Browsing by Author "Tusiime, Geoffrey"

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    Agrobacterium tumefaciens-mediated transformation of pseudocercospora fijiensis to determine the role of PfHog1 in osmotic stress regulation and virulence modulation
    (Frontiers in Microbiology, 2017) Onyilo, Francis; Tusiime, Geoffrey; Chen, Li-Hung; Falk, Bryce; Stergiopoulos, Ioannis; Tripathi, N. Jaindra; Tushemereirwe, Wilberforce; Kubiriba, Jerome; Changa, Charles; Tripathi, Leena
    Black Sigatoka disease, caused by Pseudocercospora fijiensis is a serious constraint to banana production worldwide. The disease continues to spread in new ecological niches and there is an urgent need to develop strategies for its control. The high osmolarity glycerol (HOG) pathway in Saccharomyces cerevisiae is well known to respond to changes in external osmolarity. HOG pathway activation leads to phosphorylation, activation and nuclear transduction of the HOG1 mitogen-activated protein kinases (MAPKs). The activated HOG1 triggers several responses to osmotic stress, including up or down regulation of different genes, regulation of protein translation, adjustments to cell cycle progression and synthesis of osmolyte glycerol. This study investigated the role of the MAPK-encoding PfHog1 gene on osmotic stress adaptation and virulence of P. fijiensis. RNA interference-mediated gene silencing of PfHog1 significantly suppressed growth of P. fijiensis on potato dextrose agar media supplemented with 1 M NaCl, indicating that PfHog1 regulates osmotic stress. In addition, virulence of the PfHog1-silenced mutants of P. fijiensis on banana was significantly reduced, as observed from the low rates of necrosis and disease development on the infected leaves. Staining with lacto phenol cotton blue further confirmed the impaired mycelial growth of the PfHog1 in the infected leaf tissues, which was further confirmed with quantification of the fungal biomass using absolute- quantitative PCR. Collectively, these findings demonstrate that PfHog1 plays a critical role in osmotic stress regulation and virulence of P. fijiensis on its host banana. Thus, PfHog1 could be an interesting target for the control of black Sigatoka disease in banana.
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    Genome-wide association analysis identifies resistance loci for bacterial blight in diverse East African rice germplasm
    (Academic Journals, 2023-07-31) Okello, Moses; Mildred, Ochwo S.; Lamo, Jimmy; Onaga, Geoffrey; Odong, Thomas L.; Tusiime, Geoffrey; Tukamuhabwa, Phinehas; Mukasa, Settumba B.; Wasswa, Peter; Ogwal, Jonathan; Oliva, Ricardo
    Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of rice bacterial blight disease has been extensively characterized, and loci against different races identified. Many rice cultivars have been developed and utilized to combat the disease, however, due to the rapid evolution of Xoo, several resistances have broken down. The continuous challenge of ever-evolving Xoo and the breakdown of resistance in cultivated rice varieties make it even more important to discover new loci to enable sustainable durable deployment of broad-spectrum resistance genes in elite breeding lines. African germplasm can be exploited as reservoirs of useful genetic variation for bacterial blight (BB) resistance. This study was conducted to identify loci associated with BB resistance and new genetic donors for the breeding program. To identify candidate sources of resistance for advancing breeding, four virulent strains of Xoo (PXO99, MAI1, BAI3, and Xoo3-1) were used to screen 78 East African accessions by genome-wide association studies. The diverse accessions' core genetic base exhibited high resistance to the Xoo strains. 50.63% of the accessions were highly resistant to the Philippines strain PX099, while 20.25% were highly susceptible to the virulent West African strain MAI1. Two novel resistant loci significantly associated hotspots were identified using 1901 SNPs. The two hits were located on chromosome 12 (Xa25) and Chr. 6 (Xa7, Xa27, Xa33). Novel loci were identified that gives a useful basis for more investigation and a wide core genetic pool of high resistance for broad-spectrum resistance for genetic improvement.
