Analysis of genetic diversity in a sweet potato (Ipomoea batatas L.) germplasm collection from Tanzania as revealed by AFLP

Sweet potato (Ipomoea batatas L.) is the fifth most important crop in the developing countries after rice, wheat, maize and cassava. The amplified fragment length polymorphism (AFLP) method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection of Sokoine University of Agriculture, Morogoro and Sugarcane Research Institute, Kibaha, Tanzania. AFLP analysis of 97 sweet potato accessions using ten primer combinations gave a total of 202 clear polymorphic bands. Each one of the 97 sweet potato accessions could be distinguished based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the un-weight pair-group method using arithmetic average. AFLP-based genetic similarity varied from 0.388 to 0.941, with a mean of 0.709. Cluster analysis using genetic similarity divided the accessions into two main groups suggesting that there are genetic relationships among the accessions. Principal Coordinate analysis confirmed the pattern of the cluster analysis. Analysis of molecular variance revealed greater variation within regions (96.19%) than among regions (3.81%). The results from the AFLP analysis revealed a relatively low genetic diversity among the germplasm accessions and the genetic distances between regions were low. A maximally diverse subset of 13 accessions capturing 97% of the molecular markers diversity was identified. We were able to detect duplicates accessions in the germplasm collection using the highly polymorphic markers obtained by AFLP, which were found to be an efficient tool to characterize the genetic diversity and relationships of sweet potato accessions in the germplasm collection in Tanzania.     

AFLP assessment of diversity in sweetpotato from Latin America and the Pacific region: Its implications on the dispersal of the crop

Although originally domesticated in tropical America, the sweetpotato [Ipomoea batatas (L.) Lam.] has a long history of cultivation in the Pacific region. While the post-Columbus dispersal of sweetpotato to Asia and the Pacific is well documented, the hypothesis that there was a prehistoric transfer of sweetpotato by Peruvian or Polynesian voyagers from Peru to Oceania has long been a controversial issue. The objective of this study was to assess the genetic diversity and interrelationships of sweetpotato landraces from the Pacific region and Latin America, and test the hypothesis of human transfer of this crop to the Pacific Island in prehistoric times. Seventy-five sweetpotato landraces from Peru, Ecuador, Mexico, the Philippines, Papua New Guinea (PNG) and 5 Oceania countries were analyzed using amplified fragment length polymorphism (AFLP). Multidimensional scaling (MDS) and analysis of molecular variance (AMOVA) revealed a large genetic variation in the Oceania gene pool, far greater than that in Peru-Ecuador. The Mexican cultivars were grouped together with those of Oceania. In contrast, there is little association between the Peru-Ecuador germplasm and that of Oceania. These results suggest that Peru-Ecuador may not be the source of the Oceania sweetpotato germplasm. Natural dispersal from Mesoamerica is an alternative explanation, to the Kumara hypothesis, for the origin of the Oceania sweetpotato.     

Genetic diversity in sweetpotato [Ipomoea batatas (L.) Lam.] in relationship to geographic sources as assessed with RAPD markers

Available evidence shows that sweetpotato originated from either Central or South American lowlands with subsequent dispersal to North America, Europe, Africa, Asia and the pacific islands. A total of 71 polymorphic RAPD molecular markers were used to assess the genetic relationships amongst 74 sweetpotato varieties originating from a total of 23 sweetpotato producing countries within six geographical regions, namely, South America, Central America/Caribbean, United States of America (USA), East Africa, Asia and Oceania. An Analysis of Molecular Variance (AMOVA) indicated that 93.4% of the total variance was due to the differences between genotypes within regions. The difference between regions was significant (P < 0.001) but only contributed 6.6% to the variance. Genetic distance (PhiST) calculated with AMOVA and multidimensional scaling (MDS) revealed that the South American and the Central American/Caribbean genotypes formed two separate clusters. East African varieties, which have unique characteristics from other traditional varieties, were distinct from other traditional varieties from South America and Oceania. These results support the reported hypothesis of the origin and dispersal of the sweetpotato and indicate that the primary centre of diversity probably has two distinct genepools. It is proposed that the dispersal of the sweetpotato from its origin may have mainly involved varieties from Central America/Caribbean as opposed to varieties from South America. There is an indication that new genepools may be evolving in Africa and Asia due to hybridisation and adaptation to the local environments.     

