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.     

VIRUS DISEASES OF CACAO IN WEST AFRICA: I. CACAO VIRUSES 1A, 1B, 1C AND 1D

As an introduction to this series of papers, a brief history is given of swollen-shoot disease of cacao in the Gold Coast, and the nomenclature for cacao viruses is discussed.
The symptoms of four virus diseases of cacao are described in detail, together with experimental data under the headings: cross-immunity reactions, latent period, seed transmission, staining reaction, heat inactivation and rate of spread.     

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.     

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.     

The effects of cocoa swollen-shoot virus on the growth and yield of Amelonado and Amazon cocoa (Theobroma cacao) in Ghana

The virulent strain A of cocoa swollen-shoot virus (CSSV) severely decreased the growth and yield of Amelonado cocoa (Theobroma cacao) trees kept free of capsids (Distantiella theobroma and Sahlbergella singularis) and the dieback fungus (Calonectria rigidiuscula) in Ghana. Fifteen per cent of graft-inoculated Amelonado trees showed symptoms within 4 months, and 48, 80 and 100% within 6, 12 and 20 months, respectively. Infected trees, whether shaded or unshaded, began to decline 6 months after infection, and deteriorated rapidly during the next 27 months by which time 16 % had died and most others were moribund; fertilizer applications had no significant effect on the rate at which infected trees deteriorated. Yields of pods and dry cocoa were greatly reduced 2 yr after infection and were very low after 3 yr; yields were significantly reduced by virus infection but there were no significant further effects of applying fertilizer. These results confirm that CSSV strain A alone is very damaging and often eventually lethal to Amelonado trees in Ghana, and indicate that the conflicting results obtained previously in Ghana and Nigeria were probably due to differences in the virulence of the CSSV strains tested. In contrast, the virus had much less effect on cocoa trees of the Amazon type; only 3% of graft-inoculated Amazon trees showed symptoms within 4 months, and 43, 84 and 97% after 1, 2 and 3 yr, respectively. Slight deterioration of tree canopies was first detected c. 15 months after infection and, although it continued slowly during the next 21 months, the decline was much less severe than that of Amelonado trees. Yields of both unshaded and shaded trees were apparently reduced by virus infection, but yield losses were much smaller than those of Amelonado trees. These results support the present objectives of controlling the spread of CSSV in Ghana by roguing infected trees, and selecting cultivars with greater tolerance to infection for future use.     

Occurrence of Cryphonectria canker disease of Eucalyptus in Kerala,India*

Cryphonectria cubensis, recorded for the first time in India, is responsible for causing severe stem cankers in Eucalyptus grandis in Kerala. Disease outbreaks are localised and vary in severity with host species, at times affecting up to 30% of the trees in a stand. Infection usually occurs on the main stem 1 to 1 ·5 m above ground and occasionally near ground level. There is evidence of tree to tree spread of the disease from the primary infection foci. The disease also affects coppicing by stumps after harvest. Details on the incidence and epidemiology of the disease are presented.     

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.     

Quantifying above- and below-ground growth responses of the western Australian oil mallee,Eucalyptus kochii subsp. plenissima,to contrasting decapitation regimes

Resprouting in the oil mallee, Eucalyptus kochii Maiden & Blakely subsp. plenissima Gardner (Brooker), involves generation of new shoots from preformed meristematic foci on the lignotuber. Numbers of such foci escalated from 200 per lignotuber in trees aged 1 year to 3,000 on 4- to 5-year-old trees. Removal of shoot biomass by decapitation 5 cm above ground in summer (February) or spring (October) resulted in initiation of 140-170 new shoots, but approx. 400 shoots were induced to form if crops of new shoots were successively removed until sprouting ceased and rootstocks senesced. Initially, the new shoot biomass of regenerating coppices increased slowly and the root biomass failed to increase appreciably until 1.7-2.5 years after cutting. Newly cut trees showed loss of fine root biomass, and structural roots failed to secondarily thicken to the extent shown by uncut trees. After 2 years, the biomass of shoots of coppiced plants was only one-third that of uncut control trees and shoot:root dry mass ratios of coppiced plants were still low (1.5-2.0) compared with those of the controls (average ratio of 3.1). Spring cutting promoted quicker and greater biomass recovery than summer cutting. Starch in below-ground biomass fell quickly following decapitation and remained low for a 12-18 month period. Utilization of starch reserves in naturally regenerating coppices was estimated to provide only a small proportion of the dry matter accumulated in new shoots. Results are discussed in relation to their impact on coppicing ability of the species under natural conditions or when successively coppiced for shoot biomass production.     

