Perspectives on the Application of Next-generation Sequencing to the Improvement of Africa’s Staple Food Crops

The persistent challenge of insufficient food, unbalanced nutrition, and deteriorating natural resources in the most vulnerable nations, characterized by fast population growth, calls for utilization of innovative technologies to curb constraints of crop production. Enhancing genetic gain by using a multipronged approach that combines conventional and genomic technologies for the development of stress-tolerant varieties with high yield and nutritional quality is necessary. The advent of next-generation sequencing (NGS) technologies holds the potential to dramatically impact the crop improvement process. NGS enables whole-genome sequencing (WGS) and re-sequencing, transcriptome sequencing, metagenomics, as well as high-throughput genotyping, which can be applied for genome selection (GS). It can also be applied to diversity analysis, genetic and epigenetic characterization of germplasm and pathogen detection, identification, and elimination. High-throughput phenotyping, integrated data management, and decision support tools form the necessary supporting environment for effective utilization of genome sequence information. It is important that these opportunities for mainstreaming innovative breeding strategies, enabled by cutting-edge “Omics” technologies, are seized in Africa; however, several constraints must be addressed before the benefit of NGS can be fully realized. African breeding programs must have access to high-throughput genotyping facilities, capacity in the application of genome selection and marker-assisted breeding must be built and supported by capacity in genomic analysis and bioinformatics. This chapter demonstrates how interventions with NGS-enabled innovative strategies can be applied to increase genetic gain with insights from the Consortium of International Agricultural Research (CGIAR) in general and the International Institute of Tropical Agriculture (IITA) in particular

A time series transcriptome analysis of cassava (Manihot esculenta Crantz) varieties challenged with Ugandan cassava brown streak virus

Open Access Journal; Published online: 29 August 2017A time-course transcriptome analysis of two cassava varieties that are either resistant or susceptible to cassava brown streak disease (CBSD) was conducted using RNASeq, after graft inoculation with Ugandan cassava brown streak virus (UCBSV). From approximately 1.92 billion short reads, the largest number of differentially expressed genes (DEGs) was obtained in the resistant (Namikonga) variety at 2 days after grafting (dag) (3887 DEGs) and 5 dag (4911 DEGs). At the same time points, several defense response genes (encoding LRR-containing, NBARC-containing, pathogenesis-related, late embryogenesis abundant, selected transcription factors, chaperones, and heat shock proteins) were highly expressed in Namikonga. Also, defense-related GO terms of ‘translational elongation’, ‘translation factor activity’, ‘ribosomal subunit’ and ‘phosphorelay signal transduction’, were overrepresented in Namikonga at these time points. More reads corresponding to UCBSV sequences were recovered from the susceptible variety (Albert) (733 and 1660 read counts per million (cpm)) at 45 dag and 54 dag compared to Namikonga (10 and 117 cpm respectively). These findings suggest that Namikonga’s resistance involves restriction of multiplication of UCBSV within the host. These findings can be used with other sources of evidence to identify candidate genes and biomarkers that would contribute substantially to knowledge-based resistance breeding

A global overview of cassava genetic diversity

Although numerous studies of diversity have been conducted in cassava, there is no comprehensive assessment of global genetic diversity. Here we draw on previous studies and breeders’ knowledge to select diversity sets from the International Institute of Tropical Agriculture (IITA) and the International Center for Tropical Agriculture (CIAT) genebanks and breeders’ germplasm, as well as elite germplasm and landraces from eastern, southern and central (ESC) Africa to make a global assessment of diversity in cassava, using a SNP based GoldenGate (Illumina Inc.) assay. A synthesis of results from genetic distance and ADMIXTURE analysis essentially revealed four populations (i) South American germplasm characterised by relatively higher genetic diversity with hypothetical ancestral founder genotypes from Brazil, (ii) a smaller group of African introduction germplasm which is more distantly related to all other germplasm, (iii) West Africa germplasm dominated by IITA breeding lines, containing sources of cassava mosaic disease resistance, and IITA genebank accessions from West Africa, both characterised by slightly lower diversity, and (iv) a less cohesive group of African germplasm, termed ‘Other’, with moderate levels of diversity and a majority of germplasm from ESC Africa. This study highlights opportunities for heterosis breeding, purging of duplicates in genebanks and the need for conservation of ESC Africa landraces