RAPD variation in sweetpotato (Ipomoea batatas (L.) Lam) cultivars from South America and Papua New Guinea

The island of New Guinea is considered a secondary center on diversity for sweetpotato, because of its range of isolated ecological niches and large number of cultivars found within a small area. Information of genetic diversity in Papua New Guinea (PNG) sweetpotato is essential for rationalizing the global sweetpotato germplasm collection. Using random amplified polymorphic DNA (RAPD), we compared the genetic variation and genetic diversity in 18 PNG cultivars versus 18 cultivars from South America. The analysis of molecular variance revealed large genetic diversity in both groups of cultivars. The within-group (among individuals) variation accounted for 90.6% of the total molecular variance. However, the difference between PNG and South American groups is statistically significant, although it explained only 9.4% of the total molecular variance. The PNG cultivars are also less divergent than their South American ancestors as the mean genetic distance in PNG group is significantly smaller than that of South American group. The lower level of genetic diversity in PNG cultivars was also reflected by multidimensional scaling. This study shows that PNG cultivars, after many years of isolated evolution in an unique agro-ecological environment are substantially divergent from their ancestors in South America. The genetic diversity level in PNG cultivars is significantly lower than that in South American cultivars. It thus provides a baseline for continuing studies of genetic diversity in different sweetpotato gene pools.     

Genetic diversity in grasspea (Lathyrus sativus L.)

Genetic diversity was investigated in 348 accessions and subaccessions of grasspea (Lathyrus sativus L.) from 10 geographical regions. Polymorphism for 20 isozymes of 13 enzyme systems was studied to estimate the genetic diversity. The Near East and North Africa regions included the most variability for these isozyme systems, suggesting that the center of diversity (center of origin) for grasspea is in this general area. The lowest variability was found in accessions and subaccessions from South America, followed by those from Sudan–Ethiopia. Diversity was measured for individual loci over regions and EST-1 and SKDH had the highest genetic diversity. The closest genetic diversity was observed for LAP-2, followed by AAT-1 and PGM. The closest genetic distance existed between populations from the Near East and North Africa. Populations from South Asia and Sudan–Ethiopia, though geographically widely separated, exhibited a closer genetic distance from each other than from other regions.     

Analysis of genetic diversity in Turkish sesame (Sesamum indicum L.) populations using RAPD markers⋆

The genetic diversity of 38 cultivated populations of Sesamum indicum L. from four different regions of Turkey was estimated at the DNA level with the random amplified polymorphic DNA (RAPD) technique. Sixty-one bands were obtained for all populations 78% of which were polymorphic. Analysis of molecular variance (AMOVA) was used to investigate the genetic diversity of the populations which yielded highly significant differences among populations within regions (91.9% of the total genetic diversity). According to AMOVA and Shannon's index that were performed separately for each region, the highest value of genetic variation was observed among Northwest region populations (CV = 7.7; H₀ = 0.304) and lowest in the Southeast regions' populations (CV = 2.6; H₀ = 0.068). Nei and Li's similarity index was calculated and phylogenetic tree was established using the neighbor-joining algorithm. This phenetic analysis grouped 35 of 38 accessions in six groups leaving three highly diverse accessions outside. Wagner phylogenetic method was used to assess the phylogenetic relationships among the populations. In the majority-rule consensus tree, only 7 of the 32 forks showed above 60% occurrence. Using Principal Coordinate Analysis (PCO) of the RAPD data set, the groups were clearly separated along the first three axis. These results indicate that RAPD technique is useful for sesame systematics, and should be valuable for the maintenance of germplasm banks and the efficient choice of parents in breeding programs.     

