James Legg

Cassava mosaic geminiviruses in Africa

Cassava mosaic disease (CMD) caused by cassava mosaic geminiviruses (CMGs) (Geminiviridae:Begomovirus) is undoubtedly the most important constraint to the production of cassava in Africa at the outset of the 21st century. Although the disease was recorded for the first time in the latter part of the 19th century, for much of the intervening period it has been relatively benign in most of the areas where it occurs and has generally been considered to be of minor economic significance. Towards the end of the 20th century, however, the inherent dynamism of the causal viruses was demonstrated, as a recombinant hybrid of the two principal species was identified, initially from Uganda, and shown to be associated with an unusually severe and rapidly spreading epidemic of CMD. Subsequent spread throughout East and Central Africa, the consequent devastation of production of the cassava crop, a key staple in much of this region, and the observation of similar recombination events elsewhere, has once again demonstrated the inherent danger posed to man by the capacity of these viruses to adapt to their environment and optimally exploit their relationships with the whitefly vector, plant host and human cultivator. In this review of cassava mosaic geminiviruses in Africa, we examine each of these relationships, and highlight the ways in which the CMGs have exploited them to their own advantage.

A distinct Bemisia tabaci (Gennadius) (Hemiptera: Sternorrhyncha: Aleyrodidae) genotype cluster is associated with the epidemic of severe cassava mosaic virus disease in Uganda

During the 1990s, an epidemic of cassava mosaic virus disease caused major losses to cassava production in Uganda. Two factors associated with the epidemic were the occurrence of a novel recombinant begomovirus, EACMV‐Ug, and unusually high populations of the whitefly vector, Bemisia tabaci. Here we present molecular evidence for the occurrence of two cassava‐colonizing B. tabaci genotype clusters, Ug1 and Ug2, one of which, Ug2, can be consistently associated with the CMD epidemic in Uganda at the time of collection in 1997. By contrast, a second genotype cluster, Ug1, only occurred ‘at’ or ‘ahead of’ the epidemic ‘front’, sometimes in mixtures with Ug2. Comparison of mitochondrial cytochrome oxidase I gene sequences for Ug1 and Ug2 and well‐studied B. tabaci reference populations indicated that the two Ugandan populations exhibited ≈ 8% divergence, suggesting they represent distinct sub‐Saharan African lineages. Neither Ugandan genotype cluster was identified as the widely distributed, polyphagous, and highly fecund B biotype of Old World origin, with which they both diverged by ≈ 8%. Within genotype cluster divergence of Ug1 at 0.61 ± 0.1% was twice that of Ug2 at 0.35 ± 0.1%. Mismatch analysis suggested that Ug2 has undergone a recent population expansion and may be of nonUgandan origin, whereas Ug1 has diverged more slowly, and is likely to be an indigenous genotype cluster.

Molecular biodiversity of cassava begomoviruses in Tanzania: evolution of cassava geminiviruses in Africa and evidence for East Africa being a center of diversity of cassava geminiviruses

Cassava is infected by numerous geminiviruses in Africa and India that cause devastating losses to poor farmers. We here describe the molecular diversity of seven representative cassava mosaic geminiviruses (CMGs) infecting cassava from multiple locations in Tanzania. We report for the first time the presence of two isolates in East Africa: (EACMCV-[TZ1] and EACMCV-[TZ7]) of the species East African cassava mosaic Cameroon virus, originally described in West Africa. The complete nucleotide sequence of EACMCV-[TZ1] DNA-A and DNA-B components shared a high overall sequence identity to EACMCV-[CM] components (92% and 84%). The EACMCV-[TZ1] and -[TZ7] genomic components have recombinations in the same genome regions reported in EACMCV-[CM], but they also have additional recombinations in both components. Evidence from sequence analysis suggests that the two strains have the same ancient origin and are not recent introductions. EACMCV-[TZ1] occurred widely in the southern part of the country. Four other CMG isolates were identified: two were close to the EACMV-Kenya strain (named EACMV-[KE/TZT] and EACMV-[KE/TZM] with 96% sequence identity); one isolate, TZ10, had 98% homology to EACMV-UG2Svr and was named EACMV-UG2 [TZ10]; and finally one isolate was 95% identical to EACMV-[TZ] and named EACMV-[TZ/YV]. One isolate of African cassava mosaic virus with 97% sequence identity with other isolates of ACMV was named ACMV-[TZ]. It represents the first ACMV isolate from Tanzania to be sequenced. The molecular variability of CMGs was also evaluated using partial B component nucleotide sequences of 13 EACMV isolates from Tanzania. Using the sequences of all CMGs currently available, we have shown the presence of a number of putative recombination fragments that are more prominent in all components of EACMV than in ACMV. This new knowledge about the molecular CMG diversity in East Africa, and in Tanzania in particular, has led us to hypothesize about the probable importance of this part of Africa as a source of diversity and evolutionary change both during the early stages of the relationship between CMGs and cassava and in more recent times. The existence of multiple CMG isolates with high DNA genome diversity in Tanzania and the molecular forces behind this diversity pose a threat to cassava production throughout the African continent.

