Browsing by Author "Tairo, Fred"
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Item African ancestry of New World, Bemisia tabaci-whitefly species(Scientific reports, 2018) Mugerwa, Habibu; Seal, Susan; Wang, Hua-Ling; Patel, Mitulkumar V.; Kabaalu, Richard; Omongo, Christopher A.; Alicai, Titus; Tairo, Fred; Ndunguru, Joseph; Sseruwagi, Peter; Colvin, JohnBemisia tabaci whitefly species are some of the world’s most devastating agricultural pests and plant-virus disease vectors. Elucidation of the phylogenetic relationships in the group is the basis for understanding their evolution, biogeography, gene-functions and development of novel control technologies. We report here the discovery of five new Sub-Saharan Africa (SSA) B. tabaci putative species, using the partial mitochondrial cytochrome oxidase 1 gene: SSA9, SSA10, SSA11, SSA12 and SSA13. Two of them, SSA10 and SSA11 clustered with the New World species and shared 84.8‒86.5% sequence identities. SSA10 and SSA11 provide new evidence for a close evolutionary link between the Old and New World species. Re-analysis of the evolutionary history of B. tabaci species group indicates that the new African species (SSA10 and SSA11) diverged from the New World clade c. 25 million years ago. The new putative species enable us to: (i) re-evaluate current models of B. tabaci evolution, (ii) recognise increased diversity within this cryptic species group and (iii) re-estimate divergence dates in evolutionary time.Item Cassava brown streak virus has a rapidly evolving genome: implications for virus speciation, variability, diagnosis and host resistance(Scientific reports, 2016) Alicai, Titus; Ndunguru, Joseph; Sseruwagi, Peter; Tairo, Fred; Okao-Okuja, Geoffrey; Nanvubya, Resty; Kiiza, Lilliane; Kubatko, Laura; Kehoe, Monica A.; Boykin, Laura M.Cassava is a major staple food for about 800 million people in the tropics and sub-tropical regions of the world. Production of cassava is significantly hampered by cassava brown streak disease (CBSD), caused by Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). The disease is suppressing cassava yields in eastern Africa at an alarming rate. Previous studies have documented that CBSV is more devastating than UCBSV because it more readily infects both susceptible and tolerant cassava cultivars, resulting in greater yield losses. Using whole genome sequences from NGS data, we produced the first coalescent-based species tree estimate for CBSV and UCBSV. This species framework led to the finding that CBSV has a faster rate of evolution when compared with UCBSV. Furthermore, we have discovered that in CBSV, nonsynonymous substitutions are more predominant than synonymous substitution and occur across the entire genome. All comparative analyses between CBSV and UCBSV presented here suggest that CBSV may be outsmarting the cassava immune system, thus making it more devastating and harder to control.Item Expansion of the cassava brown streak pandemic in Uganda revealed by annual field survey data for 2004 to 2017(Scientific Data, 2019) Alicai, Titus; Szyniszewska, Anna M.; Omongo, Christopher A .; Abidrabo, Phillip; Okao-Okuja, Geoffrey; Baguma, Yona; Ogwok, Emmanuel; Kawuki, Robert; Esuma, Williams; Tairo, Fred; Bua, Anton; Legg, James P.; Stutt, Richard O. J. H.; Godding, David; Sseruwagi, Peter; Ndunguru, Joseph; Gilligan, Christopher A.Cassava brown streak disease (CBSD) is currently the most devastating cassava disease in eastern, central and southern Africa affecting a staple crop for over 700 million people on the continent. A major outbreak of CBSD in 2004 near Kampala rapidly spread across Uganda. In the following years, similar CBSD outbreaks were noted in countries across eastern and central Africa, and now the disease poses a threat to West Africa including Nigeria - the biggest cassava producer in the world. A comprehensive dataset with 7,627 locations, annually and consistently sampled between 2004 and 2017 was collated from historic paper and electronic records stored in Uganda. The survey comprises multiple variables including data for incidence and symptom severity of CBSD and abundance of the whitefly vector (Bemisia tabaci). This dataset provides a unique basis to characterize the epidemiology and dynamics of CBSD spread in order to inform disease surveillance and management. We also describe methods used to integrate and verify extensive field records for surveys typical of emerging epidemics in subsistence crops.Item Real time portable genome sequencing for global food security(F1000 Research, 2018) Boykin, Laura; Ghalab, Ammar; Rossitto De Marchi, Bruno; Savill, Anders; Wainaina, James M.; Kinene, Tonny; Lamb, Stephen; Rodrigues, Myriam; Kehoe, Monica; Ndunguru, Joseph; Tairo, Fred; Sseruwagi, Peter; Kayuki, Charles; Mark, Deogratius; Erasto, Joel; Bachwenkizi, Hilda; Alicai, Titus; Okao-Okuja, Geoffrey; Abridrabo, Phillip; Ogwok, Emmanuel; Osingada, John Francis; Akono, Jimmy; Ateka, Elijah; Muga, Brenda; Kiarie, SamuelCrop losses due to viral diseases and pests are major constraints on food security