Virus diseases of cacao in West Africa; technique of insect transmission

Experiments on the technique of insect transmission of the cacao virus 1A (swollen-shoot) are described. This virus is unique in being transmitted by mealybugs (Coccoidea) and the experiments show that all stages of Pseudococcus njalensis Laing and of Ferrisia virgata Ckll. are vectors. These insects become infective after feeding for less than 4 hr. on the infected plant and transmit after less than 3 hr. on the test plant. The virus is non-persistent in the vector after 3 hr. test-feeding. The vectors can collect virus from either leaf, green shoot, bark or pod; the young symptom-bearing leaf is the best site for infection-feeding and the cotyledon of the bean for test-feeding.     

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.     

Mountain pine beetle selectivity in old‐growth ponderosa pine forests,Montana, USA

A historically unprecedented mountain pine beetle (MPB) outbreak affected western Montana during the past decade. We examined radial growth rates (AD 1860–2007/8) of co‐occurring mature healthy and MPB‐infected ponderosa pine trees collected at two sites (Cabin Gulch and Kitchen Gulch) in western Montana and: (1) compared basal area increment (BAI) values within populations and between sites; (2) used carbon isotope analysis to calculate intrinsic water‐use efficiency (iWUE) at Cabin Gulch; and (3) compared climate‐growth responses using a suite of monthly climatic variables. BAI values within populations and between sites were similar until the last 20–30 years, at which point the visually healthy populations had consistently higher BAI values (22–34%) than the MPB‐infected trees. These results suggest that growth rates two–three decades prior to the current outbreak diverged between our selected populations, with the slower‐growing trees being more vulnerable to beetle infestation. Both samples from Cabin Gulch experienced upward trends in iWUE, with significant regime shifts toward higher iWUE beginning in 1955–59 for the visually healthy trees and 1960–64 for the MPB‐infected trees. Drought tolerance also varied between the two populations with the visually healthy trees having higher growth rates than MPB‐infected trees prior to infection during a multi‐decadal period of drying summertime conditions. Intrinsic water‐use efficiency significantly increased for both populations during the past 150 years, but there were no significant differences between the visually healthy and MPB‐infected chronologies.     

Influences of the biophysical environment on blister rust and mountain pine beetle,and their interactions,in whitebark pine forests

Aim To understand how the biophysical environment influences patterns of infection by non‐native blister rust (caused by Cronartium ribicola) and mortality caused by native mountain pine beetles (Dendroctonus ponderosae) in whitebark pine (Pinus albicaulis) communities, to determine how these disturbances interact, and to gain insight into how climate change may influence these patterns in the future. Location High‐elevation forests in south‐west Montana, central Idaho, eastern and western Oregon, USA. Methods Stand inventory and dendroecological methods were used to assess stand structure and composition and to reconstruct forest history at sixty 0.1‐ha plots. Patterns of blister rust infection and mountain pine beetle‐caused mortality in whitebark pine trees were examined using nonparametric Kruskal–Wallis ANOVA, Mann–Whitney U‐tests, and Kolmogorov–Smirnov two‐sample tests. Stepwise regression was used to build models of blister rust infection and mountain pine beetle‐related mortality rates based on a suite of biophysical site variables. Results Occurrence of blister rust infections was significantly different among the mountain ranges, with a general gradient of decreasing blister rust occurrence from east to west. Evidence of mountain pine beetle‐caused mortality was identified on 83% of all dead whitebark pine trees and was relatively homogenous across the study area. Blister rust infected trees of all ages and sizes uniformly, while mountain pine beetles infested older, larger trees at all sites. Stepwise regressions explained 64% and 58% of the variance in blister rust infection and beetle‐caused mortality, respectively, indicating that these processes are strongly influenced by the biophysical environment. More open stand structures produced by beetle outbreaks may increase the exposure of surviving whitebark pine trees to blister rust infection. Main conclusions Variability in the patterns of blister rust infection and mountain pine beetle‐caused mortality elucidated the fundamental dynamics of these disturbance agents and suggests that the effects of climate change will be complex in whitebark pine communities and vary across the species’ range. Interactions between blister rust and beetle outbreaks may accelerate declines or facilitate the rise of rust resistance in whitebark pine depending on forest conditions at the time of the outbreak.     

