Conversion and validation of uniplex SNP markers for selection of resistance to Cassava Mosaic Disease in Cassava breeding programs

Open Access journal; Published online: 25 Feb 2021Cassava mosaic disease (CMD) is a major viral disease adversely affecting cassava production in Africa and Asia. Genomic regions conferring resistance to the disease have been mapped in African cassava germplasm through biparental quantitative trait loci (QTL) mapping and genome-wide association studies. To facilitate the utilization of these markers in breeding pipelines to support selections, proof-of-concept technical and biological validation research was carried out using independent pre-breeding and breeding populations. Kompetitive Allele-Specific Polymerase Chain Reaction (KASP) assays were designed from three single nucleotide polymorphism (SNP) markers linked to a major resistance locus on chromosome 12 (S12_7926132, S12_7926163) and a minor locus on chromosome 14 (S14_4626854). The designed assays were robust and easy to score with >99% genotype call rate. The overall predictive accuracy (proportion of true positives and true negatives) of the markers (S12_7926132 and S14_4626854) was 0.80 and 0.78 in the pre-breeding and breeding population, respectively. On average, genotypes that carried at least one copy of the resistant allele at the major CMD2 locus had a significantly higher yield advantage. Nevertheless, variation was observed in prediction accuracies for the major locus (S12_7926132) among sub-families from the two populations, suggesting the need for context-specific utilization, for example, by screening for co-segregation of favorable SNP alleles with resistance in the parents being used for crosses. Availability of these validated SNP markers on the uniplex KASP genotyping platform represents an important step in translational genetics toward marker-assisted selection to accelerate introgression of favorable resistant alleles in breeding populations

Validation of KASP-SNP markers in cassava germplasm for marker-assisted selection of increased carotenoid content and dry matter content

Open Access Journal; Published online: 12 Oct 2022Provitamin A biofortification and increased dry matter content are important breeding targets in cassava improvement programs worldwide. Biofortified varieties contribute to the alleviation of provitamin A deficiency, a leading cause of preventable blindness common among pre-school children and pregnant women in developing countries particularly Africa. Dry matter content is a major component of dry yield and thus underlies overall variety performance and acceptability by growers, processors, and consumers. Single nucleotide polymorphism (SNP) markers linked to these traits have recently been discovered through several genome-wide association studies but have not been deployed for routine marker-assisted selection (MAS). This is due to the lack of useful information on markers’ performances in diverse genetic backgrounds. To overcome this bottleneck, technical and biological validation of the loci associated with increased carotenoid content and dry matter content were carried out using populations independent of the marker discovery population. In the present study, seven previously identified markers for these traits were converted to a robust set of uniplex allele-specific polymerase chain reaction (PCR) assays and validated in two independent pre-breeding and breeding populations. These assays were efficient in discriminating marker genotypic classes and had an average call rate greater than 98%. A high correlation was observed between the predicted and observed carotenoid content as inferred by root yellowness intensity in the breeding (r = 0.92) and pre-breeding (r = 0.95) populations. On the other hand, dry matter content-markers had moderately low predictive accuracy in both populations (r< 0.40) due to the more quantitative nature of the trait. This work confirmed the markers’ effectiveness in multiple backgrounds, therefore, further strengthening their value in cassava biofortification to ensure nutritional security as well as dry matter content productivity. Our study provides a framework to guide future marker validation, thus leading to the more routine use of markers in MAS in cassava improvement programs

Classification of elite cassava varieties (Manihot esculenta Crantz) cultivated in Benin Republic using farmers’ knowledge, morphological traits and simple sequence repeat (SSR) markers

Published online: 23 August 2017Cassava (Manihot esculenta Crantz) is an important food security crop or resource for poor rural communities particularly in Africa. The crop’s ability to produce high yields even under poor conditions and storability of its roots underground for longer periods or until needed makes it a model ‘food security crop’. In Benin Republic, cassava has been recognized as one of the major crop contributing towards dynamic value chains generating incomes for small-holder farmers. The crop is grown all over the country, however, the increased production are mainly recorded from far south and central parts of the country. Genetic improvement of cassava in Benin Republic is limited because of poor knowledge of genetic diversity present within the country. The main objective of this study was to assess the genetic diversity and relationships among elite cassava varieties collected from different regions of Benin using fluorescently labelled simple sequence repeat (SSR) markers and to compare the results with farmer’s knowledge and morphological traits. A total of 96 cultivars collected from major cassava growing areas such as Southern and Central Benin were classified into 24 different groups using farmers’ knowledge, while classification based on 18 morphological traits resulted in five groups. In total, sixteen SSR markers were tested for molecular analysis of the ninety-six cassava varieties. Among the sixteen, twelve SSR markers gave good banding pattern and were used to genotype the varieties. An average of 3.58 and 0.47 for number of alleles and polymorphism information content respectively was observed. The observed heterozygosity (Ho) ranged from 0.23 to 1.0 with an average of 0.66 indicating moderate level of diversity among the cultivars. Based on the proportion of shared alleles and hierarchical clustering, the 96 elite cassava varieties were classified as 74 unique varieties. Principal component analysis and analysis of molecular variance revealed no significant variation between the regions thus, explaining regular exchange of planting materials among cassava farmers across various regions. The moderate level of genetic diversity in famer’s field, revealed in the present study, is a good indication of the need for broadening the genetic base of cassava in Benin Republic and establishing a formal breeding program in the country

Historical introgressions from a wild relative of modern cassava improved important traits and may be under balancing selection

Published online: 17 Oct 2019Introgression of alleles from wild relatives has often been adaptive in plant breeding. However, the significance of historical hybridization events in modern breeding is often not clear. Cassava (Manihot esculenta) is among the most important staple foods in the world, sustaining hundreds of millions of people in the tropics, especially in sub-Saharan Africa. Widespread genotyping makes cassava a model for clonally-propagated root and tuber crops in the developing world and provides an opportunity to study the modern benefits and consequences of historical introgression. We detected large introgressed M. glaziovii genome-segments in a collection of 2742 modern cassava landraces and elite germplasm, the legacy of 1930's era breeding to combat disease epidemics . African landraces and improved varieties were on average 3.8% (max 13.6% ) introgressed. Introgressions accounted for significant (mean 20% , max 56% ) portion of the heritability of tested traits. M. glaziovii alleles on the distal 10Mb of chr. 1 increased dry matter and root number. On chr. 4, introgressions in a 20Mb region improved harvest index and brown streak disease tolerance. We observed the introgression frequency on chr. 1 double over three cycles of selection and that later stage trials selectively excluded homozygotes from consideration as varieties. This indicates a heterozygous advantage of introgressions. However, we also found that maintaining large recombination-suppressed introgressions in the heterozygous state allowed the accumulation of deleterious mutations. We conclude that targeted recombination of introgressions would increase the efficiency of cassava breeding by allowing simultaneous fixation of beneficial alleles and purging of genetic load