Parasitic dodder expresses an arsenal of secreted cellulases with multi-substrate specificity during host invasion.

Cuscuta campestris is a common and problematic parasitic plant which relies on haustoria to connect to and siphon nutrients from host plants. Glycoside hydrolase family 9 (GH9) cellulases (EC 3.2.1.4) play critical roles in plant cell wall biosynthesis and disassembly, but their roles during Cuscuta host invasion remains underexplored. In this study, we identified 22 full-length GH9 cellulase genes in C. campestris genome, which encoded fifteen secreted and seven membrane-anchored cellulases that showed distinct phylogenetic relationships. Expression profiles suggested that some of the genes are involved in biosynthesis and remodeling of the parasite's cell wall during haustoriogenesis, while other genes encoding secreted B- and C-type cellulases are tentatively associated with degrading host cell walls during invasion. Transcriptomic data in a host-free system and in the presence of susceptible or partially resistant tomato hosts, showed for especially GH9B7, GH9B11 and GH9B12 a shift in expression profiles in the presence of hosts, being more highly expressed during host attachment, indicating that Cuscuta can tune cellulase expression in response to a host. Functional analyses of recombinant B- and C-type cellulases showed endoglucanase activities over wide pH and temperature conditions, and activities towards multiple cellulose and hemicellulose substrates. These findings improve our understanding of host cell wall disassembly by Cuscuta, and cellulase activity towards broad substrate range potentially explain its wide host range. This is the first study to provide a broad biochemical insight into Cuscuta GH9 cellulases, which based on our study may have potential applications in industrial bioprocessing.

Genomic analysis of Sweet potato feathery mottle virus from East Africa.

Sweet potato feathery mottle virus is a potyvirus that infect sweet potato. The genome of the virus was analysed to understand genetic diversity, evolution and gene flow. Motifs, nucleotide identity and a phylogenetic tree were used to determine phylogroup of the isolates. Gene flow and genetic diversity were tested using DnaSP v.5. Codons evolution were tested using three methods embedded in Datamonkey. The results indicate occurrence of an isolate of phylogroup B within East Africa. Low genetic differentiation was observed between isolates from Kenya and Uganda indicating evidence of gene flow between the two countries. Four genes were found to have positively selected codons bordering or occurring within functional motifs. A motif within P1 gene evolved differently between phylogroup A and B. The evidence of gene flow indicates frequent exchange of the virus between the two countries and P1 gene motif provide a possible marker that can be used for mapping the distribution of the phylogroups.

Distance and Microsphere Aggregation-Based DNA Detection in a Paper-Based Microfluidic Device.

In paper-based microfluidics, the simplest devices are colorimetric, giving qualitative results. However, getting quantitative data can be quite a bit more difficult. Distance-based devices provide a user-friendly means of obtaining quantitative data without the need for any additional equipment, simply by using an included ruler or calibrated markings. This article details the development of a quantitative DNA detection device that utilizes the aggregation of polystyrene microspheres to affect the distance that microspheres wick through filter paper. The microspheres are conjugated to single-stranded DNA (ssDNA) oligomers that are partially complementary to a target strand and, in the presence of the target strand, form a three-strand complex, resulting in the formation of aggregates. The higher the concentration of the target strand, the larger the aggregate, and the shorter the distance wicked by the microspheres. This behavior was investigated across a wide range of target concentrations and under different incubation times to understand aggregate formation. The device was then used to successfully detect a target strand spiked in extracted plant DNA.

Prevalence of sweetpotato viruses in Acholi sub-region, northern Uganda.

