病的香蕉果实组织和接种可可球二孢菌或多种香蕉病原真菌混合接种的果实组织进行检测,Bth-S和Bth-A引物对不仅能够在自然感染黑腐病果实组织中特异检测到可可球二孢菌,而且能在未显症和发病的接菌香蕉果实组织中特异检测得到可可球二孢菌。这为香蕉可可球二孢菌潜伏侵染检测提供了技术支持。     

Spatiotemporal pattern analysis of Cacao swollen shoot virus in experimental plots in Togo

In order to quantitatively analyse the spatial pattern of cacao swollen shoot disease, particularly in cases of re‐emergence, three experimental plots were installed in a diseased area of cacao cv. Amelonado in Togo. After thorough cleaning and grubbing, the three plots were planted with less susceptible, hybrid plant material. Twenty years after replanting, a survey of healthy, diseased and dead trees was carried out during 2 consecutive years. Data were analysed using Ripley’s functions and join counts analysis. The re‐emergence of the disease occurred in patches or foci: following analyses with the two statistical methods, diseased trees and dead trees were found to be clearly aggregated on the three observed plots for the 2 years. The observed progress of the disease was not the same on the three plots and seemed dependent on the disease state of the first year: the higher the attack rate of the first year, the faster the disease progression. The use of less susceptible plants helped keep the land productive for 15 years. In conclusion, uprooting of the first infection focus can extend the life of cacao plots.     

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.     

Changes in morphology and measurement of cytokinin levels during the development of witches' brooms on cocoa

The anatomy of diseased stems from brooms induced on cocoa ( Theobroma cacao ) by the fungus Crinipellis perniciosa was examined using the scanning electron microscope, and the amounts of selected cytokinins were measured by immunoassay.
Although the diseased stems were wider in diameter with a lower dry weight:fresh weight ratio than stems of healthy plants, the overall internal organization of tissues remained unchanged, and all the cell types were present. However, the vascular tissue was less differentiated in diseased stems since xylem vessels were absent and the ratio of phloem fibres and sieve tubes to phloem parenchyma was reduced. Cell sizes in diseased stems were larger for all tissues but cell numbers were unchanged except for xylem where fewer cells were seen.
Cytokinins (zeatin, zeatin riboside, isopentenyl adenine and isopentenyl adenosine) were measured in healthy plants and diseased stems at specific stages of disease development. Of these cytokinins only zeatin riboside was present in significantly greater amounts in diseased tissue. The likely role of plant growth regulator balance in stem enlargement and proliferation in witches' broom disease of cocoa is discussed.     

Diversity and pathogenicity of the Ceratocystidaceae associated with cacao agroforests in Cameroon

Knowledge of the diversity and ecology of plant pathogenic fungi in cacao agroforests and surrounding natural ecosystems can inform the development of sustainable management strategies for new cacao disease outbreaks. This study investigated the occurrence of fungi related to the Ceratocystidaceae and their nitidulid beetle vectors in cacao agroforests in Cameroon, under diverse agroecological conditions. The fungi and their vectors were collected from artificially induced stem wounds on cacao and associated shade trees. Collections were also made from abandoned cacao pod husks and other tree wounds within and around plantations. Fungal isolates were identified using DNA sequence‐based phylogenies and morphological comparisons, and two representatives of each species were evaluated for pathogenicity on cacao. Five species of Ceratocystidaceae were recovered, including Huntiella chlamydoformis sp. nov., H. pycnanthi sp. nov. and H. moniliformis, as well as Thielaviopsis cerberus and T. ethacetica. The incidence of these fungi appeared to be influenced by the prevailing agroecological conditions. Nitidulid beetles in the genus Brachypeplus were found to be their most common insect associates on cacao. Both T. ethacetica and H. pycnanthi produced extensive lesions after inoculation on branches of mature cacao trees, while T. ethacetica also caused pod rot. Although their impact remains unknown, fungi in the Ceratocystidaceae and their nitidulid beetle vectors are common and probably contribute to the parasitic pressure in Cameroonian cacao agrosystems.     

Activity of Bacillus thuringiensis toxins against cocoa pod borer larvae

Twelve Cry proteins from Bacillus thuringiensis Berliner were tested in bioassays on cacao plantations in Indonesia for activity against the larvae of cocoa pod borer (Conopomorpha cramerella (Snellen)), an insect pest of the cacao tree. Through the damage caused by their feeding, the larvae of cocoa pod borer cause the pods of the cocoa tree to ripen prematurely. They are difficult to control with conventional measures. Preliminary assays identified five toxins that were more active than others. In two subsequent bioassays the activity of selected toxins was determined more accurately. Three Cryl proteins with relatively little homology were all found to be toxic, opening perspectives for controlling cocoa pod borer by expression of Cry proteins in transgenic plants.     

