Evaluation and use of a screening method to aid selection of cocoa (Theobroma cacao) with field resistance to cocoa swollen-shoot virus in Ghana

Seeds of cocoa progenies were inoculated with cocoa swollen-shoot virus manually or by vector transfer techniques to assess the relative resistance of the progenies to infection. Assessment of progeny included in both inoculation tests and in field trials were positively correlated, indicating that the inoculation technique is suitable for the selection of progenies for testing in the field. Pure Upper Amazon progenies were more resistant than selfed Amelonado, with hybrids between Amelonado and Upper Amazons usually intermediate. Among the main Upper Amazon populations, Iquitos and Nanay clones were better sources of resistance than Parinari and Scavina clones. A survey of these populations indicated that, within populations, resistance levels do not vary greatly. A range of progenies based on the Upper Amazon female parents in existing seed gardens was screened for resistance and some were consistently more resistant than the equivalent Amazon x Amelonado hybrid now being distributed to farmers.     

Methods for assessing the tolerance and resistance of different types of cocoa to cocoa swollen-shoot virus

Different types of cocoa were compared for their resistance to infection and tolerance of infection with the virulent strain A of cocoa swollen-shoot virus, inoculated either by mealybugs or manually. The method of infection did not affect the ranking of the cocoa types for resistance, except at high percentages of infection, or for tolerance. In comparison with other methods, manual inoculation simplifies the screening of cocoa types for resistance and tolerance to virus infection, and provides a more sensitive test for resistance.     

Genetic aspects of resistance to cocoa swollen-shoot virus in Ghana

The inheritance of resistance in cocoa to infection with cocoa swollen-shoot virus was studied in seed inoculation experiments with progenies of Upper Amazon, Trinitario and West African Amelonado parents. In an eight parent diallel cross grown in two seasons and in six male × female factorial experiments, general combining abilities were much more important than specific combining abilities. The diallel cross revealed significant maternal effects but there was little evidence of specific reciprocal differences. Estimates of general combining abilities corresponded with expectation and were positively correlated with maternal effects, the latter indicating that tester parents should be used as males. The results from scaling tests with hybrids between selections from the River Nanay (a Peruvian tributary of the River Amazon) and Amelonado fitted a model with additive resistance; there was no evidence of dominance. Attempts to obtain stronger sources of resistance by hybridisation and selection were unsuccessful. In three crosses the resistance of individual seedlings was compared with that of the parents using test crosses but no evidence of transgressive segregation was obtained. In further experiments seedings of two hybrids between contrasting parents were heavily inoculated with virus so that few healthy survivors remained. Test crosses showed that these survivors and randomly selected seedlings of the same parentage did not differ as resistance sources.     

Studies on the possibility of increasing resistance to cocoa swollen-shoot virus by breeding

In tests using seed, the resistance of cocoa progenies to primary infection with cocoa swollen-shoot virus by mealybugs was stable to inoculum pressure and effective against several virus strains. The results suggested that the resistance would be effective throughout the cocoa growing areas of Ghana. There were indications that resistance factors from different cocoa populations could be accumulated to give progenies of higher resistance than presently available.     

Resistance of cocoa to swollen-shoot virus in Ghana. I. Field trials

In Ghana, field resistance to cocoa swollen-shoot virus (CSSV) in young, bearing trees was assessed by monitoring natural spread by mealybug vectors from adjacent graft-inoculated line sources. Apparent rates of virus spread were estimated by regressing proportions of infected trees on time from inoculation. In some hybrids between parents derived from Upper Amazonian material, spread was only 25–40% that in currently recommended varieties. This resistance is the most effective, feasible measure to reduce economic losses and the resistant hybrids could be made available quickly for large scale use. The results from trials in which virus spread most rapidly were the easiest to interpret. Spread of CSSV is erratic, necessitating large experiments, and in trials with equal replication the largest plots gave the greatest precision. Neighbouring plot adjustment proved unsatisfactory as it improved precision but appeared to reduce accuracy.     

