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Article

Analysis of Parent and F1 Progeny Verification in African Yam Bean (Sphenostylis stenocarpa Hochst, Ex. A. Rich. Harms) Using Cowpea SSR Markers

by
Abimbola O. Adefiranye
1,2,3,
Liasu A. Ogunkanmi
1,
Oyenike A. Adeyemo
1,
Temitope E. Shonde Olatunde
2,
Rajneesh Paliwal
2,*,
Michael T. Abberton
2,* and
Olaniyi A. Oyatomi
2
1
Department of Cell Biology and Genetics, Faculty of Science, University of Lagos, Lagos 101017, Nigeria
2
Genetic Resources Centre, International Institute of Tropical Agriculture, Ibadan 200001, Nigeria
3
Department of Biotechnology, Federal Institute of Industrial Research, Oshodi, Lagos 102215, Nigeria
*
Authors to whom correspondence should be addressed.
Crops 2024, 4(4), 480-490; https://doi.org/10.3390/crops4040034
Submission received: 3 August 2024 / Revised: 25 September 2024 / Accepted: 8 October 2024 / Published: 15 October 2024
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Abstract

African yam bean (Sphenostylis stenocarpa Hochst, Ex. A. Rich. Harms) is an important grain legume in Sub-Saharan Africa because of its nutritional value and adaptability to various agroecological zones. To foster the varietal development of improved African yam bean (AYB) genotypes with economic traits, it is necessary to validate parental polymorphism for key markers in selecting progenies from crosses between desired parents. This study aims to analyze the genetic fidelity between parents and F1 progenies in African yam bean through putative cowpea simple sequence repeat (SSR) markers. Hence, a total of seventy-seven progenies were derived from four sets of biparental crossings using drought-susceptible (TSs-96, TSs-363, and TSs-274) and drought-tolerant (TSs-417, TSs-111, and TSs-78) AYB accessions. These were validated using 120 cowpea primers targeting SSRs under optimal PCR conditions, and the size of the PCR-amplified DNA fragments was checked using gel electrophoresis. Twenty primers exhibited polymorphism in the parental lines, while ten displayed higher levels of the same polymorphism. The average polymorphism level for the surveyed SSR markers was 0.36. Given these findings, our study demonstrates that cowpea SSR markers are a reliable method for the regular testing and clear identification of AYB crosses, indicating potential AYB improvements.