Identifying and selecting for genetic diversity in Papua New Guinea sweetpotato Ipomoea batatas (L.) Lam. germplasm collected as botanical seed

A total of 141 Ipomoea batatas (L.) Lam. accessionsderived from botanical seed originally collected from 26 sites in 4 Provinces inPapua New Guinea, a secondary center of genetic diversity for sweetpotato, weregenetically analyzed. Two hundred Amplified Fragment Length Polymorphism (AFLP)markers were identified and utilized in the analysis. Relatedness amongaccessions was estimated by analyzing the AFLP data using the Dice coefficientof similarity and UPGMA methods. The molecular analysis revealed relativelylimited genetic diversity within and between sites. Genotypes collected in agiven region often displayed molecular marker variability similar to thatobserved over the entire sampled area. However, a subset of 14 genotypes derivedfrom seed collected from New Ireland island differed from genotypes collected onNew Guinea island. Estimates of genetic diversity-based similarity valuescalculated from the AFLP data indicated a moderate level of diversity (0.767mean coefficient of similarity) across all plant materials analyzed. Threemethods of selection were evaluated for their efficacy in capturing themolecular marker diversity within the plant materials in the form of a subset.They were random, stratified-random (geographic based), and marker-assistedselection (MAS). MAS was the most efficient. A Maximally Diverse Subset (MDS) of12 genotypes capturing 92% of the molecular marker diversity was identified.     

Genetic diversity in Sorghum bicolor(L.) Moench accessions from different ecogeographical regions in Malawi assessed with RAPDs

Genetic variation within and among several Sorghum populations from different agroecological zones in Malawi were investigated using random amplified polymorphic markers (RAPDs). DNA samples from individual plants were analyzed using 35 oligonucleotides of random sequence. Twenty five of these primers allowed amplifications of random polymorphic (RAPD) loci. Overall, 52% of the scored loci were polymorphic. Every accession was genetically distinct. The analysis of molecular variance revealed that the within-region (among accessions) variations accounted for 96.43% of the total molecular variance. Observed variations in allelic frequency was not related to agroecological differences. The degree of band sharing was used to evaluate genetic distance between accessions and to construct a phylogenetic tree. Further analysis revealed that the sorghum accessions analyzed were genetically close despite considerable phenotypic diversity within and among them. It is suggested that all the sorghum landraces currently available in Malawi should be conserved both ex situ and in situ to maintain the current level of genetic diversity.     

Genetic diversity and gene flow in six accessions of Meso-America teosintes

The genus Zea (teosinte) has a native distribution from Mexico to Nicaragua (Meso-America region) and studies on teosinte species may help improve our understanding of maize genetics, population genetics, genome evolution and crop evolution. Moreover, since F1 hybrids between teosinte and maize are fertile, all teosinte species can potentially contribute to maize breeding in the form of traits such as resistance or tolerance to heavy metals in the soil and other adverse environmental conditions. Gene flow between maize and teosinte occurs in Mexico and Central America. This study analysed the genetic diversity of teosintes by using the microsatellite technique on a total of 120 individuals from six different species and 21 Simple Sequence Repeats (SSR). A total of 109 alleles were found and the number of alleles per locus ranged from 2 to 7. The genetic diversity varied between all species from 0.514 to 0.572, with a mean value of 0.548 (Nei index). Rare and unique alleles were detected in all species, with the highest frequencies found in Zea diploperennis Iltis, Doebley et Guzman, Zea perennis (Hitchc.) Reeves et Mangelsdorf and Zea nicaraguensis Iltis et Benz, which had more than seven rare alleles each. Analysis of molecular variance revealed that 19.87% of the variation between species was statistically significant (P < 0.001). When the different species were compared with the Wright (Fst) value, genetic variation between species was confirmed. The data also revealed a moderate rate gene flow between the species. Genetic diversity was found to be somewhat higher in Central America than in Mexico.     

