Browsing by Author "Tusiime, Geoffrey"
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Item 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, Jaindra N.; Tushemereirwe, Wilberforce; Kubiriba, Jerome; Changa, Charles; Tripathi, LeenaBlack 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 absolutequantitative 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.Item Evaluation of Cowpea Genotypes for Resistance to Fusarium redolens in Uganda(American Journal of Plant Sciences, 2017) Wanjala Namasaka, Roy; Tusiime, Geoffrey; Orawu, Martin; Gibson, Paul; Nyiramugisha, Josiane; Edema, RichardFusarium redolens, a virulent fungus which causes damping off, leaf yellowing, wilting and root rots has recently been devastating cowpea fields in Uganda. This study aimed at identifying cowpea genotypes that are resistant to Fusarium redolens. Therefore, ninety cowpea genotypes were evaluated two times against a highly virulent Fusarium redolens (isolate from Zombo in Paidha district) in the screen house in 2016. Genotype effect was highly significant (P < 0.001) for root rot severity. Based on the Index of Susceptibility (IS), three genotypes (Asontem, Dan1 LA and IT89KD-88) remained resistant (IS < 3.5) over the two screening periods, 72 moderately resistant (3.5 ≤ IS < 6.5) and 11 susceptible (IS ≥ 6.5). Resistance was found to be enhanced by presence of lateral roots above or at the ground level. Further results suggested a difference in genetic control of resistance to root rots and seed rots caused by Fusarium redolens . All the released varieties tested (SECOW 1 T, SECOW 2 W, SECOW 3 B, SECOW 4 W and SECOW 5 T) had moderate resistance to Fusarium redolens . Correlation analysis revealed root rot severity was strongly correlated to disease incidence (+0.64, P < 0.001), to proportion of plants with lateral roots (−0.56, P < 0.001), to amount of leaf chlorophyll (−0.53, P < 0.001) and to proportion of plants that died prematurely due to Fusarium redolens infection (+0.45, P < 0.001). No significant correlation was detected between root rot severity and proportion of plants that germinated. The established resistance could be exploited for improvement of farmer preferred cowpea varieties towards Fusarium redolens resistance in Uganda.Item Genetic diversity and population structure of Peronosclerospora sorghi isolates of Sorghum in Uganda(International Journal of Environment, Agriculture and Biotechnology (IJEAB), 2018) Kumi, Frank; Agbahoungba, Symphorien; Badji, Arfang; Mwila, Natasha; Ibanda, Angele; Anokye, Michael; Odong, Thomas; Wasswa, Peter; Ochwo- Ssemakula, Mildred; Tusiime, Geoffrey; Biruma, Moses; Kassim, Sadik; Rubaihayo, PatrickSorghum is the third most important staple cereal crop in Uganda after maize and millet. Downy mildew disease is one of the most devastating fungal diseases which limits the production and productivity of the crop. The disease is caused by an obligate fungus, Peronosclerospora sorghi (Weston & Uppal) with varying symptoms. Information on the genetic diversity and population structure of P.sorghi in sorghum is imperative for the screening and selection for resistant genotypes and further monitoring possible mutant(s) of the pathogen. Isolates of P. sorghi infecting sorghum are difficult to discriminate when morphological descriptors are used. The use of molecular markers is efficient, and reliably precise for characterizing P. sorghi isolates. This study was undertaken to assess the level of genetic diversity and population structure that exist in P. sorghi isolates in Uganda.Item Genetic inheritance of resistance to Fusarium redolens in cowpea(Journal of Plant Breeding and Crop Science, 2017) Wanjala, Namasaka Roy; Tusiime, Geoffrey; Orawu, Martin; Paul, Gibson; Agbahoungba, Symphorien; Mahulé Elysé Boris, Alladassi; Edema, RichardFusarium related root rots have been associated with reduced cowpea productivity in Uganda. Sources of genetic resistance to Fusarium redolens which was found to be the most virulent have been identified but the mode of inheritance of the genes conferring the resistance is unknown. This study aims to investigate how the genes for resistance to F. redolens are inherited in cowpea. Four F. redolens root rot resistant cowpea genotypes were