1. Introduction

In developing countries, legumes are the primary plant protein source in the average home. Staple legumes include cowpea (Vigna unguiculata L. Walp), soybean (Glycine max), and groundnut (Arachis hypogaea L.). Only a few local farmers cultivate most grain legumes needed to meet the dietary needs of the rural population. The widespread underutilization of most grain legumes led to their classification as orphan crops [1], some of which include African yam bean (Sphenostylis stenocarpa) (AYB), Bambara groundnut (Vigna subterranea), and pigeon pea (Cajanus cajan).
AYB is a crop that produces both seed and tuber, having protein-rich edible seeds and underground swollen roots [2]. It belongs to the Fabaceae family and is the most cultivated species in the genus Sphenostylis [3]. AYB has been reported to contain about 29 and 19% crude protein content in its grain and tuber, respectively [4]. Regarding the amino acid content (lysine and methionine), the grain has a considerably higher amount when compared to major staple legumes like cowpea and soybean [5]. Also, its tubers are like sweet potatoes and Irish potatoes in size and appearance but contain a higher amount of crude protein content twice that of sweet potatoes and ten times higher than that of cassava roots [6].The potential of this crop, particularly in terms of nutrition, can help mitigate widespread malnutrition and chronic food insecurities in Sub-Saharan Africa (SSA) [7].
It is cultivated as a major legume in the eastern part of Nigeria, serving as an important source of food and income for rural dwellers [8]. It is indigenous to tropical Africa and can grow to a height of about 1–3m. Agroecologies ranging from savannah to rainforest support AYB cultivation, so long as there is a combination of adequate rainfall of about 100 cm or more during the growing season and good drainage. AYB seeds can be cooked as porridge or puddings, processed into flour, fermented as a soup condiment, and fortified with cassava or other cereal-based products. The milk from its seeds helps boost breast milk production in lactating mothers [2]. Some parts of Ghana prepare AYB at various local festivals, including puberty rites for maidens [9]. The serum uric acid concentration of AYB is minimal and compares well with that of soybeans [10]. Some tribes in Central Africa commonly incorporate AYB tuber into their meals and consume the leaves as cooked vegetables [11]. Despite this crop’s food and nutritional security potential, constraints such as drought, photoperiod, reduced yield potential, seed coat hardiness, laborious staking needs, long maturation period, presence of anti-nutritional factors (ANFs) or secondary metabolites, and insect infestation have greatly limited its cultivation and use [12,13].
Attempts at making crosses for hybrid generation in AYB plants have been largely unsuccessful due to the early abortion of flowers and it being predominantly a self-pollinating plant [14]; hence, no improved variety has been developed to date. All germplasm available to geneticists and breeders consists of cultivated landraces collected from gene banks. Hence, this study conducted successful biparental crossings using the hand pollination mechanism: dusting the female stigma on the pollen of another AYB plant, both having extreme traits, and protecting them with pollen-proof bags, under screen house conditions [15].
Quite a few molecular markers have been employed to obtain information on AYB at the molecular level. Previously used DNA-based molecular markers include random amplified polymorphic DNA (RAPD) [16] and amplified fragment length polymorphisms (AFLPs) [17,18], simple sequence repeats (SSRs) [19], single nucleotide polymorphisms (SNPs) [20], and, more recently, restriction site-associated DNA sequencing (RADseq) [21] and diversity arrays technology sequencing (DArTseq) [11,22].
SSR markers have been used in a wide range of applications, especially in the identification of true hybrids in breeding programs. Simple sequence repeats (SSRs), and single nucleotide polymorphisms (SNPs) have been the most extensively used marker technologies. SSRs are DNA sequences with a few (1~6) nucleotides in tandem repeats that are widely distributed in eukaryotic genomes. SSR markers have the advantages of good repeatability, high polymorphism and resolution, as well as ease of operation [23,24,25]. Diverse legume species, such as yam bean, lima bean, and mung bean, have been studied extensively using SSRs for variety identification [26].
When breeding for new crop varieties, hand pollination is the most used technique in population development. Hybridization easily produces spurious hybrids, potentially misleading the breeder in selecting the ideal candidates during the improvement phase. Before proceeding with future breeding programs, an accurate mechanism for determining the true-to-type nature of any hybrid must be established. Therefore, this study seeks to ascertain the identities of 77 AYB progenies obtained from four sets of biparental crossings and estimate the genetic diversity in the parental genotypes for varietal development and crop improvement in AYB using cowpea-derived SSR markers.

2. Materials and Methods

2.1. Plant Material and Crosses

Six parental AYB lines that are susceptible and resilient to drought were obtained from the Genetic Resources Centre, International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria. These accessions were evaluated using the Carolina II mating design, in which drought-tolerant accessions (TSs-417, TSs-111, and TSs-78) were used as male progenitors and drought-susceptible accessions (TSs-96, TSs-363, and TSs-274) were used as female progenitors; this was a preliminary work on drought gene action, with the aim of developing hybrids that are true to type (Table 1). The hybridization block was established at the research screen house of the IITA, Ibadan, and the crossings produced seventy-seven F1 progenies resulting from the four chosen sets of biparental crossings.
The young leaves of the parents and progenies were collected in liquid nitrogen 21 days after planting and stored at −80 °C for later use. Genomic DNA (gDNA) of AYB genotypes was extracted using a modified CTAB extraction method [27]. A DNA quality check was completed using electrophoresis on a 1% (w/v) agarose gel. The DNA concentration and quality were determined using NanoDrop 2000/200C spectrophotometer v1.0. (Thermo Scientific Nanodrop, Wilmington, DE USA) by measuring the absorption at 260 and 280 nm and agarose gel electrophoresis. Genomic DNA samples were diluted to a final concentration of 20 ng/µL and stored at −20 °C.