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.     

Two Different Epidemiological Scenarios of Border Disease in the Populations of Pyrenean chamois (Rupicapra p. pyrenaica) after the First Disease Outbreaks

Since 2001 several outbreaks of a new disease associated with Border disease virus (BDV) infection have caused important declines in Pyrenean chamois (Rupicapra pyrenaica) populations in the Pyrenees. The goal of this study was to analyze the post-outbreak BDV epidemiology in the first two areas affected by disease with the aim to establish if the infection has become endemic. We also investigated if BDV infected wild and domestic ruminants sharing habitat with chamois. Unexpectedly, we found different epidemiological scenarios in each population. Since the disease outbreaks, some chamois populations recuperated quickly, while others did not recover as expected. In chamois from the first areas, prevalence was high (73.47%) and constant throughout the whole study period and did not differ between chamois born before and after the BDV outbreak; in all, BDV was detected by RT-PCR in six chamois. In the other areas, prevalence was lower (52.79%) and decreased during the study period; as well, prevalence was significantly lower in chamois born after the disease outbreak. No BDV were detected in this population. A comparative virus neutralisation test performed with four BDV strains and one Bovine viral diarrhoea virus (BVDV) strain showed that all the chamois had BDV-specific antibodies. Pestivirus antibodies were detected in all the rest of analyzed species, with low prevalence values in wild ruminants and moderate values in domestic ruminants. No viruses were detected in these species. These results confirm the hypothesis that outbreaks of BDV infection only affect the Pyrenean chamois, although other wild ruminants can occasionally be infected. In conclusion, two different scenarios have appeared since the first border disease outbreaks in Pyrenean chamois: on the one hand frequent BDV circulation with possible negative impact on population dynamics in some areas and on the other, lack of virus circulation and quick recovery of the chamois population.     

ACUTE DIE-BACK OF CLOVE TREES IN THE ZANZIBAR PROTECTORATE

The most widespread die-back of clove trees ( Eugenia aromatica) in the Zanzibar Protectorate is caused by Cryptosporella eugeniae sp.nov. The most noticeable symptom is the death of a branch or a portion of a branch, or, in young saplings, of the entire tree; in mature trees infection eventually leads to the semi-moribund trees now common in almost every clove plantation. The fungus invariably enters through a wound, most often one resulting from harvest damage. Pycnidia, and later perithecia, develop near the point of entry. Internally the infected wood is clearly distinguishable from the healthy by a dark red-brown stain.
Experimental infections that reproduce typical symptoms can rarely be caused in plants aged 18 to 36 months, and not at all in young seedlings. Experimental infection of older plants becomes more certain with increasing age, and with saplings of 7–10 years old failure is rare.
The prevalence of Cryptosporella die-back is almost entirely attributable to the destructive methods of harvesting now practised, combined with the presence of much infected material present in the plantations. Suggested control measures are based on the removal of old sources of infection and prevention of future damage, combined with remedial treatment where possible.     

Early seed fall and seedling emergence: precursors to tropical restoration

We explore processes of seed immigration and seedling recruitment before an experimental rainforest restoration matures enough to affect either. Twenty-four 30 × 30-m plots were fenced in 12 ha of pasture in 2006. Seeds were collected in ninety-six 1-m⁻² seed traps; recruits were censused in ~12,000 m² in which establishment was allowed. We tested effects of distance from forest, living trees, and stumps of trees cut during site preparation on seed rain in 2007 and effects of these and soil depth on recruits through June 2008. Seed fall and recruitment were not correlated with distance to forest 90–400 m away, nor to living shade trees outside the 160 × 485-m experimental grid. Recruitment differed for animal- and wind-dispersed species in a topographically complex landscape. Recruitment of wind-dispersed species was random with respect to soil depth or distance to recent stumps. Recruitment of animal-dispersed species was multimodal; partial correlations with number of stumps within 30 m of plots were significant with soil depth held constant (P < 0.025), as were correlations of recruitment with soil depth with number of stumps held constant (P < 0.01). Animal-dispersed recruits were often not conspecifics of adults that had been cut, indicating a legacy of attraction by fruiting trees of animals bearing seeds from distant sources. Ecological implications are that recruitment in pastures released from grazing reflects a mix of widely scattered wind-dispersed pioneers and, where animal-dispersed trees exist, multi-modal and decidedly non-random recruitment of pioneer and later successional animal-dispersed trees from seed banks.     