The purpose of the study was to identify different viruses infecting sweetpotato and the level of co-infection and spatial distribution of the viruses within the Acholi sub-region of northern Uganda. Multiplex PCR was used to screen and determine level of co-infection in 380 sweetpotato plants. The PCR scores were computed to give overall frequency of occurrence of different viruses. The spatial distribution of viruses was represented on an ArcGIS map. Of all screened samples, 24% (92/380) were infected with at least one virus. Sweetpotato feathery mottle virus (65/92), sweetpotato chlorotic fleck virus (17/92) and sweetpotato mild mottle virus (8/92) were the most frequent viruses detected. Of sampled fields, 74% (28/38) had at least one virus-infected sweetpotato plant. The four viruses detected are the major viruses causing significant yield losses in major sweetpotato growing regions of Uganda and East Africa. The findings of limited distribution and low prevalence of the viruses in the region indicate it causes less burden to sweetpotato production in the sub-region compared with other parts of Uganda.

Farmers' practices and their knowledge of biotic constraints to sweetpotato production in East Africa.

Sweetpotato (Ipomoea batatas) is a vital crop for overcoming food insecurity in sub-Saharan Africa and its production is highest in East Africa where yields are high and the growing seasons are short. This cross-country study assessed farmers' local practices and their knowledge of the biotic constraints to sweetpotato production in Uganda, Rwanda, Kenya and Tanzania with the aim of providing empirical data that can ultimately be used to enhance sweetpotato production in these four countries. We collected data from 675 households using a standardized questionnaire integrated with a web-based mobile app. Survey results provided strong evidence that sweetpotato is valued as an important subsistence crop among smallholder farmers on pieces of land of less than 0.4 ha, and we observed that females were more involved than males in sweetpotato production. Sweetpotato was ranked as the second most important staple crop after cassava. Farmers noted an increase in sweetpotato production over the past five years in Uganda and Kenya but a decrease in Rwanda and Tanzania; the proportion of farmers who reported a decrease (33%) and an increase (36%) did not significantly differ. The main constraints to production were reported to be pests (32.6%), drought (21.6%), diseases (11.9%) and lack of disease-free planting materials (6.8%). Farmers recognized the signs and symptoms associated with sweetpotato diseases on leaves, root tubers, and whole plants, but most were unable to assign the disease type (bacterial, fungal or viral) correctly. We suggest that regional governments improve education, increase the provision of clean planting materials and strengthen breeding programs to improve disease resistance.

Is nodding syndrome in northern Uganda linked to consumption of mycotoxin contaminated food grains?

OBJECTIVE: Nodding syndrome (NS) is a type of epilepsy characterized by repeated head-nodding seizures that appear in previously healthy children between 3 and 18 years of age. In 2012, during a WHO International Meeting on NS in Kampala, Uganda, it was recommended that fungal contamination of foods should be investigated as a possible cause of the disease. We therefore aimed to assess whether consumption of fungal mycotoxins contributes to NS development. RESULTS: We detected similar high levels of total aflatoxin and ochratoxin in mostly millet, sorghum, maize and groundnuts in both households with and without children with NS. Furthermore, there was no significant association between concentrations of total aflatoxin, ochratoxin and doxynivalenol and the presence of children with NS in households. In conclusion, our results show no supporting evidence for the association of NS with consumption of mycotoxins in contaminated foods.

Sweetpotato seed exchange systems and knowledge on sweetpotato viral diseases among local farmers in Acholi Sub Region-Northern Uganda.

Viral infections greatly limit sweetpotato yields. Good farming practices are critical for effective disease management. 383 Ugandan farmers were surveyed to document symptom incidence, crop-management practices, and buyer preferences. Results showed that 89.27% farmers grow sweet potatoes yearly and 62.76% of these farmers were female. A total of 56.83% farmers obtained vine seeds from their previous gardens, 25.85% from neighbours, and 12.20% purchased. Only ~8% of sellers and ~4% of buyers were selected for disease-free materials. None of the farmers who used vine-cutting knives sterilised them. Almost half of farmers (47%) observed whitefly or aphids but most were unaware they are viral vectors. Most farmers (77%) observed viral symptoms, but few (<2%) recognised them as infections. Insufficient knowledge of sweetpotato viruses and their vectors is common and increases the risk of spread. Practices like vine selling, sharing of vines coupled with insufficient knowledge on sweet potato viruses and its vectors among farmers increase the risk of virus spread among different farms.