Patrick Okori

A highly conserved NB-LRR encoding gene cluster effective against Setosphaeria turcica in sorghum

BackgroundThe fungal pathogen Setosphaeria turcica causes turcicum or northern leaf blight disease on maize, sorghum and related grasses. A prevalent foliar disease found worldwide where the two host crops, maize and sorghum are grown. The aim of the present study was to find genes controlling the host defense response to this devastating plant pathogen. A cDNA-AFLP approach was taken to identify candidate sequences, which functions were further validated via virus induced gene silencing (VIGS), and real-time PCR analysis. Phylogenetic analysis was performed to address evolutionary events.ResultscDNA-AFLP analysis was run on susceptible and resistant sorghum and maize genotypes to identify resistance-related sequences. One CC-NB-LRR encoding gene GRMZM2G005347 was found among the up-regulated maize transcripts after fungal challenge. The new plant resistance gene was designated as St referring to S. turcica. Genome sequence comparison revealed that the CC-NB-LRR encoding St genes are located on chromosome 2 in maize, and on chromosome 5 in sorghum. The six St sorghum genes reside in three pairs in one locus. When the sorghum St genes were silenced via VIGS, the resistance was clearly compromised, an observation that was supported by real-time PCR. Database searches and phylogenetic analysis suggest that the St genes have a common ancestor present before the grass subfamily split 50-70 million years ago. Today, 6 genes are present in sorghum, 9 in rice and foxtail millet, respectively, 3 in maize and 4 in Brachypodium distachyon. The St gene homologs have all highly conserved sequences, and commonly reside as gene pairs in the grass genomes.ConclusionsResistance genes to S. turcica, with a CC-NB-LRR protein domain architecture, have been found in maize and sorghum. VIGS analysis revealed their importance in the surveillance to S. turcica in sorghum. The St genes are highly conserved in sorghum, rice, foxtail millet, maize and Brachypodium, suggesting an essential evolutionary function.

Two loci in sorghum with NB-LRR encoding genes confer resistance to Colletotrichum sublineolum.

The aim of this work was to identify plant resistance genes to the sorghum anthracnose fungus Colletotrichum sublineolum. cDNA-AFLP transcript profiling on two contrasting sorghum genotypes inoculated with C. sublineolum generated about 3,000 informative fragments. In a final set of 126 sequenced genes, 15 were identified as biotic stress related. Seven of the plant-derived genes were selected for functional analysis using a Brome mosaic virus-based virus-induced gene silencing (VIGS) system followed by fungal inoculation and quantitative real-time PCR analysis. The candidate set comprised genes encoding resistance proteins (Cs1A, Cs2A), a lipid transfer protein (SbLTP1), a zinc finger-like transcription factor (SbZnTF1), a rice defensin-like homolog (SbDEFL1), a cell death related protein (SbCDL1), and an unknown gene harboring a casein kinase 2-like domain (SbCK2). Our results demonstrate that down-regulation of Cs1A, Cs2A, SbLTP1, SbZnF1 and SbCD1 via VIGS, significantly compromised the resistance response while milder effects were observed with SbDEFL1 and SbCK2. Expanded genome analysis revealed that Cs1A and Cs2A genes are located in two different loci on chromosome 9 closely linked with duplicated genes Cs1B and Cs2B, respectively. The nucleotide binding-leucine rich repeat (NB-LRR) encoding Cs gene sequence information is presently employed in regional breeding programs.

Genetic variability and genomic divergence of Elymus repens and related species

SummaryIn order to study the genetic variation and phylogenetic relationship in Elymus repens, amplified fragment length polymorphism (AFLP) were used, together with sequence data for the nuclear gene phytochrome B, phyB, and the chloroplast ribosomal protein encoding gene rps4. A total of 83 collected E. repens samples, 3 E. repens reference samples and 18 related species accessions were analysed and compared with 13 GenBank sequences. AMOVA analysis revealed a moderate genetic differentiation between the populations of E. repens in the three Swedish provinces investigated, while no differentiation was observed due to landscape type. A moderate genetic differentiation was also found when samples from different fields in one province were compared to samples from a selected field. A common female origin was found in E. repens and seven other Elymus species, Pseudoroegneria spicata, Thinopyrum intermedium, T. junceum, Hordeum bogdanii and H. stenostachys. The latter two both harbour the H genome. Taken together, the data suggest that the Swedish E. repens population is slightly heterogeneous and comprises multiple origins of genome donors; a nuclear genome with contributions from Pseudoroegneria (St), Hordeum (H), Thinopyrum (E) and Y with an unknown donor together with a maternal genome donated from Pseudoroegneria.

Disease severity, incidence and races of Setosphaeria turcica on sorghum in Uganda

In order to understand the underlaying causes of new severe turcicum leaf blight outbreaks in East Africa, a survey was undertaken in Uganda to examine the sorghum—Setosphaeria turcica interaction in terms of disease severity and incidence, the overall fungal population structure, and new resistant resources. Highest disease severities were recorded on caudatum accessions, whereas kafir genotypes were most resistant. The disease was more severe in the most humid farmlands compared to moderately dry agro-ecologies. In districts with wide adoption of the Epuripur variety a very high incidence (100%) of turcicum leaf blight was found. The two S. turcica mating type genes MAT1-1 and MAT1-2 assessed on fungal isolates deriving from both sorghum and maize diseased leaves were found in 20 of 23 districts sampled and in equal proportions. Upon cross inoculation on maize differential lines, four S. turcica isolates were identified as race 1, two as race 2, and one isolate corresponded to race 0 and race 3, respectively. The remaining 10 S. turcica isolates did not cause any disease symptoms on the maize lines assessed. Highly resistant accessions originating from a regional collection were found among the five sorghum races (kafir, guinea, caudatum, bicolor and durra), and are now implemented in new sorghum disease resistance programs.

