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

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

Identification of Pseudomonas syringae pathogens of Arabidopsis and a bacterial locus determining avirulence on both Arabidopsis and soybean.

To develop a model system for molecular genetic analysis of plant-pathogen interactions, we studied the interaction between Arabidopsis thaliana and the bacterial pathogen Pseudomonas syringae pv tomato (Pst). Pst strains were found to be virulent or avirulent on specific Arabidopsis ecotypes, and single ecotypes were resistant to some Pst strains and susceptible to others. In many plant-pathogen interactions, disease resistance is controlled by the simultaneous presence of single plant resistance genes and single pathogen avirulence genes. Therefore, we tested whether avirulence genes in Pst controlled induction of resistance in Arabidopsis. Cosmids that determine avirulence were isolated from Pst genomic libraries, and the Pst avirulence locus avrRpt2 was defined. This allowed us to construct pathogens that differed only by the presence or absence of a single putative avirulence gene. We found that Arabidopsis ecotype Col-0 was susceptible to Pst strain DC3000 but resistant to the same strain carrying avrRpt2, suggesting that a single locus in Col-0 determines resistance. As a first step toward genetically mapping the postulated resistance locus, an ecotype susceptible to infection by DC3000 carrying avrRpt2 was identified. The avrRpt2 locus from Pst was also moved into virulent strains of the soybean pathogen P. syringae pv glycinea to test whether this locus could determine avirulence on soybean. The resulting strains induced a resistant response in a cultivar-specific manner, suggesting that similar resistance mechanisms may function in Arabidopsis and soybean.     

Intrinsic resistance of feline peritoneal macrophages to coronavirus infection correlates with in vivo virulence.

Cats infected with virulent feline coronavirus strains develop feline infectious peritonitis, an invariably fatal, immunologically mediated disease; avirulent strains cause either clinically inapparent infection or mild enteritis. Four virulent coronavirus isolates and five avirulent isolates were assessed by immunofluorescence and virus titration for their ability to infect and replicate in feline peritoneal macrophages in vitro. The avirulent coronaviruses infected fewer macrophages, produced lower virus titers, were less able to sustain viral replication, and spread less efficiently to other susceptible macrophages than the virulent coronaviruses. Thus, the intrinsic resistance of feline macrophages may play a pivotal role in the outcome of coronavirus infection in vivo.     

Luciferase imaging of a neurotropic viral infection in intact animals

The identification of viral determinants of virulence and host determinants of susceptibility to virus-induced disease is essential for understanding the pathogenesis of infection. Obtaining this information requires infecting large numbers of animals to assay amounts of virus in a variety of organs and to observe the onset and progression of disease. As an alternative approach, we have used a murine model of viral encephalitis and an in vivo imaging system that can detect light generated by luciferase to monitor over time the extent and location of virus replication in intact, living mice. Sindbis virus causes encephalomyelitis in mice, and the outcome of infection is determined both by the strain of virus used for infection and by the strain of mouse infected. The mode of entry into the nervous system is not known. Virulent and avirulent strains of Sindbis virus were engineered to express firefly luciferase, and the Xenogen IVIS system was used to monitor the location and extent of virus replication in susceptible and resistant mice. The amount of light generated directly reflected the amount of infectious virus in the brain. This system could distinguish virulent and avirulent strains of virus and susceptible and resistant strains of mice and suggested that virus entry into the nervous system could occur by retrograde axonal transport either from neurons innervating the initial site of replication or from the olfactory epithelium after viremic spread.     

A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A is unable to process the precursor fusion glycoprotein of Newcastle disease virus.

RPE.40, a mutant strain of CHO-K1 cells isolated for resistance to Pseudomonas exotoxin A and cross-resistant to alphaviruses, is also highly resistant to virulent strains of Newcastle disease virus. The resistance of RPE.40 cells to Newcastle disease virus results from the failure to cleave the viral envelope precursor glycoprotein Fo to fusion glycoprotein F1 at the consensus sequence (Lys/Arg)-Arg-Gln-(Lys/Arg)-Arg.     

粳稻子预44抗LP11稻瘟病菌基因Pizy6(t)的定位

稻瘟病是世界范围内严重威胁水稻(Oryza sativa)生产可持续发展的主要病害之一,每年造成10%–30%的水稻产量损失。抗瘟水稻品种的培育和育种利用是解决稻瘟病危害最经济有效的方法。对新的致病性菌株进行分离和筛选是定位与克隆抗病新基因及抗病育种的基础。选择分离自不同稻瘟病发生重灾区的单孢菌株,对广谱抗瘟水稻子预44和感病水稻江南香糯进行致病性鉴定,筛选出两材料间致病性差异明显的5个菌株;进一步利用子预44、湘资3150、9311、日本晴、丽江新团黑谷、中花11、TP309和江南香糯8个抗瘟性不同的水稻材料,对筛选的菌株进行致病性鉴定。结果显示,LP11能使广谱抗瘟籼稻湘资3150严重发病,推测其很可能是新进化出来的强致病菌株。利用子预44和江南香糯杂交构建的F2群体进行抗性遗传分析,结果表明子预44对LP11菌株的抗性是由单显性基因控制。利用SSR分子标记和图位克隆方法在子预44中定位了1个抗稻瘟病基因Pizy6(t)。研究结果不仅为抗病相关研究提供了有价值的新菌株,而且为子预44中抗稻瘟病基因Pizy6(t)的克隆奠定了基础。     

Radiation-Resistant and Radiation-Sensitive Forms of Host Resistance to Polyomavirus

Newborn mice of several inbred strains develop few or no tumors following inoculation with highly tumorigenic strains of polyomavirus. Here we show that such resistant strains can be divided into two groups based on the responses of adult mice to radiation followed by virus inoculation. Most strains show a radiation-sensitive form of resistance (R^r-s) and develop tumors following radiation and virus challenge. This type of resistance has previously been recognized as immunological, based on T-cell responses against virus-encoded neoantigen(s) expressed in tumor cells. Other strains exhibit a radiation-resistant form of resistance (R^r-r) and fail to develop tumors when treated in the same manner. Three additional properties of R^r-r mice distinguish them from R^r-s mice: (i) survival of newborns following inoculation with a highly virulent and usually lethal strain of virus, (ii) resistance to virus spread in newborns inoculated with either tumorigenic or virulent virus strains, and (iii) dominant or semidominant transmission of resistance in crosses with a highly susceptible strain. The R^r-r phenotype reflects a constitutive nonimmunological type of resistance that is targeted to the virus and blocks its dissemination.