2.2. SSR Marker Selection and PCR Amplification

This study considered 120 selected SSR markers out of 202 SSR markers from 11linkage groups, as reported from cowpea linkage mapping studies conducted by Andargie et al., 2011 [28] for genotyping purposes in validating true crosses (the list of primers can be found in Supplementary Table S1). These primers were chosen based on their ability to target simple sequence repeat (SSR) regions. Information on the nucleotide sequences was sent to Integrated DNA Technologies (IDT), Iowa, USA, for primer synthesis. Polymerase chain reactions were conducted using the Applied biosystem thermal cycler GeneAmp PCR System 9700 (Applied Biosystems, Califonia, CA, USA). DNA from the parent and progenies of AYB were fingerprinted using SSR markers in a 25 μL reaction volume of master mix containing 2.5 μL of 10X reaction buffer, 1 μL of 50 mM MgCl2, 2 μL of 10 mM of dNTPs (Deoxynucleotide Triphosphates), 0.6 μM each of forward and reverse primers, and 0.1 μL of Taq polymerase. Water was added to make the final volume. Reactions were conducted inan initial denaturation step at 94 °C for 5 min. The annealing step was completed at 65 °C for 30 s, reducing by −1 °C per cycle, 10 times, followed by 94 °C for 30 s and 55 °C for 30 s, then 72 °C for 1 min for 30 cycles, and a final extension/elongation step at 72 °C for 10 min and then the reaction was held at 4 °C. The amplified products were stored at −20 °C before gel electrophoresis.
The amplified DNA fragments were separated on 2% agarose gel at 100 V for 45 min–1 h in 1X TBE (Tris-boric ethylene diamine tetra acetic acid) visualized using agarose gel electrophoresis (sunrise 96, Biometra, Gotringen, Germany). A 6X DNA loading dye was added to the PCR products for visual tracking of DNA fragment migration during electrophoresis. A 50 bp DNA marker (New England, Biolabs, County Road, Ipswich, MA 01938, USA) was used as a reference to estimate the size of the specific DNA bands in the PCR-amplified products visualized on a UV transilluminator and photographed using a Gel Documentation System (Apelegen). The preliminary screening using 120 SSR markers on the parental genotypes for polymorphism allowed for the selection of informative loci used to confirm the identity of the 77 AYB progenies.

2.3. Gel Electrophoresis

The PCR-amplified products were resolved on a 3% agarose gel stained with ethidium bromide. Gel electrophoresis was conducted at 100 V for 2 h in 1X TBE buffer. The DNA fragments were visualized under UV light and photographed using a gel documentation system.

2.4. Polymorphism Analysis

The SSR primers exhibiting co-dominance and dominance among the parental lines and their F1 progenies were identified. Polymorphic bands were scored as present (1) or absent (0) to create a binary data matrix (Supplementary Table S2). The polymorphic information content (PIC) for each primer was calculated using the following formula:
P I C = 1 − ∑ P i 2
where Pi is the frequency of the ith allele.

2.5. Determination of True Progeny

To accurately differentiate between genuine and false hybrids, it is necessary to compare the alleles of the parents and their offspring at multiple genetic loci. A true hybrid will exhibit biparental inheritance, meaning it inherits one allele from the mother and one from the father at each locus. False or contaminated hybrids may exhibit uniparental inheritance, where all alleles originate from the same parent. This can occur due to self-pollination, pollen contamination from a different plant, or other factors. However, the absence of a paternal allele at one or two loci does not necessarily indicate a false hybrid, as genetic recombination can sometimes mask parental alleles. Therefore, a combination of genetic markers and careful consideration of breeding history is essential for accurately determining the true parentage of hybrid individuals

2.6. Statistical Analysis

The percentage of hybrid genetic purity was calculated using the scored data and purity index of Bohra et al., 2011 [29]. Hybrid purity was determined by dividing the number of true hybrids, which includes alleles from both parents, by the total number of hybrids screened, then multiplying the results by 100. We estimated genetic diversity parameters using the power marker software (version 3.25) and data generated from 20 SSR markers. The polymorphic information content (PIC), percentage of polymorphic loci, mean number of alleles per polymorphic locus, observed heterozygosity (Ho), and gene diversity (He) were used to estimate the diversity index. A simple matching dissimilarity index determines pairwise comparisons of the proportion of shared alleles between individual genotypes. The resulting genetic dissimilarity coefficient was then transformed into a distance matrix averaging over 1000 bootstraps. We generated a cluster analysis from the distance matrix using the unweighted pair group method and the arithmetic averages (UPGMA) algorithm to enhance the visualization of the genetic relationships between the parents. We used the ape package in R to cluster the progeny based on genetic distance, and pedigree information was used as a cofactor to confirm the genetic relatedness of the hybrids from respective parental crosses [30].