The virobacterial agglutination test as a rapid means of detecting cocoa swollen shoot virus

The virobacterial agglutination (VBA) test was developed as a means of detection of cocoa swollen shoot virus (CSSV). Identification of CSSV-infected Theobroma cacao in the field has only been possible by visual examination of symptoms, by virus transmission using mealybugs and by grafting to induce symptom expression in Amelonado cocoa seedlings. Detection of latent infection has not been possible even using enzyme-linked immunosorbent assays (ELISA). The VBA test successfully detected CSSV in infected sap diluted to 1/2560. Antisera to a range of mild and severe CSSV isolates were tested, and the results suggest a close relationship between seven isolates (1A, Bosomtwi, Bosomuoso, Nkrankwanta, Nsaba, Seidi-Nkawie and SS365B) while the mild isolate N1 appears to be less closely related. The VBA test was compared with both direct and indirect ELISA in the field. Only VBA detected all the cocoa trees which were known to be infected and additionally identified infection in many symp-tomless trees.     

The resistance and tolerance of different cocoa varieties to cocoa swollen-shoot virus in Ghana

The resistance and tolerance to infection with cocoa swollen-shoot virus (virulent strain A) of forty-five and thirty-eight cocoa progenies respectively was estimated using a manual inoculation method. Resistance was estimated from the proportion of inoculated seedlings which developed symptoms, and tolerance from the severity of these symptoms during a 4-month period. Resistance was found among Nanay, Iquitos and Scavina, all Upper Amazon types; and an inter-Nanay cross was outstandingly resistant and offers scope for improvement by breeding. Some Amazon-Amazon and Trinitario-Amazon progenies snowed tolerance, especially those with Trinitario T9/21 as a parent. Some progenies seem sufficiently promising for field testing to select varieties suitable for planting in those areas in Ghana where swollen-shoot disease is epidemic.     

Seed transmission of broad bean stain virus and Echtes Ackerbohnenmosaik-Virus in field beans (Viciafaba)

When grown in a glasshouse during spring or autumn field bean (Vicia faba minor) seedlings infected with seed-borne broad bean stain virus (BBSV) or Echtes Ackerbohnenmosaik-Virus (EAMV) usually showed symptoms on some leaves within 4 wk of emergence. Symptoms caused by each virus were indistinguishable. The viruses were transmitted as often through unblemished seeds as through seeds with necrotic patches or stains on the seed coat, and sometimes as often through large as through small seeds. Soaking seeds for 24 h in solutions of 8-azaguanine or polyacrylic acid did not decrease transmission. Both viruses were detected in nearly mature seeds by inoculation to Phaseolus vulgaris but neither virus was detected in fully ripened seeds by inoculation or serological tests. The percentage of seeds from field plots that produced infected seedlings when sown in a glasshouse was closely related to the percentage of parent plants that showed symptoms of BBSV and/or EAMV at the end of flowering. The relationship seemed similar in different cultivars. On average EAMV was transmitted through more seeds than BBSV, probably because more parent plants were infected with EAMV. Inspection of seed crops for symptoms of BBSV and EAMV at the seedling stage and again at the end of flowering is probably the most practicable way of identifying progeny seed lots with little or no infection.     

Determining the effectiveness of resistance to subterranean clover mottle sobemovirus in different genotypes of subterranean clover in the field using the grazing animal as virus vector

The effectiveness of resistance to subterranean clover mottle sobemovirus (SCMoV) previously identified in different genotypes of subterranean clover (Trifolium subterraneum) inoculated with infective sap in the glasshouse, was tested in two field experiments which used the grazing animal as virus vector. Replicated plots each consisting of paired test rows of 20 different genotypes were used. Clover plants infected with SCMoV were transplanted in between the paired test rows and these acted as sources of the virus for spread by grazing sheep. Although used in different years at different sites with different virus isolates, the field exposure methodology employed produced consistent results. The genotypes each behaved similarly in both experiments as regards the relative extents of SCMoV infection that developed, levels ranging from 0–98%. The previously identified resistance in six ‘highly resistant’ and three ‘partially resistant’ cultivars was effective under field conditions. However, the ‘partial resistance’ in three others was overcome, cvs Green Range and Mt Barker developing levels of infection approaching those in ‘susceptible’ cultivars, while an intermediate infection level developed in cv. Karridale. The three cultivars in which partial resistance was not effective all belonged to ssp. subterraneum. In subterranean clover breeding programmes, field screening using the grazing animal as a vector is advisable to determine whether SCMoV resistance found by sap inoculation is still effective under field conditions.     