crossed with four intermediately resistant and 2 susceptible cowpea genotypes using North Carolina mating design II. The F1 and the parents were evaluated and data were collected on resistance to seed rot, leaf chlorophyll amount, produced lateral roots, response to plant mortality and root rot severity. Results revealed that additive gene effects were significant for all evaluated traits and non-additive genetic effects were significant in resistance to seed rot and chlorophyll amount. General combining ability (GCA) effects showed that the Asontem genotype was a good combiner for increased lateral roots production and resistance to root rot. Degree of dominance estimates revealed that response to plant mortality, root rots and increased lateral root production traits were recessively inherited while seed rot and amount of leaf chlorophyll were dominantly inherited.Item Genetic variation, Heritability estimates and GXE effects on yield traits of Mesoamerican common bean (Phaseolus vulgaris L) germplasm in Uganda(Plant Genetic Resources, 2018) Okii, Dennis; Mukankusi, Clare; Sebuliba, Sulaiman; Tukamuhabwa, Phinehas; Tusiime, Geoffrey; Talwana, Herbert; Odong, Thomas; Namayanja, Annet; Paparu, Pamela; Nkalubo, Stanley; Otim, Michael; Ugen, Michael; Buah, Stephen; Gepts, PaulGermplasm of common beans from the Mesoamerican gene pool races: Durango, Jalisco, Mesoamerica and Guatemala have highest genetic variation for the crop’s improvement. The objective was to assign 50 common bean germplasm in Uganda into its gene pool races based on analyses of population structure. Secondly, to estimate heritability and effects of genotype environment (GXE) interaction on common bean agronomic and yield traits in space and time. Sample genomic DNA was amplified in 2011 with 22 Simple sequence repeat markers (SSRs) and alleles separated using capillary electrophoresis. Field evaluations were conducted in 2010 and 2011 at NaCRRI and 2015 at CIAT – Kawanda. Multivariate analyses of SSRs data identified four subgroups within the germplasm: K4.1–K4.4, with corresponding Wrights fixation indices (FST) as 0.1829 for K4.1, 0.1585 for K4.4, 0.1579 for K4.2 and least for K4.3 at 0.0678. Gene pool race admixtures in the population (14%) were notable and attributed to gene flow. Four superior parents currently used in improving resistance to major diseases grouped as; Jalisco for MLB49-89A; Mesoamerica for MCM5001 and G2333; Durango for MEXICO 54. Heritability values for yield traits estimated using phenotypic data from above fixed parents, was above 0.81. Season and location had significant effect (P < 0.05) on numbers of: flower buds per inflorescence, pod formation and weight of 100 seeds. The findings will improve understanding of co-evolutionary relationships between bean hosts and pathogens for better disease management and will broaden the germplasm base for improving other tropical production constraints.Item Morphological Identification of Phytophthora a Causal Organism of Pineapple Heart Rot Disease in Uganda(International Journal of Phytopathology, 2018) Akasairi, Ocwa; Bua, Bosco; Oculi, Jasper; Tusiime, GeoffreyPineapple (Ananas comosus L. Merr) is an economic horticultural crop in Uganda. However, pineapple production is currently being threatened by the latest outbreak of pineapple heart rot disease in Uganda. Yet, information on pineapple heart rot disease causal pathogen is unknown. Therefore, the objective of this study was to identify the pathogen causing pineapple heart rot disease (PHRD) in Uganda. Samples of pineapple leaves with symptoms of PHRD were collected from four districts of Masaka, Luwero, Mukono and Kayunga. Pathogen isolation was done using corn meal agar (CMA) amended with Pimaricin, Ampicillin, Rifampicin Pentachloronitrobenzene, Hymexazol and Benomyl (PARPHB). Macro- and micro- morphological characteristics of the isolates were assessed using Potato dextrose agar (PDA) and V8 media, respectively. Pathogenicity of the isolates was tested using healthy pineapple plants based on Koch’s postulate. The results showed a significant difference (P<0.05) in the growth rates, hyphae diameter and asexual structure dimensions of the isolates. Isolates were characterized by a dense rosette and stoloniferous