Resistance to Cassava Whitefly (Bemisia tabaci) among Eastern and Southern African Elite Cassava Genotypes.

Cassava whitefly, Bemisia tabaci, directly damages cassava leaves by feeding on phloem, causing chlorosis and abscission, leading to a yield loss of up to 50%. The pest also causes indirect damage through sooty mold formation. Most Ugandan cassava varieties resist cassava mosaic disease (CMD) and tolerate cassava brown streak disease (CBSD), but little is known about their response to whitefly infestation. The main objective of this study was to identify cassava genotypes with putative resistance to whitefly in Uganda. This was conducted on 24 improved cassava varieties in three agro-ecological zones during the second rains of 2016. Monthly data were taken for adult and nymph counts, whitefly and sooty mold damage, and CMD and CBSD severities from 2 to 9 months after planting (MAPs). The results show that the whitefly population is highly significantly (p < 0.000) amongst varieties across the three agro-ecological zones. Mkumba consistently supported the low adult numbers and nymphs. The findings demonstrate the potential of the improved cassava varieties as sources of whitefly resistance for sustainable management.

A comprehensive framework for the delimitation of species within the Bemisia tabaci cryptic complex, a global pest-species group.

Identifying cryptic species poses a substantial challenge to both biologists and naturalists due to morphological similarities. Bemisia tabaci is a cryptic species complex containing more than 44 putative species; several of which are currently among the world's most destructive crop pests. Interpreting and delimiting the evolution of this species complex has proved problematic. To develop a comprehensive framework for species delimitation and identification, we evaluated the performance of distinct data sources both individually and in combination among numerous samples of the B. tabaci species complex acquired worldwide. Distinct datasets include full mitogenomes, single-copy nuclear genes, restriction site-associated DNA sequencing, geographic range, host speciation, and reproductive compatibility datasets. Phylogenetically, our well-supported topologies generated from three dense molecular markers highlighted the evolutionary divergence of species of the B. tabaci complex and suggested that the nuclear markers serve as a more accurate representation of B. tabaci species diversity. Reproductive compatibility datasets facilitated the identification of at least 17 different cryptic species within our samples. Native geographic range information provides a complementary assessment of species recognition, while the host range datasets provide low rate of delimiting resolution. We further summarized different data performances in species classification when compared with reproductive compatibility, indicating that combination of mtCOI divergence, nuclear markers, geographic range provide a complementary assessment of species recognition. Finally, we represent a model for understanding and untangling the cryptic species complexes based on the evidence from this study and previously published articles.

In silico prediction of candidate gene targets for the management of African cassava whitefly (Bemisia tabaci, SSA1-SG1), a key vector of viruses causing cassava brown streak disease.

Whiteflies (Bemisia tabaci sensu lato) have a wide host range and are globally important agricultural pests. In Sub-Saharan Africa, they vector viruses that cause two ongoing disease epidemics: cassava brown streak disease and cassava mosaic virus disease. These two diseases threaten food security for more than 800 million people in Sub-Saharan Africa. Efforts are ongoing to identify target genes for the development of novel management options against the whitefly populations that vector these devastating viral diseases affecting cassava production in Sub-Saharan Africa. This study aimed to identify genes that mediate osmoregulation and symbiosis functions within cassava whitefly gut and bacteriocytes and evaluate their potential as key gene targets for novel whitefly control strategies. The gene expression profiles of dissected guts, bacteriocytes and whole bodies were compared by RNAseq analysis to identify genes with significantly enriched expression in the gut and bacteriocytes. Phylogenetic analyses identified three candidate osmoregulation gene targets: two α-glucosidases, SUC 1 and SUC 2 with predicted function in sugar transformations that reduce osmotic pressure in the gut; and a water-specific aquaporin (AQP1) mediating water cycling from the distal to the proximal end of the gut. Expression of the genes in the gut was enriched 23.67-, 26.54- and 22.30-fold, respectively. Genome-wide metabolic reconstruction coupled with constraint-based modeling revealed four genes (argH, lysA, BCAT & dapB) within the bacteriocytes as potential targets for the management of cassava whiteflies. These genes were selected based on their role and essentiality within the different essential amino acid biosynthesis pathways. A demonstration of candidate osmoregulation and symbiosis gene targets in other species of the Bemisia tabaci species complex that are orthologs of the empirically validated osmoregulation genes highlights the latter as promising gene targets for the control of cassava whitefly pests by in planta RNA interference.

