Browsing by Author "Kiarie, Samuel"

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    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, Samuel
    Crop 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.
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    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.