Importance of site conditions and time since abandonment for coppice vegetation on Langeland, Denmark

In 57 sample plots in neglected coppices on Langeland, Denmark (14—c. 125 years since last coppicing) the cover of trees and shrubs was analysed by the method of Domin-Krajina and the ground layer by the frequency method of Raunkiær. The data were analysed by DC A. The tree and shrub layer showed significant correlations with topsoil pH and litter depth. The ground layer was significantly correlated with inclination, area of the wood in which the coppice was situated, light, topsoil pH and the tree layer, and to a lesser extent with litter depth, but was not correlated with time since last coppicing. It is suggested that for some stands the suppression of Corγlus avellana by tree species ( Quercus robur, Fagus sylvatica, Carpinus betulus ) which form a permanent litter layer and acidify the soil is more important to the ground layer than the time elapsed since last coppicing per se. Formation of a permanent litter layer reduces species density, e-specially of some forest species without clonal growth. One main reason for the lack of correlation between the ground layer and time since coppicing may be the great environmental variation in the neglected coppices on Langeland. Another reason seems to be the absence of competitive forest species (which otherwise would benefit from non-intervention) due to the isolated position of the coppiced woods. It is concluded that neglect in excess of c. 35 years beyond the normal coppice rotation has some effect on the composition of the tree and shrub layer, but not a direct effect on the ground layer.     

Water relations of citrus exocortis viroid-infected grapefruit trees in the field

Citrus viroids (CVds) were used to limit citrus tree size ('dwarfing'). It is hypothesized that changes also occur in the hydraulic properties of the conducting system, thereby affecting water relations. Grapefruit trees (var. Star Ruby on Troyer Citrange rootstock) in northern Israel were infected with citrus exocortis viroid (CEVd). The orchard was drip irrigated. The infected (I) and apparently healthy (H) control plots were subdivided into wet (W) and dry (D) irrigation treatments receiving 100% and 70% of the recommended irrigation quantity for the region. Soil water content, soil temperature, water uptake, leaf and stem water potential, and leaf conductance were measured in addition to climatic variables. Water uptake response to increasing spring-time soil temperatures was greater in healthy than in infected trees. Leaf conductance in the infected trees was lower than the non-infected trees only in the sixth year after inoculation. Stem water potential was significantly lower in the CEVd-infected trees than in the control trees. Water uptake was lower and hydraulic resistance higher in infected than in healthy trees. It was concluded that CEVd causes a reduction in the water uptake ability of the root and canopy system. Possible mechanisms for this are discussed.     

Coppicing affects growth, root:shoot relations and ecophysiology of potted Quercus rubra seedlings

Two experiments were conducted to examine the response of Quercus rubra L. seedlings to coppicing. In a greenhouse experiment, growth, biomass distribution, leaf gas exchange, and water and carbohydrate relations were measured for 1-year-old seedlings that were either coppiced when dormant at the time of planting or left intact as controls. Coppicing induced sprouting from the base of the stem, and, in general, the physiology of sprouts and controls was similar. However, the relative growth rate (RGR) of sprouts was 9% higher than that of controls, allowing sprouts to compensate fully for the initial mass lost to coppicing. In a second experiment, in an outdoor cold frame, growth, biomass distribution, leaf gas exchange and plant water relations were measured on 1-year-old seedlings that were either coppiced at the time of planting (dormant-coppiced), coppiced soon after bud break (active-coppiced) or left intact (controls). Dormant coppicing again had little impact on seedling physiology, and dormant-coppiced plants again compensated for initial mass loss with a higher RGR. In contrast, active-coppiced seedlings did not compensate for initial mass loss, as their RGR did not differ from that of controls. By the tenth week of the study, leaf gas exchange rates of active-coppiced sprouts were higher than those of dormant-coppiced and control seedlings. Active-coppiced sprouts also had a greater soil-to-leaf hydraulic conductivity (expressed on a leaf area basis) and a lower ratio of leaf area to root surface area than did controls. Across treatments, photosynthetic rate and stomatal conductance were positively correlated with soil-to-leaf hydraulic conductivity, and gas exchange rates and hydraulic conductivity were negatively related to leaf:root area ratio. Thus, the removal of actively growing shoots may have altered subsequent leaf gas exchange largely through coppice-induced changes in leaf-root balance.     

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.