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.     

Spatio-temporal dynamics on a plot scale of cocoa black pod rot caused by <Emphasis Type="Italic">Phytophthora megakarya</Emphasis> in Cameroon

Black pod disease is caused by several species of Phytophthora. In Cameroon, the disease is mainly due to Phytophtora megakarya. The pathogen attacks cocoa pods and can lead to almost total production losses in a plot if no control measures are applied. To control the disease, several research programmes are being conducted: breeding for increased resistance, development of several biocontrol or agronomic methods. However, for better use of a specific method it is useful to have a good understanding of several epidemiological processes and more effectively know how the disease is distributed in the field. The purpose of this study was to describe the spatial development of the disease in several cocoa fields in Cameroon. In particular, we determined the spatial relation of the disease using several tools, including geostatistics models and Moran indices. The results indicated that the disease was not randomly distributed, while correlations between neighbouring cocoa trees existed. The relationships were detected up to a distance of between 7 and 9 m, revealing the wide dispersal pattern of the pathogen over short distances. No spatial structure was found in the spread of the disease in the oldest cocoa plantations and the inoculum was dispersed throughout the plot. Disease dispersal over short distances should make it possible to adapt control methods by attempting to confine the first disease foci in young plots. Research should also be undertaken to limit inoculum dispersal.     

Host specialization of Verticillium dahliae, with emphasis on isolates from cocoa (Theobroma cacao)

Isolates of Verticillium dahliae from cocoa (four Brazilian and one Ugandan) were screened against cocoa, aubergine, tomato, cotton, and pepper. Isolates originating from hosts other than cocoa were also inoculated. In general, all inoculated crops were systemically invaded by isolates from cocoa. Isolates from each host tended to be more aggressive on their original host. All isolates from cocoa were pathogenic to cocoa, but exhibited various degrees of aggressiveness to other crops. They induced severe symptoms on aubergine, but few symptoms on tomato, although colonization occurred up to the stem apex in both cases. Likewise, symptomless invasion of pepper was found with some Brazilian isolates. The Ugandan isolate was significantly more aggressive on cotton and pepper than the Brazilian isolates.
A Brazilian isolate from cocoa from the State of Bahia was also used to inoculate 12 common weed species from the same geographical area. Four species showed wilt symptoms, while V. dahliae was readily recovered from the stems of a further four species. The role of alternative hosts on disease spread in cocoa growing areas is discussed.     

山楂花腐病的研究Ⅱ.山楂花腐病的症状

 山桂花腐病菌为害叶片、新梢和幼果。叶片上,展叶后4~5天即可出现褐色小班,经6~7天后病斑可扩展到病叶的1/2~2/3。天气潮湿时,在病斑上生出灰白色霉层,即病菌的分生孢子。在新梢上,初生褐色至红褐色病斑,当病斑环绕新梢一周时,新梢上部萎蔫枯死。在幼果上,落花后10天左右幼果果面出现褐色病斑,逐渐扩大,直至整个果实变成暗褐色脱落。     

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.     

Fusarium species from the cassava root rot complex in west Africa

ABSTRACT Fusarium species are a significant component of the set of fungi associated with cassava root rot. Yield losses due to root rot average 0.5 to 1 ton/ha but losses >3 ton/ha, an equivalent of 15 to 20% yield, often occur. This paper reviews previous work on cassava root rot and summarizes a few recent studies on Fusarium species associated with the disease. Our studies in Cameroon showed that 30% of rotted tubers were infected by Fusarium spp. 12 months after planting and represented 25% of all the fungal isolates recovered. Other commonly recovered fungi were Botryodiplodia theobromae and Armillaria spp. Numerous and diverse species of Fusarium were associated with rotted cassava roots in Nigeria and Cameroon. At least 13 distinct amplified fragment length polymorphism (AFLP) groups of Fusarium were distinguishable, each group probably a distinct species, and many of them might represent previously undescribed Fusarium species. The two largest of the AFLP groups correspond to F. oxysporum and F. solani species complex. The distribution of Fusarium spp. varied among countries and among locations within a country, suggesting that germ plasm resistant at one location may not be resistant at another. Fusarium spp. also cause seedling blight of cassava and can be recovered from the stems of infected plants up to 1 m above the ground. Therefore, the pathogen can spread with stems cut as planting material. Fusarium spp. also can colonize Chromolaena odorata, the dominant weed in short fallows, which could further complicate management efforts by serving as an alternative host for strains that colonize cassava.