Quantitative Trait Loci for Resistance to Stenocarpella maydis and Fusarium graminearum Cob Rots in Tropical Maize

Stenocarpella maydis and Fusarium graminearum are the predominant species causing maize (Zea mays L.) cob rots in the tropics and sub-Saharan Africa. Developing varieties resistant to cob rots is an alternative strategy that is practical and provides better insurance for small-scale farmers. The subjectivity of scoring and the varying virulence responses of these pathogens to environmental conditions make selection for resistance difficult. The objectives of this study were to map quantitative trait loci (QTL) associated with resistance to S. maydis and F. graminearum and to analyze the possibilities of utilizing these QTL for marker-assisted selection (MAS). Stable QTL mapped were Fg_4,2 (r2 = 0.22) and Sm_4,1 (r2 = 0.16) associated with resistance to F. graminearum and S. maydis, respectively, on chromosome 4. Another QTL associated with resistance to F. graminearum was Fg_5 (r2 = 0.30) on chromosome 5. A QTL with pleiotropic effect was detected on chromosome 1, 22 cM from umc1269 marker (r2 values of 13% and 22% for resistance to S. maydis and F. graminearum, respectively). Additive effects ranged from -0.14 to -0.35 for associated QTL of both pathogens, and all mapped QTL were more than 5 cM from the nearest molecular marker utilized in the study. Therefore, there is need to utilize the maize genomic map to identify and test several markers, < 5 cM, near the detected QTL, in order to locate more reliable molecular markers for utilization in MAS.

Responses of tropical maize landraces to damage by Chilo partellus stem borer

The potential to manage insect pests using host-plant resistance exists, but has not been exploited adequately. The objective of this study was to determine the resistance of 75 tropical maize landraces through artificial infestation with Chilo partellus Swinhoe. The trial was laid in alpha-lattice design and each seedling was infested with five neonates three weeks after planting, over two seasons in 2009 and 2010. The number of exit holes, tunnel length, ear diameter, ear length, plant height, stem diameter, stem lodging and grain yield were measured and a selection index computed. GUAT 1050 was the most resistant with an index of 0.56, while BRAZ 2179 was the most susceptible with an index of 1.66. Ear characteristics were negatively correlated with damage parameters. The principal component biplot suggested that exit holes, cumulative tunnel length, leaf damage, cob diameter, stem lodging, selection index, ear and plant height contributed 71.2% of the variation in resistance. The mean number of exit holes and tunnel length for resistant landraces and resistant hybrid checks were similar; at 5.5 and 2.48 cm, respectively. The identified resistant landraces (GUAT 1050, GUAT 280, GUAT 1093, GUAT 1082, GUAT 1014, CHIS 114, and GUAN 34) could be used to develop C. partellus stem borer-resistant maize genotypes. Key words: Chilo partellus , ear length, exit holes, stem borer resistance, tunnel length.

Assessment of genetic diversity amongst Ugandan sesame (Sesamum indicum L.) landraces based on agromorphological traits and genetic markers

Sesame (Sesamum indicum L.) is one of the most important ancient oilseed crops grown throughout the tropical and subtropical regions of the world. In Uganda, most of the cultivated sesame varieties are local landraces which are frequently traded between farmers. Although these traditional landraces are an important source of genetic diversity, knowledge of their genetic diversity is still limited.Agromorphological traits and a set of published and newly developed microsatellite markers were analyzed on a collection of 121 accessions of Ugandan sesame landraces. CpSSR analysis revealed four haplotypes, whereby haplotype B was present in 96% of the individuals. The analysis of nSSR markers from 6 non-coding regions revealed a mean PIC value of 0.56 whereas the PIC value of eight selected EST-derived SSRs was 0.26. Accession-wise, the expected heterozygosity (He) varied from 0 to 0.396. AMOVA revealed that the majority of the variance occurred among the individuals accounting for 75% of the total variation, only 6% was attributed to differences among the districts, pointing towards a high gene flow (Nm = 4.476). These results are supported by the PCoA analysis as well as the NJ tree both of which revealed no clustering of the accessions according to their geographic origin. Also the statistical analysis of 10 agromorphological traits indicated no clear pattern related to the geographic origin. Such a poor grouping, indicative of considerable gene flow across geographic domains, could be explained either by a high outcrossing rate, and/or through extensive seed trading.

Upland Rice Breeding in Uganda: Initiatives and Progress

Until recently, there was limited research on breeding upland rice varieties. Moreover, there is an increasing expansion of rice production from traditional irrigated production areas to rain‐fed environments in the East African region, where drought problem is a serious challenge. To date, several initiatives aimed at increasing rice production have been made. Of the initiatives, promotion of upland rice production has been the most important in Uganda, but yield penalty due to drought continued to be a major drawback. This article traces progress in the upland rice breeding that started with improvement of late maturing varieties that had nonpreferred cooking qualities. Initially, introduced lines were evaluated and released. These varieties are the ‘New Rice for Africa’ (NERICA) that had been generated from interspecific crosses involving Oryza glaberrima and Oryza sativa. Several studies to understand the mode of gene action and modified pedigree breeding approaches for drought tolerance were conducted and used to develop new rice varieties. Up to 11 improved upland rice varieties were released and deployed in the country from 2002 to 2011 as a result of this initiative.