3. Results

3.1. Screening the Parents and Progenies Using SSR Markers

The selection of the SSR markers from the synthesized 120 SSR primers, which were used to screen the AYB parental genotypes and their hybrids, was based on their capacity to produce polymorphic bands under optimum PCR conditions and several PCR optimizations as reported by Shitta et al., 2016 [19]. The findings indicated that out of the 120 SSR markers used, 100 markers were either monomorphic or did not amplify in the AYB DNA. After screening for variation, we identified 10 of the total 20 primers as polymorphic for all six parents. This is illustrated in Figure 1. The gel fragment analysis (Table 2) revealed that 52 (68%) of the 77 hybrids were true-to-type. We obtained these hybrids from the crosses TSs-274 × TSs-78, TSs-363 × TSs-111, TSs-96 × TSs-111, and TSs-96 × TSS-417, yielding 28 (64%), 7 (100%), 15 (71%), and 12 (57%) hybrids, respectively.
The genotyping findings showed a total of 24 alleles that varied in number from twoto four across different loci and groups. The average number of alleles was 2.4. The SSR 6466 and SSR 6171 markers exhibited the greatest number of alleles, with a count of four, whereas the other primers displayed two alleles each. The mean gene diversity, also known as the expected heterozygosity (He), varied from 0.42 (SSR-6577) to 0.50 (SSR-6466), with an overall average of 0.45. The range of observed heterozygosity varied from 0.24 (SSR-6225) to 0.64 (SSR-6466), with an average of 0.44. The polymorphic information content (PIC) of loci in the two groups was highest in SSR 6466 and SSR 6171 (0.42), and lowest in SSR 6577 (0.33), with an average of 0.36 (Table 3).

3.2. Cluster Analysis and F-Statistics

Figure 2 presents a dendrogram based on the hybrid pedigree. The phylogenetic relationship showed two main clusters, or genetic groups. The first cluster has the largest number of members, combining true F1 hybrids and progenies from all four biparental crossings. This cluster grouped progenies from respective cross combinations as a genetic sub-group. However, out of the four biparental offspring, only the F1 progenies from the cross TSs-363 × TSs-111 showed no evidence of false hybrids. In the other three cross combinations, namely, TSs-274 × TSs-78, TSs-96 × TSs-111, and TSs-96 × TSs-417, the true progenies that aligned to their respective parental identities were 18, 15, and 12, respectively. Cluster two comprises two sub-groups, A and B, each containing F1 progenies identified as false F1 hybrids. These clusters’ progenies were a crosscombination involving TSs-274 × TSs-78 (10 progenies), TSs-96 × TSs-111 (6 progenies), and TSs-96 × TSs-417 (9 progenies).
The phylogenetic tree of the parental lines revealed three genetic groups, with cluster one having four parental lines (TSs-78, TSs-363, TSs-96, and TSs-417), cluster two having one line (TSs-111), and cluster three having one member (TSs-274) (Figure 3). An analysis of molecular variance (AMOVA) showed that most of the variation was partitioned among the population (57%). The variability partitioned among individuals was 7%, while within individuals it was 36%, with a remarkably high level of genetic differentiation (Table 4).