Comparative evolutionary analyses of eight whitefly Bemisia tabaci sensu lato genomes: cryptic species, agricultural pests and plant-virus vectors.

BACKGROUND: The group of > 40 cryptic whitefly species called Bemisia tabaci sensu lato are amongst the world's worst agricultural pests and plant-virus vectors. Outbreaks of B. tabaci s.l. and the associated plant-virus diseases continue to contribute to global food insecurity and social instability, particularly in sub-Saharan Africa and Asia. Published B. tabaci s.l. genomes have limited use for studying African cassava B. tabaci SSA1 species, due to the high genetic divergences between them. Genomic annotations presented here were performed using the 'Ensembl gene annotation system', to ensure that comparative analyses and conclusions reflect biological differences, as opposed to arising from different methodologies underpinning transcript model identification. RESULTS: We present here six new B. tabaci s.l. genomes from Africa and Asia, and two re-annotated previously published genomes, to provide evolutionary insights into these globally distributed pests. Genome sizes ranged between 616-658 Mb and exhibited some of the highest coverage of transposable elements reported within Arthropoda. Many fewer total protein coding genes (PCG) were recovered compared to the previously published B. tabaci s.l. genomes and structural annotations generated via the uniform methodology strongly supported a repertoire of between 12.8-13.2 × 103 PCG. An integrative systematics approach incorporating phylogenomic analysis of nuclear and mitochondrial markers supported a monophyletic Aleyrodidae and the basal positioning of B. tabaci Uganda-1 to the sub-Saharan group of species. Reciprocal cross-mating data and the co-cladogenesis pattern of the primary obligate endosymbiont 'Candidatus Portiera aleyrodidarum' from 11 Bemisia genomes further supported the phylogenetic reconstruction to show that African cassava B. tabaci populations consist of just three biological species. We include comparative analyses of gene families related to detoxification, sugar metabolism, vector competency and evaluate the presence and function of horizontally transferred genes, essential for understanding the evolution and unique biology of constituent B. tabaci. s.l species. CONCLUSIONS: These genomic resources have provided new and critical insights into the genetics underlying B. tabaci s.l. biology. They also provide a rich foundation for post-genomic research, including the selection of candidate gene-targets for innovative whitefly and virus-control strategies.

Genetic diversity, distribution, and structure of Bemisia tabaci whitefly species in potential invasion and hybridization regions of East Africa.

Outbreaks of whitefly, Bemisia tabaci species in East and Central Africa, have become increasingly prevalent during the previous 25 years and are responsible for driving the spread of plant-virus diseases, such as cassava mosaic disease and cassava brown steak disease. Epidemics of these diseases have expanded their ranges over the same period, spreading from Uganda into other sub-Saharan African countries. It was hypothesised that a highly abundant 'invader' population of B. tabaci was responsible for spreading these diseases from Uganda to neighbouring countries and potentially hybridising with the resident cassava B. tabaci populations. Here, we test this hypothesis by investigating the molecular identities of the highly abundant cassava B. tabaci populations from their supposed origin in Uganda, to the northern, central, eastern and coastal regions of Tanzania. Partial mitochondrial cytochrome oxidase I (mtCOI) barcoding sequences and nuclear microsatellite markers were used to analyse the population genetic diversity and structure of 2734 B. tabaci collected from both countries and in different agroecological zones. The results revealed that: (i) the putative SSA1 species is structured according to countries, so differ between them. (ii) Restricted gene flow occurred between SSA1-SG3 and both other SSA1 subgroups (SG1 and SG2), even in sympatry, demonstrating strong barriers to hybridization between those genotypes. (iii) Not only B. tabaci SSA1-(SG1 and SG2) was found in highly abundant (outbreak) numbers, but B. tabaci SSA1-SG3 and the Indian Ocean (IO) species were also recorded in high numbers in several sites in Tanzania. (iv) The SSA1-(SG1 and SG2) species was distributed in both countries, but in Tanzania, the B. tabaci IO and SSA1-SG3 species predominated. These data confirm that multiple, local Tanzanian B. tabaci species produce highly abundant populations, independent of the spread of the putative invasive B. tabaci SSA1-(SG1 and SG2) populations.