AFLP analysis of genetic diversity in leafy kale (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. convar. <Emphasis Type="Italic">acephala</Emphasis> (DC.) Alef.) landraces,cultivars and wild populations in Europe

AFLP markers were used to characterize diversity and asses the genetic structure among 17 accessions of kale landraces, cultivars and wild populations from Europe. The range of average gene diversity in accessions was 0.11–0.27. Several landraces showed higher levels of diversity than the wild populations and one cultivar had the lowest diversity measures. The landraces that were most genetically diverse were from areas where kales are known to be extensively grown, suggesting in situ conservation in these areas as a supplement to storage of seeds in gene banks. An analysis of molecular variance (AMOVA) showed that 62% of the total variation was found within accessions. For most accessions, genetic distance was not related to geographic distance. Similarities among accessions were probably not caused by recent gene flow since they were widely separated geographically; more likely the relationship among them is due to seed dispersal through human interactions. Our results indicate that a kale population found in a natural habitat in Denmark was probably not truly wild but most likely an escape from a cultivated Danish kale that had subsequently become naturalized.     

Genetic diversity in sorghum [<Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.) Moench] germplasm from Southern Africa as revealed by microsatellite markers and agro-morphological traits

Cultivated sorghum [Sorghum bicolor (L.) Moench] is an important food security crop in the semi-arid regions of the world including Asia and Africa. Its genetic diversity is contained mostly in traditional varieties and modern cultivars used by farmers. In this study, agro-morphological traits and molecular markers were used to assess genetic diversity in 22 accessions of cultivated sorghum from five countries (Botswana, Namibia, Swaziland, Zambia and Zimbabwe) in the Southern African Development Community (SADC) region. The study revealed a significant variation among 22 accessions in both qualitative and quantitative morphological traits, indicating the accessions’ promising potential as breeding material. For molecular analysis, 11 microsatellite primer-pairs were used, and generated a total of 70 alleles across 20 accessions. Analysis of molecular variance revealed a high level of genetic variation; 67 % among the accessions and 10 % among the five countries. The patterns of genetic diversity and the relationships observed in this study should provide insights for genetic resource conservation and utilization of sorghum germplasm in the SADC region.     

Genetic diversity of anchote (Coccinia abyssinica (Lam.) Cogn.) from Ethiopia as revealed by ISSR markers

Anchote (Coccinia abyssinica) is plant endemic in Ethiopia with a high calcium content grown for its edible tuberous roots. In spite of its importance as food security crop, there is no information available on molecular genetic diversity of this crop. Therefore, the aim of this study was to assess genetic diversity within and among 12 populations of anchote using ISSR markers. Using nine ISSR primers, a total of 87 scorable bands was generated of which 74 were polymorphic. Within population diversity based on polymorphic bands ranged from 13.8 to 43.53 % with a mean of 33.05 %, Nei’s genetic diversity of 0.04–0.156 with a mean of 0.12, Shannon information index of 0.07–0.23 with a mean of 0.175 and analysis of molecular variation (AMOVA) of 51.4 % were detected. With all diversity parameters, the highest diversity was obtained from Gimbi, Bedele and Ale populations, whilst the lowest was from Manna. AMOVA showed a 48.56 % between populations variability and significantly lower than that of within population variation. Population differentiation with F_ST was 48.56 %. From Jaccard’s pairwise similarity coefficient, Decha and Nedjo were most related populations exhibiting 0.76 similarity and Manna and Nedjo were the most distantly related populations with similarity of 0.52. The only pentanucliotide primer used in the study, Primer 880 (GGAGA)3, showed a unique band in some individuals that appeared to be associated with morphological quantitative traits (lowest seed number, high protein content, largest fruit size and smallest vine length). Illubabor and Gimbi populations exhibited highest genetic diversity so that the populations should be considered as the primary sites in designing conservation areas for this crop.     

