Browsing by Author "Runo,Steven"
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Item Engineering local Kenyan Irish potato varieties as carriers for edible cholera vaccines(Springer Nature, 2025-06-02) Siamalube, Beenzu; Ehinmitan,Emmanuel; Maina, Ngotho; Onguso,Justus; Runo,StevenCholera is a major public health concern in sub-Saharan Africa, particularly in low-resource areas where vaccine access is limited. Plant-based production of antigens, like the cholera toxin B subunit, presents a promising supplement for oral immunization. This study reviews the efciency of Agrobacterium tumefaciens gene-mediated transformation method in expressing antigenic proteins in three farmer preferred Kenyan Irish potato varieties: Wanjiku, Sherekea, and Shangi. Aimed at maximizing gene expression, facilitating a scalable production platform for potential edible vaccines. Genetically engineered Irish potatoes represent a transformative innovation at the intersection of agriculture and biotechnology, ofering a sustainable and cost-efective platform for vaccine production. Acting as bioreactors, transgenic Irish potatoes provide scalable, safe, and stable solutions to the challenges of traditional vaccine manufacturing, such as high costs, infrastructure demands, and dependency on cold chain logistics. Despite challenges like regulatory hurdles, public perception of genetically modifed organisms, and technical barriers in antigen expression, ongoing research and development hold promise for overcoming these obstacles. By harnessing the power of molecular farming, scientists can produce vaccines and therapeutic proteins in potatoes, ofering a near-user-site alternative to traditional production methods.Item Evaluation of Granular Formulated Strigolactone Analogs for Striga Suicidal Germination(John Wiley & Sons, 2024) Jamil,Muhammad; Margueritte,Ouedraogo; Yonli,Djibril; Jian You Wang; Navangi,Lynet; Mudavadi,Patrick; Patil,Rohit H.; Bhoge,Satish Ekanath; Traore,Hamidou; Runo,Steven; Al-Babili,SalimBACKGROUND: Striga hermonthica, an obligate root parasitic weed, poses a significant threat to cereal production in subSaharan Africa. Lowering Striga seed bank in infested soils is a promising strategy to mitigate infestation levels. The dependency of Striga seed germination on strigolactones opens up the possibility of a ‘suicidal germination’ approach, where synthetic germination stimulants induce lethal germination in the absence of a host. Implementing this approach requires active germination stimulants with a suitable formulation for field application. Here, we describe the development of slowreleasing granular formulation of two potent germination stimulants ‘Methyl Phenlactonoate 3’ and ‘Nijmegen-1’ and the assessment of their activity under Laboratory, greenhouse, mini-field, and field conditions. RESULTS: Under laboratory conditions, the granular formulation of either of the two germination stimulants (1.25 mg per plate, corresponding to 0.09 mg a.i.) induced Striga seed germination at a rate of up to 43%. With 10 mg granular product (0.75 mg a. i.) per pot, we observed 77–83% reduction in Striga emergence under greenhouse pot conditions. Application of the formulated stimulants under artificially or naturally infested fields resulted in approximately 56%, 60%, and 72% reduction in Striga emergence in maize, sorghum, and millet fields in Kenya and Burkina Faso, respectively. CONCLUSION: Our findings on the newly designed granular formulation of Methyl Phenlactonoate 3 and Nijmegen-1 reveal encouraging prospects for addressing the Striga problem in Africa. These findings underscore several significant advantages of the formulated stimulants, including suitability for the African agricultural context, and, most importantly, their effectiveness in reducing Striga infectionItem Recent Trends of Vibrio cholerae: Global and Regional Incidences(2025) Siamalube,Beenzu; Ehinmitan,Emmanuel; Runo,Steven; Onguso,Justus; Maina, NgothoCholera is still with us! The foodborne/waterborne dis ease caused by the bacterium Vibrio cholerae continues to pose a significant threat to the safety of public health glob ally (Mohammed et al., 2024). Despite various medical inter ventions such as the use of oral cholera vaccines (OCVs) (Zeitoun et al., 2024), hundreds of thousands of lives are lost annually worldwide to this virulent disease. Cholera mostly affects vulnerable populations in regions with inadequate access to clean water and sanitation (Bose et al., 2024). Recent outbreaks in parts of Asia (Nasr et al., 2024), Africa (Taty et al., 2024), and the Caribbean underscore the ongoing risk and the urgent need for comprehensive strategies to control its spread (Siamalube and Ehinmitan, 2024). This commentary aims to unravel the challenges caused by V. cholerae by exploring the global cholera inci dences and the multifaceted approaches required to combat this persistent pathogenItem Simple and Fail-safe Method to Transform Miniprep Escherichia coli Strain K12 Plasmid DNA Into Viable Agrobacterium tumefaciens EHA105 Cells for Plant Genetic Transformation(bio-protocol, 2025) Siamalube,Beenzu; Ehinmitan,Emmanuel; Maina Ngotho; Onguso,Justus; Runo,StevenAgrobacterium-mediated gene transformation method is a vital molecular biology technique employed to develop transgenic plants. Plants are genetically engineered to develop disease-free varieties, knock out unsettling traits for crop improvement, or incorporate an antigenic protein to make the plant a green factory for edible vaccines. The method’s robustness was validated through successful transformations, demonstrating its effectiveness as a standard approach for researchers working in plant biotechnology. It enables the introduction of foreign DNA into plant genomes. Conventionally, plant genetic transformation has relied on time-consuming, costly, and technically demanding procedures, such as electroporation and chimeric viruses or biolistic methods, which usually yield variable transformation efficiencies. This study presents a simple and fail-safe protocol that involves a modified freeze-thaw and heat-shock concoction method. This approach involves a streamlined plasmid miniprep procedure to isolate high-quality plasmid DNA from Escherichia coli K12 strain, followed by a target-specific transfer into A. tumefaciens EHA105 strain. The optimized method minimizes DNA degradation and maximizes uptake by Agrobacterium cells, making it a reproducible and accessible protocol for various genetic engineering applications. The transformation efficiency is consistently high, enhancing plasmid uptake while maintaining cell viability, requiring minimal specialized equipment and reagents. The proposed protocol offers significant advantages, including simplicity, reliability, and cost-effectiveness, positioning it as a valuable alternative to traditional techniques in the field of plant biotechnology.Item Transcriptome Atlas of Striga Germination: Implications for Managing an Intractable Parasitic Plant(ppp, 2023) Irafasha,Gilles; Mutinda, Sylvia; Mobegi,Fredrick; Hale,Brett; Omwenga,George; Wijeratne, Asela J.; Wicke,Susann; Bellis,Emily S.; Runo,StevenSocietal Impact StatementWitchweeds, parasitic plants of the genus Striga, are nicknamed “cereal killers”because of their devastating destruction of Africa's most staple cereals, includingmaize, sorghum, millets, and upland rice. The parasite relies on biomolecules emittedfrom the host roots to germinate and therefore initiate its infectious lifecycle. Somesorghum varieties have evolved to not produce effective germination stimulants,making them resistant to the parasite. Here, the genetic factors that underpin Strigagermination were assessed, followed by a discussion of how such knowledge can beused to develop new Striga management strategies through the disruption of host–parasite communication exchange.Summary• Seeds of the parasitic plant Striga are dormant. They only germinate in responseto biomolecules emitted from the host's root exudate, strigolactones (SL). But it isnow emerging that Striga germination is a much more complex process regulatedby crosstalk of hormone signaling pathways.• To further understand the genetic basis of the communication exchange betweenStriga and its host sorghum, we performed a comparative transcriptomic analysis.We sought to identify major transcriptomic changes that define the germinationprocess in Striga and a set of genes that may contribute to the differences in ger-mination rates.• Results showed that germination proceeds immediately after SL perception and ismarked by a wave of transcriptional reprogramming to allow for metabolic pro-cesses of energy mobilization. Cluster analysis using self-organizing maps revealeda time-phased and genotype-differentiated response to germination stimulation.The variation in germination was also a function of hormonal crosstalk. The earlygermination stage was associated with significant repre....