Spatio-temporal changes in endosymbiont diversity and composition in the African cassava whitefly, Bemisia tabaci SSA1.

Sap-sucking insects, including whiteflies, are amongst the most devastating and widely distributed organisms on the planet. They are often highly invasive and endosymbiont communities within these insects help them adapt to new or changing environments. Bemisia tabaci (Gennadius; Hemiptera: Aleyrodidae) whitefly species are vectors of more than 500 known plant-viruses and harbour highly diverse endosymbionts communities. To date, however, whitefly-endosymbiont interactions, community structure and their spatio-temporal changes are still poorly understood. In this study, we investigated the spatio-temporal changes in the composition and diversity of bacterial endosymbionts in the agricultural crop pest whitefly species, Bemisia tabaci sub-Saharan Africa 1-subgroup 1 and 2 (SSA1-SG1 and SSA1-SG2). 16S rRNA amplicon sequencing analysis was carried out to characterise endosymbiont compositionsin field-collected SSA1 (SSA1-SG1 and SSA1-SG2) populations infesting cassava in Uganda in 1997 and 2017. We detected Portiera, Arsenophonus, Wolbachia, Hamiltonella and Hemipteriphilus, with Arsenophonus and Wolbachia infections being predominant. Hemipteriphilus and Hamiltonella frequencies were very low and were detected in seven and two samples, respectively. Bacterial diversity based on three independent parameters including Simpson index, number of haplotypes and Bray-Curtis dissimilarity matrix was significantly higher in 1997 than in 2017. This period also coincided with the advent of super-abundant cassava-whitefly populations on cassava crops in Uganda. We discuss how endosymbionts may influence the biology and behaviour of whiteflies leading to population explosions.

Bean Leaf Beetle (Ootheca spp.) (Coleoptera: Chrysomelidae) Management via Planting Timing and Insecticides.

Bean leaf beetles (Ootheca spp.) (Coleoptera: Chrysomelidae) are one of Africa's most important pests of the common bean (Phaseolus vulgaris L.). Roots, leaves, floral parts, and young pods are all attacked, leading to a considerable loss in grain yield. In Uganda, there are no comprehensive prescribed management strategies for bean leaf beetles, but farmers typically try to control the pest by delaying bean crop sowing, and to a lesser extent, using insecticides. Although farmers have consistently implemented the two approaches, there is no information on the effects of the approaches in Uganda. To assess the impact of planting timing and insecticide spray regimes on bean leaf beetle populations, concomitant foliar damage, and grain yield, we set up trials in three agro-ecological zones with known presence of the beetles during the second rainy season of 2016 (2016) and the first rainy season of 2017 (2017). The first planting, coinciding with early planting, was conducted within one week after the onset of rains. The second planting, coinciding with mid planting, followed two weeks later, while the third planting, considered late planting in this study, was conducted one month after the second planting. A foliar application of cypermethrin commencing at 7 days after emergence (DAE), 14 DAE, 21 DAE, 28 DAE, and 35 DAE; a soil drench of imidacloprid at planting combined with a foliar spray starting at 7 DAE; and an untreated control were among the insecticide spray regimes evaluated. Higher bean leaf beetle abundance was recorded from mid-planting, while higher foliar damage was recorded from late planting in two of the three agro-ecological zones. However, higher marketable grain yield was recorded from early planting in all agro-ecological zones, suggesting that delayed planting may not be beneficial. Insecticide application reduced foliar damage and increased marketable grain yield, with a combination of soil drench and foliar spray resulting in much less foliar damage and, as a result, higher grain yield. However, this did not result in economic benefits. Furthermore, marketable grain yield was higher when insecticide spray regimes were combined with early planting in all agro-ecological zones during both seasons. Our findings suggest that the common bean should be planted early and that the control of the bean leaf beetle should target both the adults and the juvenile stages in the soil. Therefore, there is a need for farmers to be able to access less-expensive soil treatments.

A high-throughput amplicon sequencing approach for population-wide species diversity and composition survey.