4. Discussion

Twenty primers were used to screen for polymorphismsin six parental accessions; however, ten of these primers were polymorphic for these accessions (the remaining ten primers exhibited low levels of polymorphism) and were further used to screen the AYB parental accessions and their corresponding progenies in the hybrid verification process. A proportion of 68% of the progenies, specifically fifty-two, was true-to-type and could proceed to the next stage of inbreeding through selfing. Additionally, only progenies from the cross TSs-363 ×TSs-111 had perfect clustering. This population had the least number of progenies among the four crosses, which may account for why the progenies showed no contamination from another parental cross, as, was observed in the other three populations. The offspring from the crosses TSs-363 × TSs-111, TSs-96 × TSs-111, and TSs-96 × TSs-417 were not grouped together perfectly. This could be because the crosses were not labeled correctly or there were other problems with the pollination. This study reported that the set of ten SSR loci, which were highly informative and provided good coverage distribution of the cowpea genome, was adequate for genetic diversity in the cowpea accessions [19]. The cross-transferability of cowpea markers among different legume crops, particularly legume species, without reference genome information and/or genomic tools (markers) such as African yam bean increases their usefulness in diversity assessment [19,31].
The ten SSR markers showed a mean PIC of 35.6%, which is lower than some results obtained in some previous studies on African yam bean [16,17,19,32]. The markers’ nature could be the reason for the low mean PIC. Additionally, dissimilarity in the genotype and the number of accessions could be probable factors [33]. Furthermore, DArT-SNP markers in winged bean [34] and sugar beet [35] found similar PIC values, indicating their ability to distinguish between the different accessions used in this study. The most informative markers were SSR 6466 and SSR 6171, with a mean PIC of 0.43, while SSR-6701, SSR-6924, SSR-6225, SSR-6982, and SSR-6294 were the least informative (0.34). Earlier research on African yam beans using different markers, like ISSR [32], AFLP [18], and SSR [19], found that the average PICs for the accessions from Nigeria were 0.52, 0.25, and 0.78. These results are in line with those findings. The average number of alleles found at each locus (2.4) was in line with what other genetic diversity studies in winged beans using SSR markers [34,36,37] had found. The small number of alleles per locus corresponds to the African yam bean’s nature as a self-pollinating crop.
This study found a remarkably high level of genetic differentiation among the parental lines of African yam beans, indicating that these lines are extremely far apart in terms of their allele constituents and therefore suitable for breeding population development. Apart from the remarkably high genetic differentiation, the populations partitioned most of the variability among themselves. This is contrary to findings from previous genetic diversity studies in African yam beans using diverse DNA molecular markers that have shown low to medium genetic differentiation and diversity [18,19]. Both the expected and observed heterozygosity averaged 0.45 and 0.44, respectively. When using isozyme markers, this study found more heterozygosity than the average for the self-pollinating plants (He = 33%) [16], consequently, the mechanism of mutations and their high rate of occurrence are associated with high levels of polymorphism in SSR markers. Heterozygosity is an indication that two randomly selected alleles from an accession are different [38]. The expected heterozygosity observed in this study affirmed that the parental materials have moderate to high levels of heterozygosity, as expected for African yam beans.However, during hybridization, the timing and photoperiod effects were major challenges observed. The crosses were performed in the early hours of the day and late evenings to reduce the abortion of flowers and the effect of the photoperiod.
While SSR markers are a valuable tool for genetic studies, they have certain limitations such as not being informative enough to distinguish between closely related genotypes, especially in genetically homogeneous populations, not covering the entire genome, potentially missing important genetic variation and linkage disequilibrium with other loci, thus making it difficult to accurately identify the causal variants for traits of interest. To address these limitations, researchers may consider using additional marker types, such as single nucleotide polymorphisms (SNPs) or whole-genome sequencing, in combination with SSR markers to affirm their findings.

5. Conclusions

The selected SSR markers had enough polymorphism to verify the hybrids generated from the crosses involving three drought-tolerant and three drought-susceptible African yam bean genotypes. We identified fifty-two offspring as being true-to-type, allowing them to advance to the next phase of inbreeding by self-pollination. The F-statistics indicated that genetic differences account for 57% of the variability among the parental lines, thereby confirming the presence of genetic dissimilarity. This is the first study to examine biparental crossings of the African yam bean to determine the success of four distinct, unique populations. Cowpea SSR markers are tools for analyzing genetic variation in African yam bean species. Furthermore, this study displayed the effectiveness of SSR markers in confirming the parentage of hybrid offsprings, a crucial component of breeding schemes.