mycelial growth pattern in PDA media. Although, sporangia were non-caduceus, terminal, papillate and mostly ovoid, obpyriform and limoniform sporangia (32-56μm) with a mean length/breadth ratio of 1.32:1μm were observed. Chlamydospores were spherical and thick-walled (25-42μm diameter) while Sporangiophores branching was sympodial. Based on the morphological characteristics of isolates, it was concluded that Phytophthora nicotianae is the species causing pineapple heart rot disease in Uganda. This study, therefore, represents the first comprehensive information in Uganda. However, molecular identification of the pathogen is recommended to confirm the genetic identity of the species.Item Multi-Environmental Evaluation of Protein Content and Yield Stability among Tropical Soybean Genotypes Using GGE Biplot Analysis(Agronomy, 2021) Obua, Tonny; Sserumaga, Julius Pyton; Awio, Bruno; Nganga, Fredrick; Odong, Thomas L.; Tukamuhabwa, Phinehas; Tusiime, Geoffrey; Mukasa, Settumba B.; Nabasirye, MargaretThe yield and protein performance in a soybean genotype result from its interaction with the prevailing environmental conditions. This makes selecting the best genotypes under varied target production environments more complex. This study’s objectives were to determine protein content and protein stability of 30 elite soybean genotypes in major soybean-growing areas of Uganda, assess the yield performance and stability in soybeans and determine the relationship between the protein content and grain yield in soybeans. The genotypes were planted in a randomized complete block design of three replications for six seasons across eight locations in Uganda. Genotype and genotypeby- environment (GGE) biplot analyses classified the test locations into three mega-environments for soybean protein and grain yields. Genotype NII X GC 20.3 had the highest mean protein content of 43.0%, and BSPS 48A-9-2 and BSPS 48A-28 were superior for the mean grain yield (1207 kg ha1). Bulindi was the most discriminating and representative test environment for soybean yield. A weak and negative correlation (r = 0.1**, d.f. = 29) was detected between the protein content (%) and yield (kg ha1). The highest-yielding genotypes BSPS 48A-9-2, BSPS 48A-31, and Nam II GC 44.2 are recommended for further evaluation under farmers’ production conditions for selection and release as new soybean varieties in Uganda.Item New sources of sorghum resistant genotypes to downy mildew disease in Uganda(BIODIVERSITAS, 2019) Kumi, Frank; Badji, Arfang; Mwila, Natasha; Odong, Thomas; Ochwo-Ssemakula, Mildred; Tusiime, Geoffrey; Gibson, Paul; Biruma, Moses; Prom, Louis K.; Cuevas, Hugo E.; Agbahoungba, Symphorien; Rubaihayo, PatrickKumi F, Badji A, Mwila N, Odong T, Ochwo-Ssemakula M, Tusiime G, Gibson P, Biruma M, Prom KL, Cuevas HE, Agbahoungba S, Rubaihayo P. 2019. New sources of sorghum resistant genotypes to downy mildew disease in Uganda. Biodiversitas 20: 3391-3397. Sorghum downy mildew (SDM) disease is still prevalent in Uganda at varying levels of incidence and severity. In this study, a total of 100 sorghum genotypes, five (5) from (U.SA, India, and Sudan) and 95 genotypes from Uganda were evaluated for resistance to downy mildew and other agronomic traits during the second growing season of 2016 (August-December). The experiment was conducted in two locations at Makerere University Agricultural Research Institute at Kabanyolo (MUARIK) and Abi-Zonal Agricultural Research and Development Institute (Abi-ZARDI) research station at Arua. The experimental design used was 10 x 10 alpha lattice design with three replicates. Data were collected on plant disease incidence (PDI), plant disease severity (PDS), area under disease progress curve (AUDPC), days to 50% flowering, plant height, 1000 seed weight, and grain yield. Results for analysis of variance showed highly significant differences (P < 0.001) in genotypes, location, and AUDPC, yield and yield components. Disease incidence varied significantly (P < 0.001) between locations, and Arua recorded highest disease incidence and severity of 80.6 and 2.8, respectively. Results from correlation analysis showed a highly significant (P < 0.001) positive association of downy mildew disease incidence with AUDPC (0.835) which suggests that the severity of SDM disease increased with disease incidence, whiles significant (P < 0.001) negative correlation was recorded for days to 50 % flowering (-0.302), 1000 seed weight (-0.471), and grain yield (-0.585), suggesting that grain yield and yield component decreased significantly with increase in SDM incidence and severity. Two resistant (PI 656061 and PI 533831) and four moderately resistant (E 40, MAKSO 8, PI 655990 and Epuripur) genotypes were identified from this study. These genotypes were recommended for sorghum breeding program against downy mildew disease.Item PCR validation of predicted virulence factors in a collection of Xanthomonas campestris (pv. musacearum and vascolurum) strains(African Journal of Biotechnology, 2017) Wasukira, Arthur; Tusiime, Geoffrey; Kubiriba, JeromeBacterial plant pathogens belonging to the Xanthomonas genus are adapted to their host plants and are not known to colonize other environments. Xanthomonas colonize host parts such as leaves, stems and roots before entering vascular tissues and engaging in an invasive pathogenic phase. These bacterial strains have evolved strategies to adapt to life in this environment. The host-pathogen interactions of Xanthomonas vasicola (Xv) need to be well understood to properly map the target genes in the host and pathogen so as to understand the mechanism of resistance. Genotypic characterization, based on the analysis of restriction fragment length polymorphism of virulence factor fragment products was performed on members of the X. vasicola pv. musacearum (Xcm) and X. vasicola pv. vasculorum (Xvv) from varying geographical locations. The study showed that Xcm and Xvv are different from each other based on amplification of virulence factors within fragments of their DNA. Bacterial strains of similar species can have unique Type four pili (Tfp) and Tfp pilus assembly protein PilF a fimbrial biogenesis protein was amplified in all Xanthomonas strains except NCPPB1131 only. Type III effector protein RipT was confirmed to be present in all strains of Xcm and Xvv but not NCPPB1131 and NCPPB1132. All the Xcm and Xvv strains under test yielded bands of type III effector HopAF1 except Xvv206, NCPPB1131 and NCPPB1132. YopJ type III secretion system effector protein hybridizes in DNA of all Xcm strains tested but not in NCPPB1131 or NCPPB1132. This study confirmed the predicted presence or absence of virulence factors especially effectors across bacterial strains and within strains of the same species and other clusters conserved in gram negative bacteria.Item Rice Blast Prevalence in Smallholder Rice Farmlands in Uganda(Journal of Agricultural Science, 2020) Amayo, R.; Oparok, Teddy; Lamo, Jimmy; Drissa, Silue; Edema, Richard; Tusiime, Geoffreythe role of the smallholder farming system in shaping the prevalence of the disease in the country is not known. In 2015B and 2016A, we surveyed smallholder rice farmlands in 27 districts of Uganda and recorded blast incidence, severity, and symptoms expression. Infected rice samples taken from the infected plants were sub-cultured on PDA media to confirm the pathogen and obtain isolates for the establishment of a core collection for breeding work. Rice blast prevalence in the districts varied from 50-100% and the national average stood at 72.61%, higher than that recorded five years ago. Mean incidence and severity varied significantly (< 0.001) with the highest incidence (96.8%) recorded in Luwero district and the least (21.3%) was recorded in the Amuru district. However, the eastern region recorded the highest average incidence (74.5%) followed by the central, the northern, and Mid-western regions. In the rice ecologies, the highest blast incidence was recorded in the rain-fed lowland rice (72.18%) followed by irrigated lowland (59.53%) and rain-fed upland rice (47.27%). This is the first report on the prevalence of blast in smallholder rice farmlands in Uganda and showed a higher prevalence of the disease.Item 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, Jaindra N.; Chen, Li-Hung; Falk, Bryce; Stergiopoulos, Ioannis; Tushemereirwe, Wilberforce; Kubiriba, Jerome; Tripathi, LeenaPseudocercospora 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.