Management of agricultural pests requires an understanding of pest species diversity, their interactions with beneficial insects and spatial-temporal patterns of pest abundance. Invasive and agriculturally important insect pests can build up very high populations, especially in cropping landscapes. Traditionally, sampling effort for species identification involves small sample sizes and is labour intensive. Here, we describe a multiprimer high throughput sequencing (HTS) metabarcoding method and associated analytical workflow for a rapid, intensive, high-volume survey of pest species compositions. We demonstrate our method using the taxonomically challenging Bemisia pest cryptic species complex as examples. The whiteflies Bemisia including the"tabaci" species are agriculturally important capable of vectoring diverse plant viruses that cause diseases and crop losses. Our multiprimer metabarcoding HTS amplicon approach simultaneously process high volumes of whitefly individuals, with efficiency to detect rare (i.e., 1%) test-species, while our improved whitefly primers for metabarcoding also detected beneficial hymenopteran parasitoid species from whitefly nymphs. Field-testing our redesigned Bemisia metabarcoding primer sets across the Tanzania, Uganda and Malawi cassava cultivation landscapes, we identified the sub-Saharan Africa 1 Bemisia putative species as the dominant pest species, with other cryptic Bemisia species being detected at various abundances. We also provide evidence that Bemisia species compositions can be affected by host crops and sampling techniques that target either nymphs or adults. Our multiprimer HTS metabarcoding method incorporated two overlapping amplicons of 472 bp and 518 bp that spanned the entire 657 bp 3' barcoding region for Bemisia, and is particularly suitable to molecular diagnostic surveys of this highly cryptic insect pest species complex that also typically exhibited high population densities in heavy crop infestation episodes. Our approach can be adopted to understand species biodiversity across landscapes, with broad implications for improving transboundary biosecurity preparedness, thus contributing to molecular ecological knowledge and the development of control strategies for high-density, cryptic, pest-species complexes.

Distribution and Relative Abundance of Bean Leaf Beetles (Ootheca spp.) (Insecta: Coleoptera: Chrysomelidae) in Uganda.

Bean leaf beetles (Ootheca spp.) (Insecta: Coleoptera: Chrysomelidae) are one of Africa's most destructive pests of common bean and other leguminous crops. The beetles are widely distributed in Africa where they are estimated to cause annual crop yield losses of 116,400 tons of crop yields in sub-Saharan Africa. Despite their importance, little is known about the distribution, relative abundance and damage caused by bean leaf beetles in Uganda. As a result, the development of effective management methods has been hampered. We conducted surveys in six key Ugandan agro-ecological zones to determine the species distribution and relative abundance of bean leaf beetles. Findings indicate that leaf beetles belonging to 12 genera are present, including members of the genera Afrophthalma Medvedev, 1980, Buphonella Jacoby, 1903, Chrysochrus Chevrolat in Dejean, 1836, Diacantha Dejean, 1845, Exosoma Jacoby, 1903, Lamprocopa Hincks, 1949, Lema Fabricius, 1798, Nisotra Baly, 1864, Neobarombiella Bolz and Wagner, 2012, Ootheca Dejean, 1935, Parasbecesta Laboissière, 1940, and Plagiodera Dejean, 1835. We identified only three species belonging to the genus Ootheca: O. mutabilis, O. proteus, and O. orientalis. Seventy percent of all the beetles collected were O. mutabilis and these were present in all agro-ecological zones studied. The Northern Moist Farmlands (21.9%), West Nile Farmlands (12.9%), Central Wooded Savanna (4.4%) and Southern and Eastern Lake Kyoga Basin (1.4%) were the only agro-ecological zones where O. proteus was found. Only one specimen of O. orientalis was found at a single site in the Central Wooded Savanna. The Northern Moist Farmlands had a significantly (p < 0.05) higher bean leaf beetle density than the West Nile Farmlands and Southwestern Highlands. Similarly, the Northern Moist Farmlands had the highest beetle foliar damage per plant (1.15 ± 0.05), while the Southwestern Highlands had the lowest (0.03 ± 0.02). We provide the first information on Ootheca species distribution, abundance and damage in Uganda. Our findings provide a foundation for assessing the importance of Ootheca spp. as common bean pests in Uganda.

Genetic diversity of whitefly (Bemisia spp.) on crop and uncultivated plants in Uganda: implications for the control of this devastating pest species complex in Africa.