The effect of Erwinia carotovora subsp. atroseptica (blackleg) on potato plants. II. Compensatory growth

In field experiments in 1981 and 1982, uninoculated seed tubers (cv. Désirée) and those inoculated with Erwinia carotovora subsp. atroseptica at the rose (apical) or heel (stolon attachment) ends were planted at normal (35 cm) or double spacing; in additional plots, inoculated and uninoculated tubers were planted alternately. Inoculation, especially at the rose end, decreased plant height and sometimes resulted in blackleg symptoms. Individual plant yields were recorded at the end of the season. In plots of uniform seed type at normal spacing, inoculation decreased total yield compared with uninoculated by 12–13% (heel-end inoculation) or 26–40% (rose-end inoculation). At double spacing, yields increased compared with normal spacing by 44–58% (uninoculated or heel-end inoculation) or 30–39% (rose-end inoculation). When rose-end-inoculated and uninoculated seed tubers were planted alternately, inoculated plants yielded less and uninoculated plants more than in plots planted throughout with the same seed treatment. The abilities of inoculated and uninoculated plants to compensate for weak or missing neighbours were combined using equations to predict the yields of crops with different proportions of diseased or missing plants.     

Screening for Barley yellow dwarf virus-Resistant Barley Genotypes by Assessment of Virus Content in Inoculated Seedlings

The content of Barley yellow dwarf virus (BYDV) in roots and leaves of barley seedling plants differing in their level of resistance was assessed by quantitative ELISA 1–42 days after inoculation with the strain of BYDV (PAV). High virus accumulation in roots and low concentration in leaves was characteristic of the period 9–15 days after inoculation. In leaves, the differences in virus content between resistant and susceptible genotypes became significant after 15 days and resistance to virus accumulation was better expressed 30–39 days after inoculation. Roots of resistant materials exhibited evident retardation of virus accumulation and the greatest difference in virus content between resistant and susceptible plants was detected 9 days after inoculation. By these criteria, the selected winter and spring barley cultivars and lines (in total 44 materials) fell in to five groups according to field reactions and the presence or absence of the Yd2 resistance gene. There were highly significant and positive relations between ELISA values and 5‐year field data on symptomatic reactions and grain‐yield reductions due to infection. Using the described method, resistant and moderately resistant genotypes (both Yd2 and non‐Yd2) were significantly differentiated from susceptible genotypes. The possible use of this method in screening for BYDV resistance is discussed.     

The host range and vector species of viruses from Cola chlamydantha K. Schum., Adansonia digitata L. and Theobroma cacao L

Seventeen species in the Tiliaceae, Sterculiaceae and Bombacaceae were tested for susceptibility to three virus isolates from naturally infected Cola chlamydantha trees and three from Adansonia digitata trees in Ghana. Seven species of Pseudococcidae were tested as vectors of the six isolates. These studies indicate that the Cola isolates should be classified with cocoa swollen shoot virus and those from Adansonia with cocoa mottle leaf virus.     

Heritability of Phenols in the Resistance of Theobroma cacao against Phytophthora megakarya, the Causal Agent of Black Pod Disease

The black pod disease caused by Phytophthora megakarya is responsible for 80% of the cocoa production loss in Cameroon. To assess the resistance of cocoa plants against this pathogen, necrotic lesions, phenolic content and qualitative alteration of phenolics were conducted in ICS84 and ICS95 clones (two Trinitario introduced from Trinidad) and their hybrids (families F30 and F25) derived from reciprocal cross breeding between these two parental clones after inoculation. The existence of strong hybrid vigour has been shown. Ninety percentage of the hybrid's genotypes manifested a positive heterosis effect for the development of lesion size. This suggests the existence of hybrid vigour with a genetic additive effect. F3086, F2509, F2552 and F2586 hybrids were characterized by localized lesions. Those hybrids genotypes can be considered as elite clones. In relation to analysis of total phenolics and lesion size, no maternal effect was detected in the transmission of these characters. A significant and negative correlation (r = −0.683) (P < 0.01) has been observed between necrosis evolution and phenolics accumulation. The values of the heritability of lesion size and the total phenolic content in offsprings did not permit to show the maternal effect. Qualitative analyses of phenolics showed high flavonones content in cocoa leaves. Qualitative analyses of phenolics in ICS84, ICS95 clones and their hybrids showed a modification of the phenolics profiles, notably concerning apigenin and luteolin derivatives due to the inoculation. These compounds, along with others that were not identified, could have a role in the reaction and mechanism of defence of cocoa against P. megakarya.     