Supplementary Materials

The following supporting information can be downloaded at https://www.mdpi.com/article/10.3390/crops4040034/s1, Table S1: List of cowpea SSR primers used to screen for polymorphism in four sets of biparental crossings from six parents and 77 progenies of Africa yam bean, Table S2: Summary of SSR alleles for the identification of true to type hybrids in 77 progenies from four sets of biparental crossings of Africa yam bean.

Author Contributions

Conceptualization, A.O.A. and M.T.A.; data curation, A.O.A.; Formal analysis, A.O.A. and T.E.S.O.; Funding acquisition, M.T.A.; Investigation, A.O.A.; Methodology, A.O.A., L.A.O., T.E.S.O., R.P. and O.A.O.; Project administration, M.T.A. and O.A.O.; Supervision, L.A.O., O.A.A., R.P., M.T.A. and O.A.O.; Writing—original draft, A.O.A.; Writing—review & editing, A.O.A., L.A.O., O.A.A., T.E.S.O. and O.A.O. All authors have read and agreed to the published version of the manuscript.

Funding

This research is funded by the Global Crop Diversity Trust and Consultative Group on International Agricultural Research (CGIAR) through the Genetic Resources Centre, International Institute of Tropical Agriculture. Ibadan, Nigeria.

Data Availability Statement

The original contributions presented in the study are included in the article/Supplementary Material; further inquiries can be directed to the corresponding authors.

Acknowledgments

The authors express their gratitude to the members of staff seed bank GRC-IITA, Ibadan, GRC Internship students, IITA Ibadan staff in other units and the staff of cowpea breeding unit, IITA Kano, for their technical assistance. We also thank The Bioscience Centre IITA for their field and technical assistance.

Conflicts of Interest

The authors declare that there are no conflicts of interest.