Over the past three decades, highly increased whitefly (Bemisia tabaci) populations have been observed on the staple food crop cassava in eastern Africa and associated with ensuing viral disease pandemics and food insecurity. Increased whitefly numbers have also been observed in other key agricultural crops and weeds. Factors behind the population surges on different crops and their interrelationships are unclear, although in cassava they have been associated with specific populations within the Bemisia tabaci species complex known to infest cassava crops in Africa. This study carried out an in-depth survey to understand the distribution of B. tabaci populations infesting crops and uncultivated plant hosts in Uganda, a centre of origin for this pest complex. Whitefly samples were collected from 59 identified plant species and 25 unidentified weeds in a countrywide survey. Identities of 870 individual adult whiteflies were determined through mitochondrial cytochrome oxidase 1 sequences (651 bp) in the 3' barcode region used for B. tabaci systematics. Sixteen B. tabaci and five related whitefly putative species were identified based on > 4.0% nucleotide divergence, of which three are proposed as novel B. tabaci putative species and four as novel closely related whitefly species. The most prevalent whiteflies were classified as B. tabaci MED-ASL (30.5% of samples), sub-Saharan Africa 1 (SSA1, 22.7%) and Bemisia Uganda1 (12.1%). These species were also indicated to be the most polyphagous occurring on 33, 40 and 25 identified plant species, respectively. Multiple (≥ 3) whitefly species occurred on specific crops (bean, eggplant, pumpkin and tomato) and weeds (Sida acuta and Ocimum gratissimum). These plants may have increased potential to act as reservoirs for mixed infections of whitefly-vectored viruses. Management of whitefly pest populations in eastern Africa will require an integration of approaches that consider their degree of polyphagy and a climate that enables the continuous presence of crop and uncultivated plant hosts. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10340-021-01355-6.

Influence of Cassava Morphological Traits and Environmental Conditions on Field Populations of Bemisia tabaci.

High populations of species in the whitefly complex Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) were reported to cause severe damage to cassava in East and Central Africa. However, reasons for B. tabaci population increases are not well understood. We investigated the effect of cassava morphological traits, temperature, rainfall and relative humidity (RH) on the abundance of B. tabaci. Five cassava genotypes with varying levels of resistance to cassava mosaic disease, cassava brown streak disease, and B. tabaci infestation were planted in three Ugandan agro-ecological zones. The experiment was conducted in 2016 and 2017 in a randomized complete block design. Across all locations, the tallest genotype Alado alado supported the lowest number of B. tabaci adults. In areas with high B. tabaci prevalence, leaf area, leaf lobe width, and leaf lobe number exhibited significant positive effects (p < 0.001) on B. tabaci adult count. Positive effects of relative humidity and negative effects of temperature and rainfall on B. tabaci adult and nymph counts were observed in 2016 and 2017, resulting in low populations in Lira. Evidently, temperatures of 28-30 °C, rainfall of 30-150 mm and RH of 55-70%, and deployment of cassava genotypes of low plant height, large leaf area, and lobe width significantly enhanced B. tabaci population growth.

Activation and detoxification of cassava cyanogenic glucosides by the whitefly Bemisia tabaci.

Two-component plant defenses such as cyanogenic glucosides are produced by many plant species, but phloem-feeding herbivores have long been thought not to activate these defenses due to their mode of feeding, which causes only minimal tissue damage. Here, however, we report that cyanogenic glycoside defenses from cassava (Manihot esculenta), a major staple crop in Africa, are activated during feeding by a pest insect, the whitefly Bemisia tabaci, and the resulting hydrogen cyanide is detoxified by conversion to beta-cyanoalanine. Additionally, B. tabaci was found to utilize two metabolic mechanisms to detoxify cyanogenic glucosides by conversion to non-activatable derivatives. First, the cyanogenic glycoside linamarin was glucosylated 1-4 times in succession in a reaction catalyzed by two B. tabaci glycoside hydrolase family 13 enzymes in vitro utilizing sucrose as a co-substrate. Second, both linamarin and the glucosylated linamarin derivatives were phosphorylated. Both phosphorylation and glucosidation of linamarin render this plant pro-toxin inert to the activating plant enzyme linamarase, and thus these metabolic transformations can be considered pre-emptive detoxification strategies to avoid cyanogenesis.

Minimal clinically important difference in walking velocity, gait profile score and two minute walk test for individuals with lower limb amputation.