and income for millions of households in sub-Saharan Africa (SSA). Such losses can be reduced if plant diseases and pests are correctly diagnosed and identified early. Currently, accurate diagnosis for definitive identification of plant viruses and their vectors in SSA mostly relies on standard PCR and next generation sequencing technologies (NGS). However, it can take up to 6 months before results generated using these approaches are available. The long time taken to detect or identify viruses impedes quick, within-season decision-making necessary for early action, crop protection advice and disease control measures by farmers. This ultimately compounds the magnitude of crop losses and food shortages suffered by farmers. The MinION portable pocket DNA sequencer was used, to our knowledge globally for the first time, to sequence whole plant virus genomes. We used this technology to identify the begomoviruses causing the devastating cassava mosaic virus, which is ravaging smallholder farmers’ crops in sub-Saharan Africa.Item Resistance of advanced cassava breeding clones to infection by major viruses in Uganda(Crop Protection, 2019) Mukiibi, Daniel Rogers; Alicai, Titus; Kawuki, Robert; Okao-Okuja, Geoffrey; Tairo, Fred; Sseruwagi, Peter; Ndunguru, Joseph; Ateka, Elijah MiindaCassava brown streak disease (CBSD) and cassava mosaic disease (CMD) are two viral diseases that cause severe yield losses in cassava of up to 100%, thereby persistently threatening food and income security in sub-Saharan Africa. For effective management of these diseases, there is a critical need to develop and deploy varieties with dual resistance to CBSD and CMD. In this study, we determined the response of advanced breeding lines to field infection by cassava brown streak viruses (CBSVs) and cassava mosaic begomoviruses (CMBs). This aim helped in identifying superior clones for downstream breeding. In total, 220 cassava clones, three in uniform yield trials (UYTs) and 217 in a crossing block trial (CBT), were evaluated for virus and disease resistance. Field data were collected on disease incidence and severity. To detect and quantify CBSVs, 448 and 128 leaf samples from CBSD symptomatic and symptomless plants were analyzed by reverse transcription PCR and real-time quantitative PCR, respectively. In addition, 93 leaf samples from CMD symptomatic plants in the CBT were analyzed by conventional PCR using CMB species-specific primers. In the CBT, 124 (57%) cassava clones did not express CMD symptoms. Of the affected plants, 44 (55%) had single African cassava mosaic virus infection. Single Cassava brown streak virus (CBSV) infections were more prevalent (81.6%) in CBT clones than single Ugandan cassava brown streak virus (UCBSV) infection (3.2%). Of the three advanced clones in the UYT, NAROCASS 1 and NAROCASS 2 had significantly lower (P < 0.05) CBSD severity, incidence, and CBSV load than MH04/0300. In the UYT, only 22% of samples tested had CBSVs, and all showed a negative result for CMBs. The low disease incidence, severity, and viral load associated with NAROCASS 1 and NAROCASS 2 is evidence of their tolerance to both CBSD and CMD. Therefore, these two cassava clones should be utilized in CBSD and CMD management in Uganda, including their utilization as progenitors in further virus resistance breeding.Item Review and guide to a future naming system of African Bemisia tabaci species(Systematic Entomology, 2018) Boykin, Laura M .; Kinene, Tonny; Wainaina, James M.; Derssavill, A. N.; Seal, Susan; Mugerwa, Habibu; Macfadyen, Sarina; Tek Tay, Wee; De Barro, Paul; Kubatko, Laura; Alicai, Titus; Omongo, Christopher A .; Tairo, Fred; Ndunguru, Joseph; Sseruwagi, PeterOnce a pest has been correctly identified, its genus and species name can provide a link to valuable indications of its ecology, biology and life history that are critical for developing control strategies. Importantly, this link should exist even when the pest was known under other names (synonyms), or was not considered a pest at all (National Research Council, 1968). Many examples have shown that incorrect identification or classification of a pest has led to fruitless searches for biocontrol agents in the native range, incorrect assignments as disease vectors, and costly, yet misdirected, suppression measures. As new approaches for delimiting species based on molecular information become more widely used, the process of correctly identifying a species has become even more complex. Fortunately, we have good systematic frameworks and nomenclatural systems that are able to cope with these challenges. Here we review challenges associated with classification and identificationwithin the Bemisia tabaci (Gennadius) species complex.Item Tree Lab: Portable Genomics for Early Detection of Plant Viruses and Pests in Sub-Saharan Africa(Genes, 2019) Boykin, Laura M.; Sseruwagi, Peter; Alicai, Titus; Ateka, Elijah; Umar Mohammed, Ibrahim; Stanton, Jo-Ann L.; Kayuki, Charles; Mark, Deogratius; Fute, Tarcisius; Erasto, Joel; Bachwenkizi, Hilda; Muga, Brenda; Mumo, Naomi; Mwangi, Jenniffer; Abidrabo, Phillip; Okao-Okuja, Geofrey; Omuut, Geresemu; Akol, Jacinta; Apio, Hellen