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Figure 1. PCR results of ten SSR primers showing polymorphism across the six parental lines.
Figure 1. PCR results of ten SSR primers showing polymorphism across the six parental lines.
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Figure 2. Dendrogram of 77 progenies derived from four biparental crossings of African yam bean based on Euclidean genetic distance revealed a cluster of true F1 hybrids and another for false F1 hybrids.
Figure 2. Dendrogram of 77 progenies derived from four biparental crossings of African yam bean based on Euclidean genetic distance revealed a cluster of true F1 hybrids and another for false F1 hybrids.
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Figure 3. Dendrogram of the six parental lines based on Euclidean genetic distance derived from ten SSR markers revealed three clusters with cluster one having four parents while clusters two and three have one parent each.
Figure 3. Dendrogram of the six parental lines based on Euclidean genetic distance derived from ten SSR markers revealed three clusters with cluster one having four parents while clusters two and three have one parent each.
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Table 1. Accession numbers, sources, and qualitative morphological characters of the six studied parental lines of African yam bean (S. stenocarpa).
Table 1. Accession numbers, sources, and qualitative morphological characters of the six studied parental lines of African yam bean (S. stenocarpa).
S/NAccession NoSourceGTFCSCSSPDSBSTSP
1TSs-417NigeriaMalePurplish pinkG/VM/LBO/OB/RHNSHShinySmoothRed
2TSs-111NigeriaMalePale violetG/VM/LBO/R/RHNSHShinySmooth/RoughGreen
3TSs-78NigeriaMalePurplish pinkG/VMO/OB/RHNSHShiny/
Medium		SmoothRed
4TSs-363NigeriaFemalePale violetG/VM/LBO/OBNSHShinySmoothGreen
5TSs-274NigeriaFemalePurplish pinkVMOV/OB/R/RHSH/NSHShinySmoothRed
6TSs-96NigeriaFemaleViolet whiteG/VM/LBO/OB/RHNSHShinySmooth/WrinkledRed
GT = gamete type, FC = flower color, SC = seed color, SS = seed shape, PD = pod dehiscence, SB = seed brilliance, ST = seed texture, SP = stem pigmentation, G = grey, VM = variegated mosaic, LB = light brown, O = oval, OB = oblong, R = Round, RH = rhomboid, SH = shattering, NSH = non-shattering.
Table 2. Summary of SSR alleles for the identification of true-to-type hybrids in seventy-seven progenies from four sets of biparental crossings of African yam bean.
Table 2. Summary of SSR alleles for the identification of true-to-type hybrids in seventy-seven progenies from four sets of biparental crossings of African yam bean.
Hybrid CrossesTrue-to-TypeTotal Progeny
TSs-274 × TSs-781828
TSs-363 × TSs-11177
TSs-96 × TSs-1111521
TSs-96 × TSs-4171221
5277
Table 3. The genetic diversity estimates for six parents and seventy-seven progenies from four sets of biparental African yam bean crossings.
Table 3. The genetic diversity estimates for six parents and seventy-seven progenies from four sets of biparental African yam bean crossings.
MarkerMAFAllele NoHeHoPIC
SSR-67010.68720.4300.3330.338
SSR-66230.67520.4390.4530.343
SSR-64660.63940.5000.6430.424
SSR-65770.69920.4210.3210.332
SSR-69240.68720.4300.4530.338
SSR-69820.67520.4390.5660.343
SSR-62940.67520.4390.4230.343
SSR-62250.67520.4390.2460.343
SSR-61710.63940.5000.6430.424
SSR-67300.68720.4300.3560.338
Mean0.6732.40.4470.44370.356
MAF: minor allele frequency; He: expected heterozygosity/gene diversity; Ho: observed heterozygosity; PIC: polymorphic information content.
Table 4. Analysis of molecular variance (AMOVA) of the parental lines involved in the development of the biparental population as assessed through SSR markers.
Table 4. Analysis of molecular variance (AMOVA) of the parental lines involved in the development of the biparental population as assessed through SSR markers.
SourceDFSSMSEst. Var.%FstP
Among populations213.0006.5001.33357%0.5710.009
Among individual33.5001.1670.1677%
Within individual65.0000.8330.83336%
Total1121.500 2.333100%
DF: degree of freedom; SS: sum of squares; MS: mean squares; Est. Var: estimated variance; %: percentage variance; Fst: pairwise differentiation; P: significance level.
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Adefiranye, A.O.; Ogunkanmi, L.A.; Adeyemo, O.A.; Shonde Olatunde, T.E.; Paliwal, R.; Abberton, M.T.; Oyatomi, O.A. Analysis of Parent and F1 Progeny Verification in African Yam Bean (Sphenostylis stenocarpa Hochst, Ex. A. Rich. Harms) Using Cowpea SSR Markers. Crops 2024, 4, 480-490. https://doi.org/10.3390/crops4040034

AMA Style

Adefiranye AO, Ogunkanmi LA, Adeyemo OA, Shonde Olatunde TE, Paliwal R, Abberton MT, Oyatomi OA. Analysis of Parent and F1 Progeny Verification in African Yam Bean (Sphenostylis stenocarpa Hochst, Ex. A. Rich. Harms) Using Cowpea SSR Markers. Crops. 2024; 4(4):480-490. https://doi.org/10.3390/crops4040034

Chicago/Turabian Style

Adefiranye, Abimbola O., Liasu A. Ogunkanmi, Oyenike A. Adeyemo, Temitope E. Shonde Olatunde, Rajneesh Paliwal, Michael T. Abberton, and Olaniyi A. Oyatomi. 2024. "Analysis of Parent and F1 Progeny Verification in African Yam Bean (Sphenostylis stenocarpa Hochst, Ex. A. Rich. Harms) Using Cowpea SSR Markers" Crops 4, no. 4: 480-490. https://doi.org/10.3390/crops4040034

APA Style

Adefiranye, A. O., Ogunkanmi, L. A., Adeyemo, O. A., Shonde Olatunde, T. E., Paliwal, R., Abberton, M. T., & Oyatomi, O. A. (2024). Analysis of Parent and F1 Progeny Verification in African Yam Bean (Sphenostylis stenocarpa Hochst, Ex. A. Rich. Harms) Using Cowpea SSR Markers. Crops, 4(4), 480-490. https://doi.org/10.3390/crops4040034

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