BACKGROUND: Individuals with lower limb amputation are routinely assessed with a variety outcome measures, however there is a lack of published data to indicate minimal clinically important differences (MCID) for many of these outcome measures. Three such important gait-specific outcome measures include walking velocity, gait profile score (GPS) and the two minute walk test (2MWT). RESEARCH QUESTION: Determine the MCIDs for walking velocity, GPS and 2MWT for individuals with lower limb amputation. METHODS: Walking velocity and GPS (n = 60), and 2MWT (n = 119) data for individuals with unilateral transfemoral or knee disarticulation were identified retrospectively from a database held at the study centre. An anchor-based method was used with Medicare functional classification level (MFCL) acting as the impairment-related criterion, and a least-squares linear regression approach was used to calculate the gradient required for a change between MFCL levels. RESULTS: An increase of 0.21 m/s (95 % CI: 0.13,0.29) for walking velocity, a reduction of 1.7° (95 % CI: -2.449,-1.097) for GPS and an increase of 37.2 m (95 % CI: 28.8,45.5) for 2MWT were found to correspond to an increase in MFCL of one level. Walking velocity, GPS and 2MWT correlated with MFCL with R2 values of 0.333, 0.322 and 0.398 respectively (p < 0.00001). The authors propose that 0.21 m/s for walking velocity, 1.7° for GPS and 37.2 m for 2MWT be used as MCID values for individuals with lower limb amputation. SIGNIFICANCE: The results of this study can be used to help both researchers and clinicians to objectively evaluate if interventions for individuals with lower limb amputation are effective.

Evaluation of gait outcomes for individuals with established unilateral transfemoral amputation following the provision of microprocessor controlled knees in the context of a clinical service.

BACKGROUND: Microprocessor controlled knees (MPKs) have been shown to improve gait outcomes for individuals with unilateral transfemoral amputation (TFA); however, previous studies have limitations, which may affect their applicability. OBJECTIVES: To compare gait outcomes between non-MPK and MPK conditions within a large and diverse cohort of established TFAs in the pragmatic context of an operational clinical service and to identify any demographic variables that correlate with changes in gait outcomes. STUDY DESIGN: This is a retrospective cohort study. METHODS: Full-body three-dimensional motion capture and a portable breath gas analyser were used to evaluate gait-specific outcomes both pre-MPK and 6 months post-MPK provisions in TFAs (n = 32). The primary outcome measure was gait profile score, along with the following secondary outcome measures: walking velocity, step length, step length symmetry ratio, step time symmetry ratio, vertical ground reaction force symmetry index, base of support, center of mass deviation, and oxygen cost of walking. RESULTS: Data showed statistically significant improvement in gait profile score from 11.2° ± 2.2° to 10.1° ± 2.1° (p < 0.01) between non-MPK and MPK conditions. Other outcomes which showed significant improvement were walking velocity (p < 0.01), step length (p < 0.01), vertical ground reaction force symmetry index (p < 0.01), and center of mass deviation (p < 0.05). CONCLUSIONS: MPK provision offered significant gait improvements to this cohort of TFAs in an operational clinical setting. There still remains a large gap in gait outcomes between this cohort and those of healthy unimpaired adults. Improvements in socket design, prosthetic foot design, and physiotherapy intervention could further reduce this gap.

Within-Season Changes in Land-Use Impact Pest Abundance in Smallholder African Cassava Production Systems.

Cassava (Manihot esculenta Crantz), an important commercial and food security crop in East and Central Africa, continues to be adversely affected by the whitefly Bemisia tabaci. In Uganda, changes in smallholder farming landscapes due to crop rotations can impact pest populations but how these changes affect pest outbreak risk is unknown. We investigated how seasonal changes in land-use have affected B. tabaci population dynamics and its parasitoids. We used a large-scale field experiment to standardize the focal field in terms of cassava age and cultivar, then measured how Bemisia populations responded to surrounding land-use change. Bemisia tabaci Sub-Saharan Africa 1 (SSA1) was identified using molecular diagnostics as the most prevalent species and the same species was also found on surrounding soybean, groundnut, and sesame crops. We found that an increase in the area of cassava in the 3-7-month age range in the landscape resulted in an increase in the abundance of the B. tabaci SSA1 on cassava. There was a negative relationship between the extent of non-crop vegetation in the landscape and parasitism of nymphs suggesting that these parasitoids do not rely on resources in the non-crop patches. The highest abundance of B. tabaci SSA1 nymphs in cassava fields occurred at times when landscapes had large areas of weeds, low to moderate areas of maize, and low areas of banana. Our results can guide the development of land-use strategies that smallholder farmers can employ to manage these pests.