B.; Osingada, Francis; Kehoe, Monica A.; Eccles, David; Savill, Anders; Lamb, Stephen; Kinene, Tonny; Rawle, Christopher B.; Muralidhar, Abishek; Mayall, Kirsty; Tairo, Fred; Ndunguru, JosephIn this case study we successfully teamed the PDQeX DNA purification technology developed by MicroGEM, New Zealand, with the MinION and MinIT mobile sequencing devices developed by Oxford Nanopore Technologies to produce an e ective point-of-need field diagnostic system. The PDQeX extractsDNAusing a cocktail of thermophilic proteinases and cell wall-degrading enzymes, thermo-responsive extractor cartridges and a temperature control unit. This closed system delivers purified DNA with no cross-contamination. The MinIT is a newly released data processing unit that converts MinION raw signal output into nucleotide base called data locally in real-time, removing the need for high-specification computers and large file transfers from the field. All three devices are battery powered with an exceptionally small footprint that facilitates transport and setup. To evaluate and validate capability of the system for unbiased pathogen identification by real-time sequencing in a farmer’s field setting, we analysed samples collected from cassava plants grown by subsistence farmers in three sub-Sahara African countries (Tanzania, Uganda and Kenya). A range of viral pathogens, all with similar symptoms, greatly reduce yield or destroy cassava crops. Eight hundred (800) million people worldwide depend on cassava for food and yearly income, and viral diseases are a significant constraint to its production. Early pathogen detection at a molecular level has great potential to rescue crops within a single growing season by providing results that inform decisions on disease management, use of appropriate virus-resistant or replacement planting. This case study presented conditions of working in-field with limited or no access to mains power, laboratory infrastructure, Internet connectivity and highly variable ambient temperature. An additional challenge is that, generally, plant material contains inhibitors of downstream molecular processes making e ective DNA purification critical. We successfully undertook real-time on-farm genome sequencing of samples collected from cassava plants on three farms, one in each country. Cassava mosaic begomoviruses were detected by sequencing leaf, stem, tuber and insect samples. The entire process, from arrival on farm to diagnosis, including sample collection, processing and provisional sequencing results was complete in under 3 h. The need for accurate, rapid and on-site diagnosis grows as globalized human activity accelerates. This technical breakthrough has applications that are relevant to human and animal health, environmental management and conservation.Item Unusual occurrence of a DAG motif in the Ipomovirus Cassava brown streak virus and implications for its vector transmission(PLoS ONE, 2017) Ateka, Elijah; Alicai, Titus; Ndunguru, Joseph; Tairo, Fred; Sseruwagi, Peter; Kiarie, Samuel; Makori, Timothy; Kehoe, Monica A.; Boykin, Laura M.Cassava is the main staple food for over 800 million people globally. Its production in eastern Africa is being constrained by two devastating Ipomoviruses that cause cassava brown streak disease (CBSD); Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV), with up to 100% yield loss for smallholder farmers in the region. To date, vector studies have not resulted in reproducible and highly efficient transmission of CBSV and UCBSV. Most virus transmission studies have used Bemisia tabaci (whitefly), but a maximum of 41% U/CBSV transmission efficiency has been documented for this vector. With the advent of next generation sequencing, researchers are generating whole genome sequences for both CBSV and UCBSV from throughout eastern Africa. Our initial goal for this study was to characterize U/CBSV whole genomes from CBSD symptomatic cassava plants sampled in Kenya. We have generated 8 new whole genomes (3 CBSV and 5 UCBSV) from Kenya, and in the process of analyzing these genomes together with 26 previously published sequences, we uncovered the aphid transmission associated DAG motif within coat protein genes of all CBSV whole genomes at amino acid positions 52±54, but not in UCBSV. Upon further investigation, the DAG motif was also found at the same positions in two other Ipomoviruses: Squash vein yellowing virus (SqVYV), Coccinia mottle virus (CocMoV). Until this study, the highly-conserved DAG motif, which is associated with aphid transmission was only noticed once, in SqVYV but discounted as being of minimal importance. This study represents the first comprehensive look at Ipomovirus genomes to determine the extent of DAG motif presence and significance for vector relations. The presence of this motif suggests that aphids could potentially be a vector of CBSV, SqVYV and Coc- Mov. Further transmission and ipomoviral protein evolutionary studies are needed to confirm this hypothesis.