Establishment of Bradyrhizobium japonicum for soybean by inoculation of a preceding wheat crop

On fields with no history of soybean (Glycine max (L.) Merr.) production, inoculation alone is often inadequate to provide for adequate nodulation the first time this crop is grown. The objective of this study was to determine if inoculation of spring wheat (Triticum aestivum L.) seed with Bradyrhizobium japonicum would lead to an increase of B. japonicum numbers in the soil, and improve nodulation of a subsequent soybean crop. In the greenhouse, wheat seed inoculation increased B. japonicum numbers from undetectable numbers to >9000 g^–1 of soil, whereas the numbers of introduced B. japonicum declined in unseeded pots. In the field, inoculation of wheat seed increased B. japonicum numbers in the soil from undetectable levels to >4000 g^–1 the following year. When soybean seed was inoculated, but grown in soil devoid of B. japonicum, nodules formed only near the point of seed placement. The heaviest nodulation, and widest distribution of nodules in the topsoil were found whenB. japonicum was established the year before by wheat seed inoculation, plus soybean seed inoculation. Wheat seed inoculation the year before growing soybean, combined with proper soybean seed inoculation, should provide for abundant nodulation the first time soybean is grown on a field.     

Further studies on seed transmission of broad bean stain virus and Echtes Ackerbohnenmosaik -Virus in field beans (Vicia faba)

Broad bean stain virus (BBSV) and Echtes Ackerbohnenmosaik-Virus (EAMV) were detected in the seed coat and embryo sac fluid of immature seeds from infected field beans (Viciafaba minor) by inoculation to Phaseolus vulgaris; BBSV was also detected in immature embryos. The proportion of seeds infected with either virus decreased during maturation. The viruses were transmitted to seedlings as often through fully ripened seeds from which the seed coats had been removed as through intact seeds. Both viruses were detected in pollen from infected plants, but in glasshouse tests only BBSV was transmitted through pollen to seeds. Delaying fertilization in plants infected with BBSV or EAMV seemed not to affect seed transmission of either virus. In glasshouse tests BBSV was transmitted more often through seeds from plants that were inoculated before flowering than during flowering, and was not transmitted through seeds from plants inoculated after flowering; EAMV was transmitted only through seeds from plants inoculated before flowering. In tests on seed from naturally infected plants BBSV was transmitted more often through seeds from plants that developed symptoms before flowering than during flowering. Both viruses were seed-borne in all cultivars tested and there was no marked difference in the frequency of transmission of either virus among the spring-sown cultivars most common in Britain. Both viruses persisted in seed for more than 4 yr.     

Seedling disease in an annual legume: consequences for seedling mortality, plant size, and population seed production

The effect of seed and seedling mortality on plant population dynamics depends on the degree to which the growth and reproduction of surviving individuals can compensate for the deaths that occur. To explore this issue, we sowed seeds of the annual Kummerowia stipulacea at three densities in sunken pots in the field, which contained either field soil, microwaved field soil, or microwaved field soil augmented with oospores of three Pythium species. High sowing density reduced seedling establishment and seedling size, but these effects were independent of the soil treatment. In the oospore-augmented soil, seed and seedling survival was low. The surviving plants were initially smaller but, at maturity, average plant size was greatest in the oospore-augmented soil, compared to the other treatments. Total population seed production was unaffected by soil treatment, suggesting that the effect of disease was limited to the seedling stage, with surviving plants released from intraspecific competition. To test the hypothesis that the surviving plants in the oospore-augmented soil were more disease-resistant, seeds from each of the sowing density-soil type treatments were sown in a growth chamber inoculation study. No evidence for selection for resistance was found. A second inoculation experiment revealed that oospore inoculum reduced plant numbers and mass regardless of whether field or microwaved soil was used, suggesting that results from the field experiment were not dependent on the use of microwaved soil. The findings of this study indicate that the ecological effects of disease on individual plants and on plant populations are not necessarily equivalent. Received: 13 January 1999 / Accepted: 21 September 1999     

Occurrence of potato virus X strain group 1 in seed stocks of potato cultivars lacking resistance genes

Potato virus X (PVX) isolates were obtained from a simple seed potato production scheme or from ware potatoes produced by seed potatoes obtained from it. In this scheme, PVX infection is widespread in seed stocks and most of the potatoes grown lack PVX resistance genes. Thirteen PVX isolates were typed to strain group by inoculation to potato cultivars containing different combinations of hypersensitivity genes Nx and Nb. Six failed to overcome either gene and therefore belonged to strain group 1, four overcame Nb only and were placed in strain group 3 and three were mixtures of the two. All 13 isolates failed to overcome extreme resistance/immunity gene Rx. Naturally infected cultivars of genotype nx.nb contained strain group 1 alone or strain groups 1 and 3, while those of genotype nx:Nb contained only strain group 3. The widespread occurrence of strain group 1 contrasts with the predominant occurrence of strain group 3 in potatoes in the UK. However, it resembles the UK situation before sophisticated seed potato production schemes were introduced and before PVX hypersensitivity genes Nx and Nb were deliberately exploited in potato breeding. Prior infection with potato leafroll virus (PLRV) did not affect expression of hypersensitivity to PVX in inoculated leaves of an nx:Nb genotype.     