Is High Whitefly Abundance on Cassava in Sub-Saharan Africa Driven by Biological Traits of a Specific, Cryptic Bemisia tabaci Species?

In East Africa, the prevalent Bemisia tabaci whiteflies on the food security crop cassava are classified as sub-Saharan Africa (SSA) species. Economically damaging cassava whitefly populations were associated with the SSA2 species in the 1990s, but more recently, it has been to SSA1 species. To investigate whether biological traits (number of first instar nymphs, emerged adults, proportion of females in progeny and development time) of the cassava whitefly species are significant drivers of the observed field abundance, our study determined the development of SSA1 sub-group (SG) 1 (5 populations), SG2 (5 populations), SG3 (1 population) and SSA2 (1 population) on cassava and eggplant under laboratory conditions. SSA1-(SG1-SG2) and SSA2 populations' development traits were similar. Regardless of the host plant, SSA1-SG2 populations had the highest number of first instar nymphs (60.6 ± 3.4) and emerged adults (50.9 ± 3.6), followed by SSA1-SG1 (55.5 ± 3.2 and 44.6 ± 3.3), SSA2 (45.8 ± 5.7 and 32.6 ± 5.1) and the lowest were SSA1-SG3 (34.2 ± 6.1 and 32.0 ± 7.1) populations. SSA1-SG3 population had the shortest egg-adult emergence development time (26.7 days), followed by SSA1-SG1 (29.1 days), SSA1-SG2 (29.6 days) and SSA2 (32.2 days). Regardless of the whitefly population, development time was significantly shorter on eggplant (25.1 ± 0.9 days) than cassava (34.6 ± 1.0 days). These results support that SSA1-(SG1-SG2) and SSA2 B. tabaci can become highly abundant on cassava, with their species classification alone not correlating with observed abundance and prevalence.

The molecular mechanisms that determine different degrees of polyphagy in the Bemisia tabaci species complex.

The whitefly Bemisia tabaci is a closely related group of >35 cryptic species that feed on the phloem sap of a broad range of host plants. Species in the complex differ in their host-range breadth, but the mechanisms involved remain poorly understood. We investigated, therefore, how six different B. tabaci species cope with the environmental unpredictability presented by a set of four common and novel host plants. Behavioral studies indicated large differences in performances on the four hosts and putative specialization of one of the species to cassava plants. Transcriptomic analyses revealed two main insights. First, a large set of genes involved in metabolism (>85%) showed differences in expression between the six species, and each species could be characterized by its own unique expression pattern of metabolic genes. However, within species, these genes were constitutively expressed, with a low level of environmental responsiveness (i.e., to host change). Second, within each species, sets of genes mainly associated with the super-pathways "environmental information processing" and "organismal systems" responded to the host switching events. These included genes encoding for proteins involved in sugar homeostasis, signal transduction, membrane transport, and immune, endocrine, sensory and digestive responses. Our findings suggested that the six B. tabaci species can be divided into four performance/transcriptomic "Types" and that polyphagy can be achieved in multiple ways. However, polyphagy level is determined by the specific identity of the metabolic genes/pathways that are enriched and overexpressed in each species (the species' individual metabolic "tool kit").

Microbiome diversity and reproductive incompatibility induced by the prevalent endosymbiont Arsenophonus in two species of African cassava Bemisia tabaci whiteflies.