Analysis of genomic sequences from peanut ( Arachis hypogaea )

Peanut is an important legume crop across the world. However, in contrast to most legume crops, groundnut lacks taxonomic proximity to any major model genome. A relatively large number of genomic sequences were generated from groundnut as part of a microsatellite marker development project. In the current study, a total of 1312 sequences were analyzed of which 448 contained microsatellite motifs. All sequences (GenBank Accessions: BZ999351-CC000573) were analyzed after clustering for possible similarity with publicly available sequences from Arabidopsis, Lotus, soybean and Medicago. At least 39% of the sequences analyzed had significant BLAST similarities with sequences from the four databases searched, of which nearly half (47%) found significant similarity with Lotus japonicus sequences. Over one quarter (26.7%) of sequences found similarity with Arabidopsis thaliana , while the remainder aligned with publicly available sequences from the legumes soybean and Medicago truncatula . At least 17% of microsatellite containing sequences could be assigned an identity. The codon usage pattern for Arachis hypogaea most closely resembles that of L. japonicus reflecting the similarly high sequence similarity observed in BLAST searches at the protein level. The implications of these findings for the taxonomy, and comparative genomics of groundnut and its legume family relatives are discussed

Development of expressed sequence tags-simple sequence repeats (EST-SSRs) for Musa and their applicability in authentication of a Musa breeding population

Maintaining identity of clones is essential in breeding programs. New EST-SSR markers have been developed for banana and used to screen a diploid population for clonal identity. A total of 410 primer pairs were designed from an EST database, validated using polyacrylamide gel electrophoresis (PAGE) and a subset was optimized for accurate genotyping on a capillary genetic analyzer. Combining PAGE and capillary electrophoresis, about 44% of the designed primers were informative in the diploid population. The majority of markers produced two alleles as expected in a diploid population. However, some showed three to four alleles, possibly indicating closely-related members of gene families. Screening of field samples using SSR markers revealed genotype identity issues in the target population. The present study demonstrates the applicability of SSRs in the  establishment of parentage and relatedness between accessions. The newly-developed SSRs will be valuable tools in the understanding of Musa genetics, in marker-trait associations, thereby enhancing the effectiveness of breeding programs.Key words: EST, SSRs, markers, Musa, breeding

Patterns of nucleotide diversity in Meisa1 and G3pdh in wild and cultivated cassava

The distribution and frequency of single nucleotide polymorphisms (snps) is an excellent tool for discerning evolutionary relatedness between cultivated and wild plant genomes. This type of information is scanty for the genus Manihot, and thus limiting systematic approaches in the genetic improvement of cassava. Here, we present a detailed description of the comparative patterns of snps in Isoamylase1 (Meisa1) and Glyceraldehyde-3-phosphate dehydrogenase (G3pdh) in 10 accessions of wild (Manihot esculenta subsp. flabellifolia) and 12 accessions of cultivated cassava (M. esculenta). The results show that Meisa1 is more variable in cultivated cassava than that in subspecies flabellifolia, where the 954 bp sequence region differs at 1 in 111 and 250 nucleotides of cultivated and wild species, respectively. Frequency analysis shows that snp occurs once every 42 bp in cultivated and every 70 bp in wild. Tajima’s D test statistics showed that Meisa1 has been evolving under different selection pressures, diversifying in cultivated and purifying in wild. G3pdh is under diversifying selection in both populations. This may indicate the importance for isoamylase1 in starch quality traits in cassava, a trait that is likely to have been the target for artificial selection by farmers and breeders, in addition to natural selection. This study also suggests that G3pdh may be a good marker for phylogeny study while Meisa1 may be useful for intra and inter-cultivar diversity studies. The non-synonymous snps that changed the amino acid property were identified and the potential implication of the change in protein function was analyzed and discussed