Evaluation of genetic diversity in Turkish melons (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.) based on phenotypic characters and RAPD markers

The genetic relationships among 56 melon (Cucumis melo L.) genotypes collected from various parts of Turkey were determined by comparing their phenotypic and molecular traits with those of 23 local and foreign melon genotypes to investigate the taxonomic relationships and genetic variation of Turkish melon germplasm. Sixty-one phenotypic characters and 109 polymorphic RAPD markers obtained from 33 primers were used to define the genetic similarity among the melon genotypes by dendrograms or two and three dimensional scaling. There were high correlations (r ≥ 0.97) among the four resulting matrices used in molecular characterization. The correlations between phenotypic (Euclidean) and molecular Euclidean, Jaccard, Simple matching, and Nei analyses were r = 0.41, r = −0.40, r = −0.43 and r = −0.40, respectively. Related genotypes or genotypes collected from similar regions were partitioned to similar clusters. Both analyses (phenotypic and molecular) indicated that non-sweet melon types were dissimilar from sweet types and diversity of Turkish melon genotypes was higher than that of sweet foreign cultivars examined, but similar to that of the reference accessions employed. It was also observed that sweet Turkish melon genotypes belonging to groups inodorus and group cantalupensis were highly variable and could have intermated or have crossed with other non-sweet types.     

Phenotypic Variation and Relationships in Landraces and Improved Varieties of Rye (Secale cereale L.) from Northern Europe

Phenotypic diversity for agronomic characteristics was determined in an experiment with 29 landraces and 14 improved varieties of rye from the Nordic area, Germany and Poland. The accessions were scored for 12 characters. Effects of the location, year, type, and country of origin for landraces were investigated with analysis of variance. Phenotypic variations for the traits were estimated using the Shannon–Weaver diversity index. The genetic variation was high, with an average of H ₀ = 0.566. The landraces from Norway, Sweden and Finland showed the highest variation, whereas the improved varieties had the lowest. The German material also had low variation but the status of this material is uncertain. The genetic diversity showed that 70% of the variation was found within the accessions. A cluster analysis was carried out to identify the relationship between the accessions. The material grouped into eight clusters, where clusters I to V included landraces from Sweden, Finland and Norway, except for cluster III, which included one improved variety from Denmark. Cluster VI comprised a single Swedish landrace from Gotland. Most of the improved varieties were in cluster VII and the last cluster contained accessions from Germany and Sweden.     

Analyses of genetic diversity and population structure of anchote (Coccinia abyssinica (Lam.) Cogn.) using newly developed EST-SSR markers

Anchote (Coccinia abyssinica (Lam.) Cogn.) is a perennial root crop belonging to Cucurbitaceae family. It is endemic to Ethiopia and distributed over wide range of agro-ecologies. For further improvement and efficient conservation of this crop, characterization of its genetic diversity and its pattern of distribution is a vitally important step. Expressed sequence tags-simple sequence repeats (EST-SSRs) markers were developed from publicly available watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] ESTs in the GenBank database. Among those novel markers, eight were polymorphic and subsequently used for genetic diversity and population structure analyses of 30 anchote accessions collected from western Ethiopia. A total of 24 alleles were obtained across the eight polymorphic loci and 30 accessions that revealed moderate level of genetic diversity in this minor crop. Among the eight loci, locus CA_06 was the most informative with six alleles and polymorphic information content (PIC) of 0.76. The accessions showed about threefold variation in terms of genetic diversity, with expected heterozygosity (He) ranging from 0.15 (accession An) to 0.44 (accession Dg). Other accessions with higher genetic diversity include Ar and Gu (He?=?0.43 and 0.41, respectively). Analysis of molecular variance (AMOVA) revealed that the variation within accessions and among accessions accounted for 84.7% and 15.3% of the total variation, respectively. The study revealed low but significant population differentiation in this crop with no clear pattern of population structure. The EST-SSR markers developed in this study are the first of their kind for anchote and can be used for characterization of its wider genetic resources for conservation and breeding purposes.