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    Pathogenic and genetic diversity of sclerotium rolfsii, the causal agent of southern blight of common bean in Uganda
    (MDPI, 2025-12-26) Erima, Samuel; Nyine, Moses; Ssemakula, Mildred Ochwo; Tusiime, Geoffrey; Akhunov, Eduard; Akhunova, Alina; Yunusbaev, Ural; Adjei, Emmanuel Amponsah; Mukasa, Settumba B.; Otim, Michael Hilary; Odong, Thomas Lapaka; Nkuboye, Allan; Candiru, Agnes; Paparu, Pamela
    Sclerotium rolfsii Sacc. is a soil-borne fungus that causes southern blight on many crops in the tropical and subtropical regions. In 2018, southern blight was reported as the most prevalent bean root rot in Uganda. Earlier studies ascertained the morphological and pathogenic diversity of S. rolfsii, but a limited understanding of its genetic diversity exists. Knowledge of S. rolfsii genetic diversity is a critical resource for pathogen surveillance and developing common bean varieties with durable resistance. A total of 188 S. rolfsii strains from infected common bean plants were collected from seven agro-ecological zones of Uganda in 2013, 2020 and 2021, and characterized morphologically and pathogenically. The genetic diversity of the strains was assessed using single-nucleotide polymorphisms (SNPs) obtained from whole-genome sequencing. The growth rate of the strains ranged between 1.1 and 3.6 cm per day, while the number of sclerotia produced ranged from 0 to 543 per strain. The strains had fluffy, fibrous, and compact colony texture. The strains were pathogenic on common bean and caused disease severity indices ranging from 10.1% to 93.3%. Average polymorphic information content across all chromosomes was 0.27. Population structure analysis identified five genetically distinct clusters. The results of analysis of molecular variance revealed that 54% of the variation was between clusters while 46% of variation was within clusters. Pairwise comparison of Wright’s fixation indices between genetic clusters ranged from 0.31 to 0.78. The findings of this study revealed moderate genetic diversity among S. rolfsii strains, which should be taken into consideration when selecting strains for germplasm screening.
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    Silencing of the Mitogen-Activated Protein Kinases (MAPK) Fus3 and Slt2 in Pseudocercospora fijiensis Reduces Growth and Virulence on Host Plants
    (Frontiers in Plant Science, 2018) Onyilo, Francis; Tusiime, Geoffrey; Tripathi, N. Jaindra; Chen, Li-Hung; Falk, Bryce; Stergiopoulos, Ioannis; Tushemereirwe, Wilberforce; Kubiriba, Jerome; Tripathi, Leena
    Pseudocercospora fijiensis, causal agent of the black Sigatoka disease (BSD) of Musa spp., has spread globally since its discovery in Fiji 1963 to all the banana and plantain growing areas across the globe. It is becoming the most damaging and economically important disease of this crop. The identification and characterization of genes that regulate infection processes and pathogenicity in P. fijiensis will provide important knowledge for the development of disease-resistant cultivars. In many fungal plant pathogens, the Fus3 and Slt2 are reported to be essential for pathogenicity. Fus3 regulates filamentous-invasion pathways including the formation of infection structures, sporulation, virulence, and invasive and filamentous growth, whereas Slt2 is involved in the cell-wall integrity pathway, virulence, invasive growth, and colonization in host tissues. Here, we used RNAi-mediated gene silencing to investigate the role of the Slt2 and Fus3 homologs in P. fijiensis in pathogen invasiveness, growth and pathogenicity. The PfSlt2 and PfFus3 silenced P. fijiensis transformants showed significantly lower gene expression and reduced virulence, invasive growth, and lower biomass in infected leaf tissues of East African Highland Banana (EAHB). This study suggests that Slt2 and Fus3 MAPK signaling pathways play important roles in plant infection and pathogenic growth of fungal pathogens. The silencing of these vital fungal genes through host-induced gene silencing (HIG) could be an alternative strategy for developing transgenic banana and plantain resistant to BSD.