Tolerance trials with young cocoa trees infected with cocoa swollen-shoot virus

In three short-term field trials, 24 cocoa progenies were assessed for sensitivity to infection with a severe Ghanaian strain of cocoa swollen-shoot virus. Some of the progenies were derived from tolerant parents, others had themselves shown mild symptoms as young seedlings. Stem diameters, diameter increments, canopy scores and latent periods (time between inoculation and expression of symptoms) of plants infected when 12 to 22 months old were compared with similar measurements of healthy plants. During the period between 6 and 18 months after inoculation, infection reduced the stem diameter increment of the least sensitive progeny in each trial by 52, 25 and 7%, the last value increasing to 38% during the period between 24 and 30 months after inoculation. Within trials, the effects of virus infection on stem diameter increments and canopy scores were well correlated but latent periods were not significantly correlated with any other variate. In one trial, pod yield was more closely related to canopy condition at the time of cropping than to stem diameter increment 2 yr earlier. Weak correlations between pre- and post-bearing increases in diameters in these trials indicated that tolerance was not directly related to vigour. None of the progenies is considered suitable for direct use as tolerant varieties in Ghana.     

Host preferences of the cocoa pod borer,Conopomorpha cramerella,the main threat to cocoa production in Southeast Asia

Insects ensure the survival of their offspring by depositing their eggs in suitable environments. Even generalist egg-laying insects often show preferences for specific host plants. The cocoa pod borer (CPB), Conopomorpha cramerella (Snellen) (Lepidoptera: Gracillariidae), successfully infests and reproduces on relatively few host plants, but has a major economic impact only on cocoa, Theobroma cacao L. (Malvaceae). Choice tests were performed in the laboratory to compare the frequency of insect visits, the duration of the visits, and the number of eggs laid on the fruits for each combination of host plants tested – that is, cocoa clones or fruits of rambutan (Nephelium lappaceum L.), Fijian longan (Pometia pinnata JR Forst & G Forst, both Sapindaceae), and langsat (Lansium domesticum Corrêa, Malvaceae). Our laboratory study showed that, when given the choice, CPB significantly favored cocoa pods over other host fruits (rambutan, langsat, or Fijian longan). Females also deposited more eggs on unripe fruits than on ripe fruits of similar size. The preference to lay eggs on specific cocoa clones in the bioassays did not reflect the level of the clone resistance to CPB damage reported from the field. Consequently, oviposition preference of female CPB does not seem to be the main factor explaining field resistance of some cocoa clones to CPB infestation.     

Effect of cucumber mosaic virus infection on morphology,yield and phenolic contents of tomato

Ten tomato genotypes were screened for their resistance against cucumber mosaic virus (CMV) and its vector Myzus persicae under natural infection in field, using aphids M. persicae under net-house and mechanical inoculation under greenhouse. Large differences were observed among genotypes for infection percentage (IP) and severity index (SI) among the testing methods used. All genotypes showing tolerance to CMV in the field or through aphid inoculation, however, become susceptible and highly susceptible after mechanical inoculation. All the test genotypes also showed susceptibility to the aphid M. persicae population. Plants inoculated with CMV showed substantial decrease in yield and yield-contributing parameters which varied with cultivars that probably depended upon its genetic make up. All the test genotypes exhibited 0.97–30.19% decrease in plant height, 11.47–52.65% decrease in root length, 46.56–95.56% decrease in fresh plant weight, 65.78–92.84% decrease in root fresh weight, 19.97–87.65% decrease in the dry weight of plants, 75.63–95.43% decrease in dry root weight, 69.51–95.65% reduction in the number of fruits and 89.04–99.89% decrease in yield per plants. After 15 days of inoculation, the quantitative analysis using double beam spectrophotometer showed an increase in total phenolics in CMV-inoculated plants as compared to un-inoculated plants among genotypes. Similarly the thin layer chromatography (TLC) on silica gel G indicated that the number of phenolic compounds was increased in most of the inoculated genotypes while in others they were either decreased or remained same.