     

Genetic diversity of potato landraces from northwestern Argentina assessed with simple sequence repeats (SSRs)

Andean potato varieties are cultivated in the northwest of Argentina and constitute the most important staple food for the local farmers. The genetic diversity of 155 accessions conserved at the Genebank of Balcarce (INTA) was tested using four microsatellites. Three commercial potato varieties of Tuberosum group and one accession of Curtilobum group were used as outgroups. The presence of bands was scored for each microsatellite and for each accession and the data were analysed by principal coordinate analysis. The polymorphism information content was obtained for each molecular marker from banding patterns. Analysis of molecular variance was carried out with a variable number of accessions for each landrace, from different departments and sites within departments. More than one genotype was detected in the majority of the potato landraces. Some accessions within each landrace did not differentiate. AMOVA revealed that most of the genetic variation occurred among sites within departments and among local varieties. These findings are discussed considering the agricultural practices carried out in the Andean farming system.     

Identification and genetic relationships of kenaf (Hibiscus cannabinus L.) germplasm revealed by AFLP analysis

Kenaf (Hibiscus cannabinus L.) is one of the world's most economically important fiber crops. In order to identify different varieties, and investigate its diversity and genetic relationships, twenty-three kenaf accessions and two accessions of its relative, roselle (H. sabdariffa var. altissima), were analyzed by morphological characterization and AFLP fingerprinting. It is very difficult to identify kenaf accessions based merely on morphological characters, due to their limited variation. For the AFLP study, a total of 505 polymorphic markers (out of 560) were produced by six selected AFLP primer combinations. The AFLP fingerprinting was effective in identifying all kenaf accessions included in the study. Kenaf and roselle are independent species with close relationships, and great genetic diversity was also detected among the kenaf accessions with different origins, based on the analysis of the AFLP markers. The AFLP analysis strongly supports the opinion that kenaf originated in Africa. It also demonstrated that the dissemination of kenaf was from Africa through Asia to Central and North America.     

Genetic Structure of Wild and Domesticated Populations of Capsicum annuum (Solanaceae) from Northwestern Mexico Analyzed by RAPDs

Levels of genetic variation and genetic structure of 15 wild populations and three domesticated populations of Capsicum annuum were studied by RAPD markers. A total of 166 bands (all of them polymorphic) and 126 bands (125 of them polymorphic) were amplified in wild and domesticated populations, respectively. Mean percentage of polymorphism was 34.2% in wild populations and 34.7% in domesticated populations. Mean and total genetic diversity were 0.069 and 0.165 for wild populations and 0.081 and 0.131 for domesticated populations. Parameters of genetic diversity estimated from 54 bands with frequencies ≥1 − (3/n) (n = sample size) showed that 56.7% of the total variation was within and 43.3% among wild populations, whereas 67.8% of the variation was within and 32.2% among domesticated populations. AMOVA indicated that total genetic diversity was equally distributed within (48.9 and 50.0%) and among (50.0 and 51.1%) populations in both wild and domesticated samples. Wild and domesticated populations were clearly resolved in a UPGMA dendrogram constructed from Jaccard’s distances (average GD = 0.197), as well as by AMOVA (17.2% of variance among populations types, p = 0.001) and by multidimensional scaling analysis. Such differentiation can be associated with domestication as well as different origin of gene pools of the wild (Northwestern Mexico) and cultivated (more probably Central Mexico) samples analyzed. The considerable genetic distances among cultivars (average GD = 0.254) as well as the high number of diagnostic bands per cultivar (33 out of 126 bands), suggest that genetic changes associated with domestication could have resulted from artificial selection intervening in different directions, but the inclusion of more domesticated samples might clarify the nature of distinctions detected here.     

Diversity of common bean (Phaseolus vulgaris L.) germplasm from Portugal

Common bean (Phaseolus vulgarisL.) is a traditional crop in Portugal, where farmers growvarieties selected and maintained by themselves. A collection of 88landraces of common bean was evaluated for 17 quantitative andqualitative traits and the biochemical marker phaseolin to displaythe degree of variation of this germplasm. Agronomic data weresubjected to cluster analysis and several groups were identified,with three groups clustering most of the landraces. Regardingphaseolin variation the C and T banding patterns are the mostfrequent ones, so the origin of the Portuguese beans is thus probablythe Andean region of South America. These results give informationabout the origin, diversity and breeding value of the Portuguesegermplasm, that could be useful to widen the genetic base ofcurrently cultivated bean varieties in Europe.