Cassava mosaic virus disease in East Africa: a dynamic disease in a changing environment

Cassava mosaic disease (CMD), now known to be caused by cassava mosaic geminiviruses (Family Geminiviridae; Genus Begomovirus), was first reported in East Africa in 1894. Epidemics occurred in Madagascar and Uganda in the 1930s and 1940s, and more localised rapid spread of CMD was observed in parts of coastal Tanzania in the 1930s and coastal Kenya in the 1970s. During the 1990s, a major regional pandemic of an unusually severe form of CMD has expanded to affect parts of at least five countries, causing massive economic losses and destabilising food security. Mechanisms responsible for the development and progress of the pandemic have been described, and comparisons of epidemiological data for varieties grown throughout the period under review suggest that the recent pandemic has been characterised by rapid rates of CMD spread hitherto unknown in East Africa. A key factor in the genesis and spread of the pandemic has been the recombination between two distinct cassava mosaic geminiviruses to produce a novel and more virulent hybrid. Although such events may be common, the known history of CMD in East Africa suggests that the frequency with which they become epidemiologically significant is low. A corollary of this is that resistance, developed originally in Tanzania between 1934 and 1960, and utilized and supplemented at the International Institute of Tropical Agriculture, Nigeria, since 1971, is providing effective CMD control in current pandemic-affected areas of East Africa. Consequently, it is concluded that prospects for managing CMD in the 21st century are good, and that the approach adopted should build on the model of collaborative research and implementation that has been established in tackling the current CMD pandemic.

Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virus-derived transgenes from plant tissues.

BackgroundPlant viral diseases present major constraints to crop production. Effective sampling of the viruses infecting plants is required to facilitate their molecular study and is essential for the development of crop protection and improvement programs. Retaining integrity of viral pathogens within sampled plant tissues is often a limiting factor in this process, most especially when sample sizes are large and when operating in developing counties and regions remote from laboratory facilities. FTA is a paper-based system designed to fix and store nucleic acids directly from fresh tissues pressed into the treated paper. We report here the use of FTA as an effective technology for sampling and retrieval of DNA and RNA viruses from plant tissues and their subsequent molecular analysis.ResultsDNA and RNA viruses were successfully recovered from leaf tissues of maize, cassava, tomato and tobacco pressed into FTA® Classic Cards. Viral nucleic acids eluted from FTA cards were found to be suitable for diagnostic molecular analysis by PCR-based techniques and restriction analysis, and for cloning and nucleotide sequencing in a manner equivalent to that offered by tradition isolation methods. Efficacy of the technology was demonstrated both from sampled greenhouse-grown plants and from leaf presses taken from crop plants growing in farmers fields in East Africa. In addition, FTA technology was shown to be suitable for recovery of viral-derived transgene sequences integrated into the plant genome.ConclusionResults demonstrate that FTA is a practical, economical and sensitive method for sampling, storage and retrieval of viral pathogens and plant genomic sequences, when working under controlled conditions and in the field. Application of this technology has the potential to significantly increase ability to bring modern analytical techniques to bear on the viral pathogens infecting crop plants.

Survey of cassava virus diseases in Tanzania

Abstract A survey was carried out of African cassava mosaic geminivirus and cassava brown streak virus diseases (ACMD and CBSD) in Tanzania. ACMD occurred throughout the country at low to moderate incidences in the fifteen different regions surveyed (1- 64%). The incidence was generally higher along the coastalplain than in higher altitude areas in the interior. There was a significant correlation between numbers ofadults ofthe whitefly vector, Bemisia tabaci, and incidence of recently infected plants ( R = 0.77, p < 0.01), although most infection was attributable to the use of infected cuttings. CBSD was more restricted in distribution and, where present, incidence was low to moderate (1- 36%). Cassava plantings along the southern border with Mozambique link the highest incidence area on the south-east coast near Mtwara, with the other important area ofCBSD incidence along the shore ofLake Malawi. The effects of climatic parameters on ACMD and CBSD incidence are discussed. The survey data suggest that bo...

Cassava Virus Diseases: Biology, Epidemiology, and Management

Cassava (Manihot esculenta Crantz.) is the most important vegetatively propagated food staple in Africa and a prominent industrial crop in Latin America and Asia. Its vegetative propagation through stem cuttings has many advantages, but deleteriously it means that pathogens are passed from one generation to the next and can easily accumulate, threatening cassava production. Cassava-growing continents are characterized by specific suites of viruses that affect cassava and pose particular threats. Of major concern, causing large and increasing economic impact in Africa and Asia are the cassava mosaic geminiviruses that cause cassava mosaic disease in Africa and Asia and cassava brown streak viruses causing cassava brown streak disease in Africa. Latin America, the center of origin and domestication of the crop, hosts a diverse set of virus species, of which the most economically important give rise to cassava frog skin disease syndrome. Here, we review current knowledge on the biology, epidemiology, and control of the most economically important groups of viruses in relation to both farming and cultural practices. Components of virus control strategies examined include: diagnostics and surveillance, prevention and control of infection using phytosanitation, and control of disease through the breeding and promotion of varieties that inhibit virus replication and/or movement. We highlight areas that need further research attention and conclude by examining the likely future global outlook for virus disease management in cassava.

Optimization of diagnostic RT-PCR protocols and sampling procedures for the reliable and cost-effective detection of Cassava brown streak virus

Sampling procedures and diagnostic protocols were optimized for accurate diagnosis of Cassava brown streak virus (CBSV) (genus Ipomovirus, family Potyviridae). A cetyl trimethyl ammonium bromide (CTAB) method was optimized for sample preparation from infected cassava plants and compared with the RNeasy plant mini kit (Qiagen) for sensitivity, reproducibility and costs. CBSV was detectable readily in total RNAs extracted using either method. The major difference between the two methods was in the cost of consumables, with the CTAB 10x cheaper (0.53 pounds sterling=US

Cassava Virus Diseases

0.80 per sample) than the RNeasy method (5.91 pounds sterling=US

A global alliance declaring war on cassava viruses in Africa

8.86 per sample). A two-step RT-PCR (1.34 pounds sterling=US