Molecular Characterization of Solanum Species (Solanum aethiopicum complex; Solanum macrocarpon and Solanum anguivi) Using Multiplex RAPD primers

Abstract

Genetic diversity of S. aethiopicum complex, S. macrocarpon and S. anguivi was investigated using molecular approach. Seeds were randomly secured across north central locations in Nigeria and planted. Nine (9) oligonucleotide primers were initially screened for their polymorphism from where five (5) polymorphic primers were selected. A total of twenty five (25) accessions were studied. DNA was extracted from each accession using FTA PlantSaver card method following standard protocol (Whatman®, 2014). Amplification of the extracted DNA was performed on a thermal cycler (Applied Biosystem in Life Technology version 2720) through the use of a multiplex of OPP-11, B-18, OPU-13 and OPU-15 primers. A total of fifty (50) PCR reactions were carried out (25 separate reactions each for OPQ-07 primer and Multiplex primer). Amplified products were resolved on agarose gel electrophoresis. DNA band profiles generated by primers were manually scored (using zero (0) for absence of band and one (1) for presence of band) to generate binary matrices for the two band profiles. Statistical analysis was performed using the SPSS (Statistical Package for Social Scientists) software (20.0 Version). Hierarchical cluster analysis of all the individual accessions was done using the Average Linkage "Between Group" Method based on Euclidean Distance measurement, to generate two dendrograms, one for each primer set. Results revealed the polymorphic strength of each primer sets as 45%. Each primer yielded an average of 4.5 polymorphic loci per 10 DNA bands. Dendrograms showed high level of intraspecific and interspecific variability and similarities among the accessions whose clustering patterns were both location dependent and location independent. Based on the clustering patterns, there were sharp genetic dissimilarities between S. aethiopicum gilo, S. anguivi medium, S. anguivi tiny, striped gilo-kumba complex and S. macrocarpon on one hand, and similarities among the species on the other hand indicating a common ancestral origin. Having established a substantial level of variability in this study, breeding efforts of varieties may be facilitated for improvement programme and this can also be used as a template for further taxonomic studies of the unidentified seeds and fruits showing divergent characters.

Comparison of transcriptome-derived simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers for genetic fingerprinting, diversity evaluation, and establishment of relationships in eggplants

“…When considering only SSR or SNP markers, accessions from different groups of the scarlet eggplant complex (i.e., S. anguivi, the Intermediate S. anguivi-S. aethiopicum group, and the four groups of S. aethiopicum) often shared a same profile. Our data are in agreement with those of other authors who used AFLPs and SSRs (Sunseri et al 2010), RAPDs (Aguoru et al 2015), and SNPs (Acquadro et al, 2017) and also found that accessions of S. aethiopicum did not cluster according to the cultivar group. This is an additional indication that these groups, which are distinguished on the basis of morphology (Lester, 1986;Lester and Daunay, 2003;Plazas et al 2014), are not genetically differentiated.…”

Varietal Characterisation and Taxonomic Evaluation of Sweet Potato (Ipomoea batatas) Using Macro- and Micromorphological Evidence

“…Hence, polyploidy as a combination of new chromosome may lead to sympatric speciation where new species of distinct character may likely evolve. The same type of hybrid complexes was reported in eggplant (Solanaceae) which continuously gives rise to new species because of their interfertile nature [17]. Mechanism of unreduced pollen formation was proposed by [21] as responsible for the polyploidization of sweetpotato.…”

“…This is substantiated in their root colour differences (Figure 4(d)). Similar approach of classification was successfully adopted by [17] in the determination of phylogenetic relationship in eggplant molecular studies in north central Nigeria. The present study generally agrees with the views of most plant taxonomists that microanatomical features in plants are of huge taxonomic values [15] [18]- [20].…”

Comparative Optimized Protocols of DNA Extraction and Purification Using FTA PlantSaver Card and DNAzol Methods for Eggplant (Solanum Species) Studies in North Central Nigeria

“…DNA extraction is the first step in the study of plant DNA as it precedes all other molecular activities [7] [8]. The two methods compared in this report extracted sufficient quantity of DNA from leaf samples.…”