A minimum of 13 diverse whitefly species belonging to the Bemisia tabaci (B. tabaci) species complex are known to infest cassava crops in sub-Saharan Africa (SSA), designated as SSA1-13. Of these, the SSA1 and SSA2 are the predominant species colonizing cassava crops in East Africa. The SSA species of B. tabaci harbor diverse bacterial endosymbionts, many of which are known to manipulate insect reproduction. One such symbiont, Arsenophonus, is known to drive its spread by inducing reproductive incompatibility in its insect host and are abundant in SSA species of B. tabaci. However, whether Arsenophonus affects the reproduction of SSA species is unknown. In this study, we investigated both the reproductive compatibility between Arsenophonus infected and uninfected whiteflies by inter-/intraspecific crossing experiments involving the sub-group three haplotypes of the SSA1 (SSA1-SG3), SSA2 species, and their microbial diversity. The number of eggs, nymphs, progenies produced, hatching rate, and survival rate were recorded for each cross. In intra-specific crossing trials, both male and female progenies were produced and thus demonstrated no reproductive incompatibility. However, the total number of eggs laid, nymphs hatched, and the emerged females were low in the intra-species crosses of SSA1-SG3A+, indicating the negative effect of Arsenophonus on whitefly fitness. In contrast, the inter-species crosses between the SSA1-SG3 and SSA2 produced no female progeny and thus demonstrated reproductive incompatibility. The relative frequency of other bacteria colonizing the whiteflies was also investigated using Illumina sequencing of 16S rDNA and diversity indices were recorded. Overall, SSA1-SG3 and SSA2 harbored high microbial diversity with more than 137 bacteria discovered. These results described for the first time the microbiome diversity and the reproductive behaviors of intra-/inter-species of Arsenophonus in whitefly reproduction, which is crucial for understanding the invasion abilities of cassava whiteflies.

Whole-genome single nucleotide polymorphism and mating compatibility studies reveal the presence of distinct species in sub-Saharan Africa Bemisia tabaci whiteflies.

In sub-Saharan Africa cassava growing areas, two members of the Bemisia tabaci species complex termed sub-Saharan Africa 1 (SSA1) and SSA2 have been reported as the prevalent whiteflies associated with the spread of viruses that cause cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) pandemics. At the peak of CMD pandemic in the late 1990s, SSA2 was the prevalent whitefly, although its numbers have diminished over the last two decades with the resurgence of SSA1 whiteflies. Three SSA1 subgroups (SG1 to SG3) are the predominant whiteflies in East Africa and vary in distribution and biological properties. Mating compatibility between SSA1 subgroups and SSA2 whiteflies was reported as the possible driver for the resurgence of SSA1 whiteflies. In this study, a combination of both phylogenomic methods and reciprocal crossing experiments were applied to determine species status of SSA1 subgroups and SSA2 whitefly populations. Phylogenomic analyses conducted with 26 548 205 bp whole genome single nucleotide polymorphisms (SNPs) and the full mitogenomes clustered SSA1 subgroups together and separate from SSA2 species. Mating incompatibility between SSA1 subgroups and SSA2 further demonstrated their distinctiveness from each other. Phylogenomic analyses conducted with SNPs and mitogenomes also revealed different genetic relationships among SSA1 subgroups. The former clustered SSA1-SG1 and SSA1-SG2 together but separate from SSA1-SG3, while the latter clustered SSA1-SG2 and SSA1-SG3 together but separate from SSA1-SG1. Mating compatibility was observed between SSA1-SG1 and SSA1-SG2, while incompatibility occurred between SSA1-SG1 and SSA1-SG3, and SSA1-SG2 and SSA1-SG3. Mating results among SSA1 subgroups were coherent with phylogenomics results based on SNPs but not the full mitogenomes. Furthermore, this study revealed that the secondary endosymbiont-Wolbachia-did not mediate reproductive success in the crossing assays carried out. Overall, using genome wide SNPs together with reciprocal crossings assays, this study established accurate genetic relationships among cassava-colonizing populations, illustrating that SSA1 and SSA2 are distinct species while at least two species occur within SSA1 species.

Improving climate suitability for Bemisia tabaci in East Africa is correlated with increased prevalence of whiteflies and cassava diseases.

Projected climate changes are thought to promote emerging infectious diseases, though to date, evidence linking climate changes and such diseases in plants has not been available. Cassava is perhaps the most important crop in Africa for smallholder farmers. Since the late 1990's there have been reports from East and Central Africa of pandemics of begomoviruses in cassava linked to high abundances of whitefly species within the Bemisia tabaci complex. We used CLIMEX, a process-oriented climatic niche model, to explore if this pandemic was linked to recent historical climatic changes. The climatic niche model was corroborated with independent observed field abundance of B. tabaci in Uganda over a 13-year time-series, and with the probability of occurrence of B. tabaci over 2 years across the African study area. Throughout a 39-year climate time-series spanning the period during which the pandemics emerged, the modelled climatic conditions for B. tabaci improved significantly in the areas where the pandemics had been reported and were constant or decreased elsewhere. This is the first reported case where observed historical climate changes have been attributed to the increase in abundance of an insect pest, contributing to a crop disease pandemic.